i- i- m- %-'^:-^-^^j&'^W:. v r-i m% >.>i >M rn';«w- .iu.r^ >■ V ^■H / ir:> - ',' * «jip' )(Sjjm £2 >->-<.- BIOCHEMICAL BULLETIN ISSUED QUARTERLY BY THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION PRESS OF THE NEW ERA PRINTINQ COMPANY LANCASTEK, PA. BiocHEMiCAL Bulletin Edited, for the Columbia University Biochemical Association, by the EDITORIAL COMMITTEE: iJuly, igiz-June, 1913) ALFRED P. LOTHROP, Chairman, PAUL E. HOWE, Secretary, WILLIAM J. GIES, Treasurer, WALTER H. EDDY, MAX KAHN, EMILY C. SEAMAN, Oct., 1912-July, 191 3 NELLIS B. FOSTER, ARTHUR KNUDSON, CLAYTON S. SMITH, July-September, 19 12 F. G. GOODRIDGE, EDGAR G. MILLER, Jr., ETHEL WICKWIRE, TULA L. HARKEY, H. O. MOSENTHAL, LOUIS E. WISE, Oct., 1912-July, 1913 JOSEPH S. HEPBURN, JACOB ROSENBLOOM, JuIy-September, 1912 ALL OK THE StAFF OF THE BlOCHEMICAL DEPARTMENT OF COLUMBIA UnIVERSITY VOLUME II : Nos. 5-8 1912-1913 WITH EIGHT PORTRAITS, EIGHT PLATES AND TWO ADDITIONAL ILLUSTRATIONS LIBRARY NnvV YORK NEW YORK Columbia University Biochemical Association 1913 Entered as second-class matter in the Post Ofiice at Lancaster, Pa. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Members PROF, R. H. CHITTENDEN, First Director of ihe Columbia University De- partment of Biological (Physiological) Chemistry; Director of the Shef- field Scientific School of Yale University PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Switserland PROF. SAMUEL W. LAMBERT, Dean of the Columbia University School of Mediane DR. JACQUES LOEB, Member of the Rockef eller Institute for Medical Re- search; Head of the Department of Experimental Biology PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. LEON ASHER, University of Bern, Switserland PROF. FILIPPO BOTTAZZI, University of Naples, Italy PROF, ROBERT B. GIBSON, University of the Philippines, P. I. PROF. VLADIMIR S. GULEVIC, University of Moscow, Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. D. McCAY, Medical College, Calcutta, India PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden. — C. Stuart Gager. College of the City of New York. — Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Gold färb. Columbia University: Departments. — Anatomy: Alfred J. Brown, H. von W. Schulte; Bacteriology: James G. Dwyer; Biological Chemistry: Walter H. Eddy, Nellis B. Foster, William J. Gies, F. G. Goodridge, Tula L. Harkey, Joseph S. Hepburn, Benjamin Horowitz, Paul E. Howe, Max Kahn, Arthur Knudson, Alfred P. Lothrop, Edgar G. Miller, Jr., H. O. Mosenthal, Emily C. Seaman, Chris. Seifert, Ethel Wickwire, Louis E. Wise; Botany: E. R. Alten- burg, C. A. Darling, Fred D. Fromme; Cancer Research: W. H. Woglom; Chem- istry: A. M. Buswell, R. P. Calvert, Gustave Egloff, H. L. Fisher, P. W. Punnett, A. W. Thomas; Clinical Pathology: Edward Cussler, Peter Irving; Diseases of Children: Herbert B. Wilcox; Gynecology: Wilbur Ward; Mediane: T. Stuart Hart, I. Ogden Woodruff; Pathology: B. S. Oppenheimer, Alwin M. Pappen- heimer; Pharmacology: Charles C. Lieb; Physiology: Russell Burton-Opitz, Donald Gordon, Leander H. Shearer, Wm. K. Terriberry; Surgery: Hugh Auchincloss, William Darrach, Rolfe Kingsley, Adrian V. S. Lambert, F. T. Van Buren, Jr. ; Therapeutics: Maximilian Schulman; University Physician: iv Members resident in New York (con.) Wm. H. McCastline; Vanderbilt Clinic: F. Morris Class, Julius W. Weinstein; Zoology: H. B. Goodrich, John D. Haseman, H. J. Muller, Charles Packard. Colleges. — Barnard: Helene M. Boas, Ella H. Clark, Ruth S. Finch, Louise H. Gregory; College of Pharmacy: Charles W. Ballard; Teachers College: Mary G. McCormick, Mrs. A. P. McGowan, Sadie B. Vanderbilt. Students. — Graduate: Cora J. Beckwith, Sidney Born, O. C. Bowes, Helen B. Davis, Mary C. de Garmo, Frank R. Eider, Louis J. Hirschleifer, Mildred A. Hoge, Shojiro Kubushiro, Victor E. Levine, Darwin O. Lyon, W. A. Perlzweig, Edward Plaut, Geo. S. Rosenthal, Edward C. Stone, Fred L. Thompson, Jennie A. Walker, Charles Weisman, C. A. Wells, Isabel Wheeler. — Teachers College: Anna M. Connelly, Ula M. Dow, Ada M. Field, Helen McClure, Alice H. McKinney, Elizabeth Rothermel, Mary B. Stark, Helen B. Thompson. — Medical: Louis Berman, Ernst Boas, David C. Bull, Will H. Chapman, Robert T. Corry, Calvin B. Coulter, Joseph Felsen, Joseph Goldstone, Julius Gottesman, Leon M. Herbert, Martin Holzman, Walter F. Hume, Julius Hyman, M. V. Miller, Nathan Rosenthal, A. V. Salomon, Harry J. Seiflf, Jacob Shulansky, H. J. Spencer, Henry A. Sussman, Wm. W. Tracey, Grover Tracy. CoRNELL Univeesity Medical COLLEGE. — Stanley R. Benedict, Ernest D. Clark, Robert A. Cooke, Jessie A. Moore, Charles R. Stockard, Geo. W. Vandegrift. EcLECTic Medical College. — David Alperin. Harriman Research Laboratory. — Marston L. Hamlin. Hospitals. — Babies': Morris Stark; Bellevue: Edward C. Brenner, Edward M. Colie, Jr., Ralph W. Lobenstine; Beth Israel: Charles J. Brim and Alfred A. Schwartz; City: Henry H. Janeway, Louis Pine; Flower: Henry L. Weil; Flushing: Eimer W. Baker; General Memorial: Clinton B. Knapp; German: H. G. Baumgard, Alfred M. Hellman, Melvin G. Herzfeld, Frederick B. Humphries, Charles H. Sanford, Fred S. Weingarten; Jewish: Abraham Ravich; Lebanon: Samuel Gitlow, M. J. Gottlieb, William Weinberger; Lutheran: Daniel R. Lucas; Mt. Sinai: George Baehr, Samuel Bookman, Leo Buerger, Burrill B. Crohn, Simon S. Friedman, David J. Kaliski, John L. Kantor, Leo Kessel, Reuben Otten- berg, Harry Wessler; TV. Y.: James C. Greenway, Ralph G. Stillman; N. Y. Nursery and Child's: Oscar M. Schloss; Presbyterian: Herbert S. Carter, Russell L. Cecil, Arthur W. Swann; Roosevelt: J. Buren Sidbury; St. Luke's: Norman E. Ditman, Edward C. Kendali, W. S. Schley, Chas. H. Smith. Long Island Medical College. — Matthew Steel. MoNTEFioRE HoME. — Isidor GrecHwald. Museum of Natural History. — Louis Hussakof, Israel J. Kligler. N. Y. Aquarium. — Raymond C. Osburn. N. Y. Association for Improving the Condition of the Poor. — Donald B. Armstrong. N. Y. Botanical Garden. — Fred J. Seaver. N. Y. City Department of Education. — Boys" High School: Frank T. Hughes; Brooklyn Training School: C. A. Mathewson ; Commercial High School: ' W. J. Donvan, B. C. Gruenberg, Edgar F. Van Buskirk; DeWitt Clinton High School: Frank M. Wheat; Eastern District High School: Gertrude S. Burling- ham; Girls' High School: Marguerite T. Lee; High School of Commerce: Harvey B. Clough, Fred W. Hartwell; Jamaica High School: Ella A. Holmes, Charles H. Vosburgh; Manual Training High School: Anna Everson; Morris Members resident in New York (con.) High School: Charles A. Wirth; Newtown High School: Nellie P. Hewins; Wadleigh High School: Helen Gavin, Elsie A. Kupfer, Helen G. Russell, Helen S. Watt. N. Y. City Department of Health. — Charles F. Bolduan, Alfred F. Hess. N. Y. City Normal College. — Beatrix H. Gross. N. Y. Eye and Ear Infirmary. — Harold M. Hays. N. Y. Milk Committee. — Philip Van Ingen. N. Y. Polyclinic Medical School. — Jesse G. M. Bullowa, Mabel C. Little. Post Graduate Medical School. — Louis E. Bisch, Arthur F. Chace. Pratt Institute. — Grace MacLeod. Rockefeller Institute. — Alfred E. Cohn, George W. Draper, Frederic M. Hanes, Michael Heidelberger, Gustave M. Meyer. Russell Sage Institute of Pathology. — Eugene F. DuBois. TuRCK Institute. — Anton R. Rose. Vettin School. — Laura I. Mattoon. E. V. Delphey, 400 West 57th Street, Manhattan; Leopold L. Falke, 5316 Thirteenth Avenue, Brooklyn; Mabel P. Fitzgerald, 416 East 6sth Street, Man- hattan; Abraham Gross, c/o Arbuckle Sugar Co., Brooklyn; Alfred H. Kropff, 619 Kent Avenue, Brooklyn. Non-Resident Members Agnes Scott College (Decatur, Ga.). — Mary C. de Garmo. Allegheny General Hospital (Pittsburgh). — James P. McKelvy. Carnegie Institution (Cold Spring Harbor, L. I.). — Ross A. Gortner. Cornell University (Ithaca). — Jean Broadhurst. Drake University Medical School (Des Meines, la.). — E. R. Posner. Forest School (Biltmore, N. C). — Homer D. House. Iowa University Hospital (Iowa City). — Louis Baumann. Isolation Hospital (San Francisco, Cal.). — L. D. Mead. Jefferson Medical College (Phila.). — P. B. Hawk, Edward A. Spitzka. Johns Hopkins University (Baltimore). — John Howland, W. M. Kraus, Burton E. Livingston, Edwards A. Park. Lehigh University (Bethlehem, Pa.). — William H. Welker. Leland Stanford University (Palo Alto, Cal.). — Hans Zinsser. MacDonald College (Quebec). — Kathryn Fisher. Mass. Agricultural College (Amherst). — H. D. Goodale. New Mexico Agricultural College (State College). — R. F. Hare. N. J. Agricultural Experiment Station (New Brunswick). — Carl A. Schwarze, Guy West Wilson. N. Dakota Agricultural College (Agricultural College). — H. L. White. Ohio Agricultural Experiment Station (Wooster). — A. D. Selby. Princeton University (N. J.). — E. Newton Harvey. Psychopathic Hospital (Boston). — Herman M. Adler. Rensselaer Polytechnic Institute (Troy, N. Y.). — Fred W. Schwartz. Rochester A and M Institute. — Elizabeth G. Van Hörne. Secondary Schools. — Brockport State Normal School (N. Y.) : Ida C. Wads- worth; Hermon High School (N. Y.) : Sidney Liebovitz; Indiana State Normal School (Terre Haute): Roscoe R. Hyde; Ingleside School (New Milford, Conn.) : Mary L. Chase; Knox School (Tarrytown, N. Y.) : Clara G. Miller; New Bedford Industrial School (Mass.): Constance C. Hart; North Texas vi Non-resident members (con.) State Normal School (Benton) : Blanche E. Shaffer; Passate High School (N. J.) : Hazel Donham, Helene M. Pope; Rochester High School (N. Y.) : David F. Renshaw; State Normal School (Truro, N. S.) : Blanche E. Harris. Texas A and M College (College Station). — M. K. Thornton. Trinity College (Hartford, Conn.). — Max Morse, R. M. Yergason. TuLANE University (New Orleans, La.). — Allan C. Eustis. U. S. Department of Agriculture (Wash.). — Carl L. Aisberg, W. N. Berg, H. E. Buchbinder, William Salant, Clayton S. Smith. U. S. Food and Drug Inspection Laboratory (Phila.). — Harold E. Woodward. U. S. FooD- Research Laboratory (Phila.). — Joseph S. Hepburn. University of Alabama Medical School (Birmingham). — Richard A. Bliss. University of California (Berkeley). — William T. Home, University of Chicago. — Mathilde Koch. University of Georgia Medical School (Atlanta). — ^William D. Cutter. University of Illinois (Urbana). — George D, Beal, Isabel Bevier, A. D. Emmett. University of Indiana (Bloomington). — Clarence E. May. University of Kentucky (Louisville). — Mary E. Sweeny. University of Manitoba (Winnipeg, Can.). — A. T. Cameron. University of Michigan (Ann Arbor). — A. Franklin Shull. University of Montana (Missoula). — J. E. Kirkwood. University of Pennsylvania (Phila.). — A. N. Richards. University of Porto Rico (Las Pietras). — L. A. Robinson. University of Tennessee (Memphis). — Edwin D. Watkins. University of Texas (Austin). — Mary E. Gearing, Anna E. Richardson. University of Toronto (Canada). — Olive G. Patterson. i University of Utah (Salt Lake City). — H. A. Mattill. University of Wisconsin (Madison). — ^W. H. Peterson. Vassar College ( Poughkeepsie, N. Y.). — Winifred J. Robinson. Washington State College (Pullman). — Josephine T. Berry, Louise McDanell. Wesleyan University (Middletown, Conn.). — David D. Whitney. West Pennsylvania Hospital (Pittsburgh). — J. Bronfen Brenner, Jacob Rosenbloom. Williams College (Williamstown, Mass.). — John S. Adriance, Yale University (New Haven, Conn.). — Lorande Loss Woodruff. Albert H. Allen, Saranac Lake, N. Y. ; Emma A. Buehler, Newark, N. J. ; George A. Geiger, West Orange, N. J. ; Edward G. Griffin, Albany, N. Y. ; F. C. Hinkel, Utica, N. Y.; Cavalier H. Joüet, Roselle, N. J.; A. E. Olpp, West Hoboken, N. J. ; Adeline H. Rowland, Pittsburgh, Pa. ; William A. Taltavall, Redlands, Cal. ; David C. Twichell, Saranac Lake, N. Y. Vll EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the coUaboration of the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockef eller Institute for Medical Research PROF. WILDER D. BANCROFT, Cornell University, Ithaca DR. WALTER L. GROLL, Elizabeth Steel Magee Hospital, Pittshurgh, Pa. DR. CHARLES A. DOREMUS, 55 W. 53d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station, Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR, LEWIS W. FETZER, U. S. Dep't of Agriculture, Washington, D. C. PROF. MARTIN H. FISGHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. PROF. J. E. GREAVES, Utah Agricultural College, Logan DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. M. HARDISTY, McGill University, Montreal, Canada DR. J. A. HARRIS, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. K. A. HASSELBALCH, Einsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, A''. Y. Univ. and Bellevue Hospital Med. College PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan PROF. LAFAYETTE B. MENDEL, Yale University PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven DR. AMOS W. PETERS, The Training School, Vineland, N. J. PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon PROF. UMETARÖ SUZUKI, University of Tokyo, Japan MISS ANNA W. WILLIAMS, University of Illinois, Urbana, III. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switserland DR. JULES WOLFF, Pasteur Institute, Paris vm OFFICERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION 1912-1913 HONORARY OFFICERS Post President (1910-1912) : Prof. Alfred N. Richards, University of Pennsylvania, Phila. President: Prof. Philip B. Hawk, Jefferson Medical College, Phila. Vice Presidents: Dr. Herman M. Adler, Psychopathie Hospital, Boston, Mass. Prof. Allan C. Eustis, Tulane University, New Orleans, La. Miss Olive G. Patterson, Toronto University, Toronto, Can. Prof. Winifred J. Robinson, Vassar College, Poughkeepsie, N. Y. Prof. Lorande Loss Woodruff, Yale University, New Haven, Conn. ACTIVE OFFICERS President, Dr. Walter H. Eddy. Vice President, Prof. Stanley R. Benedict. Secretary, Dr. Alfred P. Lothrop. Treasurer, Prof. William J. Gies. Executive Commiftee — Prof. Stanley R. Benedict, Dr. Wal- ter H. Eddy, Dr. Nellis B. Foster, Prof. William J. Gies, Dr. Frederic G. Goodridge, Prof. Paul E. Howe and Dr. Alfred P. Lothrop. Editorial Committee: See the title page. ix SUMMARY OF CONTENTS: VOL. II, Nos. 5-8. No. 5. September, 19 12. PAGE Ernst ScHxn-ZE. Biography and Bibliography (with portrait). Ernst Winterstein. i A ReSUME OF THE LiTERATURE ON InOSITE-PhoSPHORIC AciD ("PhYTIN"), WITH Special Reference to the Relation of that Substance to Plants. Anton Richard Rose 21 A New Type of Artificial Cell Suitable for Permeability and Other BioCHEMicAL Studies. E. Ncwtou Hürvey 50 On a New Function of the Catalyzer Called Peroxidase and on the Biochemical Transformation of Orcin into Orcein. Jules Wolff . ... 53 Studies of Diffusion through Rubber Membranes : 1. Preliminary Observations on the Diffusibility of Lipins and Lipin- soluble Substances. William J. des SS 2. Dififusibility of Lipins from Ether through Rubber Membranes into Ether. Jacob Rosenbloom 64 3. Diffusibility of Protein through Rubber Membranes, with a Note on the Disintegration of Collodion Membranes by Common Ethyl Ether and Other Solvents. William H. Welker 70 4. The Comparative DiffusibiHty of Various Pigments in Different Sol- vents. George D. Beal and George A. Geiger 78 The Colloidal Nitrogen in the Urine from a Dog with a Tumor of the Breast. Max Kahn and Jacob Rosenbloom 87 General Aspects of Fasting. Paul E. Hozue 90 The Physico-chemical Basis for the Contraction of Striated Muscle: 2. Surface Tension. William N. Berg loi A Study of Some Protein Compounds. Walter H. Eddy iii Effects of Intraperitoneal Injections of Epinephrin on the Partition OF Nitrogen in Urine from a Dog. Jacob Rosenbloom and William Weinberger. 123 The Biochemical Society, England. W. D. Halliburton 128 Proceedings of the Section (II) ON DiETETic Hygiene and Hygienic Physiology OF the 15TH International Congress on Hygiene and Demography, with Abstracts of Some of the Papers. Lafayette B. Mendel, Secretary. 129 Program of the Proceedings of the Section on Biochemistry Including Pharm acology (viii, d), of the 8th International Congress of Applied Chemistry. John A. Mandel, Secretary 150 Proceedings of the Sixth Scientific Meeting of the Columbia Univer- siTY Biochemical Association. Alfred P. Lothrop, Secretary 156 Biochemical News, Notes and Comment: General 188 X PAGE Columbia University Biochemical Association 200 Columbia Biochemical Department 201 Editori ALS : Ernst Schulze 205 Important though Unknown Factors in Nutrition 205 The Coming of Age of the Babcock Test 207 Organotherapy 208 Biochemical Society, England 20g "Baustein" or " Construction Unit"? 209 " Splitting Products " or Cleavage Products ? 209 A Rare Compliment 210 X-rays 210 No. 6. January, 19 13. PAGE Carl L. Alsberg. Biography and Bibliography (with portrait). H.M.A.. 211 A Differential Chemical Study of Glucoses from a Case of Pancreatic Diabetes. Frederic Landolph 217 The Detection of Aceto-acetic Acid by Sodium Nitroprussid and Ammonia. V. J. Harding and R. F. Ruttan. 223 Ortho-tolidin as an Indicator for Occult Blood. R. F. Ruttan and R. H. M. Hardisty. 225 Synthetical Properties of Emulsin. Vernon K. Krieble 227 On the Occurrence of Nicotinic Acid in Rice Bran. U. Suzuki and S. Matsunaga. 228 A Study of the Influence of Cancer Extracts on the Growth of Lupin Seedlings. Jacob Rosenhloom 229 The Biochemistry of the Female Genitalia: 3. A Quantitative Study of Certain Enzymes of the Ovary, Uterus, and Bladder, of Pregnant and Non-pregnant Sheep. Thuisco A. Erpf-Lefkovics and Jacob Rosenbloom. 233 4. On the Absence of Certain Enzymes from the Human Chorion. Jacob Rosenbloom. 236 A Department of Biochemical Research at Vineland, New Jersey. Arnos W. Peters. 238 Biochemistry in New York Twenty Years Ago. E. E. Smith 243 Immunity in Some of its Biochemical Aspects. Charles Frederick Bolduan. 247 A Plan for the Organization of the American Biological Society. Albert P. Mathews. 261 Organization of the Federation of American Societies for Experimental BlOLOGY, COMPRISING THE AmERICAN PhYSIOLOGICAL SoCIETY, AmERICAN Society of Biological Chemists, and American Society for Pharma- COLOGY and Experimental Therapeutics. John Auer 269 Annual Meetings of the Organizations Comprising the Federation of American Societies for Experimental Biology : I. The American Physiological Society. Joseph Erlanger, Acting Secretary. 271 PAGB 2. The American Society of Biological Chemists. Alfred N. Richards, Secretary. 275 3. The American Society for Pharmacology and Experimental Thera- peutics. John Aner, Secretary 27g Meeting of the American Society of Animal Nutrition (American Society of Animal Production) . Lewis W. Fetzer 282 Proceedings of the Eighth Scientific Meeting of the Columbia Univer- siTY BiocHEMicAL ASSOCIATION. Alfred P. Lothrop, Secretary ;.. 284 Folio Microbiologica. C. A. Pekelharing 297 BiocHEMiCAL Bibliography AND Index. William J. Gies 298 BiocHEMicAL News, Notes and Comment: General 307 Columbia University Biochemical Association 321 Columbia Biochemical Department 324 Editorials : New Plan of Quarterly Issue of the Bulletin 329 Carl L. Aisberg 329 Stock Poisoning Due to Spoiled Silage. Help ! 330 Demand for Biological Chemists in the Hospitals 330 Federation of American Societies for Experimental Biology 331 Electrons 332 No. 7. April, 19 13. PAGE Heinrich Ritthausen (Portrait) 334 Appreciation. Thomas B. Osborne 335 Bibliography. Lewis W. Fetzer 339 Dinner to Professor Chittenden: Testimonial by his Pupils. '94S 349 Society for Experimental Biology and Medicine: Tenth Anniversary Meeting and Dinner. Nineteen O. Three 358 Methods for the Electrometric Determination of the Concentration of Hydrogen Ions in Biological Fluids. K. A. Hasselbalch 367 A Method for the Determination of Tryptophan Derived from Pro- teins. Jesse A. Sanders and Clarence E. May 373 Physical Chemistry of Muscle Plasma. Filippo Bottazzi 379 Fasting Studies : IL A Note on the Composition of Muscle from Fast- iNG DoGs. H. C. Biddle and Paul E. Howe 386 SoME Notes on the Form of the Curve of Carbon-dioxide Excretion Re- suLTiNG from Muscular Work Following Forced Breathing. G. O. Higley. 390 The Influence of Barometric Pressure on Carbon-dioxide Excretion in Man. G. O. Higley 393 The Relation of Acapniato Shock, andaConsideration of the Mechan- iCAL Effects of Artificial Hyper-respiration upon the Circulation. Henry H. Janeway and Ephraim M. Ewing. 403 Cleavage of Pyromucuric Acid by Mold Enzymes. Arthur W. Dax and Ray E. Neidig. 407 Analysis of the Ash of the Castor Bean. Marston Lovell Hamlin 410 Notes on the Chemical Nature of the " Tannin Masses " in the Fruit of the Persimmon. Ernest D. Clark 412 xü PAGE HisTON AND iTS Prep ARATION. Walter H. Eddy 419 DiD VON Wittich Antedate Ostwald in the Definition of Enzyme Action ? William N. Berg. 441 The Biochemical Society, England 446 Scientific Proceedings of the Columbia University Biochemical Asso- ciation. Alfred P. Lothrop, Secretary 452 Biochemical Bibliography and Index. William J. Gies 470 Biochemical News, Notes and Comment: General 476 Columbia University Biochemical Association 484 Editorials : Biochemical Society, England 487 The Bleached Flour Decision 487 Occupational Diseases in Chemical Trades 488 The Mathews Plan for an American Biological Society 490 Antigens 508 No. 8. July, 19 13. PAGE An Investigation to Determine the Accuracy of a Modified Meigs Method for the Quantitative Determination of Fat in Milk, with A Description of an Improved Form of Apparatus. Walter Lewis Croll. 509 The Occurrence of Arsenic in Soils. /. E. Greaves 519 Further Notes on the Relationship between the Weicht of the Sugar Beet and the Composition of its Juice. /. Arthur Harris and Ross Atken Gortner. 524 Note on the Relationship between Barometric Pressure and Carbon- dioxide ExcRETioN IN Man. /. Arthur Harris 530 The Bleached Flour Decision. Ross Aiken Gortner 532 Emil Chr. Hansen Fund. 5". P. L. Sörensen 535 Biological Chemistry in the Philippines. Robert Banks Gibson 536 Doctorates in Biological Chemistry. Conferred by American Univer- sities, 1912-13. P. H. D 538 Scientific Proceedings of the Columbia University Biochemical Asso- ciation. Alfred P. Lothrop, Secretary 541 Biochemical Bibliography and Index. William J. Gies 559 Biochemical News, Notes and Comment: General 567 Columbia University Biochemical Association 574 Columbia Biochemical Department 578 Editorials : Peroxides and Nitrites in Plants 582 Mathews Plan for the Organization of an American Biological Society. 582 Crystals 588 Index: Volume II. (Includes names of authors, and impersonal and per- sonal subj ects) 589 Title Page for Volume II, with Summary of Contents, List of Illus- TRATIONS, ETC i-Xvi . xiii Alphabetic list of authors named in the foregoing summary of Contents (See author index — page 589 — for additional names of authors of abstracts, quotations, comment, etc.) Adler, HM, 211 AuER, J, 269, 279 Beal, GD, 78 Berg, WN, ioi, 441 BiDDLE, HC, 386 BOLDUAN, CF, 247 BoTTAzzi, F, 379 Clark, ED, 412 Croll, WL, 509 Dox, AW, 407 Eddy, WH, III, 419 Erlanger, J, 271 Erpf-Lefkovics, TA, 233 EwiNG, EM, 403 Fetzer, LW, 282, 339 Geiger, GA, 78 GiBSON, RB, 536 Gies, WJ, 5s, 298, 349, 358, 470, 559 GoRTNER, RA, 524, 532 Greaves, JE, 519 Halliburton, WD, 128 Hamlin, ml, 410 Harding, VJ, 223 Hardisty, RHM, 225 Harris, JA, 524, 530 Harvey, EN, 50 Hasselbalch, KA, 367 HiGLEY, GO, 390, 393 Howe, PE, 90, 386 Janeway, HH, 403 Kahn, M, 87 Kribble, VK, 227 Landolph, F, 217 LoTHROP, AP, 156, 284, 452, 541 Mandel, JA, 150 Mathews, AP, 261 Matsunaga, S, 228 May, CE, 373 Mendel, LB, 129 Neidig, RE, 407 OsBORNE, TB, 335 Pekelharing, CA, 297 Peters, AW, 238 P. H. D., 538 Richards, AN, 275 Rose, AR, 21 ROSENBLOOM, J, 64, 87, 123, 229, 233, 236 Ruttan, RF, 223, 225 Sanders, JA, 373 Smith, EE, 243 Sörensen, SPL, 535 Suzuki, W, 228 Weinberger, W, 123 Welker, WH, 70 Winterstein, E, i WOLFF, J, 53 XIV LIST OF ILLUSTRATIONS Eight portraits, eight plates (inserts), and two additional illustrations No. 5- SEPTEMBER, 1912 PAGE Portrait. Ernst Schulze i Plate I. Structure of muscle fibrils (Berg) 107 Portrait. Paul E. Howe 201 No. 6. JANUARY, 1913 Portrait. Carl L. Aisberg 211 Plate 2. Receptors of three kinds (Bolduan) 254 No 7. APRIL, 1913 Portrait. Heinrich Ritthausen 335 Portrait. Russell H. Chittenden 349 Engrossed Greetings to Prof. Chittenden by his colleagues of the Governing Board of th^^effield Scientific School 351 Faces of the gol(^medal presented to Prof. Chittenden by the National Insti- tute of Social Sciences 353 Portrait (group). Testimonial dinner to Prof. Chittenden by his pupils, Mar. I, 1913 355 Portrait. Samuel J. Meltzer 359 Portrait (group). Dinner of the Society for Experimental Biology and Medicine, tenth anniversary, Feb. 19, 1913 363 Plate 3. Apparatus for the electrometic determination of the concentration of hydrogen ions (Hasselbalch) 371 Plate 4. Curve of carbon-dioxide excretion resulting from muscular work after forced breathing. (Higley) 390 Plate 5. Influence of barometric pressure on the excretion of carbon-dioxide (Higley) 396 Plate 6. Micro-Kjeldahl apparatus (Morse) 458 No. 8. JULY, 1913 Plate 7. Apparatus for use with the Meigs method for the determination of fat in milk (Croll) 517 Plate 8. Relationship between the weight of the sugar beet and the compo- sition of its juice (Harris and Gortner) 526 XV Vol. II September, 1912 No. 5 Biochemical Bulletin Edited, for the Columbia University Biochemical Association, by the EDITORIAL COMMITTEE: ALFRED P. LOTHROP, Chöirman, PAUL E. HOWE, Secretary, WILLIAM J. GIES, Treasurer, WALTER H. EDDY, EDGAR G. MILLER, JR., NELLIS B FOSTER, HERMAN O. MOSENTHAL, FREDERIC G. GOODRIDGE, JACOB ROSENBLOOM, TULA L. HARKEY, EMILY C. SEAMAN, JOSEPH S. HEPBURN, CLAYTON S. SMITH, ARTHUR KNUDSON, ETHEL WICKWIRE, all of the Staff of the Biochemical Department of Columbia University. CONTENTS PAGB Ernst Schulze. Biography and Bibliography (with portrait) Ernst Winterstein i A ReSUME of the LiTERATURE OX InOSITE-PhOSI'HORIC AcID (" PhYTIN "), WITH Special Reference to the Relation of that Substaxce to Plants. Anton Richard Rose 21 A New Type of Artificial Cell Suitable für Permeability and other Bio- chemical Studies. E. Newton Harvey . 50 On a New Function of the Catalyzer Called Peroxidase and on the Bio- chemical Transformation of Orcin into Orcein. ßiles Wolff. 53 Studies of Diffusion Through Rubber Membranes : i. Preliminary observations on the diffusibility of lipins and lipin-soluble sub- stances. Williajn J. Gies 55 2. Diffusibility of lipins from ether through rubber membranes into ether. Jacob Rosenbloom 64 3. Diffusibility of protein through rubber membranes, with a note onthe dis- integration of collodion membranes by common ethyl ether and other solvents. William H. Welker 7° 4. The comparative diffusibility of various pigments in different solvents. George D. Beal and George A. Geiger 78 The Colloidal Nitrogen in the Urine from a Dog with a Tumor of the Breast. Max Kahn and Jacob Roseitbloom 87 General Aspects of Fasting. Paul E. Hoive 90 The "Physico-Chemical Basis for the Contraction of Striated Muscle: 2. Surface tension (with plate i). William N. Berg lOi A Study of some Protein Compounds. Walter H. Eddy iii Effects of Intraperitoneal Injections of Epinephrin on the Partition of Nitrogen IN Urine from A Dog. Jacob Rosenbloom and William Weinberger 123 The Biochemical Society, England. W. D. Halliburton 128 PrOCEEDINGS of the SeCTION (II) ON DiETETIC HYGIENE AND HYGIENIC PhYSI- ology of the 15TH International Congress on Hygiene and Demog- raphy, with Abstracts of some of the Papers. Lajayette B. Mendel, Secretary 129 Program of the Proceedings of the Section on Biochemistry Including Pharmacology (viii, d), of the Sth International Congress of Applied Chemistry. John A. Mandel, Secretary 150 Proceedings of the Sixth Scientific Meeting of the Columbia University Biochemical Association. Alfred P. Lothrop, Secretary 156 Biochemical News, Notes and Comment 188 Editorials 205 NEW YORK Columbia University Biochemical Association. Entered as second-class matter in the Post Office at Lancaster, Pa. Honorary Members of the Columbia University Biochemical Association PROF. R. H. CHITTENDEN, First Director of the Columbia University De- partment of Biological (Physiological) Chemistry; Director of the Shef- field Scientific School of Yale University PROF. SAMUEL \V. LAMBERT, Dean of the Columbia University School of Medicine PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Editors of the Biochemical Bulletin EDITORIAL COMMITTEE (See names on the outside of this cover) CONTRIBUTING EDITORS OF THE BIOCHEMICAL BULLETIN PROF. LEON ASHER, University of Bern, Switserland PROF. FILIPPO BOTAZZI, University of Naples, Italy PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. C. A. PEKELHARING, University of Utrecht, Holland DR. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark SPECIAL CONTRIBUTORS TO THE CONTENTS OF VOLUMES I AND II PROF. WILDER D. BANCROFT, Cornell University, Ithaca DR. CHARLES A. DOREMUS, 55 W. 52d St., New York City PROF. MARTIN H. FISCHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, A^. Y. Univ. and Bellevue Hospital Med. College PROF. ALBERT P. MATHEWS, University of Chicago PROF. LAFAYETTE B. MENDEL, Yale University PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. E. E. SMITH, 50 Last 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon MISS ANNA W. WILLIAMS, University of Illinois, Urbana, III. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switserland DR. JULES WOLFF, 26 Rue Dutot, Paris ASSOCIATE EDITORS DAVID ALPERIN, Eclectic Medical College EDGAR ALTENBURG, Department of Botany, Columbia University HUGH AUCHINCLOSS, Department of Surgery, Columbia University GEORGE BAEHR, Mount Sinai Hospital ELMER W. BAKER, Flushing Hospital CHARLES W. BALLARD, College of Pharmacy, Columbia University HANS G. BAUMGARD, German Hospital Dispensary CORA J. BECKWITH, Department of Zoology, Columbia University Associate editors (continued) STANLEY R. BENEDICT, Cornell University Medical College LOUIS E. BISCH, Manhattan State Hospital HELENE M. BOAS, Barnard College, Columbia University CHARLES F. BOLDUAN, Health Department of New York City SAMUEL BOOKMAN, Mount Sinai Hospital SIDNEY BORN, Department of Chemistry, Columbia University WILLIAM BALLANTINE BOYD, College of the City of New York EDWARD C. BRENNER, Bellevue Hospital JACOB J. BRONFENBRENNER, Rockefeiler Institute for Medical Research LEO BUERGER, Mt. Sinai Hospital JESSE G. M. BULLOWA, New York Polyclinic Medical School GERTRUDE S. BURLINGHAM, Rastern District High School, Brooklyn RUSSELL BURTON-OPITZ, Department of Physiology, Columbia University HERBERT S. CARTER, Presbyterian Hospital RUSSELL L. CECIL, Presbyterian Hospital ARTHUR F. CHACE, New York Post-Graditate Medical School ERNEST D. CLARK, Cornell University Medical College ALFRED E. COHN, Rocke feller Institute for Medical Research EDWARD M. COLIE, Jr., Bellevue Hospital BURRILL B. CROHN, Mt. Sinai Hospital LOUIS J. CURTMAN, College of the City of New York EDWARD CUSSLER, Department of Clinical Pathology, Columbia University CHESTER A. DARLING, Department of Botany, Columbia University WILLIAM DARRACH, Department of Surgery, Columbia University NORMAN E. DITMAN, St. Luke's Hospital GEORGE DRAPER, Hospital of the Rockef eller Institute BENJAMIN G. FEINBERG, College of the City of New York HARRY L. FISHER, Department of Chemistry, Columbia University SIMON S. FRIEDMAN, Mt. Sinai Hospital C. STUART GAGER, Brooklyn Botanic Garden HELEN GAVIN, Wadleigh High School SAMUEL GITLOW, Lebanon Hospital Dispensary A. J. GOLDFARB, College of the City of New York DONALD GORDON, Department of Physiology, Columbia University MARK I. GOTTLIEB, Fordham University ISIDOR GREENWALD, Montefiore Home Laboratory LOUISE HOYT GREGORY, Barnard College, Columbia University ABRAHAM GROSS, Arbuckle Sugar Co., Brooklyn BEATRIX H. GROSS, N. Y. City Normal College BENJAMIN C. GRUENBERG, Brooklyn Commercial High School MARSTON L. HAMLIN, Harriman Research Laboratory, Roosevelt Hospital FREDERIC M. HANES, Rockef eller Institute for Medical Research JOHN D. HASEMAN, Department of Zoology, Columbia University HAROLD M. HAYS, New York Eye and Ear Infirmary MICHAEL HEIDELBERGER, Rockefeller Institute for Medical Research ALFRED M. HELLMAN, German Hospital MELVIN G. HERZFELD, German Hospital ALFRED F. HESS, Health Department of New York City NELLIE P. HEWINS, Newtown High School, L. I. ELLA A. HOLMES, Jamaica High School, L. I. Associate editors (continued) FRANK T. HUGHES, Boys High School, Brooklyn FREDERICK B. HUMPHRIES, Gertnan Hospital LOUIS HUSSAKOF, American Museum of Natural History PETER IRVING, Department of Clinical Pathology, Columbia University HENRY H. JANEWAY, City Hospital, New York CAVALIER H. JOÜET, Roselle, N. J. DAVID J. KALISKI, Mt. Sinai Hospital JOHN L. KANTOR, Mt. Sinai Hospital EDWARD C. KENDALL, St. Lukc's Hospital LEO KESSEL, Mt. Sinai Hospital ROLFE KINGSLEY, Department of Surgery, Columbia University ISRAEL J. KLIGLER, American Museum of Natural History CLINTON B. KNAPP, General Memorial Hospital ALFRED H. KRÖPFE, Hoffman and Kropff Chemical Co., Brooklyn ELSIE A. KUPFER, Wadleigh High School ADRIAN VAN S. LAMBERT, Department of Surgery, Columbia University MARGUERITE T. LEE, Girls High School, Brooklyn CHARLES C. LIEB, Department of Pharmacology, Columbia University MABEL C. LITTLE, New York Polyclinic Hospital RALPH W. LOBENSTINE, Bellevue Hospital DANIEL R. LUCAS, St. Joseph's Hospital CHESTER A. MATHEWSON, Brooklyn Training School for Teachers LAURA I. MATTOON, Vcttin School, i6o W. 74th Street WILLIAM H. McCASTLINE, University Physician, Columbia University MARY G. McCORMICK, Teachers College, Columbia University MRS. ELLEN BEERS McGOWAN, Teachers College, Columbia University GUSTAVE M. MEYER, Rockefeiler Institute for Medical Research JESSIE A. MOORE, Loomis Laboratory, Cornell University Medical College HERMANN J. MULLER, Cornell University Medical Collegs B. S. OPPENHEIMER, Department of Pathology, Columbia University RAYMOND C. OSBURN, New York Aquarium REUBEN OTTENBERG. Mt. Sinai Hospital CHARLES PACKARD, Department of Zoology, Columbia University ALWIN M. PAPPENHEIMER, Department of Pathology, Columbia University F. W. PUNNETT, Department of Chemistry, Columbia University ABRAHAM RAVICH, Jewish Hospital, Brooklyn ANTON R. ROSE, Department of Chemistry, Columbia University CHARLES H. SANFORD, German Hospital WINFIELD S. SCHLEY, St. Luke's Hospital OSCAR M. SCHLOSS, New York Nursery and Chlld's Hospital MAX SCHULMAN, Department of Applied Therapeutics, Columbia University H. VON W. SCHULTE, Department of Anatomy, Columbia University FRED J. SEAVER, Netv York Botanical Garden LEANDER H. SHEARER, Department of Physiology, Columbia University JAMES B. SIDBURY, Roosevelt Hospital MORRIS STARK, Babies Hospital MATTHEW STEEL, Long Island Medical College RALPH G. STILLMAN, Nezv York Hospital CHARLES R. STOCKARD, CorncU Univcrsilv Medical College ARTHUR W. SWANN, Presbyterian Hospital Associate editors (continued) WM. K. TERRIBERRY, Department of Physiology, Columbia University F. T. VAN BEUREN, Jr., Department of Surgery, Columbia University GEORGE W. VANDEGRIFT, Cornell University Medical College SADIE B. VANDERBILT, Teachers College, Columbia University CHARLES H. VOSBURGH, Jamaica High School WILBUR WARD, Department of Gynecology, Columbia University HELEN S. WATT, Wadleigh High School WILLIAM WEINBERGER, Lebanon Hospital FRED S. WEINGARTEN, German Hospital JULIUS W. WEINSTEIN, Vanderbilt Clinic, Columbia University HARRY WESSLER, Mt. Sinai Hospital H. B. WILCOX, Department of Diseases of Children, Columbia University LOUIS E. WISE, Standard Varnish Works, Staten Island, N. Y. WILLIAM H. WOGLOM, Dep't. of Cancer Research, Columbia University I, OGDEN WOODRUFF, Department of Mediane, Columbia University (Locol members of the Columbia University Biochemical Association) ASSISTANT EDITORS HERMAN M. ADLER, Psychopathie Hospital, Bos.ton, Mass. JOHN S. ADRIANCE, Williams College, Williamstozvn, Mass. CARL L. ALSBERG, Bureau of Plant Industry, U. S. Dep't. of Agriculture D. B. ARMSTRONG, Massachusetts Institute of Technology, Boston LOUIS BAUMANN, University Hospital, Iowa City, Iowa GEORGE D. BEAL, University of Illinois, Urbana, III. WILLIAM N. BERG, Bureau of Animal Industry, U. S. Dep't of Agriculture JOSEPHINE T. BERRY, State College, Pullman, Washington ISABEL BEVIER, University of Illinois, Urbana, III. A. RICHARD BLISS, Birmingham Medical College, Birmingham, Ala. JEAN BROADHURST, Cornell University, Ithaca, N. Y. WILLIAM D. CUTTER, Medical College of Georgia, Augusta, Ga. A. D. EMMETT, University of Illinois, Urbana, III. ALLAN C. EUSTIS, Tulane University, Nezv Orleans, La. KATHARINE A. FISHER, MacDonald College, Quebec, Canada MARY E. GEARING, University of Texas, Austin, Texas GEORGE A. GEIGER. Marcus Hook, Fa. H. D. GOODALE, Carnegie Sta'n. for Exp. Evolut'n, Cold Spring Harhor, L. I. R. A. GORTNER, Carnegie Sta'n for Exp. Evolu'tn, Cold Spring Harbor, L. I. R. F. HARE, New Mex. Coli, of Agric. and Mech. Arts, Agric. College, N. M. E. NEWTON HARVEY, Princeton University, Princeton, N. J. BLANCHE R. HARRIS, State Normal School, Truro, Nova Scotia CONSTANCE C. HART, New Bedford Industrial School, New Bedford, Mass. P. B. HAWK, JeffersoH Medical College, Philadelphia WILLIAM T. HÖRNE, University of California, Berkeley, Cal. HOMER D. HOUSE, Forest School, Bilfmore, N. C. J. E. KIRKWOOD, University of Montana, Missoula, Mont. MATHILDE KOCH, University of Chicago, Chicago, III. W. M. KRAUS, Johns Hopkins Medical School, Baltimore, Md. SIDNEY LIEBOVITZ, Hermon High School, Hermon, N. Y. BURTON E. LIVINGSTON, Johns Hopkins University, Baltimore, Md. J. P. McKELVY, Allegheny General Hospital, Pittsburgh, Pa. H. A. MATTILL, University of Utah, Salt Lake City, Utah CLARENCE E. MAY, Indiana University, Bloomington, Ind. Assistant editors (continued) L. D. MEAD, Isolation Hospital, San Francisco, Cal. CLARA G. MILLER, Knox School, Tarrytczvn, N. Y. MAX W. MORSE, Trinity College, Hartford, Conn. EDWARDS A. PARK, Johns Hopkins Medical School OLIVE G. PATTERSON, Toronto University, Toronto, Canada W. H. PETERSON, University of Wisconsin, Madison, Wis. E. R. POSNER, Drake University Medical School, Des Maines, la. DAVID F. RENSHAW, West High School, Rochester, N. Y. ALFRED N. RICHARDS, University of Pennsylvania^ Philadelphia ANNA E. RICHARDSON, University of Texas, Austin WINIFRED J. ROBINSON, Vassar College, Poughkeepsie, N. Y. WILLIAM SALANT, Bureau of Chemistry, U. S. Department of Agriculture CARL A. SCHWARZE, N. J. Agricultural Experiment Station, New Briuiswick FREDERICK W. SCHWARTZ. Rensselaer Polytechnic Institute, Troy, N. Y. A. D. SELBY, Ohio Agricultural Experiment Station, Wooster, Ohio BLANCHE E. SHAFFER, North Texas State Normal School, Benton, Texas A. FRANKLIN SHULL, University of Michigan, Ann Arbor, Mich. EDWARD A. SPITZKA, Jcfferson Medical College, Philadelphia EDWARD C. STONE, Trinity College, Hartford, Conn. MARY E. SWEENY, University of Kentucky, Lexington, Ky. WILLIAM A. TALTAVALL, Redlands, Cal. IDA C. WADSWORTH, Brockport State Normal School, Brockport, N. Y. WILLIAM H. WELKER, Red Hill, Pa. DAVID D. WHITNEY, Wesleyan University, Middletown, Conn. LORANDE LOSS WOODRUFF, Yale University, New Haven, Conn. HAROLD E. WOODWARD, U. S. Food and Drug Inspection Lahoratory, Philadelphia HANS ZINSSER, Leland Stanford University, Palo Alto, Cal. {Non-resident members of the Columbia University Biochentical Association) ^r^^'i^ ^^>^^^Sz^ BiocHEMiCAL Bulletin Volume II SEPTEMBER, 191 2 No. 5 IN MEMORIAM ERNST SCHULZE Born July 31, 1840 Died June 15, 19 12 The death o£ Ernst Schulze is an irreparable loss to biology. Wherever the biochemistry of plants is appreciated, Schulze's death causes profound sorrow. Schulze was one of the founders of our present exact biochemical investigation. His researches in phyto- chemistry are classical and they have been charged with funda- mental ideas that continue to influence research in this great field. Dr. Ernst Schulze, professor of agricultural chemistry in the Eidgenössischen Technischen Hochschule at Zürich, was born July 31, 1840, in the hamlet of Bovenden, near Göttingen. In 1858 Schulze studied chemistry under Wöhler in Göttingen and also spent a Semester with Bunsen in Heidelberg. In 1861 he was assistant to Lehman, and subsequently to Geuther, at the Chemical Institute in Jena. His scientific activity began at the Agricultural Experi- ment Station in Weende, under the direction of Henneberg. In 1871 Schulze was appointed director of the newly founded Agri- cultural Experiment Station in Darmstadt. Even while he was at Weende, his ability had attracted the attention of the Eidgenös- sischen institution. In June, 1872, he was called to Zürich, where his activities continued fruit fully for forty years. Schulze's first important research was published with the collab- oration of his friend Märcker, in 1870, in the Journal für Land- zmrtschaft. In this paper it was shown that the principles of pro- 2 Ernst Schuhe [Sept. tein metabolism, as they were stated by Voit on the basis of experi- ments on carnivorous animals, applied to the ruminants as well. In Zürich, Schulze brought his researches in animal physiology to an end by a thorough investigation of wool-fat. He succeeded in preparing typical cholesterol in a pure State and in isolating an isomer, isocholesterol. Since 1872 Schulze had concerned himself exclusively with phy- tochemical research ; and forty years of activity in this field f ortified the conclusion that plants and animals contain the same classes of substances and that the chemical composition of animals is in many ways identical with that of plants. Schulze developed new methods for the quantitative determina- tion of nitrogenous substances and showed how to separate them in pure forms from the complex mixtures in plant Juices and extracts. With his collaborators Schulze made classical discoveries of the fol- lowing nitrogenous Compounds and, by masterly methods, estab- lished their Constitution : Glutamin, an amide of glutamic acid ; Arginin, guanido-ot-aminovalerianic acid ; Phenylalanin, yS-phenyl-ct-aminopropionic acid ; Vernin (identical with the guanosin subsequently obtained by Levene from nucleic acid) ; Stachydrin, the dimethylbetain of a-prolin ; Lupinin, an alkaloid from lupins. Schulze found the following nitrogenous substances in different plant materials and studied their role in plant metabolism: amino- valerianic acid, leucin, isoleucin, prolin, glutamin, asparagin, Phenyl- alanin, tyrosin, arginin, histidin, lysin, vicin, convicin, xanthin, hy- poxanthin, guanidin, vernin, allantoin, cholin, betain, trigonellin, and stachydrin. Schulze was working with the betains during his last illness but, unfortunately, he was unable to complete this re- search. Considerable interest was aroused by his discoveiy of the presence of allantoin in plants. Schulze was the first to make a successful investigation of phyto- lecithins ( Phosphatids) and their cleavage products. He found that the lecithin in many seeds can be extracted only with hoiling alcohol. For this reason he believed that lecithin exists in such 1912] ■ Ernst Winterstein 3 seeds in some sort of combination with proteins. In this connection he investigated the plant cholesterols, or phytosterols. Schulze then began his thorough studies of the carbohydrates and nitrogen-free reserve materials in plants. A paper entitled: "Untersuchungen über die stickstofffreien Reservestoffe der Samen von Lupinus Intens und über die Umwandlung derselben während des Keimungsprozesses," was given a prize by the Königlichen Gesellschaft der Wissenschaften in Göttingen. In this connection Schulze studied the constituents of cell mem- branes in plants. He showed that the walls of various plant-cells contain carbohydrates which resemble cellulose to a certain extent but differ from it by dissolving easily in warm dilute Solutions of acids and alkalis. These cell-wall constituents proved to be xylans, arabans, galactans, and mannans. They play the part of food reserves in seeds. Schulze called them " hemi-celluloses." He showed, further, that ordinary cellulose on hydrolysis yields other glucoses besides dextrose. Stachys tubers were found to contain stachyose, a tetrasaccharid. All these researches yielded data and experience that proved useful to Schulze in his discussions and de- velopment of analytical methods for phytochemical research. The role of asparagin and glutamin in the protein metabolism and synthesis in plants greatly interested Schulze to the end of his life. Although he was not able fully to explain the process of protein synthesis, he made fundamental contributions to the subject. He clarified our knowledge of protein metabolism in seedlings. What chemist or biologist has not heard of the investigations which were begun in 1876, and whose results were usually published in Prus- sian agricultural year books and also in the Zeitschrift für physio- logische Chemie? Even in his second paper on the subject, in 1878, Schulze showed the importance of the characteristic composition of etiolated seedlings and their high asparagin content. He concluded from his observations that the protein decomposition products do not persist, in seedlings, in the proportions in which they were originally produced from protein, hut, that after such protein cleav- age, these nitro genotis suhstances seem to he changed for the most part into asparagin. Schulze prepared a great many plant proteins and studied their 4 Ernst Schulze [Sept decomposition products. After his pupils, working outside our Institute, had taken an active part in the study of protein metabolism in seedlings, and after it had been shown in our laboratory that protein decomposition in seedlings is an enzymic process, Schulze came to the following general conclusions regarding the protein transformations in seedlings: Asparagin is formed in seedHngs at the expense of proteins and arises from the same material in etio- lated young green plants, and in young leaves and shoots; arginin also results in seedlings from direct decomposition of protein. Perhaps the individual amids arise in the leaf-buds in the propor- tions of their production from protein by hydrolysis with acid^ and other agcnts outside the organism, but probably with the differ- ence that, in the leaf-buds, neither aspartic acid nor glutamic acid is produced, the amids of these amino acids, viz., asparagin and glutamin, resulting instead. Amino acids, however, do not occur in plants in such proportions, since they are consumed in the plant metabolism, some more rapidly, it seems, than others. The accu- mulation of asparagin in seedlings is caused by the formation of this amide from other products (amino acids) of the trans formation of protein. One of the best arguments for this conception is the Observation, made by Schulze in his experiments and repeatedly em- phasized in his papers, that in many cases asparagin is produced abundantly even after the processes of protein decompositions in the plant have ceased. An admirable outcome of Schulze's investigations is his great compilation on the composition of cultivated plants, where he re- views briefly the methods of research and gives abundant data on the chemical constituents of these plants. The later years of Schulze's life were spent in close retirement because of a serious and long standing eye-disease that prevented him from appearing in public. He lived, at the end, only for his science and for his family. His colleagues often wondered how, with his weak eyes, he was able to do any experimental work what- ever. It was pathetic to see with what extreme care and patience. he had to tax himself in order to proceed with his work. When Schulze celebrated his seventieth birthday, two years ago, we all hoped that the twilight of his life might be long and happy, I9I2] Ernst IV int erst ein 5 but in vain, for pitiless death took him from iis. His pupils mourn, a beloved friend and guide; and science, a distinguished inves- tigator.^ Ernst Winterstein. Agriculturchemischen Laboratorium der Eidgenössischen Technischen Hochschule, Zürich. PUBLIICATIONEN VON PROF. DR. E. SCHULZE I. In den " Landwirtschaftlichen Versuchsstationen " Ueber die Elementarzusammensetzung der tierischen Fette, insbeson- dere der Fette vom Schaf, vom Rind und vom Schwein. E. Schulze und A. Reinecke. 9: 97-119 (1867). Ueber die sensiblen Stickstoff. Einnahmen und Ausgaben des voll- jährigen Schafes. E. Schulze und M. MÄRCKER. 11:201(1869). Ueber die Zusammensetzung und die Verdaulichkeit des im Wiesenheu enthaltenen Fettes. E. Schulze. 15: 81-90 (1872). Beiträge zur Kenntnis des Nährwerts und der Zusammensetzung der Rüben. E. Schulze. 15: 170-181 (1872). Zur Frage über die Verdauung des Heufetts. E. Schulze. 16 : 329-335 (1873)- Notiz über den Aspargingehalt von Lupinen Keimlingen. E. Schulze und W. Umlauft. 18: 1-3 (1875). Ueber die stickstoffhaltigen Bestandteile der Futter-Rüben. E. Schulze und A. Urich. 18: 296-324 (1875). Notiz betreffend das Vorkommen des Betains in den Futter-Rüben. E. Schulze und A. Urich. 18: 409 (1875). Ueber Schwefelsäurebildung in den Keimpflanzen, E. Schulze. 19 : 172-176 (1876). Einige Bemerkungen über die Sachsse-Kormannsche Methode zur Be- stimmung des in Amid-Form vorhandenen Stickstoffs. E. Schulze. 20: 1 17-123 (1877). Ueber die stickstoffhaltigen Bestandteile der Futterrüben. E. Schulze und A. Urich. 20: 194-245 (1877). Ueber den Gehalt der Kartoffelknollen an Eiweissstoffen und an Amiden. E. Schulze und J. Barbieri. 21: 63-92 (1878). * The foregoing biographical communication was translated from Prof. E. Winterstein's manuscript, in German, by Dr. Ernest D. Clark. Prof. Winter- stein's manuscript of the appended bibliography is reproduced verbatim. [Ed.] 6 Ernst Schulze [Sept Ueber ein neues Glukosid (Bestandteil von Lupinus luteus). E. Schulze und J. Barbieri. 24: i-ii (1880). Ueber das Vorkommen von Leucin und Tyrosin in den Kartoffelknol- len. E. Schulze und J. Barbieri. 24: 167-169 (1880). Ueber die Bestimmung der Eiweissstoffe und der nicht eiweissartigen Stickstoffverbindungen in den Pflanzen. E. Schulze. 24: 358- 365 (1880); 25: lyz--^?^ (1880). Zur Bestimmung der Eiweissstoffe und der nicht eiweissartigen Stick- stoffverbindungen in den Pflanzen. E. Schulze und J. Barbieri. 26:213-283 (1881). Neue Beiträge zur Kenntnis der stickstoffhaltigen Bestandteile der Kartoffelknollen. E. Schulze und E. Eugster. 27 : 357-373 (1882). Zur quantitativen Bestimmung der Eiweissstoffe und der nicht eiweiss- artigen Stickstoffverbindungen in den Pflanzen. E. Schulze. 27 : 449-465 (1882). Ueber das Vorkommen von Hypoxanthin im Kartoffelsaft. E. Schulze. 28: 111-115 (1883). Ueber das Glutamin. E. Schulze und E. Bosshard. 29: 295-307 (1883). Zur quantitativen Bestimmung des Asparagins, des Glutamins und des Ammoniaks in den Pflanzen. E. Schulze und E. Bosshard. 29 : 399-412 (1883). Zur Kenntnis der Methoden, welche zur Bestimmung der Amide in Pflanzenextrakten verwendbar sind. E. Schulze. 30 : 459-467 (1884). Ueber einige Bestandteile des Emmentaler Käses. B. Rose und E. Schulze. 31: 115-137 (1885). Ueber das Vorkommen von Glutamin in den Zuckerrüben und über das optische Verhalten desselben. E. Schulze und E. Bosshard. 32: 129-136 (1887). Untersuchungen über die stickstoffhaltigen Bestandteile einiger Rauh- futterstoffe. E. Schulze, E. Steiger und E. Bosshard. 33: 8^123 (1887). Ueber die Methoden, welche zur quantitativen Bestimmung der stick- stoffhaltigen Pflanzenbestandteile verwendbar sind. E. Schulze. 33: 124-145 (1887). Ueber das Vorkommen von Rohrzucker in unreifen Kartoffelknollen. E. Schulze und Th. Seliwanow. 34: 403 (1887). Ueber den Nachweis von Rohrzucker in vegetabilischen Substanzen. E. Schulze. 34: 408-413 (1887). I9I2] Ernst Winterstein 7 Ein Beitrag zur Erklärung der Veränderungen, welche die stickstoff- haltigen Bestandteile eingesäuerter Grünfutterstoffe erleiden. E. Schulze. 35: 195-208 (1888). Ueber die Zersetzung von Proteinstoffen in verdunkelten grünen Pflanzen. E. Schulze und E. Kisser. 36: 1-8 (1889). Ueber das Vorkommen eines unlöslichen, Schleimsäure gebenden Kohlenhydrats in Rotklee und Luzerne- Pflanzen. E. Schulze und E. Steiger. 36: 9-13 (1889). Untersuchungen über die stickstofffreien Reservestoffe der Samen von Lupinus luteus und über die Umwandlungen derselben während des Keimungsprozesses. E. Schulze und E. Steiger. 36 : 391- 476 (1889). Untersuchungen über die chemische Zusammensetzung einiger Legumi- nosen-Samen. E. Schulze, E. Steiger und W. Maxwell. 39 : 269 (1891). Ueber einige Bestandteile der Wurzelknollen von Stachys tuberifera. A. VON Planta und E. Schulze. 40: 277-298 (1892). Bestimmung des Stachyose-Gehalts der Wurzelknollen von Stachys tuberifera. A. von Planta und E. Schulze. 41 : 123-129 ( 1892) . Zur Kenntnis der in den Leguminosensamen enthaltenen Kohlenhy- drate. E. Schulze. 41:207-229 (1892). Ueber den Lecithingehalt einiger vegetabilischer Substanzen. E. Schulze und S. Frankfurt. 43: 307-318 (1894). Untersuchungen über die zur Klasse der stickstoffhaltigen organischen Basen gehörenden Bestandteile einiger landwirtschaftlich benutz- ter Samen, Oelkuchen und Wurzelknollen, sowie einiger Keim- pflanzen. E. Schulze in Verbindung mit S. Frankfurt und E. Winterstein. 46: 23-77 (1896). Zur Kenntnis der stickstoffhaltigen Bestandteile junger grüner Pflanzen von Vicia sativa. E. Schulze. 46: 383-397 (1896), Ueber das Vorkommen von Arginin in den Wurzeln und Knollen ein- iger Pflanzen. E. Schulze. 46: 451-458 (1896). Ueber die Verbreitung des Glutamins in den Pflanzen. E. Schulze. 48: 33-55 (1897). Ueber den Lecithingehalt einiger Pflanzensamen und einiger Oelkuchen. E. Schulze. 4g: 203-214 (1898). Die Notwendigkeit der Umgestaltung der jetzigen Futter- und Nahr- ungsmittel-Analyse. E.Schulze. 49:419-441 (1898). Ueber die Verbreitung des Glutamins in den Pflanzen. {Zweite Mitteilung.) E.Schulze. 49:442-446(1898). 8 Ernst Schulze [Sept. Ueber die Bestandteile der Samen von Pinus cemhra (Zierbeikiefer oder Arve). E. Schulze und N.Rongger. 51:189-204(1899). Ueber die Rückbildung der Eivveissstoffe aus deren Zerfallsprodukten in der Pflanze. E. Schulze. 55: 33-44 (1901). Ueber die Zusammensetzung einiger Koniferen-Samen. E. Schulze. 55:267-307 (1901). Können Leucin und Tyrosin den Pflanzen als Nährstoffe dienen? E. Schulze. 56:97-106(1902). Ein Nachtrag zu der Abhandlung über die Frage ob Leucin und Tyrosin den Pflanzen als Nährstoffe dienen können. E. Schulze. 56: 293-296 (1902). Zur Kenntnis der kristallisierten Stachyose. E. Schulze. 56 : 419- 423 (1902). Ueber das Vorkommen von Hexonbasen in den Knollen der Kartoffel {Solanum tuberosum) und der Dahlie {Dahlia variabilis). E. Schulze. 59: 331-343 (1904)- Ueber Methoden, die zur Darstellung organischer Basen aus Pflanzen- säften und Pflanzenextrakten verwendbar sind. E. Schulze. 59: 344-354 (1904). Zur Kenntnis des Glutamins. E. Schulze. 65: 237-246 (1906). Ueber die Bestandteile der Samen von Pinus cemhra. E. Schulze. 67: 57-104 (1907). Zur Kenntnis des Glutamins. (Zzveite Mitteilung.) E. Schulze und Gh. Godet. 67: 313-319 (1907). Ueber die chemische Zusammensetzung der Samen unserer Kultur- pflanzen. E.Schulze. 73:35-170(1910). Zur Kenntnis des Glutamins. (Dritte Mitteilung.) E. Schulze und G. Trier. 77: 1-12 (1912). 2. In den landwirtschaftlichen Jahrbüchern Untersuchungen über einige chemische Vorgänge bei der Keimung der gelben Lupine. E. Schulze, W. Umlauft und A. Urich. 5 : 821-862 (1876). Die stickstoffhaltigen Bestandteile der vegetabilischen Futtermittel und ihre quantitative Bestimmung. E.Schulze. 6:157-175(1877). Ueber die Prozesse, durch welche in der Natur freier Stickstoff in Stickstoffverbindungen übergeführt wird, E. Schulze. 6 : 695- 707 (1877). Ueber die Zersetzung und Neubildung von Eiweissstoffen in Lupinen- keimlingen. E.Schulze. 7:411-444(1878). jgi2] Ernst Winterstein 9 Ueber den Eiweissumsatz im Pflanzenorganismus. E. Schulze. 9: 689-748; 12: 909-920; 14: 713-729; 21: 105-130 (1880-1892). Untersuchungen über den Emmentaler Käse und über einige andere schweizerische Käsesorten. E. Benecke und E. Schulze. 16: 317-400 (1887). Ueber die Bildungsweise des Asparagins und über die Beziehungen der stickstofffreien Stoffe zum Eiweissumsatz im Pflanzenorganismus. E. Schulze. 17: 683-711 (1888). Ueber die stickstofffreien Bestandteile der vegetabilischen Futtermit- tel. E.Schulze, 21:79-103(1892). Zur Kenntnis der in den pflanzlichen Zellmembranen enthaltenen Kohlenhydrate. E. Schulze. 23: 1-26 (1894). Ueber die Bildungsweise des Asparagins in den Pflanzen. E. Schulze. 30: 287-297 (1901). Ueber den Abbau und den Aufbau der organischen Stickstoff Verbind- ungen in den Pflanzen. E. Schulze. 35: 621-666 (1906). 3. Im Journal für Landwirtschaft Welchen Einfluss haben die Zubereitung des Futters und die Futter- mischung auf den Nährwert des Futters? Mit welchen Futter- stoffen sind bei den gegenwärtigen Marktpreisen Futterrationen mit angemessenem Gehalt an Nährstoffen am billigsten herzu- stellen. E. Schulze, 17: 33-48 (1869). Untersuchungen über die sensiblen Stickstoff-Einnahmen und -Aus- gaben des volljährigen Schafs und die Ausnutzung einiger Futter- stoffe durch dasselbe. E. Schulze und M. Märcker. 18: 1-39; 19: 202-222, 285-326, 347-362; 20: 46-76 (1870-1872). Fütterungsversuche mit Schafen, E. Schulze und M. Märcker. 23: 141-174 (1875)- Ueber die Zusammensetzung einer pechschweissigen Schafwolle und des daraus gewonnenen Wollfetts. E. Schulze und J. Barbieri. 27: 125-144 (1879). Ueber die zur Gruppe der stickstofffreien Extraktstoffe gehörenden Pflanzenbestandteile. E. Schulze. 52: 1-30 (1904). Ueber die in den landwirtschaftlichen Kulturpflanzen enthaltenen, nicht proteinartigen Stickstoffverbindungen. E. Schulze. 52 : 305- 336 (1904). Ueber den Nährwert der in den Futtermitteln enthaltenen nichtprote- inartigen Stickstoffverbindungen. E.Schulze. 54:65-81(1906). 10 Ernst Schicke [Sept. 4. In dem Landwirtschaftlichen Jahrbuch der Schweiz Ueber die Entstehung der Salpetersäuren Salze im Boden. E. Schulze. 1890 : 109-121 ; 1891 : 82-86. Ueber die in den Futtermitteln enthaltenen Fettsubstanzen und über die Bedeutung derselben für die tierische Ernährung. E. Schulze. 1892 : 1-9. Ueber den Humus und seine Beziehung zum Leben der Pflanze. E. Schulze. 1901 : 1-13. Die Nährstoffnormen und die Beurteilung des Nährwertes der Futter- bestandteile nach ihrer Verbrennungswärme. E. Schulze. 1902 : 1-19. Ueber die chemische Zusammensetzung des Holzes und über einige aus demselben darstellbaren Produkte. E. Schulze. 1904: i-io. 5. In der Zeitschrift für physiologische Chemie Untersuchungen über die Amidosäuren, welche bei der Zersetzung der Eiweissstoffe durch Salzsäure und durch Barytwasser entstehen. E. Schulze, J. Barbieri und E. Bosshard. 9: 63-126, 253-259 (1885). Zur Kenntnis des Vorkommens von Allantoin, Asparagin, Hypoxanthin und Guanin in den Pflanzen. E. Schulze und E. Bosshard. 9: 420-444 (1885). Notiz betreffend die Bildung von Sulfaten in keimenden Erbsen. E. Schulze. 9: 616 (1885). Ueber einen neuen stickstoffhaltigen Pflanzenbestandteil. E. Schulze und E. Bosshard. ig: 80-89 (1886). Untersuchung über die Amidosäuren, welche bei der Zersetzung der Eiweissstoffe durch Salzsäure und durch Barytwasser entstehen. Zzveite Abhandlung. E. Schulze und E. Bosshard. 10:134-145 (1886). Ueber das Vorkommen von Vernin im Blütenstaub von Corylus avellana und von Pinus sylvestris. E. Schulze und A, von Planta, ig : 326-330 (1886). Ueber das Arginin. E. Schulze und E. Steiger, ii : 43-65 (1887). Zur Kenntnis der beim Eiweisszerfall entstehenden Phenylamidopro- pionsäure. E. Schulze und E. Nägeli. ii : 201-206 (1887). Ueber das Vorkommen von Cholin in Keimpflanzen. E. Schulze. 11:365-372 (1887). Ueber einige stickstoffhaltige Bestandteile der Keimlinge von Soja hispida. E.Schulze. 12:405-415 (1888). 1912] Ernst Winterstein ii Ueber den Lecithingehalt der Pflanzensamen. E, Schulze und E. Steiger. 13:365-384(1889). Zur Chemie der Pflanzenzellmembran. E. Schulze, E. Steiger und W. Maxwell. 14: 227-273 (1890). Bilden sich Cholesterine in Keimpflanzen, welche bei Lichtabschluss sich entwickeln? E. Schulze. 14: 491-521 (1890). Ueber die Farbenreaktion des Isocholesterins mit Essigsäureanhydrid und Schwefelsäure. E. Schulze. 14: 522-523 (1890). Ueber die basischen Stickstoffverbindungen aus den Samen von Vicia sativa und Pisum sativum. E. Schulze. 15: 140-160 (1891). Ueber das Lecithin der Pflanzensamen. E. Schulze und A. Likier- NiK. 15: 405-414 (1891). Zur Chemie der pflanzlichen Zellmembranen. (Zweite Abhandlung.) E. Schulze. 16: 387-438 (1892). Ueber einige stickstoffhaltige Bestandteile der Keimlinge von Vicia sativa. E. Schulze. 17: 193-216 (1893). Ueber die Konstitution des Leucins. E. Schulze und A. Likiernik. 17: 513-535 (1893). Zur Chemie der pflanzlichen Zellmembranen. (Dritte Abhandlung.) E. Schulze. 19: 38-69 (1894). Ueber die Bestimmung des Lecithingehaltes der Pflanzensamen. E. Schulze. 20:225-232(1895). Ueber das wechselnde Auftreten einiger krystallinischen Stickstoffver- bindungen in den Keimpflanzen und über den Nachweis derselben. E. Schulze. 20: 306-326 (1895). Ueber das Vorkommen von Glutamin in grünen Pflanzenteilen. E. Schulze. 20:327-334(1895). Ueber die Verbreitung des Rohrzuckers in den Pflanzen, über seine physiologische Rolle und über lösliche Kohlenhydrate, die ihn be- gleiten. E. Schulze und S. Frankfurt. 20: 511-555; 21: 108 (1895). Ueber die Zellwandbestandteile der Cotyledonen von Lupinus luteus und Lupinus angustifolius und über ihr Verhalten während des Keimungs Vorganges. E. Schulze. 21: 392-411 (1895). Ueber das Vorkommen von Nitraten in Keimpflanzen. E. Schulze. 22: 82-89 (1896). Ueber einen phosphorhaltigen Bestandteil der Pflanzensamen. E. Schulze und E. Winterstein. 22 : 90-94 (1896). Ueber das wechselnde Auftreten einiger krystallisierbaren Stickstoff- verbindungen in den Keimpflanzen. (Zzveite Abhandlung.) E. Schulze. 22:411-434(1896). ^ 12 Ernst Schuhe [Sept. Ueber die beim Umsatz der Proteinstoffe in den Keimpflanzen einiger Coniferenarten entstehenden Stickstoffverbindungen. E. Schulze. 22:435-448 (1896). Ueber den Umsatz der Eiweissstoft"e in der lebenden Pflanze. E. Schulze. 24: 18-114 (1898). Ueber die Spaltungsprodukte der aus den Coniferensamen darstell- baren Proteinstoffe. E. Schulze. 24 : 276-284 ; 25 : 360-362 (1898). Ueber die Bildung von Ornithin bei der Spaltung des Arginins und über die Konstitution dieser beiden Basen. E. Schulze und E. Winterstein. 26: 1-14 (1898). Ueber den Eiweissumsatz und die Bildungsweise des Asparagins und des Glutamins in den Pflanzen. E. Schulze. 26 : 41 1-426 ( 1899) . Ueber die Verbreitung des Rohrzuckers in den Pflanzen, über seine physiologische Rolle und über lösliche Kohlenhydrate, die ihn be- gleiten. (Zweite Abhandlung.) E.Schulze. 27:267-291(1899). Nachweis von Histidin und Lysin unter den Spaltungsprodukten der aus Coniferensamen dargestellten Proteinsubstanzen, E. Schulze und E. Winterstein. 28: 459-464 (1899). Ueber das Vorkommen von Histidin und Lysin in Keimpflanzen. E. Schulze. 28: 465-470 (1899). Einige Bemerkungen über das Arginin. E. Schulze. 29 : 329-333 (1900). Ueber den Umsatz der Eiweissstoffe in der lebenden Pflanze. (Zweite Abhandlung.) E. Schulze. 30: 241-312 (1900). Ueber die Ausbeute an Hexonbasen, die aus einigen pflanzlichen Ei- weissstoffen zu erhalten sind. E. Schulze und E. Winterstein. 33: 547-573 (1901)- Beiträge zur Kenntnis des Arginins und Ornithins. E. Schutlze und E, Winterstein. 34: 128-147 (1901). Ueber die Trennung des Phenylalanins von anderen Aminosäuren. E. Schulze und E. Winterstein. 35: 210-220 (1902). Beiträge zur Kenntnis einiger aus Pflanzen dargestellten Aminosäuren. E. Schulze und E. Winterstein. 35: 299-314 (1902). Beiträge zur Kenntnis der Hemicellulosen. E. Schulze und N. Castoro. 37: 40-53 (1902). Beiträge zur Kenntnis der Zusammensetzung und des Stoffwechsels der Keimpflanzen. E. Schulze und N. Castoro. 38: 200-258 (1903)- Beiträge zur Kenntnis der Hemicellulosen. E. Schulze und N. Castoro. 39: 318-328 (1903). igi2] Ernst Wintersfein 13 Zur Kenntnis der aus Pflanzen darstellbaren Lecithine. (Erste Mit- teilung.) E. Schulze und E. Winterstein. 40:101-119(1903). Ein Nachtrag zur Abhandlung über einen phosphorhaltigen Bestand- teil der Pflanzensamen. E. Schulze und E. Winterstein. 40 : 120-122 (1903). Beiträge zur Kenntnis der in ungekeimten Pflanzensamen enthaltenen Stickstoflfverbindungen. E. Schulze und N. Castoro. 41 : 455-473 (1904). Einige Notizen über das Lupeol. E. Schulze. 41: 474-476 (1904). Findet man in Pflanzensamen und in Keimpflanzen anorganische Phos- phate? E. Schulze und N. Castoro. 41: 477-484 (1904), Beiträge zur Kenntnis der Zusammensetzung und des Stoflfwechsels der Keimpflanzen. (Zzveite Mitteilung.) E. Schulze und N. Cas- toro. 43: 170-198 (1904). Ueber das Vorkommen von Ricinin in jungen Keimpflanzen. E. Schulze und E. Winterstein. 43: 211-221 (1904). Ueber das Verhalten des Cholesterins gegen das Licht. E. Schulze und E. Winterstein. 43: 316-319 (1904). Ueber die aus den Keimpflanzen von Vicia sativa und Lupinus albus darstellbaren Monoaminosäuren. E. Schulze und E. Winter- stein. 45: 38-60 (1905). Ueber das spezifische Drehungsvermögen einiger aus Pflanzen darge- stellten Tyrosinpräparate. E. Schulze und E. Winterstein. 45:79-83(1905)- Neue Beiträge zur Kenntnis der Zusammensetzung und des Stoff- wechsels der Keimpflanzen. E. Schulze. 47: 507-569 (1906). Ueber den Tyrosingehalt der Keimpflanzen von Lupinus albus. E. Schulze und N. Castoro. 48: 387-395 (1906). Bildet sich Homogentisinsäure beim Abbau des Tyrosins in den Keim- pflanzen? E. Schulze und N. Castoro. 48: 396-411 (1906). Ueber das Verhalten des Cholesterins gegen das Licht. (Zweite Mit- teilimg.) E. Schulze und E. Winterstein. 48:546-548(1906). Ist die bei Luftzutritt eintretende Dunkelfärbung des Rübensaftes durch einen Tyrosin- und Homogentisinsäuregehalt dieses Saftes bedingt? E. Schulze. 50: 508-524 (1907). Ueber den Phosphorgehalt einiger aus Pflanzen dargestellter Lecithin- präparate. E. Schulze. 52: 54-61 (1907). Zum Nachweis des Rohrzuckers in Pflanzensamen. E. Schulze. 52:404-411 (1907). Ueber die zur Darstellung von Lecithin und anderen Phosphatiden 14 Ernst Schulze [Sept. aus Pflanzensamen verwendbaren Methoden. E. Schulze. 55 : 338-351 (1908). Einige Bemerkungen zu den Arbeiten über den Nährwert der in den Pflanzen enthaltenen Amide. E.Schulze. 57:67-73(1908). Ueber den Calcium- und Magnesiumgehalt einiger Pflanzensamen. E. Schulze und Ch. Godet. 58: 156-161 (1908). Ueber das Stachydrin. E. Schulze und G. Trier. 59:233-235(1909). Ueber die zur Darstellung von Cholin, Betain und Trigonellin aus Pflanzen verwendbaren Methoden und über die quantitative Be- stimmung dieser Basen. E.Schulze. 60:155-179(1909). Untersuchungen über die in den Pflanzensamen enthaltenen Kohlen- hydrate. E. Schulze und Ch. Godet. 61: 279-350 (1909). Ueber das Vorkommen von Betain in den Knollen des Topinamburs (Helianthus tuberosus). E. Schulze. 65: 293-294 (1910). Studien über die Proteinbildung in reifenden Pflanzensamen. E. Schulze und E. Winterstein. 65: 431-476 (1910). Ein Beitrag zur Kenntnis des Vernins. E. Schulze. 66: 128-136 (1910). Ueber die in den Pflanzen vorkommenden Betaine. E. Schulze und G. Trier. 67: 46-58 (1910). Ueber das Stachydrin und über einige neben ihm in den Stachysknollen und in den Orangenblättern enthaltene Basen. E. Schulze und G. Trier. 67: 59-96 (1910). Ueber das Vorkommen von Hemicellulosen in den Samenhülsen von Pisum sativum und von Phaseolus vulgaris. E. Schulze und U. Pfenninger. 68: 93-108 (1910). Erwiderung auf R. Engelands Bemerkungen zu den Abhandlungen über die pflanzlichen Betaine und das Stachydrin. E. Schulze und G. Trier. 6g: 326-328 (1910). Ein Beitrag zur Kenntnis der in den Pflanzensamen enthaltenen Kohl- enhydrate. E. Schulze und U. Pfenninger. 69: 366-382(1910). Ueber die Identität des Vernins und des Guanosins, nebst einigen Be- merkungen über Vicin und Convicin. E. Schulze und G. Trier. 70: 143-151 (1910)- Studien über die Proteinbildung in reifenden Pflanzensamen. {Zweite Mitteilung.) E. Schulze. 71: 31-48 (1911). Untersuchung über die in den Pflanzen vorkommenden Betaine. "E. Schulze und U. Pfenninger. 71: 174-185 (1911). Zur Frage der Identität des aus Melasse dargestellten Guaninpentosids mit dem Vernin. E. Schulze und G. Trier, 76 : 145-147 (1912). I9I2] Ernst Winterstein 15 Untersuchungen über die in den Pflanzen vorkommenden Betaine. (Zzueite Mitteilung.) E. Schulze und G. Trier. 76: 258-290 (1912). Dasselbe. (Dritte Mitteilung.) E. Schulze und G. Trier. 79:235- 242 (1912). 6. Im Journal für praktische Chemie Ueber die Zusammensetzung der rohen Schafwolle. M. Märcker und E. Schulze. 108: 193-207 (1870). Ueber die Zusammensetzung des Wollfetts. E. Schulze. 7 (n. f.) : 1-16 (1873). Dasselbe. E. Schulze und A. Urich. 9: 321-339 (1874). Ueber die Eivveisszersetzung in Kürbiskeimlingen. E. Schulze und J. Barbieri. 20: 385-418 (1880). Ueber das Vorkommen von Allantoin und Asparagin in jungen Baum- blättern. E. Schulze und J. Barbieri, 25: 145-158 (1882). Zur Kenntnis der Cholesterine. E. Schulze und J. Barbieri. 25: 159-180 (1882). Ein Nachtrag zu der Abhandlung: "Zur Kenntnis der Cholesterine." E. Schulze. 25: 458-462 (1882). Ueber Phenylamidopropionsäure, Amidovaleriansäure und einige andere stickstoffhaltige Bestandteile der Keimlinge von Lupinns luteus. E. Schulze und J. Barbieri. 27: 337-362 (1883). Zur quantitativen Bestimmung des Asparagins und des Glutamins. E. Schulze. 31:234-246 (1885). Zur Kenntnis der stickstoffhaltigen Bestandteile der Kürbiskeimlinge. E. Schulze. 32: 433-460 (1886). 7. In den Berichten der Deutschen Chemischen Gesellschaft Über Maltose. E. Schulze. 7: 1047 (1874). Über die Zusammensetzung des Wollfetts. E.Schulze. 8:570(1875). Selenoidodiglykolsäure. A. Urich und E. Schulze. 8: yyT, (i875)- Die stickstoffhaltigen Bestandteile der Rüben. E. Schulze und A. Urich. 9:80 (1876). Keimung der Lupinensamen. E. Schulze und W. Umlauft. 9: 1314 (1876). Über die stickstoffhaltigen Bestandteile der Runkelrüben (Glutamin). E. Schulze und A. Urich. ig: 88 (1877). Über das Vorkommen eines Glutaminsäureamids in den Kürbiskeim- lingen. E. Schulze und J. Barbieri. ig: 199 (1877). Eiweisszersetzung in Keimpflanzen. E. Schulze, ii: 520 (1878). l6 Erfist Schuhe [Sept. Bildung von schwefelsauren Salzen bei der Eiweisszersetzung in Keim- pflanzen. E. Schulze, ii: 1234 (1878). Asparagin und Tyrosin in Kürbiskeimlingen. E. Schulze und J. Bar- BiERi. 12 : 710 (1879). Leucin aus Kürbiskeimlingen. E. Schulze und J. Barbieri. 12 : 1233 (1879). Über ein Glucosid aus Lupinus luteus. E. Schulze und J. Barbieri. 12: 2200 (1879). Über das spezifische Drehungsvermögen des Isocholesterins. E. Schulze. 13: 249 (1880). Über ein neues Glucosid. E. Schulze und J. Barbierl 13:681(1880), Amidosäuren in Lupinenkeimlingen. E. Schulze und J. Barbieri. 13: 1924 (1880). Über die Eiweisszersetzung in Kürbiskeimlingen. E. Schulze und J. Barbieri. 13: 2386 (1880). Über das Vorkommen von Allantoin im Pflanzenorganismus. E. Schulze und J. Barbieri. 14: 1602 (1881). Über das Vorkommen von Phenylamidopropionsäure unter den Zersetz- ungsprodukten der Eiweissstoffe. E. Schulze und J. Barbieri. 14: 1785 (1881). Zur Kenntnis des Cholesterins. E. Schulze und J. Barbieri. 15: 953 (1882). Über das Vorkommen von Allantoin und Asparagin in jungen Baum- blättern. E. Schulze und J. Barbieri. 15:955(1882). Beiträge zur Kenntnis der stickstoffhaltigen Bestandteile der Kartoffeln. E. Schulze und E. Eugster. 15: 1090 (1882). Über das optische Verhalten einiger Aminosäuren. E. Schulze und E. Bosshard. 17: 1610 (1884). Über die Bildung von Phenylamidopropionsäure beim Erhitzen von Eiweissstoffen mit Salzsäure und Zinnchlorür. E. Schulze und J. Barbieri. 17: 171 1 (1884). Über das optische Verhalten einiger Aminosäuren. E. Schulze und E. Bosshard. 18: 388 (1885). Über das Vorkommen von Glutamin in den Zuckerrüben und über das optische Verhalten desselben. E. Schulze und E. Bosshard. 18 : 390 (1885). Über einen neuen stickstoffhaltigen Bestandteil der Keimlinge von Lupinus luteus. E.Schulze. 19:1177(1886). Über Paragalactan. E. Schulze. 20: 290 (1887). Bilden sich Nitrate im Organismus lebender Pflanzen? E. Schulze. 20: 1500 (1887). I9I2] Ernst Winterstein 17 Über das Vorkommen von Cholin in Keimpflanzen. E. Schulze. 21 : 21 (1888). Über das Vorkommen von Rohrzucker in unreifen Kartoffeln. E. Schulze und Th. Seliwanow. 21 : 299 (1888). Über den Nachweis von Rohrzucker in vegetabilischen Substanzen. E. Schulze und Th. Seliwanow. 21 : 299 (1888). Ein Beitrag zur Veränderung, welche die stickstoffhaltigen Bestand- teile eingesäuerter Grünfutterstoffe erleiden. E. Schulze. 21 : 668 (1888). Über das Vorkommen eines unlöslichen Schleimsäure gebenden Kohlen- hydrats in Rotklee und Luzerne. E. Schulze und E. Steiger. 22:345 (1889). Über die Zersetzung der Proteinsubstanzen in verdunkelten grünen Pflanzen. E. Schulze und E. Kisser. 22:350(1889). Über einige stickstoffhaltige Bestandteile der Keimlinge von Soja his- pida. E. Schulze. 22: 599 (1889). Zur Kenntnis der chemischen Zusammensetzung der Pflanzenzellmem- branen. E. Schulze. 22: 1192 (1889). Betain und Cholin in den Samen von Vicia sativa. E. Schulze, 22 : 1827 (1889). Untersuchungen über die stickstofffreien Reservestoffe der Samen von Lupinus luteus und über die Umwandlung derselben während des Keimprozesses. E. Schulze und E. Steiger. 23: 405 (1890). Über ein Krystallisieren des Kohlenhydrats. A. v. Planta und E. Schulze. 23: 1692 (1890). Zur Kenntnis der chemischen Zusammensetzung der pflanzlichen Zell- membran. E.Schulze. 23:2579(1890). Darstellung von Lecithin aus Pflanzensamen. E. Schulze und A. LiKiERNiK. 24:71 (1891). Über den Lecithingehalt der Pflanzensamen. E. Schulze und E. Steiger. 24: 327 (1891). Zur Chemie der Pflanzenzellmembran. E. Schulze, E. Steiger und W. Maxwell. 24: 530 (1891). Über die Konstitution des Leucins. E. Schulze und A. Likiernik. 24: 669 (1891). Bilden sich Cholesterine in Keimpflanzen, welche bei Lichtabschluss sich entwickeln? E. Schulze. 24: 670 (1891). Über die Farbenreaktion des Isocholesterins mit Essigsäureanhydrid und Schwefelsäure. E. Schulze. 24: 671 (1891). Über die Bildung von stickstoffhaltigen Basen beim Eiweisszerfall im Pflanzenorganismus. E. Schulze. 24: 1098 (1891). i8 Ernst Schuhe [Sept. Zur Kenntnis der chemischen Zusammensetzung der pflanzlichen Zell- membran. E. Schulze. 24: 2277 (1891). Über die Bildung von Harnstoff bei der Spaltung des Arginins. E. Schulze und A. Likiernik. 24: 2701 (1891). Zur Kenntnis des Stachydrins, A. v. Planta und E. Schulze. 24: 2705 (1891). Über basische Stickstoffverbindungen in den Samen von Vicia sativa und Pisum sativum. E. Schulze. 25: 84 (1892). Über das Lecithin der Pflanzensamen. E. Schulze und A. Likiernik, 25:85 (1892). Zur Chemie der pflanzlichen Zellmembranen. E. Schulze. 25 : 434 (1892). Über das Vorkommen von Guanidin im Pflanzenorganismus. E. Schulze. 25:658(1892). Über einen stickstoffhaltigen Bestandteil der Keimlinge von Vicia sativa, E. Schulze. 25: 869 (1892). Zum Nachweis des Guanidins. E. Schulze. 25: 2213 (1892). Über das Vorkommen von Betain und Cholin in Malzkeimen und in den Keimen des Weizenkorns. E. Schulze und S. Frankfurt. 26: 2151 (1893). Über die Verbreitung des Rohrzuckers in Pflanzen. E. Schulze und S. Frankfurt. 27: 62 (1894). Über das Vorkommen von Raffinose im Keime des Weizenkorns. E. Schulze und S. Frankfurt. 27: 64 (1894). Über krystallisiertes Lävulin. E. Schulze und S. Frankfurt. 27: 65 (1894). Über das Vorkommen von Trigonellin in den Samen von Pisum sati- vum und Cannabis sativa. E. Schulze und S. Frankfurt. 27 : 769 (1894). Über ;8-Lävulin. E. Schulze und S. Frankfurt. 27: 3525 (1894). Vorkommen von Arginin in Knollen und Wurzeln einiger Pflanzen. E, Schulze. 29: 352 (1896). Verbreitung des Glutamins in den Pflanzen. E. Schulze. 29: 1882 (1896). Stickstoffhaltige Bestandteile der Keimpflanzen von Ricinus communis. E. Schulze. 30: 2197 (1897). Über die Spaltungsprodukte des Arginins. E. Schulze und E. Win- terstein. 30:2879(1897). Bestandteile des Wollfetts. E. Schulze. 31: 1200 (1898). Konstitution des Arginins. E. Schulze und E. Winterstein. 32 : 3191 (1899). I9I2] Ernst Winterstein 19 Über das spezifische Drehungsvermögen des Glutamins. E. Schulze. 39: 2932 (1906). Die Konstitution des Stachydrins, E. Schulze und G. Trier. 42 : 4654 (1909). 8. In verschiedenen Zeitschriften Ueber die Elementarzusammensetzung der tierischen Fette, insbeson- dere der Fette vom Schaf, Rind und Schwein. E. Schulze und A. Reinecke : Annalen der Chemie und Pharmacie, 142 ; 191-218 (1867). Untersuchungen über die Respiration des volljährigen Schafes bei Erhaltungsfutter. W. Henneberg, E. Schulze, M. Märcker und L. Busse : Zentralblatt für die medizinischen Wissenscliaften, 1870; 353-356; 369-370. Ueber Stickstoffausscheidung im Harn der Wiederkäuer. E. Schulze und M. Märcker: Zeitschrift für Biologie, 7; 49-62 (1871). Ueber die Bestimmung des aus Amiden abspaltbaren Ammoniaks in Pflanzenextrakten. E. Schulze. Zeitschrift für analytische Chemie: 21 : 1-26 (1882). Ueber das Stachydrin. E. Schulze: Archiv der Pharmacie, 231 ; 305 (1893). ^ Zur quantitativen Bestimmung der Kohlenhydrate. E. Schulze: Chemikerzeitung, 1894; 527. Ueber die Analyse der Pflanzensamen. E. Schulze: Ibid., 1894; No. 43- Ueber die Cellulose. E. Schulze: Ibid., 1895; No. 65. Inwieweit stimmen der Pflanzenkörper und der Tierkörper an ihrer chemischen Zusammensetzung überein und inwiefern gleicht der pflanzliche Stoffwechsel dem tierischen? E. Schulze: Viertel- jahr sschrift der Naturforschenden Gesellschaft in Zürich, 1894; 243- (A) Verbreitung des Glutamins in den Pflanzen. (B) Die in den Keimpflanzen der Coniferen enthaltenen Stickstoffverbindungen. E. Schulze. Verhandl. d. Schweiz. Naturf. Gesellsch., Zürich (1896), 126-127. Ueber die Zellwandbestandteile der Cotyledonen von Lupinus Intens und Lupinus angustifolius und über das Verhalten während der Keimungsvorgänge. E. Schulze : Berichte der Deutschen Bo- tanischen Gesellschaft, 14; 66-71 (1896). ^ Ueber den Eiweisszerfall und Eiweissbildung in der Pflanze. E. Schulze : Ibid., 18 ; 36-42 ( 1900) . 20 Ernst Schulze [Sept. Ueber Tyrosinbildiing in den keimenden Samen von Lupinus albus und über den Abbau primärer Eiweisszersetzungsprodukte in den Keimpflanzen. E. Schulze: Ibid., 21; 64-67 (1903). Ueber die Argininbildung in den Keimpflanzen von Lupinus luteus. E. Schulze: Ibid., 22 ; 381-384 (1904). 9. Dissertationen Ueber die Eiweisssubstanz der Kürbissamen und über die Zersetzungs- produkte, welche während des Keimprozesses aus derselben ent- stehen. J. Barbieri. 1878. Zur Kenntnis des Glutamins. Ueber Ammoniakbestimmung in Pflanz- ensäften und Pflanzenextracten. E. Bosshard. 1880. Ueber die chemische Zusammensetzung der Samen von Lupinus luteus und über ein in denselben enthaltenes dextrinartiges Kohlenhy- drat. E. Steiger. 1886. Ueber das pflanzliche Lecithin und über einige andere Bestandteile der Leguminosenchalen. A. Likiernik. 1891. Zur Kenntnis des pflanzlichen Amyloids und über einige andere Be- standteile der pflanzlichen Zellmembranen. E. Winterstein. 1892. Ueber die Zusammensetzung der Samen und etiolierten Keimpflanzen von Cannabis sativa und Helianthus annuus. S. Frankfurt. 1893. Ueber die Zusammensetzung der Samen und der etiolierten Keim- pflanzen von Lupinus angustifolius. Miron Merlis. 1897. Ueber die Bestandteile der Samen von Picea excelsa und über die Spaltungsprodukte der aus diesen Samen darstellbaren Protein- stoffe. N. Rongger. 1898. Versuche zur quantitativen Bestimmung der bei der Zersetzung der Eiweisskörper durch Säuren entstehenden Basen. O. Meyer. 1900. Versuche zur Bestimmung des Gehaltes einiger Pflanzen und Pflanzen- teile an Zellwandbestandteilen, Hemicellulosen und Cellulosen. A. Kleiber. 1900. Beiträge zur Kenntnis der Cholesterine und der Methoden, die zu ihrer Abscheidung aus den Fetten und zu ihrer quantitativen Bestim- mung verwendbar sind. E. Ritter. 1902. Beiträge zur Kenntnis der in den Pflanzensamen enthaltenen Kohlen- hydrate. Ch. Godet. 1909. Ein Beitrag zur Kenntnis der pflanzlichen Betaine und ihre Bedeutung. Das Stachydrin, seine Konstitution und seine Synthese. G. Trier. 1910. A RESUME OF THE LITERATURE ON INOSITE- PHOSPHORIC ACID, WITH SPECIAL REFER- ENCE TO THE RELATION OF THAT SUBSTANCE TO PLANTS ANTON RICHARD ROSE (Laboratory of Biological Chemistry of Columbia University, at the College of Physicians and Surgeons, New York) Contents. — Discovery: by the microscopist, 21; by the chemist, 22. Occur- rence, 2^; preparation, 25; properties, 26; Constitution, 31; terminology and Clas- sification, 35 ; analytical methods, 37 ; role in plants, 39. Bibliography, 46. DISCOVERY OF INOSITE-PHOSPHORIC ACID SALTS BY THE MICROSCOPIST In 1854 Hartig/ engaged in a microscopic study of the seeds o£ various plants, noted in all his sections small particles similar to the starch grains of the potato, which at that time were absorbing the interest of plant physiologists.^ These grains were obviously not starch, as they did not give the characteristic blue color with iodine in potassium iodid Solution. They are more commonly present in seeds than starch, the latter being frequently replaced by fat. Hartig considered them an essential reserve product designed to play an important part in the germination of the seed and the growth of the plant. He first called them " Klebermehl " but within a year renamed them " aleurone grains " f rom the Greek aXevpov (wheat fiour), a term still in use. Not only did he consider them significant for the plants in which they are found but also for the animals which eat them. The method which he employed for separating them from the other parts of the seed is the one usually followed by the investigators who have since worked with these ^ The papers ref erred to in the text and not accompanied by f ootnote ref er- ences are those which pertain specially to the literature of inosite-phosphoric acid and are given at the end of this review in the alphabetical order of the names of the authors. ^The starch grains were minutely studied by Nägeli and his coworkers. Their work (collected in Die Stärkekörner, 1858) probably afforded the Stimulus for the efforts which led to the discovery of inosite-phosphoric acid so early. 21 22 Literature on Inosite-Phosphoric Acid [Sept. grains, namely, extraction of the macerated seeds with ether and removal of the aleuron grains from the cellular debris by Sedimenta- tion in this medium. They are insoluble in both alcohol and ether, somewhat soluble in water, and quite soluble in dilute acids. In the years following Hartig's discovery several other botanists turned their attention to the aleuron grains and came to conflicting conclusions as to their nature. Von Holle considered them protein carriers and referred to them as " Proteinkörner." Both Sachs and Gris looked upon the particles as fat concentrates. By far the most important study in this field was the comprehensive work of Pfeffer in 1872. He differentiated the grains described by Hartig into three groups : (i) crystals of calcium Oxalate, (2) a protein sub- stance, and (3) a Compound giving no reactions for protein, fat, or inorganic salts. This last type was found in all of the one hundred different seeds which he examined. These particles he described as having rounded surfaces, assuming spheroidal shapes and fre- quently twinning, so as to present a convoluted appearance. Enough of the grains were obtained for a chemical examination, which was made for him by his colleague Brandon. The solubilities were the same as those reported by Hartig. Nitrogen could not be detected. Positive tests were obtained for calcium, magnesium, and phos- phorus. Organic matter was noted and the Suggestion made that the substance was a phosphate combined with a carbohydrate. These phosphorus-bearing spheroidal bodies occurring with or in the aleuron grains Pfeffer named "Globoid." DISCOVERY OF INOSITE-PHOSPHORIC ACID BY THE CHEMIST Palladin, while engaged on a study of the proteins of Sinapis niger in 1893, observed an unusual phenomenon. After extraction of the fat-free finely ground seeds with ten per cent. sodium chlorid Solution and heating the extract, he obtained a voluminous precipi- tate which partly dissolved on standing. A few trials showed that he had a substance soluble in cold but insoluble in hot water. By filtering off the permanent coagulum, reheating the filtrate and filtering while hot, he obtained a fairly pure product rieh in phos- phorus, containing calcium and magnesium, but no nitrogen. It proved non-reducing when tested with Fehling Solution, both before I9I2] Anton Richard Rose 23 and after hydrolysis with acids. It was soluble in water and aclds, and precipitated by the alkali earths and the heavy metals. Subsequently Schulze and Winterstein published a paper con- firming the observations of Palladin, and also noting that the phos- phorus was not precipitated by ammonium molybdate. These authors expressed the opinion that the Compound thus discovered by chemical procedure is identical with Pfeffer's " globoid." The fol- lovving year (1897), a more detailed paper was published by the junior author, in which the identity and properties of the substance were more fully revealed, and " inosite-phosphoric acid " was sug- gested as the proper name for the Compound, inasmuch as it yielded inosite and phosphoric acid on hydrolysis. The most complete study of this substance has been made by Posternak; his findings are embodied in eight papers and several applications for patents. He discarded the name suggested by Winterstein and proposed a structural formula which does not include the inosite ring. He gave to the substance the name "phytin" (from the Greek vT7]v) and under this trade name it is now placed on the market by a chemical firm in Basel. OCCURRENCE OF INOSITE-PHOSPHORIC ACID As already noted, phytin was first thought to be a storage product in seeds ; and this early Impression has been confirmed by subsequent investigation, no case having been reported of a seed in which it is completely lacking. The accompanying table (i) lists the plants specially mentioned in the literature in connection with the study of inosite-phosphoric acid. The relative data in the table are not in close accord, but no true comparison can as yet be drawn between the species, for most of the data were obtained at periods when adequate and uniform analytical methods were unavailable. The figures quoted in the table give an approximate idea of the quantitative signijficance of this important Compound, in relation to other forms of phosphorus available for seedling growth and the phosphorus requirement of man and beast. Posternak makes the Statement that seeds rieh in fat carry the largest amount of phosphorus, which is in harmony with the micros- copist's observations that the aleuron grains are particularly numer- 24 Literatur e on Inosite-Phosphoric Acid [Sept. TABLE I Recorded analytic data on the occurrence of inosite-phosphoric acid Plant. Total P Per Cent. Spruce fir 0.66 Pea (Pisum sativum).. 0.37 Pea, yellow Bean, white {Phaseolus vulgaris) 0.51 Bean, brown Hemp (Cannabis sativa) 1.46 0.76 Rice (Ory::a sativa)... 0.95 Rice flour Rice bran 2.22 Rice germ 6.20 Wheat ( Triticum sativa) 0.45 Wheat bran i.ii i.il Graham flour Sesame (Sesamum in- dicum) 0.77 Corn (Zea mays) 0.29 0.35 Oats (Avena sativa) . . 0.46 Barley (Hordium sati- vutn) 0.50 0.47 0.54 Barley bran Sunflower {Helianthus annuus) 0.83 Rye (Seeale cereale) . . 0.43 Ratio of the P in phyto- phosphate to Total P Per Cent. 91-5 70.8 19.0 81.6 58.0 91.4 15-0 45-9 69.0 74.1 89.2 29.9 84.0 52.0 29.0 16.3 54.0 48.9 48.0 87.4 Plant. 38.0 36.S 44.0 60.4 86.3 90.3 28.9 Total P Per Cent. Ratio of the P in phyto- phosphate to Total P Per Cent. 25.0 Rye flour Rape (Brassica napus olifera) 0.98 80.0 I-I9 44-5 0.54 38.0 Rape cake 49.5 Soy bean (Soja hispida) 0.57 58.0 Lentil (Lens esculenta) 0.30 82.6 0.30 9.3 Cocoanut (Cocus nuci- fera) 88.4 695 Cottonseed (Gossipium herhaceum) 93.6 Pine : Pinus cembra . . . 0.47 14.39 Pinus excelsa ... 0.63 21.6 Castor bean (Ricinus communis) 0.26 41.6 Millet (Panicum millia- ceum) 0.77 44-97 Vetch (Vicia faba mi- nor) 0.47 4.4 Red Clover (Hay) 0.24 70.0 Radish : Root (Rapha- nus vulgaris) 0.02 15.0 Turnip : Root (Bras- sica esculenta?) 0.02 15.0 Dahlia : Tuber (Dahlia variabilis) ..." Spheroids of phjrtin " Potato : Tuber (Alliuni cepa ) " Spheroids of phytin " The analytic results in the above table are those for seeds of the plants, except in the last five cases. They are compiled from a number of sources. Among the plants studied for their phyto-phosphate content, in which the rela- tive amount of this substance is not given, are the following: Beta vulgaris, Brassica campestris, Cucurbita pepo, Ervum lens, Lupinus albus, L. angustifolius, L. luteus, Pinus laricio, P. maritima, Sinapis nigra, Solanum tuberosum, and the tubers of AI Hunt cepa and Dahlia variabilis. In only two materials reported, namely, rutabaga root and alfalfa hay, could no phyto-phosphate be found. In several instances the total phosphorus was not reported. Where there is a close agreement between two or more results, only one figure is given above. 1912] Anton Richard Rose 25 ous in oily seeds. He also remarks that the smaller seeds such as cereals are the richest in " phytin." This Compound is not entirely confined to seeds, its presence having also been noted in the potato near the eye and as characteristic spheroids in the tubers of Allium and Dahlia. Roots functioning as storage organs, such as those of the Brassicae, contain small amounts. None was found by Totting- ham and Hart in the mature stems and leaves of the common fodder plants, but it occurs in clover leaves and in millet during the late flovvering period, and also in tender shoots. PREPARATION OF INOSITE-PHOSPHORIC ACID AND ITS SALTS To prepare phytin or its closely related Compounds from seeds, they should be finely ground and, if fat is present in large amounts, it should be removed by extraction with ether and alcohol. Most of the preparations reported in the literature have been obtained from cereals by leaching with 0.2 per cent. hydrochloric acid Solu- tion. Acetic acid has also been used in i per cent. Solution, and in a few cases acid Solutions of greater concentration have been employed. To remove the soluble proteins from the extract, Levene used picric acid; other investigators have coagulated them by heating and filtration after cooling; but when acidulated water is used the proteins do not seem to interfere appreciably with the preparation of pure phytin. The reserve proteins of the seeds are of the globulin type and are soluble only in the presence of salt in the extracting agent. Precipitation of inosite-phosphoric acid from its Solutions can be accomplished by several methods, such as the use of the acetates of the heavy metals, barium chlorid in ammoniacal Solution or magnesia mixture. In these cases the precipitated Com- pound is obviously in a form different from that in which it occurs in the original material. To obtain the salt more nearly in the form in which it is found in the seed, it may be precipitated by heating the Solution to almost boiling and filtering while hot; or better, by adding four volumes of ninety-five per cent. alcohol. In obtaining pure preparations of inosite-phosphoric acid or its salts, a number of reprecipitations are necessary. These have been made alternately with copper, lead, and barium. Salts of the first two are decom- posed by suspending in distilled water and bubbling hydrogen sul- 26 Literatlire on Inosite-Phosphoric Acid [Sept. phide through the liquid ; the third is removed by adding dilute sul- phuric acid. In all cases, the lead or copper salt is the last pre- cipitated in this manner of purification; and when the product is carefully washed, and the metal removed by hydrogen sulphide, the filtrate from the lead or copper sulphide is evaporated at a low tem- perature, leaving the inosite-phosphoric acid.^ The various salts which have been studied were made from this acid. In obtaining the acid for the preparation of pure Compounds, the greatest difficulty lies in removing the last traces of magnesium. Rising overcame this difficulty by taking up the syrupy acid with absolute alcohol, and adding ether until droplets of the acid formed. He then filtered off the acid magnesium inosite-phosphate and again evaporated. The commonest impurity in phytin is inorganic ortho- phosphate which, however, is easily removed. Starkenstein uses the calcium salts of the mixed acids and washes with glacial acetic acid, which dissolves the inorganic part but not the organic phosphorus Compound. Forbes precipitates with magnesia mixture, removes the excess of this reagent by washing with ammonia water, washing again with alcohol, and extracting with 95 per cent. alcohol con- taining 0.2 per cent. mineral acid, which also dissolves all the in- organic phosphorus and none of the " phytin." Attempts to prepare these salts synthetically will be referred to in a later section. PROPERTIES OF INOSITE-PHOSPHORIC ACID AND ITS SALTS The substance widely known as "phytin," and described in the middle of the last Century by the microscopists as " spheroid bodies," frequently assumes the globular shape when forced out of Solution, In most cases, the precipitate comes down as a flocculent amorphous mass. Inosite-phosphoric acid has not as yet been obtained in crys- talline form. At room temperature, it is a syrup of light straw color, which becomes very viscid on cooling to — 20° C, and darkens on heating to 100° C. Vorbrodt found that this coloration could not be prevented by replacing the air with an inert gas during the heating, and from this concludes that the change is not due to oxidation. If the heat is allowed to reach 125° C, an insoluble dark char is produced (cf. Posternak). Inosite-phosphoric acid may 'For details of the method of preparing the acid see Hart and Patten (page48). 1912] Anton Richard Rose ^y form neutral salts, acid salts, double salts, or acid double salts. The acid, neutralized with alkali and evaporated to dryness, gives a brownish horny mass; but if an alkali earth is also present, double salts are formed, which crystallize in fine needles with eight mole- cules of water. The magnesium salts crystallize in small and uni- form spherules, while the copper salts form large and irregulär spherules. Twin forms are frequently produced in the copper pre- cipitates, resembling the globoids of which drawings appear in Pfeffer's paper. These spheroid masses may be Clusters of needles of approximately equal lengths, as is suggested by the regularly pitted surfaces sometimes seen, and the term spherocrystal can accordingly be applied to them. The copper Compounds are green ; the others, as far as reported, are white. Occasionally a faint pinkish cast has been noticed in pure preparations. The acid is miscible in all proportions with water. It is soluble in alcohol but not in the other common lipoid solvents. Ether added to an alcoholic Solution precipitates the acid in droplets. According to Posternak, the acid-alkali and acid-magnesium salts are soluble in alcohol and water. The double salts are soluble in water, forming opalescent Solutions from which they are precipitated by chlorid and acetate of potassium, redissolving if these are added in excess. The decrease in solubility of the salts of inosite-phosphoric acid is in the following Order : alkali, alkali earth and heavy metal. The magne- sium Compounds are more soluble than the calcium salts and the latter more soluble than those of barium or Strontium. The same Order of solubility also holds for acid salts, double salts and normal salts. These phytophosphates are more soluble in cold than in hot water, and heating frequently precipitates them, even in the presence of dilute acetic acid. This precipitation is largely influenced by other Compounds in Solution, halogens and sulphates inhibiting, and phosphates facilitating the reaction. Posternak noted that the precipitates thus formed by heating were not always completely dis- solved on cooling; also that the phytophosphates not readily soluble in cold water were changed to more soluble forms by dissolving in dilute acid and precipitating with alcohol. Dilute mineral acids are solvents for all of these Compounds. Acetic acid does not dissolve the salts of inosite-phosphoric acid with the heavy metals, barium. 28 Literature on Inosite-PJwsphoric Acid [Sept. calcium and Strontium ; but the magnesium and alkali salts, and the double salts, are very soluble in this reagent. Posternak says that the alkali salts of this acid are solvents for the Compounds with alkali earths, and that on standing, crystals of double salts form in these Solutions, tending to arrange themselves in rosettes — a further Suggestion as to the mode of formation of the characteristic sphero- crystals mentioned above. Inosite-phosphoric acid Solutions do not polarize light, and pass through semi-permeable membranes com- paratively slowly. All the salts of inosite-phosphoric acid, except the alkali salts and the acid magnesium salts, are precipitated f rom aqueous Solution by four volumes of alcohol. The acid and its salts in alcoholic Solu- tion are precipitated by ether. The addition of neutral Solutions of silver, lead, copper, cadmium, iron, uranium, Strontium, barium and calcium precipitates the acid f rom its Solutions ; so also do magnesia mixture and albumin. According to Posternak, precipitation with copper is prevented by the presence of fat. The copper salts are soluble in ammonium hydroxid Solution. Ammonium molybdate in nitric acid does not cause precipitation in dilute Solutions of inosite phosphates, but in concentrated Solutions white needles are formed on long Standing which are insoluble in nitric acid and soluble in water. Preparations f rom seeds retain persistently small amounts of magnesium, several reprecipitations being necessary to get a salt containing a single metal. It is equally difficult to get a preparation free from the hydrogen ion, and it may be said in general that the property of forming acid salts and double salts is very characteristic of inosite-phosphoric acid. The most important contribution to our knowledge of the nature and properties of its salts has been made recently by Anderson. From acid purified by means of barium pre- cipitation and the method described by Hart and Patten, the follow- ing Compounds have been prepared : tri-barium, penta-barium, penta- barium di-ammonium, penta-magnesium, penta-magnesium di-am- monium, tetra-cupric di-calcium, tetra-calcium, penta-calcium, hexa- cupric, octa-silver, and hepta-silver salts. Most of them are white amorphous powders, but the tri-barium and tetra-calcium salts can be reprecipitated in irregulär crystalline form. Pure preparations have been made and analyzed by several other investigators. The results of their work are given in Table 2. I9I2] Anton Richard Rose 29 TABLE 2 Analytic data pertaining to inosite-phosphoric acid (Compüed from results reported in the literature of the subject) Name of Author Anderson. Contardi. Hart, Patten. Hart. Tottingham. Horner. Levene. Elements Carbon Hydro- gen Phosphorus Barium Magne- sium Calcium Other Metals Ratios' C=6 P=x P=6 M'=x (Prepared from the purified commercial product.)^ 6.42 4-59 10.56 10.76 1-44 1-15 3-21 3.22 37-21 48.87 46.99 14-13 42-9 13-03 22.46 17.66 14.69 16.87 13.46 14.07 21.29 26.37 26.16 19.07 20.62 21.75 22.53 16.88 11.94 13-02 (Synthesized by means of inosite and ortho-phosphoric acid.) 6.42(K) 33-54(Cu) 55-98(Ag) 52.43(Ag) 6 6 6 6 6 6 6 6 6 6 6 6 6 14.24 14-23 9.16 9.60 ( 12.62 8.17 3-45 3-61 1.64 1.68 24.09 24.31 16.34 16.05 35-57 34-48 4 4 4 4 (Synthesized by means of inosite and pyro-phosphoric acid.) 3-24 1.58 (acid) 26.51 16-55 ( 26.08 21.08 35-90 (Prepared from rice bran.) 9.0 13-8 10.89 10.63 17-30 3-00 3-40 3-63 28.10 12.50 15.60 21.40 (Preparations from wheat bran.) 25-98 56.2 23.2 8.9 13-5 22.1 (Cu) 16.38 26.08 5-8 I-13 2.6 (K) 6 2 (Similar preparations.) (The commercial preparation.)- 20.32 1 I 1.45 I 11-96 I 9-84 5.66 17.90 7-70 1.47 3-57 23-00 13-93 13-16 11-95 (Preparations from hemp.) 44-50 40.14 5-5 6 1-5 2 6 10 12 12 12 10 8 IG 12 6 4 (Synthesized by means of inosite and ortho-phosphoric acid.) 12 12 12 12 63 - 1 7 8.5 4 9-5 ^ The empirical formulas have been calculated from the analytic results, and the relations between the carbon and phosphorus, and the phosphorus and base (M'), are given in these two columns. The carbon and the phosphorus are, in each case, assumed to be six atoms per molecule. ^ Placed on the market by the Gesellschaft für Chemische Industrie in Basel. ^ A 0.2 per cent. HCl extract of wheat bran several times reprecipitated from weakly acid Solution by alcohol. * The first substance; obtained by extracting with sodium chlorid Solution and, after removing protein with picric acid, precipitating with copper and removing the copper. 30 Literature on Inosite-Phosphoric Acid [Sept. TABLE 2 (coniinued) Name of Author Elements Carbon Hydro- gen Phosphorus Barium Magne- Calcium Other Metals Ratiosl C=6 P=x P=6 M'=x Posternak. Rising. Suzuki, Yoshimura, Takaishi. Vorbrodt. Winterstein. Winterstein, Schulze. (A large number of preparations were raade in which several kinds of seeds were used.) 9-97 4-79 7-25 7-44 6.51 26.08 25.89 12.70 19-42 19-73 6 6 6 6 3-70 I.OO 1-34 1.49 50-45 8.41 i9.02(Na) 5-37 1 1-03 I (Preparations from barley bran.) 13-08 I I I l52.6s(Ag)| (Preparations from rice bran.) 1.21 I 23.48 I I S-81 I 17-48 I (Preparations from corn.) 15.18 I42.62 I I (Preparations from black mustard.) 18.42 I I 7-8 I I 9.65 I 2.83 I 15-20 I I I 5-5 I - I 3.7 II 4 12 I 7 1 6 I 10.6 8 6.5 — ?* The alkali salts are hygroscopic, but the others do not change in weight under ordinary conditions. Posternak assigns to the double alkali salts of the alkali earths eight molecules of water. The barium salt prepared by Vorbrodt lost 9.33 per cent. of its weight in the presence of phosphorus pentoxid, but regained it when exposed to a moist atmosphere. At 110° C, 11.5 per cent. was lost. Ander- son reports for his tri-barium salt five molecules of water, for his tetra-calcium salt twelve, and for his penta-magnesium salt twenty- four. Inosite-phosphoric acid is easily decomposed by heating with strong acids in sealed tubes but does not spontaneously break down into its cleavage p'roducts. Mendel and Underhill kept a Solution of the acid for many months and found at the end of the time no apparent change. Posternak states that heating phytin in alkaline Solution to 100° C. causes no decomposition, but Winterstein found that if a twenty per cent. Solution of alkali were used (sodium hydroxid) and the temperature raised to 230° C, cleavage occurred. Contardi states that cleavage does not occur when these salts are * Extracted with sodium chlorid Solution, precipitated by heating, and fil- tered hot. I9I2] 'Anton Richard Rose 31 heated in water to 200° C. under pressure. According tc Giacosa it is more readily hydrolyzed than lecithin. From the fact tbat the products of hydrolysis are inosite and phosphate, Winterstein came to the conclusion that the Compound is inosite-phosphoric acid. CONSTITUTION OF INOSITE-PHOSPHORIC ACID Posternak, who has done more work on this substance than any other one investigator, did not agree with Winterstein in the con- clusion set forth above. From his analyses he first constructed the following formula: HC(OH)OP .(OH) (X) I HC(OH)0 P^ (OH) but, as benzoyl chlorid gave no positive test for the hydroxyl group, a second formula was proposed (anhydro-oxymethylene-diphos- phoric acid) : CH,-0-P(' / ^(OH), («) o: \ /^ \cHj-o— ?<;■ %(0H), He was of the opinion that inosite is synthesized from the products of hydrolysis when the "phytin" is heated under pressure with mineral acids. A number of chemists have expressed doubt concern- ing the probability of such a formation of inosite, either from this organic group or any part of it that might result from the action of the acid thereon. In 1907 Suzuki and his co-workers obtained inosite from "phytin" by the action of an enzyme, from which they concluded that inosite is an integral part of the "phytin" molecule and constructed the following formula to represent their view : 32 Litcrature on Inosite-Phosphoric Acid [Sept. HO-^P— O— Q Ho/ H -C— 0-P' /^ -OH \0H (3) P_0— CH HC— 0— Pf HO O^ =0 v\0H sOH hq// ;P— O— c- h -C— O— P= h =0 \\0H HO/ \0H M. W., 660; C = 10.91%; P = 28.18%. Neuberg came to similar conclusions the following year when he obtained inosite and furfurol on mixing " phytin " with phosphoric acid and distilling under reduced pressure, and also showed that furfurol can be obtained from inosite. He proposed the following f ormula : H -c- (4) HO. H0\ H HO>P-0-(; HO^. HO-7P— O— CH HC— O HO/ H /OH O— ^<0H /OH -P.^OH \0H t -CH HO— P\ /P— OH HO/ ^0^ \0H M. W., 714;C = 10.085% ; P = 26.05%. Levene, working with a preparation from hempseed, was led to believe that the "phytin" of this grain contained in its molecule phosphate, inosite and a carbohydrate of the pentose group. His work was criticized by Neuberg, who claimed that there were impurities in the preparation. In view of the known intimate asso- ciation of the phytin with protein and carbohydrate in the aleuron grain, and the possible occurrence of a chemical combination of both phyto-phosphate and carbohydrate with protein, it is conceiv- able that Levene had a product holding pentose as an integral part igi2] 'Anton Riclmrd Rose 33 and not as an impurity, though in view of all the available evidence Neuberg's criticism seems at the present time somewhat justifiable. Starkenstein also refused to accept the simple formula proposed by Posternak and offers the f ollowing : HO-^P— OH.HO— C-C— OH.HO— P^OH ^\ j i /^ HO— P— OH. HO— C C— OH.HO— P^H o/ 11 \) (5) HO— C— C— OH HO OH 0=P P=( /\/\ OH O OH M. W., 714 ; C = 10.985^ ; P = 26.05 He argues that the phosphoric acid is in the pyro-form^ from the fact that its silver salt is the same color as silver pyro-phosphate, and that its behavior when titrated with Standard uranium acetate Solution is also like that of pyro-phosphoric acid. That it is not combined in the usual form of an ester but held loosely in a complex "addition form," he maintains from the fact that an increase of inosite and inorganic phosphate resulted from heating some of the calcium salt for an hour at ioo° C. These arguments are not alto- gether convincing. Anderson has prepared a silver salt of the ortho- tetra-phosphoric acid ester with inosite and reports it as being white like the pyro-phosphoric acid salt. In the quantitative titration of pyrophosphoric acid with a uran- ium acetate Solution, standardized by ortho-phosphate and using fer- rocyanide as indicator, only one half of the phosphorus value is obtained. Starkenstein explains this phenomenon by the assump- tion that one half of the more reactive ions of the phosphoric acid have been removed in the dehydration. Now if two phosphoric- acid groups had formed esters with one polyalcohol, analogous con- ditions would have resulted as far as the ions are concerned, and bivalent ions would be expected to connect the two phosphoric acid * That phosphorus occurs in plants in the pyro form may seem stränge to many, but this is not the first time that such an occurrence has been suggested. In 1892 Hardin (5". C. Exp. Sta. Bull., N. S., No. 8) reported his finding both pyro- and meta- phosphate in cottonseed meal, when he sought, in this feeding material, a substance toxic to cattle. 34 Literature on Inosite-Phosphoric Acid [Sept. radicles. This may be illustrated by these two graphic represen- tations : .HO: OH^^^* \ /OH«-^ \ / 0=:P OH 0=P— OH ^ -f— ua. The idea of this type of reaction for ortho and pyro-forms is in harmony with the fact that when inosite-phosphoric acid is precipitated in acid Solutions by divalent metals, the tri-metal salt is the more readily formed. The activity of the hydrogen ions is relatively greater in the inosite-phosphoric acid than the sum total of the ions of the ortho-forms would be, probably due to the influ- ence of increased negative electric charges in the many phosphorus atoms held in one molecule, so that, altho six having been eliminated in the assumed ester formation, the very reactive ions are eight in number. The last column in Table 2 is interesting in this con- nection. Finally, the Statement that the inosite-phosphoric acid is decomposed by dry heat has been shown to be erroneous by both Anderson and the writer. That phytin is a salt of inosite-phosphoric acid seems to be con- clusively demonstrated by the synthetic work of Contardi, whose preparations from rice bran gave analyses identical with a synthetic preparation obtained by heating anhydrous inosite with ortho- phosphoric acid (sp. g. 1.7). Other workers have attempted to substantiate this result, but so far without success. Carre could obtain only a mixture of the two chemicals ; Anderson was able to produce tetra-ortho and di-pyro-phosphoric acid esters. It does not follow that these preparations of inosite-phosphoric acid are identical in form with the organic phosphorus Compound occurring in plants. The writer calls attention elsewhere to a differ- ence of behavior between the phytophosphate in seeds and in prepa- rations.^ Certain data, not as yet published, as well as differences in the products described in the literature, fall in with those suspicions.^ What inosite-phosphoric acid is, in terms of a definite chemical 'Rose: Technical Bulletin 20 of the New York Agricultural Experiment Station. ' Cf . Preparations analyzed by Patten and Hart, Winterstein and Schulze, and Levene, in table 2, p. 29. I9I2] Anton Richard Rose 35 structure, is an open question. It is probably an ester of phosphoric acid with inosite, in which six phosphoric acid groups are united with each inosite molecule. This ratio, Cß : Pg, is indicated for the vast majority of the pure preparations analyzed; exceptions are the preparations by Levene, Vorbrodt and Rising (Table 2), these authors giving the ratio Cq : P5.5. Anderson has pointed out that bis and Rising's silver salts are probably identical, Rising's analysis being equally well adapted to the formula C6Hi7027P6Ag7. It seems probable that the molecular weight when accurately deter- mined will be reported as 714 or will differ from this by the molec- ular weight of three molecules of water, The molecule seems to contain twelve hydrogen atoms readily separated in ionization, six of which are exceedingly reactive ; the remaining hydrogen atoms gradually diminish in reactivity by twos, the last four being slow to enter into an exchange with bases. The most readily formed salts are therefore those corresponding to an octavalent acid and the other common ones are in six and tenvalent combinations. TERMINOLOGY AND CLASSIFICATION The investigations of which this paper is a brief review have brought to the biological chemist and plant physiologist a type of phosphorus Compound from the plant world which is relatively new and probably of prime importance. The phosphorus which occurs in acid and water extracts was formerly considered inor- ganic phosphate, and awakened no especial interest, and the mention of organic phosphorus immediately brought to mind nucleoproteins and lecithins. In the organic laboratories combinations of phos- phorus with various organic radicals have been made and recently- prepared phosphoric acid esters'^ resemble " phytin " in some respects, and so may be considered of special concern to the biological chemist, as possibly bearing on problems in his field. A cleavage product of inosinic acid, c?-arabinose phosphoric acid,^ is significant as showing that carbohydrate esters are not confined to those pro- duced by the synthetic Operations of the laboratory. Even more 'v. Lebedew: Biochem. Zeit., 1909, 20, 114; Neuberg and his coworkers : Ber., 43, 2060; Biochem. Zeit., 23, 515; 26, 115 and 529; 36, 5; Langheld, Ber., 44, 2076. 'Levene and Jacobs, Ber., 191 1, 44, 746. 36 Literatur e on Inosite-Phosphoric Acid [Sept. striking are Iwanow's experiments® in which, when yeast was allowed to ferment sugar in the presence of sodium phosphate, there was noted a disappearance of the inorganic phosphorus, amounting to f rom eighty to ninety-three per cent. ; and in the liquors there was foiind an organic phosphorus Compound optically active and giving reactions for aldehydes and ketones. Biochemical syntheses of this class have also been successfully made by other investigators.^*' Of special interest are the inosite esters with ortho- and pyrophos- phoric acid prepared by Anderson. Mention may also be made of the spherocrystals discovered by Hansen" in the parenchyma cells of the Euphorbia caput medusae, which he describes as amorphous masses of calcium and magnesium phosphate, but which Belzug^^ later has shown to be salts of a new organic acid, phosphomalic acid. We may reasonably expect that additional phosphorus-bearing sub- stances of this kind will be discovered in nature by the phyto- chemist for which a rational System of nomenclature will be required. Rising, in his paper on inosite-phosphoric acid, refers to soluble phosphorus Compounds obtained from grains, which he promises to discuss in later contributions. These substances he considers closely related to "phytin," and proposes classifying them as a single group with the generic name " phyto-phosphoric acid." This term we may profitably adopt to indicate the acid radicals of those organic phos- phorus Compounds which may be found in the water and dilute acid extracts of plant materials. In this group will be included the gly- cerophosphates, phosphomalates, such hexose and pentose phos- phates as may be discovered in plants, and the phytin-like substances. The term " phytin " as used at present seems to designate that substance which is extracted from seeds by leaching with dilute acids, reacting positively in the tests for calcium, magnesium, and, after hydrolysis, for phosphoric acid and inosite. The multiplica- tion of trade names for definite chemical Compounds is not desirable. There are many students and other workers who must of necessity 'Iwanow: Zeit, für physiol. Chein., 1107, 50, 281-288. "Young and Hardin : Biochem. Zeit., 191 1, 32, 173-188; Proc. Chem. Soc, 21, 23, 24; Proc. Roy. Soc., London, 77, 80, 81, 82; Euler and Ohlsen: Biochem. Zeit., 191 1, 37, 313. " Hansen : Arbeit, des bot. Inst., Würzburg, 1888, 92-122. "Beizug: Jour. de bot., 1893, 7, 211-229. I9I2] Anton Richard Rose 37 carry in memory more names of organic Compounds than they can reasonably be expected to define in terms of chemical formulae, if the common names do not in themselves off er suggestions of chem- ical structure. Unsystematic naming is contrary to the modern spirit of chemical nomenclature. Winterstein's " inosite-phosphoric acid" has priority over Posternak's "phytin" and the further ad- vantage of being a chemically descriptive term. The preference of several authors for this latter designation is evidenced by the fact that the name phytin is not adhered to or is given in parenthesis after the name " inosite-phosphoric acid." In this particular case the probability of confusion is very miich increased by the fact that the term " phytine " is already applied to Chlorophyll preparations whose chemical composition we cannot hope to know for some time and for convenience must perforce carry a non-chemically descrip- tive appellation. The word " phytin " seems to have all the Psycho- logie requirements of a really good trade name and the substance which it designates in the market is widely advertised in the Euro- pean medical Journals for its therapeutic properties, which are more than likely of questionable character, and the term will undoubtedly persist. It can be readily conceived that this may not be the only inosite- phosphoric acid in plants and we should look for other combinations in which the phosphorus may be in the ortho or pyro form — even the meta phosphate — and be present as the hexa, tetra, di, or mono phosphoric acid. Various incidents have suggested to the writer that some of these forms occur in preparations from seeds when certain treatment other than those described above is used. ANALYTICAL METHODS^ The quantitative estimation of phytin phosphorus has been effected only by determining the difference between the total soluble phosphorus and the inorganic phosphorus. Phytin research in animal and plant metabolism is therefore very largely dependent upon the accuracy of the determination of inorganic phosphorus. " The analytical methods are here treated very briefly, for their development is as yet imperfect and the literature conflicting. Many papers have not been mentioned and the reader is referred for these to the bibliography on page 46. A more complete Statement with experimental data will be published later. 38 Literature on Inosite-Phosphoric Acid [Sept The term soluble phosphorus above and elsewhere means of course the phosphorus Compounds which dissolve in cold acidulated water ; the amount is obtained by evaporating the extract and destroying the organic matter with sulphuric and nitric acids according to the method of Neumann,^^ after which the phosphorus is determined by the usual ammonium molybdate and magnesia mixture method as described by Sonnenschein and later modified by Woy. As ex- tracting agents both acetic acid and hydrochloric acid have been used. The first method to approximate an accurate determination of inorganic phosphorus in the presence of soluble organic phosphorus was that used by Hart and Andrews in 1903. Their extracting agent was 0.2 per cent. hydrochloric acid Solution, a solvent which has since been used by most investigators. Hart and Andrews noted that ammonium molybdate did not precipitate the phytin phosphorus, and used this fact to devise a method for separating the two kinds of phosphorus combination in Solution. They had some apprehension lest the strong acid in the usual molybdate Solutions would hydrolyze some of the organic phosphorus Com- pounds and thus yield high results for the inorganic portion. They determined the minimum amount of nitric acid necessary to give a rapid, complete, and crystalline Separation of the yellow precipitate (2 c.c. of nitric acid, specific gravity 1.2, in each 250 c.c. of Solu- tion) and added to the liquid of this acidity neutral ammonium molybdate Solution. Vorbrodt, in his excellent monograph on " phytin," developed a method which is based on a triple precipitation of the inorganic phosphorus, first precipitating in general by means of magnesia mixture and dissolving the precipitate in the least amount of nitric acid. This is diluted to 50 c.c, heated to 100° C, and treated with an equal volume of ammonium molybdate Solution. The yellow precipitate is dissolved in ammonia water, 25 c.c. of 5 per cent. barium chlorid are added,^^ and the precipitate after being washed and dried is weighed; or the phosphorus may be precipi- tated with magnesia mixture and weighed as magnesium pyro- phosphate. "Neumann: Zeit, für physiol. Chetn., 1902, 37, 115. "Riegler: Zeit. Anal. Chem., 1902, 41, 675. 1912] 'Anton Richard Rose 39 Stutzer in Germany and Forbes in this country, working inde- pendently, introduced a new idea in the determination of inorganic phosphorus, namely, the use of acid alcohol. Forbes and bis asso- ciates make an acidulated water extract and precipitate with mag- nesia mixture; the precipitate is then washed successively with ammonia water and alcohol, and the inorganic phosphorus separated f rom the phytins by digesting in cold 95 per cent. alcohol containing 0.2 per cent. of nitric acid. This alcoholic Solution is finally filtered, the filtrate evaporated, and phosphorus determined in the residue in the usual way. Starkenstein has studied in some detail the application of titra- tion methods to this problem, and his results point to the possibility of determining quantitatively these different forms of phosphorus in the same Solution. He found that titration of a Solution con- taining ortho-phosphate, pyro-phosphate and inosite-phosphate with uranyl acetate standardized by ortho-phosphate, using cochineal as an indicator, gave in each case true values for total phosphorus; that with ferrocyanide as an indicator, the total phosphorus was equivalent to all of the phosphorus as ortho-phosphate, one half of that as pyro-phosphate and inosite-phosphate, the glycero-phosphate not entering into the reaction at all. Anderson notes that pyro- phosphoric acid can be converted into the ortho form by heating with dilute acids, while the inosite-phosphoric acid is not affected by this treatment. With these facts in mind a Volumetrie process may readily be devised. THE ROLE OF INOSITE-PHOSPHORIC ACID AND ITS SALTS IN PLANTS The literature on phosphorus metabolism in animals has become voluminous, but the botanists have published comparatively little on the changes of these Compounds and their probable significance in a plant's life history. That cell functioning is impossible in the absence of phosphorus is again emphasized in the recent work of Frouin,^^ which shows its absolute necessity in the growth of micro-organisms. The study of the role of phytin in plant life in- volves an investigation of the changes and distribution of all the ^'Frouin: Compt. Rend. Soc. Biol., 1910, 68, 801-803. 40 Literature on Inosite-Phosphoric Acid [Sept. pliosphorus Compounds and of inosite in the several stages of plant development. Since the methods of differentiating between the various combinations of phosphorus in plant substances are becom- ing highly perfected, we may expect rapid developments in our knowledge of their functions in plant processes. The universal presence of phytin in propagating and growing parts must be highly significant. This constant occurrence led Starkenstein to assume that "phytin" plays a specific role in the mechanism of growth of both plants and animals. If this be so, its biochemical reactions must be closely linked with carbohydrate and protein formation, and its occurrence with these substances in the aleuron grain must be more than a mere coincidence. In this connection it may be well to review briefly the literature regarding the aleuron grains. The best summary was found in Vines's text book of plant physiology (1886) but this is too brief to be satisfactory. From Pfeffer's comprehensive description, it appears that these grains form in the vacuoles during the ripening and desiccation of the seed; that the forms assumed are globular, which are less distorted and attain a larger size in the more fatty seeds. They consist morphologically of three parts: the large pro- tein particle, Pfeffer's globoid, and a membrane. Crystals of cal- cium Oxalate are sometimes present. Weyl^'^ isolated the grains from the " Paranuss " employing the method of all the previous inves- tigators, and made an extensive study of their proteins. This was in the days of the vegetable vitellins (globulins), and the chief protein of the aleuron grain having been shown to belong to this group, Weyl thought that the membrane was a modified form of the same protein, an albuminate. Three or four years later, Vines^^ under- took a study of these proteins and from his observations on mate- rial from a large variety of seeds, grouped them into five classes: vegetable peptone (water soluble), vegetable myosin, crystalloid, vit ellin (all three soluble in sodium chlorid Solution), albuminate (soluble in sodium carbonate Solution). These are described as plastic proteins, in part transported to the cells of the seed from other portions of the plant. According to Posternak, these pro- ^ Weyl : Zeit, für physiol. Chem., 1S77, i, 84-96. "Vines: Proc. Roy. Soc. London, 28, 218; 30, 387; 31, 59, 62; see also Lundtke : Jahrb. wiss. Bot., 1890, 21, 62. I9I2] Anton Richard Rose 41 teins constitute from fifty to seventy-five per cent. of the aleuron grain. It is doubtful whether they are simple proteins; more likely they contain both phosphorus and bases in their molecules. Besides protein, Posternak found carbohydrates which were not free, but combined with some other substances of the grain; also ash, to the extent of twenty-five to fifty per cent. The following analytic data were recorded : Per Cent. Per Cent. Phosphorus O.11-3.83 Magnesium 0.28-1.27 Sulphur 0.64-0.81 Calcium 0.11-0.37 Silicon 0.01-0.36 Iron 0.03-0.09 Potassium 2.29-2.71 Manganese Trace These results were obtained from aleuron grains of sunflower, white lupin, hemp, and red fir. The author calls attention to the interesting fact that all the elements essential to plant growth are present in these bodies. These results are notable when com- pared with Bernardini's analyses of rice embryo: P2O5, 0.95; SO3 (not given); SiOs, 0.25; K2O, 1.691; MgO, 1.389; CaO, 0.279; FesOg, 0.06; Mn, trace; NagO, trace. One would like to know whether the Silicon in these two substances is present in organic combination. The globoid or "phytin" is a calcium-magnesium salt of inosite-phosphoric acid. Phyto-phosphate is also combined in the protein granules, possibly in the form of the potassium salt, as Posternak believed, inasmuch as he could not separate the potassium salt from the globulin, although it is soluble in alcohol and globulin is not soluble in this liquor. He concluded that it is chemically attached to the protein. In germination, the aleuron grains swell up, forming a granulär viscid mass; both globoid and crystalloid go into Solution, enzymatic action sets in, and both phytin and pro- tein are hydrolyzed. The presence of an enzyme having the power to decompose phy- tin into inosite and an inorganic phosphate was first demonstrated by Suzuki and his associates in the bran of rice. It has also been found by Vorbrodt in other small grain, including wheat, rye, and barley ; likewise in larger seeds, as vetch and lentils. An extract of the kemel of indian corn gave no evidence of the presence of a phy- tase, but it was shown to develop during the germination of the grain. 42 Literatlire on Inosite-Phosphoric Acid [Sept. In the earlier analyses of seeds, the inorganic phosphorus was not given a very prominent place and was usually reported as that por- tion obtained by subtracting the sum of the protein and lecithin phos- phorus from the total phosphorus; but as this dement began to re- ceive special attention its direct determination was attempted, and consequently the amount of inorganic phosphorus reported was lessened. Thus Umikoff^^ estimates the inorganic portion as fully half of the total phosphorus, but the more recent workers report it in very small percentages. With the germination of the seed, the inorganic phosphorus gradually increases at the expense of the organic form. This was at first attributed to the breaking up of the phosphorus-bearing proteins and lecithins. Tammann,^'' one of the earliest investigators to make direct determinations of inorganic phosphorus in seeds and their sprouts, found during germination an increase of this form which, in terms of P2O5, was from 0.324 per Cent, to 0.443 P^^ cent. in a period of only twelve days. Ac- cording to the work of Prianischnikow^^ and Merlis,^^ lecithin de- creased one half during fifteen days' germination of Vicia sativa and Lupinus angustifolius. Iwanow found that the inorganic phos- phorus increased from a very small amount to 93.7 per cent. of the total phosphorus in germinating seeds of Vicia faba. He held that lecithin is the most stable of the organic forms and is altered very little. Phytin, owing to the presence of phytase, is practically all changed by this process. Vorbrodt has shown that the phosphorus Compounds, especially inosite-phosphoric acid, are particularly abundant in the germ. When the seed begins to sprout, this supply is increased by trans- portation of phosphorus from other parts of the grain, as is indi- cated by Zalesky's^^ Observation that in the sprouts of Lupinus angustifolius the total phosphorus increased in twenty-five days from 0.302 gram to 0.514 gram; the inorganic phosphorus doubled in amount, while the protein and lecithin phosphorus remained prac- "Umikoff: Russian Dissertation, 1895. (Cited by Zalesky: Ber. bot. Ges., 1902, 20, 426-433.) ^Tammann: Zeit, für physiol. Chem., 1885, 9, 416-418. " Prianischnikow, 1895. (Cited by Zalesky. See footnote 19.) ^'Merlis: Landw. Vers. Stat., 1897, 18. (Cited by Zalesky. See footnote 19.) ^Zalesky: Ber. bot. Ges., 1902, 20, 426-433. I9I2] Anton Richard Rose 43 tically unchanged. Bernardini found that the phytin decreased also in the germination of wheat, but the lecithins increased. In the early stages of plant development subsequent to germina- tion, inorganic exceeds organic phosphorus. Staniszkis could find no trace of inosite-phosphoric acid in millet during this period. The phosphorus is now drawn f rom the soil and its increase in the plant is proportional to the increase in dry matter. The organic forms of phosphorus are synthesized from this supply, according to Stan- iszkis, very slowly until the heads are formed. Hart and Totting- ham found no phytin phosphorus in the dried forage plants. Balicka-Iwanowska, working with barley, found the phosphorus Compounds at the end of the fourth week present in the same pro- portion as that in the seed ; thereafter there was a constant decrease in the inorganic phosphorus. In the seventh week the protein phos- phorus had doubled and the inosite-phosphoric acid had increased to seven times the amount present in the fourth week. As the barley seeds began to form, in the ninth week, the increase of organic phosphorus occurred mostly through the synthesis of nucleoproteins. The small increase which Staniszkis reports was more in the form of lecithins and protein phosphorus Compounds than of inosite-phos- phoric acid. At the period of flowering, the lecithins reach their maximum, which may be, according to Stoklasa, 71.6 per cent. of the total phosphorus. During the formation of millet seed, the syn- thesis of phytin goes on energetically at the expense of both inor- ganic and protein phosphorus. In the barley, Balika-Iwanowska found an increase of inosite-phosphoric acid and also one of nucleo- protein which was even greater than that of the phytophosphate. In the ripening of the seeds of millet the formation of inosite phos- phates and nucleoprotein ran parallel, but in barley the inosite phos- phate increased at the expense of some of the protein phosphorus. As the panicle grew and matured in both of these plants, there was a transportation of both the phosphorus and the protein to this part of the plant. The mobilisation of phosphorus and changes in its form in Vicia faha and other plants were also studied by Iwanow. The most interesting part of his contribution is the relation between sunlight and changes in the form of phosphorus. The plants which remained in a dark room contained more inorganic phosphorus than 44 Literature on Inosite-Phosphoric Acid [Sept. those which had snnlight. Opaque shields on the leaves produced the same results in the protected part of the leaf, hence the change of inorganic into organic phosphorus may be attributed in part if not altogether to photosynthesis. Stoklasa and his pupils^'* consider phosphorus an integral part of Chlorophyll, existing in a form which does not give the HPO4 ion. Schimper^^ in his rather comprehen- sive study of the assimilation of the ash constituents in plants also noted the decrease of the inorganic phosphorus through the action of light. Posternak, attempting to account for the formation of "phytin," which he then thought to be anhydro-oxymethylene-di- phosphoric acid, assumed that it was formed simultaneously with the reduction of carbon dioxide by a direct combination of the orod- ucts of the photo-chemical action and inorganic phosphates, an hypothesis suggested by Schimper's experiments. If Posternak's assumption is true, even in part, phosphorus may play a very signi- ficant röle in carbohydrate anabolism. Several authors have ex- pressed doubt about this explanation of "phytin" synthesis, advanc- ing the argument that inosite-phosphoric acid is not found in the early stages of growth, and when formed later is not uniformly produced, as would be expected if it were due entirely to the action of the chloroplasts. There is still the possibility that it is so syn- thesized and instantly broken down in the formation of other Com- pounds. Soave could find no inosite in dormant seeds unless they were first boiled in strong acid, but after they began to sprout its pres- ence could be easily demonstrated until the reserve material of the cotyledons was almost exhausted. It was also present in unripe seeds, indicating that the inosite-phosphoric acid is formed in the seed by the combination of the inorganic phosphorus, abundantly present at this stage, with the inosite. The occurrence of inosite in the unripe seed and the green parts of the mature plant, and the later disappearance of this substance as inosite-phosphoric-acid -forms, indicate that phytin is probably produced by the reversible action of an intracellular phytase, and that Posternak's explanation is in- correct. Rising suggests that inosite-phosphoric acid may be an interme- ** Stoklasa, Brdlika and Ernest : Ber. d. deut. bot. Ges., 1910, 27, i, "Schimper: Flora, 1890, 23, 207-261; Bot. Zeifg., 1888, 46, 81. igi2] 'Anton Richard Rose 45 diary product in the formation and destruction of the lecithins, and in this way may play an exceedingly important part in the life cf the plant. The possible relation between the two Compounds is shown in the following graphic representation : HjOjPOHC— CHOPO3H, — CHOPOjHj HjOgPOHg (JHOPOjH, ^ CHOPOjH, HjOjPOHC— CHOPOjHj — CHOPO,H, 2 The most striking Suggestion as to the functions of inosite-phos- phoric acid is contributed by Starkenstein, who thinks that the inosite is in itself inert and incidental and functions only in its com- bination with phosphorus. He has demonstrated that, in the body, inosite yields lactic acid, an interesting fact in view of its possible significance in carbohydrate formation. He assigns to inosite- phosphoric acid, as its special function, some part in the process of growth, basing his view on his experiments with animals. In har- mony with this view is the distribution of phosphorus in the seed, the greater part being localized in the germ; according to Bernardini over eighty per cent. of the total phosphorus in the rice germ is in the form of inosite-phosphoric acid. It is interesting to note in this connection the Observation of Iwanow that there is a tendency to concentration of phosphorus in the parts of the plant where growth is most active, and also that when the phosphorus supply of the Sub- strate is insufficient, the phosphorus of the other parts of the plant is rapidly transported to the growing shoots. As previously stated the phosphorus of the seed is in the form of the calcium-magnesium Salt of inosite-phosphoric acid, but, according to Posternak, the phosphorus, in transportation, is in the potassium salt of this acid. Phyto-phosphoric acids, whether they are inosite esters or other Compounds, undoubtedly play very significant roles in all higher plants, but as their specific functions have not as yet been ascertained, even the chemical structures being as yet uncertain, nearly all State- ments on the subject must be pure conjectures. The chief sug- gestions from experimental work are that these acids are concerned in the process of photo-synthesis or in the changes of the photo-syn- thetic products, for example, the formation of carbohydrates and fats; that it is an intermediary step in the synthesis of phospho-pro- teins and lipoids; and that it acts as a specific Controlling factor in 4^ Literature on Inusite-Phosphoric Acid [Sept. growth. The varioiis functions as thus outlined are probably over- estimated, but those who have worked in this field seem to be strongly of the opinion that inosite-phosphoric acid is more than a reserve material. It is attracting considerable attention and as the necessary analytical methods are perfected, we may expect to see, in increasing number, valuable contributions that will eluci- date in detail the part which this interesting Compound plays in nature. BIBLIOGRAPHY OF INOSITE-PHOSPHORIC ACID ("PHYTIN") Anderson. N. Y. Agr. Exp. Sta., Geneva, N. Y. ; A''. Y. Agr. Exp. Sta. Tech. Bull, ig, 21, 1912; /. Biol. Chem., 1912, 11, 471-487; 12, 97-113, and 447-464. Important contribution on the salts of inosite-phosphoric acid, also impor- tant synthetic work. Aso and Yoshida. Imp. Univ., Tokio; /. Coli. Agr., Imp. Univ., Tokio, 1909, i, 153-168. Compares the value of " phytin " with other forms of phosphorus as a fertilizing material. Balika-Iwanowska. Agr. Chem. Inst., Krakow; Ras. Akad. Univ., 1906, 2° ser, 6, B, 24. (From Maly's Jahrsb., 1907, 36, 741.) Gives the influence of the HPO4 ion on plant growth; also the changes in form and distribution of phosphorus at the various stages of development of barley plants. Bernardini. Chemica Agr. Scuola, Portici ; Atti. Acc. Lincei, 1912, 21, 1°, 283-289. Bernardini and Morelli. Atti. Acc. Lincei, 1912, 21, 1°, 357-362. Boorsma. Batavia ; Von Bemmelen Festschrift, 210-213 ; (see Chem. Centr., 19H (I), 296). Preparation and properties of the inosite-phosphoric acid from rice bran. Carte. Bull. soc. chim., 1901, (4), 9, 195-199. (From Chem. Zentr., 1911 (I), 1196.) Repeats the work of Contardi and reports negative results. Collison. Ohio Agr. Exp. Sta., Wooster, O. ; /. Biol. Chem., 1912, 12, 65-73; /. Ind. and Eng. Chem., 1912, 4, 606-608. Modified method of determining inorganic phosphorus in the presence of phyto-phosphates. Cf. Forbes. Contardi. Lab. di chim. org. della Reale Scuola Agr., Milan; Atti. Acc. Lincei, 1909, 5 ser., 18 (1° sem.), 64-67; 1910, 5 ser., 19 (1° sem.), 823-827. Prepa- ration of inosite-phosphoric acid from rice bran, analyses of the pure prepa- ration, synthesis of phytin by heating inosite with phosphoric acid. Syn- thetic product gave same analytic data as the preparation from rice bran. Important papers. Cook. U. S. Dept. Agr., Washington, D. C; Bur. of Chem., U. S. Dept. Agr., Bull. 123, 1909. Feeding experiments on rabbits. Dox and Golden. la. Agr. Exp. Sta., Ames, la. ; /. Biol. Chem., 191 1, 10, 183- 186. Demonstration of the presence of phytase in certain common fungi. Donath. Wien. klin. Woch., 191 1, 24, 1192-1197. Therapeutic. Fingerling and Hecking. Versuchsstation, Hohenhain; Biochem. Zeit., 1912, 37. 452. Contains a note on Stutzer's method. Forbes, Lehmann, Collison and Whittier. Ohio Agr. Exp. Sta., Wooster, 1912] Anton Richard Rose 47 Ohio; Ohio Agr. Exp. Sta. Bull., 215, 1910. Gives a good method for the determination of inorganic phosphorus. See Collison. Fürst. Centr. Kinderheilk., 1904, 409. Therapeutic. Giacosa. Giorn. della Real. Accad. di med. di Torina, 1905, 68, 369-374; 1907, 70, 290-295. (From Maly's Jahrsb., 1906, 35, 124 ; 1908, 37, 473 ; also Biochem. Centr., 6, 573.) Pharmacological study on man. Gilbert and Posternak. L'Oeuvre Medico-chirurgical, 1903, No, 36. (From Maly's Jahrsb., 1904, 34, 729. Therapeutic. Gilbert and Lippmann. La Presse Medicale, 1904, Aug. 2y and Sept. 10. Phar- macological studies on rabbit and guinea pig. Hardin. S. C. Exp. Sta., Fort Hill, S. C. ; 5". C Exper. Sta. Bull., n. s. 8, 1892. Pyrophosphoric acid in cottonseed meal; probably associated with phytin. Cf. Crawford, /. Pharm, and Exp. Ther., 1910, i, 519. Hart, McCoUum and Humphrey. Wis. Agr. Exp. Sta., Madison, Wis. ; Wis. Agr. Exp. Sta. Research Bull., 5, 1909; Am. J. Physiol., 1909, 23, 86-102; 24, 246-277. Feeding experiment with the cow. Cf. Jordan. Hart and Andrews. N. Y. Agr. Exp. Sta., Geneva, N. Y. ; A''. F. Agr. Exp. Sta. Bull., 238, 1903; Am. Chem. Jour., 1903, 30, 470-486. Gives the first approximately reliable method of determining inorganic phosphorus in the presence of phytin and other organic forms of phosphorus. Hart and Tottingham. Univ. of Wis., Madison, Wis.; /. Biol. Chem., 1909, 6, 431-444. Determination of the amounts of phytin in certain feeding stuffs. Hartig. Braunschweig; Bot. Ztg., 1855, 13, 881-882; 1856, 14, 257-355. Micro- scopic study of seeds; discovery of the substance later known as phytin. See also "Lehrbuch der Anatomie und Physiologie der Pflanzen," 1891, 48. Horner. Pathol. Inst., Univ. of Berlin ; Biochem. Zeit., 1906, 2, 428-434. Phar- macological studies on dog and rabbit. Iljin. Russ. Wratsch., 1906, No. 13, from Maly's Jahrsb., 1907, 36, 54. Com- pares the properties of phytin, lecithin and nucleoprotein. Iwanoff. Jahrb. wiss. Bot., 1901, 36, 355-379; /. Exp. Agr. (Russian), 1902, i (cited by Zaleski) ; Ber. bot. Gesell., 1902, 20, 366-372; Zeit, physiol. Chem., 1907, 50, 281-288. Studies on the changes of the forms of phosphorus in germinating vetch seeds and in the growing plant. Production of phyto- phosphates by yeast-fermentation of sugar in the presence of di-sodium phosphate. Jegorow. Landw. Inst. Petrowskoje-Rasumowskoje, Moskow; Biochem. Zeit., igi2, 42, 432-439. Study on stability of inosite-phosphoric acid and disput- ing the existence of phytase. Jordan, Hart and Patten. N. Y. Agr. Exp. Sta., Geneva, N. Y. ; N. Y. Agr. Exp. Sta. Tech. Bull, i, 1903; Am. J. Physiol., 1906, 16, 268-313. Feeding experiments with the milk cow. Cf. Hart. Korolev. Moscow. Izv. Moscov. Selsk. Khoz. Inst., 1910, 16, 1-98. (From Chemical Abstracts, 191 1, 1962.) Studies on organic phosphorus in the soil. LeClerc and Cook. U. S. Dept. Agr., Washington, D. C. ; /. Biol. Chem., 1906, 2, 203-217. Feeding experiments on rabbits. Levene. Rockefeller Inst., N. Y. ; Biochem. Zeit., 1909, 16, 399-405- Studies of phytin preparations made from hemp. Obtained a Compound consisting of three groups : inosite, pentosan and phosphate. Maestro. Lo Spermentale, 1904, 59, 456-458. (From Maly's Jahrsb., 1905, 35, 91.) Pharmacologic. 48 Literature on Inosite-Phosphoric Acid [Sept. McCoUum and Hart. Univ. of Wis.. Madison, Wis. ; /. Biol. Chem., 1908, 4, 497-500. Showing the presence of phytase in animal tissues. Mendel and Underhill. Yale University; Am. J. PhysioL, 1906, 17, 75-88. In- fluence of phytin on bacteria; pharmacological studies on dog and rabbit. Nagaoka. Imp. Univ., Tokio; Bull. Coli. Agr., Tokio, 1906, 6, No. 3. Value of inosite-phosphoric acid and other phosphorus Compounds in plant waste products as fertilizers, as compared with animal wastes, showing that the latter are the more efficient. Neuberg. Pathol. Inst., Univ. of Berlin; Biochem. Zeit., 1908, 9, 557-560; 1909, 16, 405-410. Analyses of several preparations of phytin. Concludes that the substance is inosite-phosphoric acid and does not contain a carbohy- drate group. Novi. Pharm. Lab. Bologna; Mem.r. accad. sei. Bologna, igii,^,s^r. 6. (From Zentr. Biochem. u. Biophys., 1911,11,871.) Influence of phytin and glycero- phosphates on muscle reaction. Palladin. Zürich Polytechnicum ; Zeit. Biol., 1894, 31, 191-203. Discovery of inosite-phosphoric acid by chemical procedure. Patten and Hart. N. Y. Agr. Exp. Sta., Geneva, N. Y. ; A''. Y. Agr. Exp. Sta. Bull., 250, 1904; Am. Chem. Jour., 1904, 31, 564-672. An important contri- bution on the properties and composition of inosite-phosphoric acid. Peters. Allg. med. Centralzeitg., 1908, No. 9. (From Centr. Nervenheilk. u. Psychiatrie, 1908, 31, 1081.) Therapeutic. Pfeffer. Marburg; Jahrb. wiss. Bot., 1872, 8, 429-574. Comprehensive study of aleuron grains, identification of "globoid," and approximation of its chem- ical nature. Polacci. Royal Bot. Inst., Univ. of Pavia; Malphigia, 1894, 8, 361-379. Study of phosphorus in the aleuron grain. Posternak. Zürich Polytechnicum; Pasteur Inst.; Basel; Compt. Rend., 1905, 140, 322-324; 1903, 137, 202-203; 2>37-2>29; 439-441; Bul. soc. chim., 1904, 33» 116; Rev. Gen. Bot., 1900, 12, 5-24; 65-73; Compt. Rend. Soc. Biol., 1903, 55, 1190-1192. German Patents, Kl. 12, Nos. 155798, 159749, 160470, 164298; Münch. med. Woch., 1907, p. 827. The first and most comprehensive study of the physical and chemical properties of phytin, giving methods of prepa- ration, results of analyses of pure products, and speculations on the Consti- tution and biological function of this product. Rising. St. Albonvorstadt, Basel; Svensk Kern. Tidskrift, 191 1, 22, 143-150. Study of organic phosphorus Compounds in food materials ; the name phyto- phosphoric acid is suggested, methods are given for analysis of the various forms of phosphorus, and analyses are reported for a preparation of the silver salt of inosite-phosphoric acid. Rogosinski. Anz. Akad. Wiss., Krakow, 1910, 260-310. (From Chem. Centr., 1910 (II), 1549; Chem. Abstr., 1911, 5, 1476.) Feeding experiment with the dog. Rose. N. Y. Agr. Exp. Sta., Geneva, N. Y. ; N. Y. Agr. Exp. Sta. Tech. Bull., 20, 1912. Feeding experiment with the milk cow; Dept. Biol. Chem., Colum- bia Univ., N. Y., BiocHEMiCAL Bulletin, 1912, i, 428-438. Influence of phytin on the growth of lupin seedlings. Sechert. These de Paris, 1904. (From Maly's Jahrsb., 1904, 34, 729.) Therapeutic. Schulze and Castoro. Zürich Polytechnikum; Zeit, physiol. Chem., 1904, 41, I9I2] Anton Richard Rose 49 477-484. Presents a method of analysis, and data on content of inosJte- phosphoric acid in various seeds. Schulze and Winterstein. Zürich Polytechnikum; Zeit, physiol. Chem., 1903, 40, 120-122. Phytin prepared and analyses of it made. Scofne. Pharm. Inst., Turin; dorn, della Real. Acc. di med. dt Torino, 190S, 56, 630. (From Biochem. Zentr., 1905.) Fate of inosite-phosphoric acid in the animal organism and paths of elimination. Soave. Sias, spernt. agrar. Ital., 1906, 39, 413-427, 434-438. (From Chem. Zentr., 1906, 1726.) Ann. R. Accad. di Agr. di Torino, 1906, 49, p. i et seq. (From Centr. Physiol., 1906, 772.) Shows the relation between inosite and phytin in seeds. Staniskis. Jagel Univ., Krakow ; Ans. Akad. Wiss., Krakow, 1909, 95-123. (From Chem. Zentr., 1909 (II), 114.) Determination of the distribution of the forms of phosphorus in millet at various stages of development. Starkenstein. Pharm. Inst., Univ. of Prague; Zeit. exp. Path. u. Therapie, 1908, 5, 378-389; Biochem. Zeit., 1910, 30, 56-98; 191 1, 32, 234-265. A study of inosite and its relation to animal and plant life; the relation between inosite and inosite-phosphoric acid — chemical, biochemical and biological ; the nature and Constitution of phytin; its significance in animal and plant growth ; toxicity ; reaction to common indicators ; and estimation by Volu- metrie methods. A noteworthy contribution. Streffer. Zentr. ges. Therapie, 1908, 25, 135. Therapeutic. Stutzer. Biochem. Zeit., 1908, 7, 471-487. Methods of analysis. Suzuki, Yoshimura and Takaishi. Imp. Univ., Tokio; Bull. Coli. Agri., Tokio, 1907, 7, 495-502, 503-512. Preparation of inosite-phosphoric acid from rice bran; discovery of phytase; Suggestion of a new formula, containing the inosite nucleus. Tsuda. Imp. Univ., Tokio; /. Coli. Agri., Tokio, 1909, i, 167-168. Study of the forms of phosphorus in vegetable wastes. Tyshnjenko. Therap. klin. militarmed. Akad., St. Petersburg; Dissertation (Russian), 1909, p. 117. (From Maly's Jahrsh., 1909, 39, 589.) Feeding experiments on man. Vorbrodt. Univ. Krakow; Bull, de l'Acad. des Sei. de Cracovie, 1910, ser. A, 414-511. A study of the general reactions of phytin; comparison of methods of analysis, and Suggestion of a desirable modification; the phytin content of a number of seeds reported; comparative study of phytases; ultimate composition of phytin and a proposed empirical formula. An excellent paper. Weismann. Therap. Monatshefte, 1908, 22, 470. Therapeutic. Windisch. Jahrb. Vers. u. Lehrs. f. Brauerei, Berlin, 1907, 10, 56-58. (See also Wochschr. Brauw., 1906, 23, 516; Chemical Abstracts, 1907, i, 81.) Showing that the inosite-phosphoric acid of barley does not pass into beer but disappears in the malting process. Winterstein. Zürich Polytechnikum ; Ber., 1897, 30, 2299-2302. The first inten- tional preparation of phytin; analyses of the material; cleavage products; introduction of the name inosite-phosphoric acid. Zeit, physiol. Chem., 1908, 58, 118-121. Discusses the Constitution of phytin. A NEW TYPE OF ARTIFICIAL CELL SUITABLE FOR PERMEABILITY AND OTHER BIOCHEMICAL STUDIES E. NEWTON HARVEY (Physiological Lahoratory, Princeton University) Research on the permeability of membranes has been largely confined to a study of the properties of what may be termed macro- scopic membranes; composed of parchment, collodion, rubber, or silk impregnated with various substances. The best example of membranes of a type and size comparable to the surface film of cells and yet available for permeabiHty studies are the precipitation membranes of Traube, investigated by Waiden, Tamann, and Meerburg. Protein membranes of exceeding fineness are formed at the surface of various non-miscible fluids shaken with protein Solutions, such as the surface film of oil globules in protein-oil emulsions, or the films formed on Chloroform or benzol when shaken with albu- men Solutions.^ Such membranes are useless for permeability studies so long as they Surround fluids that do not mix with water. However, it is an easy matter to replace the fluid within the mem- brane by a watery Solution, provided the former fluid is readily volatile and slightly soluble in water. Chloroform conforms to these conditions. When Chloroform is shaken with egg albumen Solutions, the globules, in the course of 10-15 minutes, shrink in size and their membranes become crumpled, due to the passage of Chloroform from water to air and from globule to water. Lecithin, if previously dissolved in the Chloroform, will take up water as the Chloroform passes out. In the course of one to two hours, in an open vessel, all the Chloroform disappears and we obtain, instead of a Chloroform Solution of lecithin, a water Solution of lecithin enclosed in a fine protein membrane, the whole of a size comparable with cell sizes. The diameter of the droplets may be varied at will ^ Robertson : Journal of Biological Chemistry, 4, p. i, 1908. 50 igi2] E. Newton Harvey 5^ according to the degree of shaking. The role of the lecithin is to hold the water as the water replaces the Chloroform. The protein membrane is impermeable to lecithin. These artificial lecithin cells are stable, persisting until destroyed by bacteria. In many ways — in shape, in general appearance and in consistency — they resemble, to a very remarkable degree, sea-tirchin or star-fish eggs. Some of their properties have been described in Science (n. s.), V'ol. 36, p. 564, 1912. The point I wish to emphasize here, however, is not that we can prepare artificial cells closely resembling real cells, but that a Solution of lecithin may be obtained within a protein membrane, the whole of known composition and of a size comparable with cell sizes. Much can be inferred concerning the living cell from a knowledge of the properties of such artificial cells where compo- sition is definitely known. As Chloroform is exchanged for water, some of the lecithin separates in the form of granules, most of which agglutinate in a dense clump. The cell as a whole, but more particularly these granules, take up neutral red from dilute Solution, becoming red in color. (Chloroform alone takes up only the yellow base of neu- tral red. When lecithin is dissolved in Chloroform it unites with the yellow base, forming a red salt.) If the permeability for alkalies of such red-stained cells is studied, a marked difference from that of living cells is noted. Both ammonium hydroxid and sodium hydroxid in w/2000 con- centration enter rapidly and at the same rate. It will be remem- bered that all living cells are very easily permeable to ammonium hydroxid, but very slightly so to sodium hydroxid.^ The surface membrane of living cells is evidently of quite different composition from the protein film which condenses on Chloroform droplets. Living cells behave toward alkalies as though they were sur- rounded by a layer of a fat solvent, as postulated by Overton. Lipoid-soluble alkalies (ammonium hydroxid) penetrate readily, lipoid-insoluble alkalies (sodium hydroxid) do not. The lipoid solubility of ammonium hydroxid can be readily demonstrated by means of a benzol-lecithin Solution shaken with egg albumen solu- ^ Harvey, E. N. : Journal of Experiniental Zoology, 10, p. 507, 1911 and BiocHEMicAL Bulletin, i, p. 227, 1911. 52 New Type of Artificial Cell [Sept. tion. The same type of protein-film is formed on these globules but they differ from chloroform-lecithin globules in that the benzol is not replaced by water. If the benzol-lecithin globule is stained in neutral red Solution and placed in 7?/iooo ammonium hydroxid Solution, the color change from red to yellow takes place almost instantly. But it is only after 15-20 minutes that sodium hydroxid in relatively high (;i/io) concentrations can enter. Ammonium hydroxid is readily soluble in the benzol droplet, while sodium hydroxid is not; and in this fact lies the explanation of the differ- ence in penetrability. When stained in neutral red Solution, prac- tically all living cells behave as though they were protected from alkali by a benzol-lecithin surface layer. It is a simple matter to introduce various substances into these cells by dissolving or suspending the material in the chloroform- lecithin Solution before it is shaken with the protein Solution. Thus, oil may be dissolved by Chloroform and will separate in the cell in several large droplets much like those in a Nereis egg. Or cholesterol, starch grains and finely divided protein particles can be likewise included ; or substances to be used as indicators in study- ing the permeability of the protein film. Such cells, regarded as complex Systems of biological sub- stances, offer exceptional advantages for interpreting phenomena observed in living cells under special conditions; for example, dur- ing the passage of an electric current. Movements and disintegra- tions take place which I have as yet only partially investigated. In the near future I intend to describe these phenomena and shall give more complete data upon the permeability of the film which sur- rounds the cells. ON A NEW FUNCTION OF THE CATALYZER CALLED " PEROXIDASE " AND ON THE BIOCHEMICAL TRANSFORMATION OF ORCIN TO ORCEIN^ JULES WOLFF In a recent publication I have described the influence which peroxidase exerts on certain phenols in the presence of various salts and alkalies.^ When dissolved in a weak sodium carbonate Solu- tion freely exposed to the air, orcin, for example, fixes from four to five times more oxygen in the presence of peroxidase than in its absence. In this note I wish to call attention to the fact that perox- idase has other powers than the fixation of atmospheric oxygen. In determining the combined influence of ammonia and perox- idase upon aqueous Solutions of orcin, I have studied conditions which favor the transformation of orcin^ into orcein, the beautiful coloring matter which is one of the principal constituents of com- mercial orseille. My observations are interesting from many points of view. They show that (i) if a 2 per cent. Solution of orcin is exposed to the air in a thin layer and subjected to the influence of different proportions of ammonia, orcein is not formed even after a month under such conditions, but, instead, there is produced a substance which imparts a brownish-red color to the liquid. (2) If, how- ever, a portion of the same Solution is put in a narrow tube, so that the reaction takes place in a deeper layer, and the surface of con- tact with air is limited (other conditions being equal), one observes a very slow but regulär formation of orcein. (3) If (everything eise being equal) one repeats the first experiment (i), but adds to the ammoniacal Solution of orcin a suitable quantity of peroxidase, orcein fails to appear, just as in the first experiment. (4) If, how- * Translated from the author's manuscript, in French, by Dr. J. J. Bronfen- brenner. [Ed.] ''Wolff: Comptes rendus, 1912, clv, p. 618. 'Orcin has been the subject of interesting work by Robiquet, Dumas, Liebig, and Laurent and Gerhardt. 53 54 A New Function of " Peroxidase" ever (all other conditions being eqiial), one repeats the second ex- periment (2), but adds to the ammoniacal Solution of orcin a siiitable amount of peroxidase, the transformation into orcein takcs place very rapidly and is quite advanced in four or five days^ Compar- ing the coloration intensities of products 2 and 4, one sees that in five days 4 contains more than twice as mtich orcein as 2. This gain is due to the action of the peroxidase. By boiling the perox- idase Solution for f rom 5 to 6 minutes, before adding it to the ammo- niacal Solution of orcin in experiment 4, there is no acceleration in the formation of orcein, evidently because the peroxidase, as the active agent in the transformation, is thus destroyed. In Order to determine the rate of oxidation in the different ex- periments, I measured the volumes of absorbed oxygen. In experi- ment 2, eight to nine times more oxygen was absorbed than in i, in the course of 48 hours. In experiments 3 and 4 there was a similar difference but, because of the presence of peroxidase, the proportions of absorbed oxygen were larger. Without discussing the nature of the combined action of am- monia, oxygen, and peroxidase upon orcin, we may conclude that in dilute Solutions of orcin, slozv oxidation by ammonia is the pri- mary condition for the formation of orcein. If, however, to this condition is added the accelerating influence of peroxidase, the action is directed toward formation of coloring matter rather than toward increased absorption of oxygen. These facts may possibly be helpful in the commercial preparation of orseille. Paris, France. * My first and third experiments could easily be performed in flasks with flat bottoms. A small test tube would be satisfactory for experiments 2 and 4. The proportions of materials indicated below are well adapted for the purposes of the experiments : ^ , ( 2 c.c. of 2.8 per Cent. Solution of ) tnm x j -.i For I and 2 < , , , ^j^ > Diluted with water to 3.5 c.c. ( orcm and 50 mg. of NH3 ) 2 c.c. of 2.8 per Cent. Solution of For 3 and 4 ■{ orcin, 50 mg. of NH3 and \- Diluted with water to 3.5 c.c. I c.c. of active peroxidase Solution STUDIES OF DIFFUSION THROUGH RUBBER MEMBRANES I. Preliminary observations on the diffusibility of lipins and lipin-soluble substances WILLIAM J. GIES (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) CONTENTS I. Introduction c :- IL On the diffusibility of biological substances through rubber cg III. A demonstration of osmotic pressure exerted by fat 50 IV. A demonstration of the diffusion of pigments from fat through rubber into fat 60 V. Comparative dialysis experiments, with demonstrations 61 VI. Experiments on the diffusibility of alkaloids through rubber 62 I. INTRODUCTION When I proposed to my biochemical associates in Cancer re- search, in 1909, that we undertake a study of intracellular chem- istry/ I realized that new analytic methods and unconventional experimental procedures were prerequisites for material progress in this as in any other chemical relation. The greatest obstacle in the path of progress in intracellular chemistry is the evident lability of the essential intracellular constituents. Our best chemical methods increase this predicament because each is essentially anti-biological in character. Biochemical discoördinations are enforced whenever any of our present chemical processes is efifectively applied to proto- plasmic material. In reflecting on the properties and possible coördinations of intracellular lipins, it seemed probable that such lipins might be separated from protoplasmic material with the least chemical vio- * Gies : Studies in cancer and allied subjects, conducted under the auspices of the George Crocker Special Research Fund; Volume III, Department of Bio- logical Chemistry, Introduction (in press). 55 56 SUidies of Diffusion through Rubber Membrancs [Sept. lence, and isolated with the least possible alteration of their qualities, if they cotild be removed by dialysis.^ When this idea first came to mind, however, execution of its essential feature appeared to be im- possible. I believed that the diffusion of a solute depends very largely on chemical affinity between the separating membrane and the solvents on both sides of the partition. In that view, it seemed highly improbable that any of the ordinary membranes, except possibly collodion, could be of Service in the dialysis of lipins under any circumstances. Collodion appeared to possess favorable quali- ties because of its solubility in common lipin solvents and its pos- sible affinity for the latter under conditions of dialysis.^ Collodion is the only one of the available membranes which, while soluble in ether-alcohol Solutions, freely permits the passage of salins, extractives, carbohydrates, and proteins from aqueous Solutions to water, or to aqueous Solutions, outside, and vice versa, At first thought this suggested special availability of collodion for the work in mind. On the other hand lipins could not be expected to dialyze through collodion in the presence of much water and, as preliminary dehydration seemed an inevitable necessity for the dialysis of lipins from cellular matter, the permeability of collodion membranes to zoater-soluble substances did not appear, after all, to imply any practical advantages for the diffusion of lipins. I also recalled the fact that, in some experiments in another relation, we found that collodion was occasionally rendered defective by ether when the latter was used as a preservative of aqueous Solutions undergoing dialysis.^ Continuing actively to consider these matters from one view- point and then another, I thought of rubber as a possible choice of membrane. Recalling the well-known fact that rubber swells very markedly in ether and even in ether vapor, I assumed that the rub- ber expands in ether under such conditions because ether dissolves in the rubber or combines with it. This was but the prelude to the ' After preliminary desiccation by treatment with anhydrous sodium sulfate or other suitable process. ° Collodion is a serviceable membrane for such purposes. See page 70. * In some experiments which Professor Welker has conducted at my request, we have found that the disintegrative effect of ether on collodion membranes may be due to contained alcohol and other impurities. (See page 70.) igi2] William J. Gies 57 deduction that if ether dissolves in or combines with rubber, ether would also carry dissolved lipins with it into a rubber membrane; and if ether were on the opposite side of such a membrane, to work inwardly under such conditions, ether currents would develop; and lipins would pass from the Solution o£ higher concentration to that of the lower, and there accumulate until an equilibrium was estab- lished. This conception was so attractive that I proceeded at once to State it to Dr. Rosenbloom and, with his Cooperation, immediately tested it. The solid residue from an evaporated ether extract of egg yolk offered the greatest advantages for a preliminary test. We accordingly made an ether Solution of such a yellow residue, transferred the deep yellow Solution to a rubber condom, immersed and supported the latter in ether in a stoppered bottle, and almost immediately observed diffusion currents as well as the rapid egress of lipochrome. Fat and cholesterol were easily detected in the diffusate. Assuming that this prompt positive result might be due to defects in the rubber, we made many tests to satisfy ourselves that the ob- servations were or were not what they appeared to be. Dr. Rosen- bloom gave very earnest attention to this phase of the matter for some time and established the fact that we were dealing, except in a few cases of obviously imperfect membranes, with true diffusion phenomena. The original experimental observations were made on March i, 1910. At that time I was ignorant of similar results of previous work with rubber membranes, although I recalled rather vaguely the fact that Kahlenberg had made use of such membranes in another connection. The references to Kahlenberg's work which are given in the Chemisches Zentralblatt [1906 (2), pp. 1391 and 1772], the only ones we could find on this subject at that time, satisfied us that if we extended these experiments, the observations of a previous observer would not be repeated.^ A month or two after the work "The references to which I allude gave the substance of a paper in the Transactions of the Wisconsin Academy of Sciences, Arts, and Letters, 1905, xv (i), pp. 209-272, entitled: " On the nature of the process of osmosis and osmotic pressure, with observations concerning dialysis." The results with which our own could be directly compared were the f ollowing ones : Copper oleate was $8 Stiidies of Diffusion through Rubber Membranes [Sept. was inaugiirated we also saw a late reference to the well-known fact, regarding the swelling of rubber in lipin solvents, on which our work was based.*^ Ten months later we demonstrated these findings at a meeting of the American Society of Biological Chemists (see page 64). The succeeding sections of this paper present reprinted prelim- inary reports on various portions of the studies which thus far have developed from the observations described above. It is my Inten- tion to discuss in detail each seotion of the work, and additional ex- periments, at the earliest opportunity, when I hope to dwell more particularly on the significance of such results for the Student of the functions of cell membranes, and for the investigator of the co- ordinations and equilibria in intracellular affairs. IL ON THE DIFFUSIBILITY OF BIOLOGICAL SUBSTANCES THROUGH RUBBER' The writer and his associates have f ound that many ether-soluble substances of biological origin, such as fat and cholesterol, pass readily from ether Solutions through rubber membranes into ether when the mechanical conditions for such diffusions are favorable. Lecithans appear to be wholly indiffusible. Many substances which are soluble in fatty oils, Chloroform, al- cohol, acetone, ethyl acetate, and other solvents of similar powers, or in mixtures of such solvents, promptly diffuse through rubber under suitable conditions. Collodion is one of the products which appears to be indiffusible under such circumstances. When an ordi- nary ethereal Solution of collodion (containing 24 per cent. of alco- hol) is dialyzed in a rubber Condom against ether in a closed vessel, the alcohol rapidly passes to the exterior and the collodion gradually gelatinizes. Liquid accumulates in the bag under these conditions. Various inorganic substances diffuse through rubber under the found to diffuse from benzene through a rubber membrane into benzene; and camphor diffused from pyridin, alcohol, and toluol through rubber membranes into the same solvents, respectively. A recent study of Kahlenberg's paper in the original makes it evident that our results may be explained on the theory of diffusion which Kahlenberg has done much to render convincing. 'Flack and Hill: Journal of Physiology, 1910, xl, p. xxxiii. ' Gies : Proceedings of the Biological Section of the American Chemical Society: Science, 1911, xxxiv, p. 223; Biochemical Bulletin, 1911, i, p. 125. igi2] William J. Gies 59 conditions mentioned above. Ferric sulphocyanate readily passes from ether Solution through rubber into ether. The writer inaugurated these studies, with Dr. Rosenbloom's Cooperation,^ in the hope of devising improvements in the methods. for the isolation of Hpins. The work is progressing along several lines, especially with reference to methods of isolation and purifi- cation, and to osmosis (see page 64). III. A DEMONSTRATION OF OSMOTIC PRESSURE EXERTED BY FAT' In the first of two demonstrations, a cylindrical rubher bag (Con- dom), 13^ inches in diameter and 8 inches long, was lowered into an oiled miislin bag of aboiit the same dimensions. The rubber bag was then filled to overflowing with olive oil. The rubber bag ex- panded, as the oil filled it, to the füll length and width of the muslin sheath. The sheath prevented further extension of the rubber bag and imparted rigidity to the Osmometer that was ultimately con- structed. The double bag, füll of oil and with its mouth wide open, was then raised so as to inclose about an inch of the lower end of a long glass tube which was firmly supported vertically above the demonstration table. The glass tube was 5 feet long and its bore was 4 mm. in diameter. Ligatures were tightly secured around the neck of the double bag against the immersed lower end of the verti- cal tube. The bag then hung directly from the end of the tube. The bag and its sheath were in a tightly distended condition and a stationary column of oil an inch high in the tube was visible above the protruding edge of the sheath. The tube and bag were then lowered into a salt-mouth liter bottle on the table until the bag almost touched the bottom of the bottle. The height of the bottle and the length of the bag were nearly equal. The tube was then marked with a label on the plane of the oil meniscus just above the neck of the bag, and enough ether was poured into the bottle to pro- vide Immersion for the bag to the depth of an inch. For a mo- ment no change in the volume of oil was apparent, and the lateral ^Rosenbloom and Gies: Proceedings of the American Society of Biological Chemists, 191 1, ii, p. 8; Journal of Biological Chemistry, 191 1, ix, p. xiv. •Rosenbloom and Gies: Proceedings of the Society for Experimental Biology and Mediane, 191 1, viii, p. 71. 6o Studies of Diffusion through Rubber Memhranes [Sept. pressure of the ether was obviously without mechanical effect. But in a miniite or two downward diffusion currents were visible along the surface of the bag and oil rose rapidly in the tube. After the initial effects of the ether had been shown, the bottle was filled with ether containing Sudan III, and a 5-foot vertical extension of the same bore was added to the upright glass tube. In a moment the upward movement of the liquid was markedly accelerated. The demonstration was started at about 9 p. m. At 10 p. m. the osmotic pressure had carried the column of oily fluid to the top of the lo-foot tube, and liquid continued to run rapidly from the Upper orifice until the apparatus was dismantled after the adjourn- ment of the meeting, at about 11.30 p. m. During the progress of the demonstration, Sudan III diffused rapidly from the exterior, through the rubber, to the very top of the rising column of fluid, before any of the liquid passed out of the Upper opening. Oil diffused rapidly through the rubber into the ether. The second demonstration was essentially the same in principle and technic as the first. Instead of a lo-foot upright tube, however, the authors substituted an L tube with an inside diameter of 6 mm. The vertical extension of the tube was 17 inches, the horizontal ex- tension was only 3 inches. The latter extension was drawn out to a narrow bore in an inclined plane, to facilitate direct delivery of any liquid that might pass through that end of the tube. When partial immersion of the bag first occurred there was no visible response, but, in a minute or two, oil began to rise in the tube. The bag was then completely covered with ether. The up- ward movement proceeded rapidly ; and in about an hour nearly 200 c.c. of liquid passed through the upper orifice into a graduated cylin- der which was supported underneath the outlet to catch the overflow. IV. A DEMONSTRATION OF THE DIFFUSION OF PIGMENTS FROM FAT THROUGH RUBBER INTO FAT»" The writer has found that many fat-soluble pigments, such as Sudan III and Scarlet R, diffuse readily from solid and liquid fats " Gies : Proceedings of the Society for Experimental Biology and Mediane, 191 1, viii, p. 73. 1912] William J. Gies 6i through rubber into various solid and liquid media, among them both solid fat and oil. Thus, when Sudan III is dissolved in melted lard, the red liquid poured into a rubber bag, the bag supported in melted lard in a bottle, and the apparatus promptly immersed in ice water — the fatty matter will congeal before any sign of pigmentary diffusion occurs but, in a few hours, a reddish tinge will develop outside of the bag, and on each successive day for several weeks further extension of the pigmented matter may be witnes'sed, until the whole of the external lard is deeply suffused with red. This process takes place quite rapidly when the lard and apparatus are kept in a thermostat at 40° C. The demonstrations were intended to exhibit a few instances of such pigmentary diffusions as occur speedily enough at room tem- No. Contents of the Rubber Bag Nature of the Liquid in which the Bag was Suspended Oil Pigment I 2 3 4 S Olive oil Cocoanut oil Lard oil Paraffin oil Olive oil Scarlet R Scarlet R Sudan III Sudan III Sudan III Olive oil Cocoanut oil Lard oil .Paraffin oil Ether Visible diffusion of the pigment oc- curred promptly Visible diffusion of the pigment oc- curred promptly Visible diffusion of the pigment oc- curred promptly Visible diffusion of the pigment oc- curred promptly Visible diffusion of the pigment oc- curred almost immediately perature to yield positive results within an hour. The accompany- ing summary indicates briefly the precise nature and results of the demonstrations (including two control tests — 4 and 5), which were made with thin rubber bags in ordinary glass bottles. The bags were securely supported in the bottles, and the mix- tures were shaken occasionally during the demonstration. The bags were found, after the adjournment of the meeting, to be with- out defects. V. COMPARATIVE DIALYSIS EXPERIMENTS, WITH DEMONSTRATIONS" When dry bags of rubber, gold-beater's skin, parchment, and collodion, each containing olive oil and Sudan III, are separately " Goodridge and Gies : Proceedings of the Society for Experimental Biology and Mediane, 191 1, viii, p. 74. 62 Studies of Diffusion through Rubber Membranes [Sept. immersed in olive oil, and the remaining conditions of the environ- ment are uniform, diffusion of the pigment promptly occurs through rubber, but does not take place at all through any of the other three membranes. When the bags are lifted from the oil, washed ex- ternally with ether, and then immersed in ether,^^ the pigment quickly passes through the rubber, but diffuses very slowly if at all through the remaining membranes. Successive immersions of nioist impermeable membranes (gold- beater's skin and parchment) in alcohol and ether, for different periods of time, fail to render the treated membranes more perme- able to Sudan III than before. The authors demonstrated the general facts in this connection pertaining to rubber and gold-beater's skin. VI. EXPERIMENTS ON THE DIFFUSIBILITY OF ALKALOIDS THROUGH RUBBER^" Various ether-soluble substances, when dissolved in ether and placed in rubber bags immersed in ether, readily pass through the rubber membranes thus imposed (I-V). We have found that various alkaloids and some related substances readily diffuse through rubber under such conditions. Our experiments were conducted as follows : A moderate quan- tity of the pure ether-soluble substance was mixed with 15 to 25 c.c. of ether.^^ This mixture was poured through a funnel into a new air-tight rubber condom in such a way as to preclude the possibility of overflow upon the external surface. The bag was then immersed in about 50 c.c. of ether in a narrow salt-mouth bottle 7 inches high. With the bag suspended at füll extension in this position, its mouth was about an inch above the opening in the bottle. The protrud- "In experiments which the senior author has been conducting with Prof. Welker's Cooperation, it has been found that collodion bags are disintegrated by ether containing more than about 1.5 per cent. of alcohol. Pure ether does not dissolve or in any way disorganize collodion membranes. A collodion bag con- taining pure ether may be immersed for a week or more in pure ether without undergoing any appreciable deterioration. (See page 70.) ^ Sidbury and Gies : Proceedings of the Society for Experimental Biology and Medicine, 191 1, viii, p. 104. " Substances which did not dissolve readily were triturated with ether in a mortar. igi2] William J. Gies 63 ing Condom was supported in the neck of the bottle by a tightly fitting cork Stopper, which also served to keep the bag closed. After a diffusion period of convenient length (sometimes 2 to 5 days)/^ the Condom was cautiously removed from the bottle, the ether diffus- ate was poured into a porcelain dish, and the ether completely re- moved by evaporation on a steam bath. At least one appropriate test was then applied to the residue.^^ Meanwhile, the ether Solution in the Condom was removed. A large volume of water was then poured into the suspended bag, which, during its distention by the water, was carefully examined for signs of leakage. In a few instances defective membranes tem- porarily rendered the outcome doubtful. All results with such bags were ignored, of course. Each of the tests, even after reliable pos- itive responses, was repeated at least once with a nezv rubber bag. The substances named below (the complete list of those already tested in this connection) are readily diffusible under the conditions of these experiments : — A. Apomorphin, atropin, brucin, caffein, Cocain, codein, col- chicin, coniin, morphin, narcein, narcotin, nicotin, physostigmin, quinin, strychnin, veratrin. B. Acetanilid, antipyrin, phenacetin, picric acid, picrotoxin, pyramidon, salicylic acid. Experiments with other solvents, and with additional substances of alkaloidal type, will be added to this series. " Some of the alkaloids pass through rubber almost immediately under the conditions of these experiments. *' In the experiments with nicotin, the " tobacco odor " of the concentrated liquids was very pronounced. STUDIES OF DIFFUSION THROUGH RUBBER MEMBRANES 2. Diffusibility of lipins from ether through rubber membranes into ether JACOB ROSENBLOOM (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) I. INTRODUCTION Many experiments, in completion of the diffusion work I have been doing in collaboration with Dr. Gies/ have been performed to determine the diffusibihty or non-diffusibihty of hpins and similar substances (page 57). Such data must obviously be obtained in detail, if any attempt to devise methods for the isolation and purifi- cation of hpins by dialysis through rubber can be successful. I present here briefly the essential resuks of the work aheady completed in this connection. II. DIFFUSION EXPERIMENTS Methods. In the experiments described below, ordinary rubber Condoms were used as diffusion membranes.^ Various kinds of "sheet rubber," known as "pure Mexican plantation rubber," and fumed with carbon-disulfid, were found to be good membranes for this kind of work, but besides allowing fats, fatty acids, soaps, cho- lesterol and hpochrome to diffuse through it, this sheet rubber also permits the passage of lecithans under the conditions to be described, although the lecithans pass through the sheet rubber very slowly compared with other lipins such as fat and cholesterol. Condoms ^ Rosenbloom and Gies : Proceedings of the American Society of Biological Chemists, 191 1, ii, p. 8; Journal of Biological Chemistry, 191 1, ix, p. xiv. ° Bef ore the Condoms were used for this purpose, they were placed in f resh portions of ether daily for several days, to free them from the powder adherent to them. This is especially important when one proposes to test the dialysates for phosphorus, since the adherent powder has been found to contain phosphorus. 64 I9I2] Jacoh Rosenbloom 65 do not permit the diffuslon of lecithans under the conditions of the tests to be described, and they were preferred for this work for that reason. The cause of the observed difference in permeability is unknown to us, but will soon be made the subject of special inquiry. The substances to be tested in the diffusion experiments were dis- solved in 100 to 200 c.c. of ether (anhydrous and distilled over sodium), the concentrations of the Solutions varying from 0.5 to 5 per cent, The Solution or Suspension was carefully poured through a funnel into a new air-tight rubber condom in such a way as to preclude the possibility of overflow upon the external surface. The bag was then immersed in from 100 to 200 c.c. of pure ether in a wide-mouthed bottle of convenient size, and suspended loosely by a thin cord held securely between the stopper and neck of the tightly stoppered bottle. The bottle was kept well stoppered throughout the whole of each test to prevent egress of ether and ingress of dust and other extraneous matter. 1. Ether extract of egg yolk. Within five minutes after ether extract of egg yolk is subjected to the diffusion treatment described above, the lipochrome appears in the dialysate, diffusion currents being visible about the same time. The following substances can be detected in the dialysate after short periods of dialysis: fat, fatty acid, cholesterol, and lipochrome. The lecithans do not pass through the Condoms, even during prolonged periods of dialysis. We tested for lecithans in the dialysate by analyzing the evapora- tion residue for phosphorus by the fusion and Neumann methods. and by seeking an " acetone precipitate " in the concentrated ether Solution after the addition of electrolyte (sodium chlorid). Some- times a positive phosphorus test was obtained from a dialysate which did not yield an " acetone precipitate." In such cases, it was found that this result was due to the presence of glycerophosphoric acid in the dialysate. If to a Solution of lecithans, which, after dialysis in a condom, does not yield a phosphorus Compound to the dialysate, one adds some glycerophosphoric acid, and then dialyzes this Solution through the same rubber condom, glycerophosphoric acid appears in the diffusate. 2. Ether extract of brain. The dialysate from ether extracts of brain contained fat, fatty acid, and cholesterol. Lecithans failed to dialyze. 66 Studies of Diffusion through Rubber Membranes [Sept. 3. Ether extract of heart muscle (ox). Fat, fatty acid, lipo- chrome, and cholesterol were detected in the dialysate from ether extracts of the heart muscle of oxen. Lecithans did not diffuse. 4. Ether extract of kidney and liver (dog). Fat, fatty acid, lipochrome, and cholesterol appeared in the diffusates from ether extracts of dog kidneys and livers. Lecithans did not dialyze. 5. Ether extract of blood (dog). Fat, fatty acid, lipochrome, and cholesterol occurred in the dialysates from ether extracts of dog blood. No lecithans dialyzed. 6. Ether extract of carrots. The coloring matter dialyzed very rapidly from ether extracts of carrots. A small amount of fat was also present in the dialysate. 7. Ether extract of Xanthoma (skin). The yellow coloring matter dialyzed very quickly from an ether extract of xanthomatous skin, but it faded in twelve hours. 8. Ether extract of cerumen. Cholesterol, fat, and fatty acid were present in the dialysates from ether extracts of cerumen. Neither the coloring matter nor the lecithans diffused. g. Ether extract of yeast. The dialysates from ether extracts of yeast exhibited a peculiar opalescence, even at the end of six weeks' dialysis. A small amount of fat dialyzed, but lecithans did not diffuse. 10. Ether Solutions of individual substances er special prod- ucts. The following substances or special products, when subjected to diffusion by the method described above, were found to be diffusible : Acetic acid Ethyl butyrate Palmitic acid Acetone Formic acid Potassium palmitate* Beta-hydroxy-butyric acid GlyceroP Potassium stearate* Butter (fresh and rancid) Lactic acid Propionic acid Butyric acid Lead oleate Sodium palmitate* Cholesterol-acetate Mutton tallow Sodium stearate* ' When ether-alcohol Solutions of glj'cerol are dialyzed against ether-alcohol, and alcohol Solutions of glycerol are dialyzed against ether, the dialysates contain glycerol. * Treated with water, then with alcohol to the point of precipitation, then with ether until a precipitate was produced. The filtrate was dialyzed against water, alcohol, and ether in identical proportions. 912] Jacob Rosenhloom 6y Cholesterol-benzoate Oleic acid Stearic acid Cholesterol (from brain, Olive oil Sudan III egg yolk, and gall-stones) Olive oil stained with Urochrome' Ethyl acetate Sudan III Valerianic acid In some special experiments we found^ that cholesterol benzoate, cholesterol stearate, cholesterol oleate and cholesterol palmitate, when dissolved in ether, readily diffuse through rubber into ether. Cholesterol stearate with a molecular weight of 652.61 diffuses, whereas the various lecithans, with molecular weights considered to be 770 to 785, do not. If we assume that the diffusion of a sub- stance depends on the size of its molecules, the above facts strengthen Hiestand's conclusion that the molecular weight of ^gg- yolk lecithin is 1446, which figure he obtained by a molecular weight determination, II. Indiffusible substances. The following substances, when subjected to diffusion by the method described above, were found to be indiffusible.'^ Sodium chlorid Lecithans from yeast Lecithans from brain Lecithans from wheat embryo Lecithans from egg yolk Kephalin from brain Lecithans from heart muscle Cuorin from heart muscle Lecithans from pig testicle Compound of lecithin with platinic chlorid Lecithans from liver and kidney Koch^ has lately described the preparation of various Compounds with lecithans, but it is uncertain whether these Compounds are colloidal adsorptions, mechanical mixtures, or true chemical Com- pounds. It seemed of interest to study the behavior of these sub- stances in ether Solution, when subjected to dialysis in rubber bags suspended in ether. The preparations used in these experiments were made accord- ' Ether-alcohol Solution (equal amounts) dialyzed against ether-alcohol. ' Boas and Rosenbloom : Proceedings of the Society for Experimental Biology and Medicine, 191 1, viii, p. 132. ''We have found that lecithans prepared by the Zuelzer, Bergeil, or Roaf and Edie method, when dialyzed, always yield traces of cholesterol to the dialysate; and often fat. * Koch and collaborators : Journal of Pharmacology and Experimental Therapeiitics, 1910, xii, 239-269. 68 Studies of Diffusion through Rubber Membranes [Sept. ing to the method described by Koch. For the dialysis tests the Solutions of the lecithan Compounds were evaporated to dryness at 38° and the residues triturated with ether. The extracts were fil- tered, and the filtrates placed inside of rubber bags and dialyzed against ether for thirty-seven days. The dialysates were tested weekly to see if the substance combined with the lecithan had diffused. Compounds of lecithin with glucose, lactic acid, strychnin, digi- tonin, salicin, urea, creatin, Creatinin, and caffein were prepared. It was found that the glucose and lactic acid dialyzed completely, the strychnin, digitonin, and salicin dialyzed partially, while urea, creatin, Creatinin, and caffein did not dialyze at all.^ It was thought that some of the various substances which did not diffuse might do so in the presence of a considerable amount of dif- fusible material, but on dialyzing various mixtures of the above- named indiffusible substances with varying amounts (up to 15 grams), of neutral fat, fatty acid, cholesterol, or olive oil, no diffu- sion of lecithan occurred. When Solutions of lecithans are subjected to dialysis by the method described above, they take up a great deal of ether, and the volume of liquid in the bag is greatly increased. We have demon- strated that lipins exert strong osmotic pressure. ( See page 59. ) We have also placed ether Solutions of lecithans with cholesterol and fat in closed rubber bags suspended in Soxhlet extractors. Soxhlet extraction in the usual way failed to remove lecithan from the bag under these conditions. These findings favor the develop- ment of a method for the thorough removal of impurities from lecithan Solutions. It is perhaps superfluous to add that the results already mentioned may be obtained by placing the Solution to be tested outside the rubber bag and allowing dialysis to take place into pure ether con- tained in the bag. III. SUMMARY OF GENERAL CONCLUSIONS I. Most lipins, chief among them, fat, fatty acid, soaps, cho- lesterol, cholesterol-esters, lipochrome, and various other ether-sol- • Boas and Rosenbloom : Loc. cit. I9I2] Jacob Rosenhloom 69 üble substances, diffuse from ether Solution through rubber membranes into ether, 2. Sodium Chlorid, lecithans prepared from various sources, kephalin, cuorin, and the Compound of platinum with lecithin, do not diffuse under such conditions. 3. One or more of the diffusible substances in these experiments may be dialyzed from Solutions containing them, together with one or more of the indiffusible ones, without inducing any of the latter to pass through the membrane. STUDIES OF DIFFUSION THROUGH RUBBER MEMBRANES 3. Diffusibility of protein through rubber membranes, with a note on the disintegration o£ coUodion membranes by common ethyl ether and other solvents WILLIAM H. WELKER (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) l. INTRODUCTION In the course of our studies of proteins, under the auspices of the George Crocker Special Research Fund, we obtained a protein product which is soluble in a mixture of equal parts of absolute alcohol and absolute ether, and which responds to the biuret, xantho- proteic, Millon and Hopkins-Cole tests. The material was prepared by the following method : 25 grams of Witte peptone were dissolved in 500 c.c. of 0.2 per cent. hydrochloric acid Solution. The liquid was evaporated to a thick syrup on a water bath. This syrup was thoroughly extracted with absolute alcohol and the resultant yellow liquid filtered. The filtrate was treated with an equal volume of absolute ether, which produced a white flocculent precipitate. After the Sedimentation of the precipitate, the supernatant liquid was de- canted and filtered. When five volumes of absolute ether were added to this filtrate, a white flocculent precipitate was produced. This product was isolated by filtration, washed with absolute ether, and exposed to the air in a thin layer on a watch glass, where it solidified as yellowish and somewhat hygroscopic granulär material, which could easily be pulverized. The product dissolved promptly in absolute alcohol. From concentrated alcoholic Solution the prod- uct may be precipitated by the addition of an equal volume of abso- lute ether. Whether the product is peptone or a much simpler Poly- peptid has not yet been determined. Dr. Gies and his collaborators have lately given much attention to 70 1912] William H. Welker 71 the diffusibility of lipins and other substances throtigh rubber mem- branes. The solubility of the above mentioned product in alcohol- ether Solution led Dr. Gies to propose a study of the comparative diffusibility of the protein through membranes of rubber, parchment and collodion. Such an investigation was accordingly conducted by the diffusion process described on page 55. The data in the accompanying tables indicate the conditions and the results of the tests in this connection.^ IL COMPARATIVE DIFFUSION EXPERIMENTS Experiments with rubber membranes. The results of the tests in the first four series show collectively [Table i (1-15)] that biuret-reacting matter appeared in the diffusates; that the rubber itself did not yield such substance; and that the occurrence of biuret- TABLE I Results of experiments with rubber membranes A. Data showing the dißusibility of biuret-reacting material First Series. With Rubber Condoms. Results of Results of the Duration of Liquid outside the biuret test for leaks at Exp. No. the experi- Contents of the bag of the bag test in the the end of the ment, days diffusate experiment I 4 r 30 c.c. ether-alc. sol. \ 10 c.c. abs. ether Abs. ether + + + Small hole in the bag 2 4 f 20 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + No leak 3 4 f 10 c.c. ether-alc. sol. l 30 c.c. abs. ether Abs. ether + No leak Second Series. With Rubber Condoms. 4 5 / 30 c.c. ether-alc. sol. \ 10 c.c. abs. ether Abs. ether + + + Small hole very high up in the bag 5 5 f 20 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + No leak 6 5 f 10 c.c. ether-alc. sol. \ 30 c.c. abs. ether Abs. ether + No leak * In the tables, " ether-alc. sol." indicates the protein Solution as it was made available by the above mentioned process before final precipitation with five volumes of ether. Such precipitation was efifected only when the solid was desired for special reasons. See the data pertaining to the eighth series of tests, Table i. 72 Studies of Diffusion through Rubber Membranes [Sept. TABLE I (continued) Third Series. With Rubber Condoms. Exp. No. Duration of the experi- ment, days Contents of the bag Liquid outside of the bag Results of the biuret test in the difTusate Results of the test for leaks at the end of the experiment 7 S / 30 c.c. ether-alc. sol. 1 lo c.c. abs. ether Abs. ether + + + No leak 8 5 f 20 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + No leak 9 5 f lo c.c. ether-alc. sol. \ 30 c.c. abs. ether Abs. ether + No leak lO« 5 Abs. ether ("control") Abs. ether — No leak II» 5 Abs. ether ("control") Abs. ether — FouRTH Series. With Bags of Sheet Rubber.* 12 30 / 60 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + + No leak 13 30 f 40 c.c. ether-alc. sol. 1 40 c.c. abs. ether Abs. ether + + No leak 14 30 / 20 c.c. ether-alc. sol. L 60 c.c. abs. ether Abs. ether — No leak IS 30 Abs. ether ("control") Abs. ether — No leak B. Data showing that the diffusible biuret-reacting material ii-15) was true protein Fifth Series. With Rubber Condoms. Duration of the experi- ment, days Contents of the bag Liquid outside of the bag Results of the tests for protein in the diffusate Results of the test for Exp. No. Biuret Hop- kins- Cole Xan- thopro- teic leaks at the end of the experiment 16 17 3 3 / 30 c.c. ether-alc. sol. \ 10 c.c. abs. ether 30 c.c. ether-alc. sol. Abs. ether Abs. ether + + + + + + No leak No leak SixTH Series. With Bags of Sheet Rubber.* 18 3 / 60 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + + No leak 19 3 r 60 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether + + + No leak Seventh Series. With Rubber Condoms. 20 31 3 3 40 c.c. ether-alc. sol. 40 c.c. ether-alc. sol. Abs. ether Abs. ether + + + + + + No leak No leak •This control experiment (10) was carried out in duplicate, with negative results in each case. *In this experiment (11) a new, clean, empty condom, with the bottom removed, was suspended in absolute ether, for " control " purposes. * " Pure Mexican plantation rubber." I9I2] William H. Welker 73 TABLE I (continued) C. Data showing the eßect of water on the diffusion phenomena (.r-21) EiGHTH Series. With Rubber Condoms.' Exp. No. Duration of the experi- ment, days Contents of the bag Liquid outside of the bag Results of the biurei test in the diffusate Results of the test for leaks at the end of the experiment 3 C.C. abs. alc. sol. 6 c.c. abs. alc. 22 5 ' 9 c.c. abs. ether 6 c.c. H2O 3 c.c. abs. alc. 3 c.c. abs. alc. sol. 18 c.c. abs. ether . 1.2 c.c. H2O 9 c.c. abs. alc. •" No leak 23 5 21 c.c. abs. ether I c.c. H2O 6 c.c. abs alc. 21 c.c. abs. ether I c.c. HsO — No leak 24 5 \ r 3 c.c. abs. alc. sol. l 3 c.c. abs. ether Equal volumes of abs. ether + No leak and abs. al- cohol D. Data showing the effect of fat on the diffusion phenomena {1-24) NiNTH Series. With Rubber Condoms. 25 10 Olive oil and Witte Pep- tone (solid) Olive oil _e 26 10 Olive oil, Witte peptone (solid) and H20^ Olive oil ^" Tenth Series. With Rubber Condoms. 27 28 30 30 Olive oil and Witte pep- tone (solid) Olive oil, Witte peptone and HjO^ Olive oil Olive oil Eleventh Series. With Rubber Condoms. 29 10 f Ether-alc. sol. iLard Abs. ether _ 30 10 f Ether-alc. sol. "iLard Abs. ether — Twelfth Series, With Rubber Condoms. 31 1 / Ether-alc. sol. LLard Abs. ether + No leak 32 I / Ether-alc. sol. iLard Abs. ether + No leak Thirteenth Series. With Rubber Condoms. 33 2 f 30 C.C. ether-alc. sol. ILard 30 c.c. abs. ether + No leak 34 3 / 30 C.C. ether-alc. sol. ILard 30 c.c. abs. ether + No leak 'For this series of tests, we used 0.2 gram of the protein material dissolved in 10 c.c. of absolute alcohol. • On the water-soluble extract of the oil. ^ Water sufficient to make a paste of the Witte peptone was used. The paste was triturated into the oil. 74 SUidies of Diffusion throngh Rubber Membranes [Sept. reacting material in the diffusate was not due to perforations of the bags. The diffusion of biuret-reacting material was always greatest in degree through the bags containing the largest proportion of protein. The results of tests 1-15 show that biuret-reacting material dif- fiised through the rubber membranes under the conditions imposed. In Order to determine more definitely, however, whether protein dif- fused through the rubber, we repeated the essential features of tests 1-15, but applied additional tests to the diffusates [Table i (16- 21)]. The results of tests 16-21 (Table i) confirm the findings of tests 1-15, and also show definitely that the dififusible biuret-reacting ma- terial was triie protein. The data of tests 1-2 1 suggest that osmosis depends upon affin- ities between the membrane, and the solvent or solute, or both. We made a direct test of this mattter in a preliminary way by adding water to the solvent and thus disturbing its affinities with the mem- brane without decreasing the solubility of the solute. The findings are given in the summary pertaining to the eighth series (Table i ). The results of tests 22-24 show that water exerted a disturbing osmotic influence and that diffusion of the protein was entirely pre- vented by the water. We extended these experiments to determina- tions of the influence of associated, readily diffusible lipins, in the presence or absence of water. The results are given in tests 25-34. In the tenth series the oil in the diffusates was emulsified with a little soap Solution and then repeatedly extracted with ether until all the fat was removed. The water containing the soap, and the aqueous extract of the oil, were evaporated to dryness and the biuret test applied to a concentrated Solution of the residue. Experiments with parchment-paper bags. The foregoing re- sults with rubber membranes naturally increased our desire to make comparative observations with bags of parchment and collodion. The results of the tests with parchment are given in tests 35-42, Table 2. That osmosis depends upon accord between the solvent and the membrane is obvious from these results also, for the protein substance, which is readily diffusible through parchment from aqueous Solution, does not dialyze through such a membrane from an alcohol-ether Solution. I9I2] William H. Welker n TABLE 2 Results of experiments with bags of parchment paper FOURTEENTH SeRIES. Duration of the Results of the Exp. No. experiment, Contents of the bag Liquid outside of biuret lest in days the bag the diffusate 35 I f 10 c.c. ether-alc. sol. \ 30 c.c. abs. ether Abs. ether 36 2 / 3 c.c. ether-alc. sol. \ 3 c.c. abs. ether Abs. ether — FiFTEENTH SERIES.« 37 10 / 30 c.c. ether-alc. sol. \ 10 c.c. abs. ether Abs. ether ^ 38 10 f 20 c.c. ether-alc. sol. 1 20 c.c. abs. ether Abs. ether — 39 IG f 10 c.c. ether-alc. sol. \ 30 c.c. abs. ether Abs. ether — SiXTEENTH SERIES.» 40 10 f 30 c.c. ether-alc. sol. \ 10 c.c. abs. ether Abs. ether _ 41 10 f 20 c.c. ether-alc. sol. \ 20 c.c. abs. ether Abs. ether -" 42 10 / 10 c.c. ether-alc. sol. \ 30 c.c. abs. ether Abs. ether — III. ON THE UTILITY OF COLLODION.BAGS IN EXPERIMENTS OF THE KIND DESCRIBED IN THE FOREGOING SECTIONS Experiments like those in the sixteenth series (Table 2) were attempted with bags made of collodion, but in each case the bags were perforated and in part dissolved, by the Contents, before the experiment could be fairly started. Several years ago, Dr. Gies observed, in some dialysis experi- ments with collodion membranes, that ethyl ether could be kept on the aqueous Contents of collodion bags, for preservative purposes in such tests, without inducing distintegration of the bags. In repeti- tions of the experiments a year or two later, however, it was found that ether under such circumstances often caused deterioration of *The results in this series, while apparently negative in each case, were somewhat doubtful owing to the fact that the paper contained some soluble material, which rendered the biuret test more or less uncertain. See the results of the tests in the sixteenth series. *The parchment paper was washed free from soluble matter before the beginning of the tests. 76 Studies of Diffusion through Rubber Membranes [Sept. the membrane, but that occasionally it did not. The reason for such variations in the action of the ether could not be conveniently ascer- tained at the time. The prompt Perforation of the collodion bags in our several attempts, as stated above, to determine the diffusibihty through col- lodion of the alcohol-ether soluble protein, recalled Dr. Gies' pre- vious experiences and led him to suspect that the alcohol, in the Solu- tions employed by us, was responsible for the observed destructive effects on the collodion membrane in these experiments. He be- lieved, also, that the previous variations in the action of ether on collodion in dialysis experiments, as already related, were due to differences in the degrees of purity of the ether employed. At Dr. Gies' request, therefore, I made direct tests of the solvent action of ether containing alcohol, and various other substances related in one way or another to alcohol and ether. Collodion bags were made, in test tubes, from U. S. P. col- lodion.^*^ It was found that such bags were not perforated by abso- lute ether when it was poured into them 10 minutes after their re- moval from the tubes, i. e., after fairly complete evaporation of the residual alcohol. The time required for the evaporation of the residual alcohol is dependent on the prevailing temperature. At low temperatures the alcohol disappears from the collodion membrane very slowly. Common ether (Merck's 0.720 sp. gr.), however, when poured into such bags, passed through them almost immedi- ately, with general Solution of the collodion, even after 2 hours of preliminary exposure of the bag outside the mould. In the first tests of the effects of alcohol it was found that absolute ether con- taining 1.5 per Cent, or more of added absolute alcohol promptly penetrated the bags. In a series of more careful tests of absolute ether containing various percentages of added absolute alcohol, it was found that the bags were penetrated promptly by ether containing more than 1,25 per Cent, of alcohol, but that the mixture containing 1.25 per cent. of alcohol acted more slowly. Ether containing less than 1.25 ^r cent. of alcohol exhibited no destructive action. Qualitative tests showed that acetone, acetaldehyde, ethyl acetate, methyl alcohol and glacial acetic acid attack and penetrate collodion "An ether Solution containing alcohol. igi2] William H. Welker 77 bags^^ immediately, toluol slowly, whereas formic acid, formalde- hyde (40 per cent.), Chloroform, petroleum ether, carbon tetra- chlorid, carbon bisulfid and paraffin oil were without distinguishable solvent action, even after long periods of contact. Acetone (5 per cent.) in absolute ether attacks collodion bags slowly, while a 10 per cent. Solution acts rapidly. Acetaldehyde (4 per cent.) in absolute ether attacks the bags slowly, but a 5 per cent. Solution acts rapidly. Methyl alcohol (3 per cent.) in absolute ether dissolves the bags, but a 2 per cent. Solution does not. Glacial acetic acid (2 per cent.) in absolute ether attacks the bags slowly, a 3 per cent. Solution acts rapidly but a i per cent. Solution appears to be inert. Five per cent. Solutions of Chloroform, toluol, petroleum ether, carbon tetra- chlorid, carbon bisulfid, benzol, ethyl acetate, and paraffin oil, in absolute ether, were without visible effect on collodion bags. Five per cent. Solutions of formic acid (sp. gr. 1.2) and formalde- hyde (40 per cent.), in absolute ether, immediately attacked and penetrated collodion bags. Further work along these lines is in progress. My cordial thanks are due Dr. Gies for his kind direction and assistance in these experiments. "Bags practically free from residual alcohol were used. STUDIES OF DIFFUSION THROUGH RUBBER MEMBRANES 4. The comparative diffusibility of various pigments in different solvents GEORGE D. BEAL and GEORGE A. GEIGER {Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) I. INTRODUCTION Dr. Gies and his associates have shown that many biological substances diffuse through rubber under suitable conditions (page 55). Inorganic as well as organic substances exhibit this capacity and various colloids share it with crystalloids. Lipochrome and ferric sulfocyanate are among the colored substances which, in the early experiments, were found to be diffusible from ether Solution through rubber membranes into ether. At Dr. Gies' Suggestion we undertook a similar study of the diffusibility of common pigments, especially " food colors." Fol- lowing his advice we also sought data which might be of service in devising methods for the purification of pigments, and for their Separation and detection under various circumstances. Our diffusion tests were conducted by the following general method: A moderate quantity of the pigment was mixed with 15- 25 c.c. of the solvent. The Solution, or Suspension, was carefully poured into a rubber Condom in such a way as to preclude the pos- sibility of overflow upon the external surface. The bag was then immersed in about 50 c.c. of solvent in a narrow salt-mouth bottle 7 inches high. With the bag suspended at füll extension in this Posi- tion, its mouth was about an inch above the opening in the bottle. The protruding condom was supported in the neck of the bottle by a tightly fitting cork stopper, which served to keep both the bag and the bottle closed. The diffusion periods varied from a few minutes \o a week or more, according to the obvious requirements in each case for a definite conclusion regarding diffusibility. 78 I9I2] George D. Beul and George A.. Geiger 79 Most of the original tests were made with new Condoms. Many tests were repeated with Condoms which had previously been em- ployed by us in pigment-diffusion experiments but which, prior to being used again, had been thoroughly washed with portions of the solvent to which they were soon to be subjected in the new diffusion tests. Defects in the rubber could easily be detected. All doubtful results were ignored. Numerous repetitions prevented erroneous deductions. In the accompanying summary we present an outline of the various tests and the main results of each. For the sake of con- venience we use in the summary the following abbreviations : D, diffusion; Di, pigment appears in the diffusate within lo minutes; D2, pigment does not diffuse within 10 minutes, but appears in the diffusate within 30 minutes; D3, pigment does not diffuse within 30 minutes, but appears in the diffusate within i hour; D4, pigment does not diffuse within i hour, but appears in the diffusate before the lapse of 2 hours ; D5, pigment cannot be seen in the diffusate before the third hour of diffusion, but appears before the fourth hour; D6, pigment cannot be seen in the diffusate before the sixth hour of diffusion, but appears before the eighth hour; D7, pigment cannot be seen in the diffusate before the tenth hour of diffusion, but appears before the twelfth hour; D8, pigment appears in diffusate in about 24 hours; O, no visible diffusion at any time within a week. IL SUMMARY OF DIFFUSION DATA Inside Outside solvent Result Solvent Pigment Remarks I Ether. . . Sudan III Ether Ether + alco- hol (25%). Ether + alco- hol (50%). Ether -f alco- hol (75%). Alcohol (100%)... Chloroform . . Di Di Di Di Di Di D very rapid. 2 Ether. . . Sudan III 3 Ether . . . 4 Ether. . . 5 Ether. . . 6 Ether . . . Sudan III D rapid though slower than i. Sudan III D slower than 2. Sudan III D slower than 3. Sudan III Ether was withdrawn, leaving a concentrated Solution of Sudan III. Currents very distinct. CoUection of color near top very marked. Color zone on top. No diffusion currents downward. 8o Studios of Diffusion through Rubber Membranes [Sept. II. SUMMARY OF DIFFUSION DATA {continued) Inside Solvent 7 Ether. 8 Ether . . . 9 Ether. . . 10 Ether . . . 11 Alcohol. . 12 Chloro- form . . . 13 Alcohol . . 14 Ethyl acetate . . 15 Acetone.. 16 Gl. acetic acid .... 17 Ether. . . 18 Ether. . . 19 Ethyl acetate . . 30 Acetone.. 21 Alcohol. . 27 Ether. . . 23 Ether. . . 24 Ether . . . 25 Ether. Pigment 26 Ether . . 27 Ether. . 28 Ether. . 29 Ether. . 30 Ether . . 31 Ether. . 32 Ether. . 33 Ether. . 34 Ether . . 35 Ether. . 36 Ethyl acetate . 37 Ethyl acetate . Sudan III Sudan III Sudan III Sudan III Sudan III Sudan III Sudan III Sudan III Sudan III Sudan III Picric acid Hematoxylin Methyl violet Methyl violet Methyl violet Methyl violet Magenta Naphthol yellow . . . Methyl violet and Sudan III Outside solvent Methyl alco- hol Acetone . Petroleum ether. . . . Gl. acetic acid Ether Chloroform . Alcohol . . . . Ethyl acetate Acetone .... Gl. acetic acid Ether Ether Ethyl acetate Acetone . Alcohol . Ether. . . Ether. . . Ether... Resnlt Ether. Chlorophyll Ether. Annatto Alcannin Metanil yellow. . Martius yellow. . Scarlet R Malachite green. Brazil wood . . . . Chrysoldin Turmeric 'Ether. Ether. Ether. Ether. Ether. Ether. Ether. Ether. Ether. Annatto . Chlorophyl . Ethyl acetate Ethyl acetate Di Di Di Di Di Di Di Di Di Di Di D3 DS D6 O D5 Ds O Di Di Di Di O Di Di D2 Di D2 Di D2 Di Remarks Ether was withdrawn, leaving layer of dye inside. Ether withdrawn. Solution of pigment in bag concentrated. Moderate diffusion. Diffusion slow. Rapid diffusion. Diffusion slow. Slow diffusion. Moderate diffusion. Slow diffusion. Rubber yellow; color not re- moved by ether. Denser Solution in bag than outside. (See 36.) Rapid diffusion of the Sudan III. Ether changed three times in 2 hours after which practically all Sudan III had been removed, leaving the methyl violet in the bag. Vary slight diffusion. Very slight diffusion. Very rapid diffusion. Rapid diffusion. Very rapid diffusion. Bag colored green. Slow diffusion. Very slow diffusion. Rapid diffusion. Pigment Solution inside concen- trated.« Pigment Solution inside concen- trated. • " Fat-soluble " chlorophyl was used in all the chlorophyl tests. * In these cases (36-44) the solvent diffused more rapidly than the solute. 1912] II. George D. Beal and George A. Geiger SUMMARY OF DIFFUSION DATA (continued) 8i Inside Outside solvent Result T< ATVl 0 f l^e Solvent Pigment JxCularKa 38 Ethyl acetate . . Alcannin Ethyl acetate Di Pigment Solution inside concen- trated. 39 Ethyl acetate . . Martius yellow .... Ethyl acetate Di Pigment Solution inside concen- trated. 40 Ethyl acetate . . Scarlet R Ethyl acetate Di Pigment Solution inside concen- trated. 41 Ethyl acetate . . Brazil wood Ethyl acetate Di Pigment Solution inside concen- trated. 42 Ethyl acetate . . Chrysoidin Ethyl acetate D3 Pigment Solution inside concen- trated. 43 Ethyl acetate . . Turmeric Ethyl acetate D3 Pigment Solution inside concen- ^-'O trated. 44 Ethyl acetate. . Metanil yellow Ethyl acetate D3 Pigment Solution inside concen- trated. 45 Ethyl acetate . . Malachite green. . . . Ethyl acetate D6 46 Ethyl acetate . . Naphthol yellow . . . Ethyl acetate 0 47 Ethyl acetate . . Sudan I Ethyl acetate Di Rapid diffusion. 48 Ethyl acetate . . Sudan G Ethyl acetate Di Moderate diffusion. 49 Ethyl acetate . . Rhodamin Ethyl acetate D in about 2 days. 50 Ethyl acetate . . Fast red A Ethyl acetate D3 Diffusion very slight in each of 51 Ethyl tests 50-56 inclusive. acetate . . Rose bengal Ethyl acetate D2 52 Ethyl acetate. . Erythrosin Ethyl acetate D2 53 Ethyl acetate . . Methylene violet . . . Ethyl acetate D5 54 Ethyl acetate. . Phloxin red Ethyl acetate D3 55 Ethyl acetate . . Auramine Ethyl acetate D5 56 Ethyl acetate . . Orange G Ethyl acetate D in about 5 hours. 57 Methyl alcohol. . Gold orange Methyl alcohol .... D8 Color of diffusate very slight i week later. 58 Methyl alcohol. . Naphthol yellow . . . Methyl alcohol. . . . No appearance of color in 8 hours. 59 Meth alcohol. . Carmosin B Methyl alcohol .... D8 Color of diffusate very slight i week later. 82 Studics of Diffusion through Ruhher Memhranes [Sept II. SUMMARY OF DIFFUSION DATA (continued) Inside Solvent 60 Methyl alcohol . 61 Methyl alcohol . 62 Methyl alcohol . 63 Methyl alcohol . 64 Amyl alcohol . 65 Amyl alcohol . 66 Amyl alcohol . 67 Amyl alcohol . 68 Amyl alcohol . . 69 Amyl alcohol. . 70 Amyl alcohol. . 71 Amyl alcohol. . 72 Amyl alcohol . , 73 Amyl alcohol. , 74 Amyl alcohol . . 75 Amyl alcohol. , 76 Amyl alcohol . , 77 Acetone. 78 Acetone. 79 Acetone. 80 Acetone. 81 Acetone. 82 Acetone. 83 Acetone. 84 Acetone. 85 Acetone. 86 Acetone. 87 Acetone. 88 Acetone. 89 Acetone. Pigment Ponceau, G. A.. Ponceau, 2 R. . Naphthol red S. Curcumin S . . . Fast red A . Safranin . . . Eosin A. Phloxin . Rose bengal . Rhodamin . . Erythrosin . . Chrysoldin Sudan I . . . Sudan G. . . Sudan III. Alcannin. . Chlorophyl . Alcannin . . , Auramine. . Barwood . . , Chlorophyl . Chrysoldin . Fast red A . Methylene violet. . Malachite green. . . Martins yellow. . . . Metanil yellow. . . . Naphthol yellow S. Picric acid Rhodamin Outside solvent Result Methyl alcohol. . . . Methyl alcohol. . . . Methyl alcohol. . . . Methyl alcohol. . . . Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Amyl alcohol Acetone. . . Acetone. . . Acetone . . . Acetone . . . Acetone . . . Acetone. . . Acetone . . . Acetone. . . Acetone . . . Acetone . . . Acetone. . . Acetone. . . Acetone. . . O O O O D7 D7 O O D7 D8 D3 D2 D2 D2 D3 D3 Di Di Di D3 D5 D4 D4 D5 D3 D8 O Dl D7 Remaxks Color of diffusate very slight I week later. D in about 3 days. Color of diffusate very shght i week later. Color of diffusate very slight I week later. Very rapid diffusion. Color of diffusate not very strong I week later. I9I2] George D. Beal and George A. Geiger 83 II SUMMARY OF DIFFUSION DATA {continued) Inside Outside solvent ^Result Remarks Solvent Pigment 90 Acetone.. Sudan I Acetone Acetone Acetone Acetone Dl Di 0 D8 91 Acetone.. Sudan G 92 Acetone.. Fustic 93 Acetone.. Rose bengal Color of dif?usate very slight i week later. 94 Acetone . . Phloxin Acetone . D4 Color of diffusate very slight I week later. A ^^^ V.. W^^ 4 Jk^h^ • • ■ • a 95 Acetone.. Eosin W. gelblich . . . Acetone D7 Color of diffusate very slight I week later. 96 Acetone.. 97 Acetone.. Eosin A Acetone Acetone 0 D4 Cape aloes 98 Gl. acetic acid .... Sudan G Gl. acetic acid Di Slow diffusion. 99 Gl. acetic fc..^**.^ TT *.A**Ä 1.4V^ A^..r AA # acid .... Sudan III Gl. acetic acid Dl Rapid diffusion. 100 Gl. acetic acid. . . . Sudan I Gl. acetic acid Di Slow diffusion. loi Gl. acetic acid. . . . Alcannin Gl. acetic acid D3 102 Gl. acetic acid .... Chlorophyl Gl. acetic acid D6 103 Gl. acetic acid .... Rose bengal Gl. acetic acid DB 104 Gl. acetic acid .... Phloxin Gl. acetic acid D in about 2 days. 105 Gl. acetic acid. . . . Malachite green. . . . Gl. acetic acid 0 106 Gl. acetic acid .... Methyl violet Gl. acetic acid D8 107 Gl. acetic acid. . . . Scarlet R Gl. acetic acid D2 108 Gl. acetic acid .... Methylene violet . . . Gl. acetic acid D in about 10 days. 109 Gl. acetic acid .... Martius yellow Gl. acetic acid D8 iio Gl. acetic acid .... Biebrich Scarlet. . . . Gl. acetic acid D8 III Gl. acetic acid. . . . Erythrosin Gl. acetic acid D in about 4 days. 112 Gl. acetic acid .... Oranee G Gl. acetic acid 0 113 Gl. acetic ^.—1' ^ v***^^ ^- ^-* ••••»• •«• ■ ^t^H« * ■ fc*^-v* W^V^ ^.l>VrA^.J acid. . . . Tropeolin OO Gl. acetic acid Very slight color after 2 days, which did not increase after Standing about four days. 114 Gl. acetic acid. . . . Aurainine Gl. acetic acid 0 115 Gl. acetic acid. . Rhodamin Gl. acetic acid D8 116 Gl. acetic ^^.AA V.' X.A%4* AA * A^ ■«•■*«*«■ acid .... Eosin A. gelblich . . . Gl. acetic acid D8 117 Gl. acetic acid .... Chrysoidin Gl. acetic acid D in about s days. 118 Gl. acetic acid .... Eosin W Gl. acetic acid D in about 4 days, which did not increase during the suc- M rf\J\^i**-* »■• •••••■••• ceeding 3 days. 84 Stndics of Diffusion through Rubber Membranes [Sept. II. SUMMARY OF DIFFUSION DATA (continued) Inside Solvent 119 Gl. acetic acid. . . . 120 Gl. acetic acid .... acetic acetic acetic 121 Gl. acetic acid 122 Gl acid . . 123 Gl acid. 124 Gl acid. . . . 125 Gl. acetic acid .... 126 Alcohol. . 127 Alcohol. . 128 Alcohol. . 129 Alcohol. . 130 Alcohol. . 131 Alcohol. . 132 Alcohol. . 133 Alcohol. . 134 Alcohol. . 135 Alcohol. . 136 Alcohol. . 137 Alcohol. . 138 Alcohol. . 139 Alcohol. . 140 Alcohol. . 141 Alcohol. . 142 Alcohol. . 143 Alcohol. . 144 Alcohol. . 145 Alcohol. . 146 Alcohol. . 147 Alcohol. . 148 Alcohol.. 149 Alcohol. . 150 Alcohol. . 151 Alcohol. . 152 Alcohol. . 153 Alcohol. . 154 Alcohol. . 15s Alcohol. . Pigment Fast Red A Safranin Turmeric Metanil yellow Barwood Annatto Picric acid Auramine ChrysoTdin. .._..... Eosin A Eosin W ' Fast Red Methyl violet Methylene violet. . . Malachite green. . . . Martins yellow Metanil yellow Rose bengal Rhodamin Sudan G Sudan I Bismarck brown . . . Benzopurpurin Tropeolin OO Phloxin Safranin Naphthol yellow . . . Alcannin Chlorophyl Barwood Fustic Turmeric Cape aloes Curcumin S Naphthol green. . . . Orange G Carmosin B Outside solvent Gl. acetic acid Gl. acetic acid Gl. acetic acid Gl. acetic acid Gl. acetic acid Gl. acetic acid Gl. acetic acid Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Alcohol Result D3 O D7 O D7 D7 O D7 D8 D7 DB D7 D7 O O D2 D2 O O D7 O DS D7 O O O O O Remarks D in about 4 days, which did not increase during the suc- ceeding 3 days. D in about 2 days. D in about 6 days. D in about 3 days. Color of diffusate very slight I week later. Color of diffusate very slight I week later. Color of diffusate very slight I week later. Color of diffusate very slight I week later. D in about 3 days. D in about 5 days. Slight I week later. Color of diffusate very slight I week later. D in about 2 days. D in about 2 days. I week later. D in about 2 days. I week later. Very slight Very slight I9I2] George D. Beal and George A. Geiger 85 III. ATTEMPTS TO SEPARATE PIGMENTS BY DIALYSIS The outcome of Test 25 encouraged us to ascertain whether two dissimilar pigments like scarlet R and malachite green might be wholly separated from each other by dialysis thru rubber in a suit- able solvent, e. g., ethyl acetate (see tests 40 and 45). A mixture of the two pigments dissolved in ethyl acetate was accordingly sub- jected to the usual mechanical treatment, but the diffusate was re- peatedly replaced with fresh solvent. The results are indicated in the f ollowing summary : Continuous differential diffusion of scarlet R and malachite green. March 28 — ist diffusate ii-i p.m. Bright red. March 28 — 2nd diffusate 4 p.m. Bright red. March 28 — 3rd diffusate 11 p.m. Deep red. March 29 — 4ith diffusate 12.30 a.m. Deep purplish red. March 29 — 5th diffusate i a.m. Light purplish red. March 29 — 6th diffusate 10.45 ^■^- Deep red with decidedly bluish tinge. March 29 — 7th diffusate 12.30 p.m. Light blue. March 29 — 8th diffusate 9.15 p.m. Blue green. March 30 — 9th diffusate 11.50 a.m. Blue green. March 31 — loth diffusate 9.15 a.m. Blue green. April I — iith diffusate i p.m. Blue green. April 2 — I2th diffusate 11.50 p.m. Light green. April 3 — I3th diffusate 11.50 p.m. Light green. Altho scarlet R and malachite green showed widely different rates of diffusion when they were treated separately, the results detailed above made it evident that it would be difficult if not im- possible to obtain all the scarlet R from mixtures like the one em- ployed without removing some of the malachite green with the red pigment. By subjecting Solutions of scarlet R and malachite green of simi- lar concentrations independently to diffusion in the usual way, we duplicated the blue and green effects with malachite green and the red effects with scarlet R, but the purplish colorations could not he ohtained under such circumstances. That these purplish effects were due to early diffusion of the malachite green with scarlet R, and that the red pigment facilitated the passage of the green one, are clearly indicated by the results. 86 Studies of Diffusion through Rubber Membranes [Sept Repeated removals of the diffusate in the independent scarlet R experiment, and replacements with fresh solvent for a period of about a weck, led to the Separation from the original pigment- product of all its red diffusible matter. The bag, at that stage of the treatment, contained considerable brownish-red, indifTusible material, which evidently was not scarlet R. This result, and simi- lar observations with other pigments, emphasized Dr. Gies' opinion that it might be possible to purify pigment preparations in this way and that their value as coloration agents, for histological staining especially, might thus be considerably enhanced. It will be noted that those pigments which diffused most rapidly were the so-called " fat colors," i. e., those soluble in, or staining, the common fats and oils. Again, with these pigments the dif- fusion is the most rapid, and therefore the most satisfactory, when the solvents are those which, in the State of vapor, soften rubber. It will thus be seen that apparently the membrane, as well as the solvent, exert selective action. This is true to a far greater extent in experiments of this kind than in the ordinary dialyses in aqueous media. When we arrived at this point in these experiments, to which we could give but a few hours weekly, our period of residence at Columbia University was about to close and, after completing some repetitions of previous observations in this connection, we were obliged to discontinue the work. It is Dr. Gies' intention to pro- ceed along lines suggested by the results already obtained in this preliminary investigation. THE COLLOIDAL NITROGEN IN THE URINE FROM A DOG WITH A TUMOR OF THE BREAST MAX KAHN AND JACOB ROSENBLOOM (Biochetnical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) In 1892 Töpfer^ found that the urine of patients suffering from Cancer contained a very large amount of "extractive substance." This " extractive substance " was calculated by first determining the total nitrogen, and then subtracting from this amount the sum of the nitrogen values for the urea, uric acid, and ammonia contained in the same urine. Bondzynski and Gottlieb,^ five years later, reported that the nitrogen in oxyproteic acid was 2 to 3 per cent, of the total urinary nitrogen. Salkowski,^ and Hess and Saxl,* using different procedures in their researches, came to the conclusion that the oxy- proteic acid or the alcohol-precipitable substances are increased in the urine of human beings suffering from Carcinoma. Salkowski and Kojo,^ in a preliminary communication, recently suggested several methods for the determination of colloidal nitro- gen in the urine. A year later Kojo published the results of a com- parative study of the various procedures suggested in this connec- tion.^ Einhorn, Kahn and Rosenbloom^ studied the zinc sulfate- precipitable, colloidal, nitrogenous material from the urine of nor- mal subjects as well as from the urine of carcinomatous patients, and came to the conclusion that the amount of colloidal nitrogen was invariably increased in subjects with carcinomatous growths. * Töpfer: Wiener klin. Wochenschrift, 1892, v, p. 49. 'Bondzynski and Gottlieb: Zentralbl. f. d. med. Wissenschaften, 1897, xxxv, P- 577- * Salkowski : Berliner klin. Woch., 1910, xlvii, p. 1746. *Hess and Saxl: Beiträge zur Carcinomforschung, 1910, Part II. 'Salkowski and Kojo: Berl. klin. Woch., 1910, xlvii, p. 2297. 'Kojo: Zeitschr. f. physiol. Chem., 191 1, Ixxiii, p. 416. ^Einhorn, Kahn and Rosenbloom: Amer. Journ. of Gastro-enterology, 1911, i, p. 2; and Archiv f. Verdauungs-Krankheiten, 191 1, xvii, p. 557. 87 88 Colloidal Nitrogen in Urinc from a Dog [Sept. The writers lately embraced an opportunity to study the colloidal nitrogen Output in the urine of a dog with a large tumor. The dog upon which this study was made had a hard calcified growth about the size of an orange in one of the breasts. The tumor involved the nipple and the breast tissue for some distance around the nipple. Several metastatic deposits were present along the "breast lines." Microscopic examination of sections of the original growth and of the metastatic infiltrations, according to several pathologists who examined them, indicated that the tumor was a chondroma which had undergone carcinomatous degenera- tion. Other pathologists, on the contrary, believed the growth to be of a benign nature, with the histological structure of a chon- droma. For the determination of colloidal nitrogen the alcoholic pre- cipitation method of Salkowski was used, with modifications, as follows :^ The total nitrogen was determlned in 5 c.c. of the urine by the Kjeldahl process. Two portions of 100 c.c. each of the urine were evaporated in a porcelain dish over a gently steaming water bath tili they were of the consistency of thin syrup. The residues were then taken up in 100 c.c. of alcohol (98.5 per cent.) and thoroughly stirred. The alcoholic extracts were then filtered through ashless filter papers, and the precipitates washed with alcohol. We determined the effect of dialysis upon this alcohol-precip- itable, so-called "colloidal," nitrogenous material. Most colloidal substances fail to dialyze through the very best grade of parchment paper. Only that fraction of the alcoholic precipitate which would remain indiffusible under suitable conditions of dialysis could be called "colloidal," at the present stage of our knowledge of the subject. Accordingly, the two precipitates on the ashless filter paper were treated as follows : The precipitate on one filter paper, together with the filter, was placed in a Kjeldahl flask, digested with sulfuric acid, and the nitrogen determined in the usual way. The second precipitate and *Before subjection to analysis the urine was first tested for protein, which, if found, was removed by means of heat coagulation aided by the addition of a few drops of dilute acetic acid Solution. I9I2] Max Kahn and Jacob Rosenbloom 89 filter paper were placed with water in a bag of the finest grade of parchment paper and dialyzed for forty-eight hours. The liquid in the bag was then analyzed quantitatively for nitrogen. The appended summaries present the results obtained for urine from the dog with the breast tumor and also for urine f rom several normal dogs. In the Salkowski method for the determination of " colloidal " nitrogen (as the results in the summary show), diffusible nitrog- enous substance is precipitated as "colloidal" nitrogen. It has not yet been shown that such diffusible nitrogenous matter in the colloidal precipitate is true colloidal material. A. Data pertaining to the urine of normal dogs ßpecimen No. Total Nitrogen in 100 c.c. of Urine Colloidal Nitro- gen in 100 o.e. of Urine Percentage of Total Nitrogen as Colloidal Nitrogen Indiffusible Colloidal Nitrogen Percentage of Total Nitrogen as Indif- fusible Colloidal Nitrogen I 2 3 4 Grams 2.304s 3-2051 0.8590 1.6436 Gram 0.0437 0.0314 0.0172 0.0214 1.85 0.98 2.00 1.28 Gram 0.02775 0.01634 0.01202 0.01841 1.2 O.S 1.4 i.l B. Date pertaining to the urine of the dog with a tumor of the breast Sa. 4.0088 0-3392 8.40 0.0939 2.3 5 b. 6.3034 0-3897 6.10 0.2293 3.6 SC. 4-4591 0.3210 7.10 0.0767 1-7 5d. 3.6862 0.3294 8.10 0.0817 2.2 Se. 3-1414 0.0958 3-04 0.0867 2.7 5 f. 3.9642 0.3566 8.90 0.1175 2.8 5 g- 2.5139 0.4617 13.10 0.1342 3.6 The results demonstrate that the " colloidal " nitrogen, both before and after dialysis, was greater in amount in the urine of the dog with the tumor than that in the urines from normal dogs. It is desirable to study the effect of dialysis upon the " colloidal " nitrogenous substances in the urine of Cancer patients. GENERAL ASPECTS OF FASTING^ PAUL E. HOWE (Department of Physiological Chemistry, University of Illinois, Urbana, III.) Fasting (starvation or inanition) is a State in which the dietary elements are withheld, either wholly or in part, so that the organism is compelled to draw upon its own resources to maintain its exist- ence. In discussing this subject it is my purpose to make a rather general survey of the changes which take place as the result of fast- ing; to show briefly how such results have been used to elucidate other scientific problems; and, also, to touch upon the therapeutic value of fasting, with relation to man. A distinction is made between physiological and experimental fasting. The first form is illustrated by the hibernation of mam- malia (hedge hog and bear), and cold blooded animals (frog), by the normal condition of the salmon during the spawning season and by the period of metamorphosis of the insects, these being natural phenomena for which the organism has made suitable preparation. In experimental fasting the animal is forced to live without sus- tenance, of one kind or another. Under this last State we may con- sider pathological fasting as a special case in which the organism is forced to fast as a result of impairment of some organ or of a gen- eral diseased condition. These forms of inanition present certain differences as evidenced in the effect upon the organism; yet it is quite probable that they are chiefly phylogenetic and we can conceive that any of the animals which do not experience these periodical physiological fasts might do so under the proper adverse circum- stances. In our discussion we will consider only the phenomena which take place as the result of experimental fasting. Here, too, we must distinguish between a number of forms of fast; such as the com- ^ A lecture delivered at the College of Physicians and Surgeons, New York, May I, 1912, under the auspices of the Columbia University Biochemical Asso- ciation. 90 I9I2] Paul E. Howe 91 plete fast in which there is total abstinence from both food and water; a modification of this, in which the subject is permitted to take water "ad libitum" or caused to ingest a uniform quantity from day to day ; and the incomplete fast, in which one or more of the food principles or chemical elements contained therein is with- held, such as a diet lacking in protein, fat, carbohydrate, water, salt or certain amino acids. There is not a marked distinction between complete fasting and fasting with water taken "ad libitum," for under the latter con- ditions the quantity of water taken decreases as the fast progresses until finally there is a natural abstinence from water. Some hold that the desire for water returns just before death. The ingestion of water causes a lengthening of the life of the animal and the severity of the fast is lessened. If at any time the quantity of water given is increased there will be for a time an increase in the metab- olism (14). This condition also holds for the well nourished animal (8), i. e., under all conditions when the water ingestion of the animal is sufficiently increased the general metabolic processes of the organism are stimulated. The length of a fast which would result in death depends upon the size, the species, the age, the nutritive condition, the external surroundings (e. g., temperature, humidity, etc.) and the intrinsic rate of metabolism. In general, we may say that the smaller the subject the shorter will be the time it can live without food; but this does not hold in all cases, for certain of the lower animals can fast much longer than the higher forms, e. g., the Salamander, which is about 3-4 inches long, has been fasted for more than 125 days (19). Adult organisms can fast longer than the young of the same species. Thus, a young pup can fast but a few days, while a füll grown dog will fast from 20 to 60 days. Of the fasts on man and other warm blooded mammals, the longest on record is one of 117 days (15). This experiment was conducted in our laboratory, a Scotch collie dog being the subject. Subsequent to this long fasting interval the dog was fed, and it returned to its normal condition. A comparison of the results obtained by various investigators shows that death does not ensue until there is a loss of between 40- 50 per Cent, of the original body weight. The real cause of death 92 General Aspects of Fasting [Sept. from fasting has not been determined. The probable reason is the failure of some organ or life process (27) and not the depletion of all possible nutritive material. From our experiments (10) it would appear that a certain definite minimal proportion of nitrogen- holding substance must be present in the body for life to exist. Fasts have been reported upon men covering periods of from 2-50 days, upon dogs as long as 117 days and Salamanders for 125 days. In each of the extreme cases, the subjects were subsequently fed and they returned to normal. The influenae of repeated fasting upon the resistance of the animal to subsequent fasts is a phenome- non which appears to be intimately associated with hibernation. As has been shown by Russian investigators (20), and more recently in our laboratory (10), repeated fasting decreases the rate of metab- olism in each succeeding fast. A French investigator (21) has shown that repeated fasting, in which the subjects were alternately fasted and fed during equal periods of about a week each, resulted in the ultimate death of the animals. From the experimental data Et band it seems that where the animal is permitted to recover completely from a fast before it is subjected to another, there will be an increased resistance to the ravages of the succeeding fast. The number of men who have made a study of the changes which take place as the result of fasting is so great that it is difficult to name those who have made the most important contributions upon this subject without doing an injustice to others. The inves- tigations of Cathcart (4) in England and of Benedict (2) in this country, upon men, are the most extensive that have been con- ducted with the more refined methods of analysis which we possess today. The names of Succi, Cetti and Breithaupt stand out in the literature as the subjects of important experimental fasts. What changes take place in an organism as the result of a fast? Outwardly the subject becomes emaciated, his body weight de- creases, he becomes weak and apathetic and, should the fast proceed long enough, he would probably die in a State of coma. In man it has been demonstrated that the brain retains its activities unimpaired during a fast and that hunger is evident only at the beginning of the ordeal. These facts are substantiated in the populär writings upon fasting and also by an experiment made by us (11) in which the I9I2] Paul E. Howe 93 subject prepared for the preliminary examination for the degree of Doctor of Philosophy during a seven-day fast. The fasting State is indicated in the body by certain changes; such as a general decrease in the body metaboHsm, represented by variations in the nitrogen excretion and the respiratory exchange, a decrease in the fat and glycogen Stores, a decrease in the volume of muscle and in the size and weight of certain organs. The tempera- ture remains normal, for a time at least, but shows a tendency to decrease toward the end of the fast. The decrease in the general metaboHsm is well Illustrated by the data obtained from the respiration calorimeter experiments. It has been shown by the earlier investigators and more recently by Bene- dict (2) that, in a well fed man, the quantities of protein and fat which were utiHzed, and the energy change (calories) per day, decreased very gradually and tended toward a constant minimum. In addition to the excreted carbon dioxide, Benedict determined the amount of oxygen consumed. From these data it was shown that the glycogen consumption, which is most rapid on the first day, decreases as the fast progresses. It is probable that the glycogen Store is never depleted and that even in fasting there is a resynthesis of glycogen from the protein material present in the body. Decreased metaboHsm in fasting is also shown by the quantities of nitrogen-containing substances eliminated in the excreta. We are particularly concerned with the losses of nitrogen, for it is the protein material which is the most fundamental nutritive substance and which the body strives to protect. In fasting, the nitrogen- containing substances in the urine or feces arise from the tissues and hence the total nitrogen excretion is a measure of the quantity of muscular or organ tissue catabolized. The excretion of total nitro- gen in the urine decreases rapidly at the beginning of the fast and soon reaches a minimum, which is maintained for some time. This minimum of nitrogen excretion, representing a minimum protein disintegration, is Held to represent the " maintenance " metaboHsm of the individual, i. e., that amount of protein substance which if supplied, with sufficient fats or carbohydrates, in the form of food would sustain life. This minimum has variously been shown to be greater or less than the metaboHsm as represented by fasting ex- periments. 94 General Aspects of Fasting [Sept The muscular disintegration is influenced by the factors already mentioned; the nutritive condition and the experience of previous fasts, or repeated fasting. The diet just before the fast influences the nitrogen excretion for a number of days. This has been demon- strated in the classical experiments of Voit (26), in which he fed varying amounts of meat and bread to a dog and showed that, when fasted the rate of nitrogen excretion varied, but that in each case the animal came to the same level of catabolism on about the seventh day of fasting. The fat available in the body exerts a marked efTect upon the protein metabolism and the Hfe of an animal. So long as there is sufficient fat in the organism to supply the energy requirements, the protein metabolism will remain at a minimum. When, how- ever, the fat deposits are depleted, the body is forced to use protein to furnish the necessary energy. The result is a more rapid protein consumption and an earlier death. This increased protein con- sumption, is, of course, accompanied by an increased nitrogen excre- tion, which has been designated as the " premortal rise." The feed- ing of carbohydrate or fat sufficient to supply the energy require- ment of the body would prevent this increased consumption of pro- tein and thus lengthen the life. Repeated fasting will also modify the rate of metabolism. This point is well illustrated by the results obtained on a subject in a repeated fast (10), in which there was a rapid and increasing con- sumption of the protein reserves of the body during the first fast, but a more gradual and uniform consumption during the second fast. The total body weight and nitrogen losses were practically identical in the two fasts and the data f rom the intermediate feeding period would indicate that an increased fat störe was not the cause of the more gradual utilization of the body resources. A study of the differential distribution of the nitrogen in the urine serves to bring out certain points with regard to the protein metabolism of fasting animals. The percentage of total nitrogen occurring as urea-nitrogen decreases in man and is accompanied by an increased ammonia-nitrogen excretion. This has been explained as due to the condition of acidosis, which may result, at least in part, from the accelerated utilization of the fat deposits and the decreased oxidative powers of the animal. I9I2] Paul E. Howe 95 In the case of dogs there is a difference o£ opinion as to the relation between the urea-nitrogen and the total nitrogen. Schön- dorf (22) and others hold that the percentage of urea-nitrogen decreases, while in all of our experiments it has remained nearly constant, which fact, coupled with the failure to find marked quan- tities of organic acids in the urine, would show that dogs are better able to utilize their body Stores. This may be due to the fact that the dog is naturally a "high-protein" animal. The daily Creatinine excretion, which is a constant for any indi- vidual under normal conditions of feeding and is generally believed to be a function of the muscular metabolism, decreases gradually as the fast progresses and in correspondence with the decreasing amounit of protoplasm. Creatine, which does not occur in normal urines, or is found only in cases associated with muscular disintegration, appears dur- ing fasting and ordinarily becomes a constant constituent. It has recently been shown that the feeding of carbohydrate causes the excretion of creatine to stop (5, 17) ; while ingested fats may even cause an increase in the excretion of this form of nitrogen. In one of our dog experiments (15) there was a disappearance of urinary creatine from the iQth to the 59th fasting days. This phe- nomenon might be explained on the above basis. It is improbable, however, that the body could synthesize sufficient glycogen at this stage of the fast to cause the disappearance of the creatine. The real explanation is therefore not apparent. In connection with the repeated fast, previously mentioned, it is interesting to note that the excretions of creatine as well as of total nitrogen were practically the same during each of the two fasts, notwithstanding the fact that the second fast was twice as long as the first. • This would indicate an intimate relation between the total-nitrogen excretion and the quantity of creatine excreted. When the data representing the creatine and Creatinine excretions of a fasting animal are examined, it is seen that there is generally a progressive increase in the creatine Output and an accompanying decrease in the Creatinine elimination, until the output of creatine exceeds that of the Creatinine. In other words, when expressed graphically, the curve representing the course of the creatine excre- 96 General Aspects of Fasting [Sept. tion crosses that representing the Creatinine Output. This phenome- non has been termed by us the "creatine crossing" and is believed to be very significant. It occurs with great uniformity a few days previous to the decrease in the total nitrogen excretion that precedes the pre-mortal rise of excreted nitrogen. By means of the " creatine crossing" the length of the subsequent Hfe tenure of the animal may be quite closely estimated. Certain pathological constituents, other than creatine, may ap- pear in the urine as the result of fasting, such as acetone, diacetic acid, lactic acid, bile pigments, albumin, etc. The processes in the large intestine during fasting have received but httle attention. Various authorities contend that it is difficult to make a Separation (2, 18) of fasting feces. Müller (19a) has shown that indican, which is now considered as an index of intes- tinal putrefaction, disappeared upon the third day of fasting. We have been able to make an undoubted Separation of fasting feces and have found indican present during the whole of a seven day fast on man {22,), Fasting feces are distinct from those of the normal individual in that they are of a peculiar brown color and are pasty in consistency. The percentage of nitrogen present is higher than in normal feces. The bacterial content of feces has received but little attention and only recently have results upon the bacterial content of fasting feces been determined. The results indicate a lovver percentage content of bacteria (3). There is not an equal wasting of all of the organs and tissues of the body, those organs most necessary for the maintenance of life show only a slight decrease in size and weight, while others are reduced to but a fraction of their original proportions. Thus the heart, lungs, and nervous System exhibit but little change while the muscles and fatty tissues exhibit a marked reduction of both volume and weight. The organs of regeneration are also resistant to the ravages of a fast. This fact is of especial significance for it demon- strates the tendency of nature to preserve the species. A histological examination of the tissues and organs of fasting animals shows a decrease in the volume of the cells as a whole and of the nuclei. MorguHs (19) has shown that the decrease in the volume of the cells of the Salamander is greater than that of the igi2] Paul E. Howe 97 nuclei and further that the nuclei become elongated. In the case of the liver, the cell walls finally begin to disappear and the small masses of pigment to clump together. Such a condition does not necessarily result in death, for Salamanders of the same size have been caused to fast for even a longer time than those whose tissues demonstrated these changes ; and, af ter feeding, it was f ound that the cell walls again appeared and the liver returned to a normal condition. We will not take up the question of the localization of the degenerative changes, i. e., as to whether they occur in an organ as a whole or in localized portions. It is an interesting fact, however, that even when the organism is undergoing the degenerative effects of a fast, there are still evidences of mitotic division of the nuclei. What changes take place which enable one organ to waste away while another retains its normal condition? The explanation most generally accepted is that of the nourishment of the more vital Organs by transference of the nutritive material from the less im- portant tissues. Thus the less resistant tissues gradually give up their stores of fat and protein to the blood stream which in turn furnishes them to the actively functioning organs. This idea has received further proof from the researches of Hottes.^ This botanist (9) worked with beans and has shown that upon removing the cotyledons, and thus the food supply, from seed- lings, the meristematic tissue which would normally go to produce lateral roots is transferred to the tip of the root (meristem) and there used for growth ; and that at the end of from three to f our weeks all the cells in the Upper part of the root have lost the major portion of their protoplasm and the only actively functioning cells are those at the tip. Hottes has also shown that the decrease in size of the root is due rather to a reduction in the number of the cells than to a mere decrease in size. This is in Opposition to the findings of certain zoölogists who hold that the reduction in the weight and volume of the organs is due more to reduction in size than in number, altho they admit a small decrease in the number of contained cells. 'I am indebted to Professor Hottes of the University of Illinois for this Information which was taken from some of his unpublished work. 98 General Aspects of Fasting [Sept The blood of fasting subjects which are ingesting water shows in general a decrease in the number of erythrocytes and leucocytes, and of the percentage of hemoglobin. The differential distribution of leucocytes varies with the species. In the dog (12) there is a decrease in the percentage of polymorphonuclear leucocytes and a corresponding increase in the small lymphocytes. The changes in the other forms of cells are but nominal. Fasting studies have been of great importance in the study of the minimum of food necessary to maintain life and upon which to base the calculation of dietary Standards. Such studies have also been utilized in the explanation of phenomena occurring in patho- logical States and of metabolism in general. Underhill and Rand (25) have explained certain anomalies in the urinary changes which occur in pernicious vomiting of preg- nancy from their knowledge of fasting metabolism. Two agriculturalists have recently made use of the results of fasting studies to elucidate problems of importance to both the scientist and the farmer. McCollum (16), fed a nitrogen-free diet to pigs and studied the efficiency of individual grains as feeding stuffs, as well as the nature of the repair processes in protein metab- olism. He shows that the difference in the nutritive values of the wheat, oat and corn kerneis is not so great as would be expected from the difference in the chemical composition; and further, that the repair processes of the cell are of a different character from those of growth, and that the cellular catabolism and repair do not involve the destruction and resynthesis of entire protein molecules. This last Statement is not in entire accord with the most widely accepted theories of metabolism. Certain zoölogists have also shown that the changes of regeneration are unlike those which occur in growth. Dietrich^ shows (7) that fasting so reduces the plane of metab- olism that the quantity of food which was insufficient for main- tenance before fasting was afterward sufficient, not only for main- tenance, but to produce a positive nitrogen balance ; in other words, the animals were more efficient machines after fasting. 'I wish to thank Professor Dietrich of the University of Illinois for per- mission to refer to his unpublished data. I9I2] Paul E. Howe 99 Aron (i), in his studies upon nutrition and growth, subjected dogs to incomplete fasts. The results showed that a growing animal, receiving only enough food to provide for little or no in- crease in weight, " is in a condition of severe starvation." Under such conditions the skeleton grows at the expense of the flesh, the Organs retain their weight and the brain reaches its normal size. The fat and protein of the muscles are largely used up, altho this loss of material is balanced by gain of water and by the growth of the skeleton. The biologists have made use of the fasting subject in the study of the Problems of degeneration, of regeneration, and of growth, The work of Morgulis and of Hottes already considered was of this nature. The therapeutic value of fasting is realized in the preliminary treatment of some digestive disorders and in the partial fasts of obesity eures. These latter eures consist in supplying only the protein requirements of the body and thus forcing the individual to utilize the surplus fat deposits to make good the energy require- ments. The increasing populär literature upon fasting and the tendency to fast on the part of certain people, especially the pronounced physical culturists, and their general good health, would seem to indicate that there are some beneficial results to be obtained from fasting. The various books upon fasting, of which the superficial, yet interesting book of some six hundred pages by Carrington (6) upon " Vitality, Fasting and Nutrition " is the most complete, lend strength to the idea that fasting as a therapeutic measure is impor- tant. The chief contention of this fasting cult is that by depriving the body of food the digestive organs are given a chance to recu- perate and the body is enabled to rid itself more effectively of the waste products and toxic substances. Fasting for short and widely-separated periods may be a bene- ficial procedure in some individuals. This conclusion is supported by the observed effects on dogs, which acquire increased resistance from repeated fasting. This view is strengthened, also, by the foregoing data pertaining to pigs as well as by Seeland's (24) results on pigeons and chickens, which show that repeated fasts. loo General Aspects of Fasting [Sept. for periods of f rom one to two days, were foUowed by better growth and greater strength. It is probable, then, that fasting under proper conditions may be advantageous. Long fasts, however, seem to be devoid of benefit and may endanger health. BIBLIOGRAPHY 1. Aron. The Philippine Journal of Science, 6, i, 1911, 2. Benedict. Carnegie Publications, 77, 1907. 3. Blatherwick, Sherwin and Hawk. Proc. Amer. Soc. of Biological Chem- ists, 2, 42, 191 1. 4. Cathcart. Biochemische Zeitschrift, 6, 109, 1907. 5. Cathcart. Journal of Physiology, 39, 311, 1909. 6. Carrington. Vitality, Fasting and Nutrition. Rebman Co., N. Y. 1908. 7. Dietrich. Unpublished data. Illinois Agricultural Experiment Station. 8. FowLER and Hawk. Journal of Experimental Medicine, 12, 388, 1910. 9. HoTTEs. Publication of the Carnegie Institution. (In press.) IG. HowE AND Hawk. Journal of the American Chemical Society, 33, 215, 1911. 11. HowE AND Hawk. Proc. Amer. Soc. of Biological Chemists, 2, 65, 191 1. 12. HowE AND Hawk. American Journal of Physiology, 30, 174, 1912. 13. HowE, Mattill and Hawk. Journal of the American Chemical Society, 33, 568, 191 1. 14. Howe, Mattill and Hawk. Journal of Biological Chemistry, 10, 417, 1911. 15. Howe, Mattill and Hawk. Journal of Biological Chemistry, 11, 103, 1912. 16. McCoLLUM. American Journal of Physiology, 29, 215, 191 1. 17. Mendel and Rose. Journal of Biological Chemistry, 10, 213, 191 1. 18. Mendel and Fine. Journal of Biological Chemistry, 11, 5, 1912. 19. MoRGULis. Archiv für Entwicklungsmechanik der Organismen, 32, 169, 1911. 20. Quoted by Pashutin. Pathological Physiology, 1902. 21. Richet. Comptes rendus de la Societe de Biologie, 61, 546, 1906. 22. Schöndorf. Archiv für die gesammte Physiologie, 117, 257, 1907. 23. Sherwin and Hawk. Journal of Biological Chemistry, 11, 169, 1912. 24. V. Seeland. Biologisches Centralblatt, 7, 145, 1887. 25. Underhill and Rand. Archives of Internal Medicine, 5, 61, 1910. 26. Voit. Zeitschrift für Biologie, 2, 307, 1866. 27. Voit, E. Zeitschrift für Biologie, 41, 188, 1901. THE PHYSICO-CHEMICAL BASIS OF STRIATED MUSCLE CONTRACTIONi 2. Surface tension WILLIAM N. BERG (WITH PLATE l) If the physico-chemical basis of muscle contraction is ever to be understood or explained, it is almost certain that it will be brought about thru speculation and experiment of a quantitative, rather than of a qualitative nature. The mere Statement that muscle con- traction is caused by surface tension, or thru osmotic action, etc., unless accompanied by quantitative data of an experimental or theoretical nature, can add little toward the Solution of the problem of the transformation of energy by muscle. It is, perhaps, regret- able that so many of the " theories of muscle contraction " which have appeared in the recent literature belong to the qualitative class. Occasionally someone attempts to treat the subject quantitatively. From this point of view the works of Bernstein,^ and of Zuntz^ are particularly meritorious, even if the problem has not yet been solved by them. Among the latest qualitative contributions to the theory of muscle contraction, is that of Strietman and Fischer.^ They studied the contraction and relaxation of catgut strings immersed in various Solutions. By attaching the strings to the usual arrangement of iever and recording drum, they found that when a catgut string is immersed in water or physiological salt Solution, even for some time, no changes in length take place (p. 66). But if the string be immersed in Solutions of hydrochloric or lactic acids (w/8o to ^ Berg, W. N. : Biochemical Bulletin, 1912, i, 535. ^ Bernstein, J. : Arch. f. d. ges. Physiol., 1901, 85, 271-312. ^Zuntz, N. : Die Kraftleistung des Tierkörpers. Festrede; Berlin, 1908. * Strietman, W. H., and Fischer, M. H. : Ztschr. f. Chemie und Industrie der Kolloide, 19 12, 10, 65-77. lOI I02 Physico-Chemical Basis of Striatcd Musclc Contraction [Sept n/20) it contracts. On replacing the acid Solution by water, the string relaxes. The relaxation is faster, however, when the acid is replaced, not by water, but by a Solution of some salt such as sodium bicarbonate, which can neutralize the acid. From their diagrams it would seem that a minute or more may be required for a Single contraction or relaxation, depending upon the strength of the acid, etc., etc. These observations are, no doubt, interesting in themselves. But before connecting them with muscle contraction, might it not be well to consider whether the conditions under which a catgut string can contract and relax are at all similar to those existing in muscle ? Strietman and Fischer State that because lactic acid is formed in a working muscle and because a catgut string will contract when immersed in a lactic acid Solution and will relax when the acid is removed, therefore, in the working muscle, the contraction is brought about by the formation of lactic acid. They quote several other investigators who have stated their belief in the same idea of the connection between lactic acid formation and contractility with- out, however, making any of the simple calculations that would naturally suggest themselves. Their theory is open to the following objections: (i) It is not likely that there is any free lactic acid in the working muscle, it is probably neutralized at once by the phosphates present in lymph. At least this would be inferred from the work of Henderson^ who showed that the mixture of phosphates and other substances in blood and various tissue fluids was such as to enable them to maintain an absolute neutrality in spite of the formation of even considerable quantities of acid or of alkali. This point was not overlooked by Zuntz (p. 20, 1. c.) when calculating the amount of energy made available by the transformation of dextrose into lactic acid. The heat of neutralization of the lactic acid by sodium, as well as the heat required to separate the sodium from its presumable combina- tion with protein, are given due consideration by Zuntz, who cal- culated that the heat liberated in the formation of lactic acid from dextrose is equivalent to 3.4 per cent. of the heat of combustion of dextrose. * Henderson, L. J. : Ergebnisse d. Physiologie, 1909, 8, 254-325. I9I2] William N. Berg 103 A repetition of the experiments of Strietman and Fischer, in which catgut strings would be immersed in Solutions comparable with lymph containing lactic acid (not exceeding the maximal amount possible if all of the muscle glycogen were changed at once to lactic acid), would probably give results more decisive than those in which free acids were used. (2) And even if free lactic acid existed in muscle, or if com- bined lactic acid could induce proteins to swell, one such Observa- tion is only one of very many that are needed for a rational theory of muscle contraction. The Statement that lactic acid swells protein adds very little to our knowledge of the mechanism in muscle by which the potential energy of the food is transformed into the kinetic energy of the moving muscle and its load. It is to be regretted that the work of Brod^ on the swelling of fibrin in acid Solutions has received practically no attention in the recent literature. The paper can be profitably studied by those con- templating studies on protein swelling. A brief resume of Brod's results is given by Berg."^ A good example of a quantitative theory of muscle contraction is the calculation of Bernstein^ on the possible changes in the sur- face energy resident on the muscle fibrils. The method of making the calculations is, perhaps, unnecessarily complicated and, in one or two instances, the mathematical equations are of doubtful correct- ness. Bernstein finds that in order that a muscle may lift an ordi- nary load, the surface tension between fibril and sarkoplasm must have an improbably great magnitude. He nevertheless concludes that the principle, that energy is transformed in muscle thru changes in surface energy, is correct. There are several reasons why, to the Student at least, a proper understanding of some of the recent applications of physical chem- istry to biology should be so difficult, if not altogether impossible. First: The indefiniteness of certain Statements that the writer has ' Brod : Beiträge zu der Lehre von der Eiweissverdauung. Dissertation, Würzburg, 1892. "Berg, W. N. : Amer. Jour. Physiol., 1909, 23, 427. Brod's method has recently been used by Tracy and Gies, Biochemical Bulletin, 1912, i, 468. * Bernstein, J. : Arch. f. d. ges. Physiol, 1901, 85, 271-312. 104 Physico-Chcniical Basis of Striated Muscle Contraction [Sept. frequently seen in the literature. This is an example taken from Freundlich's Kapillarchemie, p. 4: Stirface energy = surface tension X area of surface. A similar Statement is made by Michaelis,^ and others. Nothing further was stated that would enable the reader to use such a formula in making calculations were it desired. Expressing the surface tension in dynes per centimeter and the area of the surface in cm.2, what is the surface energy? The answer is very simple after one has taken the time to look the matter up. After a formula such as the above, a numerical example ought to be given, so that it means more than so many words to the average reader. Suppose it is desired to calculate the amount of energy required to form a water-surface (in contact with air) of i sq. cm. area? Or, what is the same thing, how much energy is liberated or is available for external work when the above water-surface diminishes by i sq. cm.? According to Michaelis (1. c, p. 14), this will require (or liberate) 70 ergs or 7 X lo"'^ kilogram-meters. The method of using the formula to obtain this result, simple as it is, was not given by Michaelis, altho at least one example of the use of a formula is desirable because it will enable the reader to make many other cal- culations. Following is a numerical example of the kind mentioned above. How much energy is required to form a water-surface (against air) of I sq. cm.? In the formula it is assumed that the surface tension remains constant during the change in area : surface energy required ^surface tension X increase in area, or surface energy liberated = surface tension X decrease in area, (ergs) = (dynes per cm.) X (cm.^). Since the surface tension of water-air is about 70 dynes per cm., it is evident that-^^ X i cm.2^70 ergs = the amount cm. of energy required. The erg is a unit of work (or energy) and is the work done when a mass is moved i cm. by a force of i dyne. ' Michaelis : Dynamik der Oberflächen, p. 13. Dresden, 1909. I9I2] William N. Berg 105 The element of time does not enter into the definition of the erg. The vvork done (ergs) is equal to the product of the force (dynes) times the distance (cm.) thru which the force acts. Of course, other Units may be used. The surface tension may be expressed in grams per cm., and the area in Square cm. The work then is ex- pressed in gram cm. But on account of the unfortunate use of the Word ' gram ' to designate a certain mass or quantity of matter and also to designate a force, it is better, for the present, to use the erg and the dyne, and later to convert ergs into kilogram-meters, or any other of the customary units for expressing muscular work. It is, of course, absolutely necessary that the terms used in such calculations be consistent. Here is the second reason why some so-called appHcations of physical chemistry to biology are not easily followed. An equation will sometimes be given that is not correct in its dimensions. To State that 2 sq. cm. = 2 cubic cm. is obviously incorrect. Such an inconsistency is to be found in one of Bern- stein's^" equations : 'Wir werden daher in dem Falle des isometri- schen Tetanus, in welchem alle chemische Energie als Wärme er- scheint, dp — ar = c. Wp setzen können, wenn Wp die in einer Zeit- einheit erzeugte Wärmemenge, c eine Constante und «p und oLt die Oberflächenspannung im Tetanus und in der Ruhe bedeuten. Da wir nun oben (S. 296) gesehen haben, dass ctr gegen CLp verhältniss- mässig sehr klein ist, so können wir annähernd ap=:c. Wp an- nehmen. ' Here are two equations in which surface tension is equated with work (or heat). It makes no difference what units are used, on one side there is a force (surface tension expressible in dynes per cm.) and on the other a quantity of energy or work (ergs, or dynes X cm.). The constant above referred to is probably meant to be the mechanical equivalent of heat. These equations are interesting for another reason. It is true that in isometric tetanus, a muscle does work in the physio- logical sense of the word. But not in the physical sense. In physics (or mechanics) work is defined as a product of force times distance thru which the force has acted. If either factor is zero, the product, work, is zero. The columns that support ^^ Bernstein, J. : Loc. cii., p. 307. io6 Physico-Chemical Basis of Striatcd Muscle Contraction [Sept. a biiilding do no work in the physical sense, n'or would a man who took the place of one of them; altho physiologically he would do a great deal of work. In this case the distance thru which the force acts (the weight supported or the upward thrust of the man's Shoulders) is zero and hence the work is zero. The above equa- tions of Bernstein could be made consistent if there were two factors on the left-hand side; one, the surface tension or change in surface tension (expressed in dynes per cm.), the other, the area or change in area (expressed in cm.^). The product (ergs) could be calculated to calories (gram-degrees C.) by dividing by 4.2 X 10''^, since i small calorie (gram-degree C.) is equivalent to 4.2 X 10'^ ergs. It is difficult to see what the other factor (omitted by Bern- stein) can be. In an isometric tetanus the muscle does not change its length. In what way can an internal diminution in area take place? If the contractu units — whatever their shape may be — do not change in length, how do their areas diminish? This difficulty does not arise in the case of the ordinary (isotonic) contraction. Here one can assume a decrease in the areas of contact between contractil unit and sarkoplasm caused by an increase in the surface tension between the same surfaces. The product of these two quantities, according to the theory, should be an amount of work sufficient to account for the external work done and perhaps also for the heat liberated at the same time. We have stated before that Bernstein's calculations on the magnitude of the surface energy changes in muscle are probably unnecessarily complex, involving, as they do, several pages of cal- culus. The same result is obtained in the following calculations, in which two simple quantities are calculated and then compared : (i) the amount of energy liberated in a working muscle thru in- crease of surface tension times diminution of area of contractil Units; and (2) the external work done in lifting a weight a known distance. Assume that in i c.c. of muscle a right section contains (as Zuntz assumes, 1. c, p. 24) 62 million rods, and that there are 800 such layers, making a total of very nearly 5 X 10^^ rods in i c.c. of muscle. Assume the general structure of muscle to be that de- scribed by Hürthle (see diagram), and that the muscle rod is the LU I- < _l Q. o > I- UJ CD _l < Ü LU I o g m s I i I ^1 «- W6 ü ' ni H 9 ^• O ^ ■< 1) o c o c 'oidO-CJ.QS'IUir w b hJ fci u «+H cn P 1^ ^ o -Ci, w H ^ < 1 — 1 ■^ Pi — I CO < < u (LI O j= 's; S öl -*-t u W u I o u I— I CO >^ Ph o -^ J9fi-VJ 07doJ}0S-IUV- ü 2 P3 O igi2] William N. Berg 107 CONTRACTIL UNIT (Hürthle, p. 157). From the dimensions 011 the accompanying diagram, it is evident that the lateral area of a rod diminishes from 4.8 /a^ when relaxed, to 2.8 /^^ when contracted. (We omit the simple geometrical calculation.) The base areas are increased, but the energy apparently required for this work will for the present be disregarded — it is an additional load on a probably overloaded theory. Since the total area of the relaxed rod (5.2 /a^) is greater than that of the contracted rod (3.8 fi^), it follows that there is an increase in the surface tension immediately preceding the contraction, according to the requirements of the theory. To calculate the energy liberated as being equivalent to the diminution in area times the surface tension of water is probably incorrect, for without an increase in surface tension there seems to be no reason why the rods should contract against the external resistance — the downward pull of the weight lifted. The rod contracts presum- ably, because in the relaxed State the surface tension on the rod surface is low. How low can it be ? It may be as low, perhaps, as 23 dynes/cm. if we assume the rod to be covered with a layer of pure acetic acid, and that the acid has the same surface tension in con- tact with the water that it has when in contact with air. Other fatty acids also give low values for the surface tension of their Solutions, and they have still lower surface tensions in the pure State. Pres- ently, the surface tension is raised, presumably by the removal of the fatty acid or other agent causing low surface tension, by instan- taneous combustion, let us say. How high can the surface tension be raised? It might be raised" to 85 dynes/cm. if we imagine the rod now to be covered with a layer of saturated sodium chlorid Solu- tion. The surface tensions of aqueous Solutions of salts cannot be raised^^ very muchbeyond that of pure water, which varies between y2 to y6 dynes/cm. (at 18° C), according to the method of measure- ment. The upper limit for any Solution that possibly could exist in the muscle might be assumed, then, to be the value for saturated sodium chlorid Solution, or any other concentrated salt Solution that might probably occur in living muscle. Of course, the existence of the films of pure acetic acid and of strong salt Solution over the " Freundlich, H. : Kapillarchemie, p. 27 and 62. Leipzig, 1909. *^ Heyd Weiler, A. : Ann. Physik., 1910, 33, 145-185. io8 Physico-Chemical Basis of Striatcd Musclc C ontraction [Sept. muscle rods is purely hypothetical. The surface tension theory requires that changes in surface energy take place, and from what follows it is apparent that these changes must be great — greater, in fact, than the probable actual change on the rod surface. For it seems hardly possible that such great changes in concentration and in surface tension could take place. The values for the surface tensions of pure acetic acid and concentrated salt Solution have been taken from the literature ; whether such limiting values are ever reached in living muscle is, for the present, purely hypothetical. If it be assumed that, during the chemical changes taking place in a working muscle, the inorganic ions in the rod-surface film rapidly change their concentrations, the film might be regarded as an electrical double layer or Helmholtz double layer. Without a doubt, changes in surface tension would result from the changes in ion concentration. The more ions in one of these layers covering a rod, the more they repel one another and the lower is the surface tension, and vice-versa. But as has been pointed out before,^^ it is not certain that such a double layer really exists between living particles and their surrounding medium. And even if there were such a layer, the total change in surface tension in such a layer is hardly significant for the present purpose. The small Variation in surface tension when the Variation is caused only by ions was prob- ably overlooked by Robertson^"* and others who advocated a capil- lary electric theory of muscle contraction. It is really a special case of surface tension in which the variations in surface tension are caused by the mutual repulsion of the ions in each of the layers. But insofar as small amounts of certain organic substances, such as fatty acids, can affect (depress) the surface tension of water very much more than even improbably large amounts of inorganic salts, the surface tension theory is given the benefit of the greatest possibilities by assuming the changes in concentration from pure acetic acid (23 dynes/cm.) to saturated sodium chlorid Solution (85 dynes/cm.). This is as large a difference as can be assumed from the experimental data on the surface tensions of Solutions. "Berg, W. N. : New York Med. Journal, 1907, July 20 and 27; and Ion, 1910, 2, 161-188. "Robertson, T. Brailsford: Trans. Royal Soc. South Australia, 1905, 29; and Quarterly Jottr. Exper. PhysioL, 1909, 2, 303-316. I9I2] William N. Berg 109 If in I c.c. of muscle there are 5 X lo^*' rods, the lateral area of each of which diminishes from 4.8 /a^ to 2.8 /^^ when the muscle con- tracts, the total reduction in area is 5 X 10^" X 2 /x,2__io^i fx^ = lO'^ cm. 2 (i ju. = 0.001 mm.). The calculations will be simplified if it be assumed that the increase in surface tension is instantaneous, giving the contracting muscle the largest surface tension during the entire contraction phase. Then since surface energy liberated = diminution in area X surface tension, (e.,s) (c..^) {^) the energy liberated is 1000 X 85 ergs. Let it be assumed that all of this is transformed into external work — lifting a weight — and that the resultant heat arises from the activity of a different mechanism ; in short, that the muscle is an engine having an efficiency of 100 per Cent. How great a weight will this i c.c. of muscle lift? Since there are 800 layers of rods, and each layer shortens by 3 /-i during the contraction (see Plate i herewith), the muscle shortens by 2400 /a or 2.4 mm., lifting a mass of W grams 2.4 mm. The energy (ergs) expended in lifting a mass of W grams thru the distance D (cm.) is PF X ■C' X 981 ergs, since gravity = 98i dynes. Therefore the 8s,ooo 85,000 ergs will hft ^^3^ = 361 grams. According to Zuntz (1. c, p. 23) i gram of muscle substance can do 0.002 kilogram-meter of work in one contraction under favorable conditions. If this muscle shortened 0.24 cm. as the above muscle did, it would lift a trifle more than 800 grams. Bern- stein^^ mentions 600 grams at least, as the pull of i cm.^ of frog muscle in an isometric contraction. Insofar as i cm.^ of many kinds of muscle can support without lengthening (but not lift) several kilograms — about 6 kilograms for human, and probably more for certain types of insect muscle — the above figure of 361 grams, as the weight a muscle could lift, is small, especially when it is borne in mind that it is an improbable maximum. The foregoing discussion may be summarized as f oUows : I. Too often there is a general lack of definiteness in the mathe- " Bernstein, J. : Arch. f. d. ges. PhysioL, 1905, 109, 326. HO Physico-Chemical Basis of Striated Muscle Contraction [Sept. matical treatment of a biological problem. Formulae are stated vvith no information as to their use or application to the problem under disciission. 2. Bernstein's calculations on the surface energy changes in working muscle are criticized. A much simpler method of calcula- tion is used with a result similar to Bernstein's, namely, the energy expended by a working muscle is much greater than the probable changes in surface energy can furnish, Of course, future investi- gations may bring to light sources of surface energy within muscle as yet unknown. Washington, D. C. A STUDY OF SOME PROTEIN COMPOUNDS WALTER H. EDDY (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) Contents. (A) Morphin mucoid, 112; (B) strychnin mucoid, 114; (C) conin mucoid, 115; (D) piperidin mucoid, 115 ; (E) anilin mucoid, 115; (F) morphin nucleoprotein, 115; (G) morphin caseinogen, 116, strychnin caseinogen, 116, calcium caseinogen, 116; (H) strychnin ovo-mucoid, 117; (I) histon mucoid, 118; (J) histon nucleoprotein, 121; (K) histon ovo-mucoid, 121. I. INTRODUCTION When I began Ph.D. work in this laboratory, six years ago, Dr. Gies was actively engaged in studies of the properties of various protein Compounds which he had prepared as early as 1904.^ He inaugurated that work from the Standpoint of his interest in the chemical composition of protoplasm, and the nature of the struc- tural and dynamic relationships of cell constituents and products. He believed that the knowledge gained from studies of artificial protein Compounds would pave the way for successful inquiry into the nature of the protein correlations in the cells — relationships of the most fundamental biological character. At his Suggestion, and in furtherance of this object, I have conducted the experiments de- scribed in this paper.^ The general plan of the research was: (A) The production of protein salts by combining organic hases, such as strychnin, mor- phin, conin, piperidin, etc., with acid-reacting proteins, such as tendo-mucoid, ovo-mucoid, yeast nucleoprotein, etc.; and {B) the production of protein salts by combining hasic proteins, such as ^Gies: Proc. See. Path. and Physiol., Amer. Med. Assn., 1906, p. 121. ''The detailed results of this work have been described in the writer's dissertation, On the synthesis of some protein salts, Columbia University, 1909 (pp. 61). A preliminary report was published by Eddy and Gies in the Pro- ceedings of the Society for Experimental Biology and Medicine, 1907, iv, pp. I4S-6. III 112 Some Protein Compounds [Sept. histons and protamins, with the acid-reacting proteins eiiumerated above. In developing the latter part of this plan certain anomalies arose in connection with the preparation of thymus histon, which led to a collateral investigation of histons. The results of the latter studies will be embodied in a future paper. (See page 169.) IL EXPERIMENTAL I. Salts of various proteins with organic bases. 'Ä. Mor- phin MUCOiD. Purification of the materials. The first step in the preparation of a typical product was the removal of free alkali f rom the base — and free acid from the protein. Chemically pure, pul- verized, morphin was washed with distilled water until the wash- ings were entirely neutral to litmus. Tendo-mucoid was prepared after the manner of Cutter and Gies^ but dehydration with alcohol and ether was omitted. The dry scales w^ere soaked in distilled water until they softened. The protein was then washed with dis- tilled water until the washings were entirely neutral to litmus. Union of base and protein. The base and the protein were then triturated together in a mortar, a very little water being added to ensure an intimate mixture. A mechanical excess of the base was used in every case. Evidence of chemical action was seen in the peculiarly viscid, smeary character of the mixture. Mucoid and w-ater give a thick, milky mixture, but it is not viscid or smeary. The mixture w^as finally treated with sufficient water in excess to dissolve the product. The viscid liquid was filtered through a wet, fluted, hardened, filter paper, but the first portions of iiltrate were returned to the paper until a clear opalescent liquid appeared. This filtrate was neutral to litmus. Purification of the product. The filtrate, preserved with tol- uene, was subjected to continuous dialysis in a parchment bag, im- mersed in frequently renewed distilled water, until the dialysate, even when concentrated to a very small volume at 40° C, gave no test for the base (morphin). The contents of the bag were then evaporated to dryness at 40° C, toluene being used and frequently renewed during the process. The resultant dry produot was then ' Cutter and Gies : Amer. Journ. PhysioL, 1902, vi, pp. 155-6. I9I2] Walter H. Eddy . 113 pulverized in a mortar and extracted three times with a large excess of ether for the removal of traces of admixed free base (morphin). Isolation of the product. The powder was next dissolved in a small amount of water and this Solution poured into a mixture of Yz ether and Yz alcohol. A copious precipitate resulted. The pre- cipitate was gelatinous and dissolved easily in water. After dissolv- ing the precipitate in water and filtering the Solution, the filtrate was precipitated with alcohol-ether. This process was repeated sev- eral times. The final product was dehydrated in the usual manner with alcohol and ether. Special difficulties in the preparation of morphin mucoid. The first Solutions of the Compound filtered very slowly. It was found that this was due to excess of mucoid. When a large excess of morphin was used there was less insoluble mucoid residue and filtra- tion became correspondingly more rapid. Precipitation of the purified product with alcohol became in- creasingly difficult with the increasing purity of the product. Am- monium Sulfate, in excess, precipitated the product from its aqueous Solution, but long dialysis was required to remove the salt. The purified product failed to respond to the iodic acid test for morphin^ This fact was carefully investigated. The results showed that the failure was not due to the quality of the iodic acid used nor to interference with the test by the mucoid. In the puri- fication of the product there seemed to be continuous loss of mor- phin. This was presumably due to hydrolytic dissociation. Evidence of the compound-naturc of the product. The product was water-soluble, demonstrating that it was neither mucoid nor morphin, nor a mechanical mixture of the two. The aqueous Solu- tion of the product frothed strongly on shaking and gave a good biuret test, indicating its protein character. Addition of a few drops of 0.2 per cent. hydrochloric acid Solution yielded a flocculent precipitate of mucoid. Conclusions regarding morphin mucoid. Morphin and mucoid *For the detection of morphin, the iodic acid test was applied as follows: I c.c. of the Solution to be tested was added to an equal volume of dilute sulfuric acid Solution. To this was added a few c.c. of iodic acid Solution and finally a little Chloroform. After vigorous shaking, the presence or absence of a violet coloration served to indicate the presence or absence of morphin. 114 Some Protein Compounds [Sept. react to form a water-soliible protein Compound which, in aqneous Solution, yields mucoid on treatment with 0.2 per cent. hydrochloric acid. The morphin enters into combination in a proportion so small as to be incapable of responding to the iodic test, or is united in such a way as to fail to respond to the test. B. Strychnin mucoid. In view of the extreme delicacy of the dichromate test for strychnin, this base was selected for the second series of preparations. Care was taken to insure purity of the original materials as in the preparation of the morphin-mucoid products. Preparation. The method of preparation was identical with that for morphin mucoid (page 112) except in the following details: The final water-solution of the product was again evaporated to dry- ness at 40° C. and the dry matter, after pulverization, was extracted with Chloroform. Twenty-six voluminous washings were neces- sary to free the powder from admixed strychnin and to obtain a strychnin-free washing. In view of the insolubility of strychnin in water and its ready solubility in ether this result seemed difficult to explain on any other basis than partial dissociation by the Chloroform. Evidence of chemical combination. The protein character of the Compound was established by the following results : The water- solution gave a strong biuret test; was precipitable by Saturation with ammonium sulfate or magnesium sulfate; gave a flocculent precipitate with 0.2 per cent. hydrochloric acid and 4 per cent. acetic acid Solutions ; and f rothed strongly on shaking. The aqueous Solu- tion of the product was neutral to litmus. The presence of strychnin was shown by the intense bitter taste and by strong "dichromate tests." Filtrates from precipitates formed by addition of 0.2 per cent. hydrochloric acid Solution yielded, in every case, strong "di-chromate tests " for strychnin. Four physiological tests were also made to establish the presence of the strychnin. The results and methods follow : The lethal dose of strychnin sulfate is about 2.5 mg. per kilo of weight for frogs and 7.6 mg. per kilo of weight for dogs. Vol- umes of aqueous Solution of strychnin mucoid (0.575 ^S- P^^* ^■^•) containing quantities equal to the lethal dose of strychnin sulfate were injected subcutaneously in frogs and dogs. I9I2] Walter H. Eddy ' 115 In the first of two experiments on frogs, the initial dose failed to produce any strychnin effects. An effect followed the second injection of an equal dose, but it required three doses to produce Opisthotonus. Recovery was complete. In the second frog, each of two doses injected successively produced Opisthotonus. The frog recovered. For the first dog a double dose was required to produce hyperes- thesia and tetanus. The results with the second dog duplicated those with the first. In all these physiological tests the strychnin appeared to be liberated slowly in the animal, the effects Coming on gradually and extending over a period of 3-5 hours, with complete recovery. Concliisions regarding strychnin mucoid. The results seemed to leave no doubt regarding the compound-nature of this product. Whether it is a true salt or an adsorption Compound can not be de- cided from the available data, but its neutrality, its water-solubility, and its power to yield both strychnin and mucoid, strongly suggest the production of a salt by a process directly comparable to the neutralization of base by acid. The physiological tests show that the Compound evidently con- tains a much smaller proportion of strychnin than that in the com- mon Sulfate. The quantitative examinations have not yet been completed. C. Conin mucoid. Conin combines with mucoid very rapidly and yields a Solution which filters easily. The product, af ter purifica- tion by dialysis and alcohol precipitation, is water-soluble and biuret- reacting. As in the case of morphin mucoid, however, it was im- possible to demonstrate the presence of the alkaloid. All tests were negative with the potassio-mercuric iodid and phospho-tung- stic acid reagents. D. PiPERiDiN MUCOID. The purified piperidin product gave the protein tests and also a test for piperidin with platinic chlorid. E. Anilin mucoid(?). A water-soluble product of mucoid and anilin was obtained but the anilin disappeared early in the puri- fication process. F. Morphin nucleoprotein. Two attempts were made to produce a Compound of morphin with yeast nucleoprotein. The ii6 Sonic Protein Compounds [Sept. method of preparation was similar to that described on page 112. Neither attempt was successful in establishing the presence of mor- phin in the final product. A water-soluble protein of different char- acter from the nucleoprotein resulted in each case. G. Morphin caseinogen, strychnin caseinogen, and cal- cium CASEiNOGEN. Studies were made of the effects of morphin, strychnin and calcium hydroxid on caseinogen. In each case water- soluble, biuret-reacting products were obtained. Rigorous purifi- cation was not attempted. Salts of ovomucoid. Neumeister^ investigated a glucoprotein in eggs which he named " pseudopeptone." This Compound was studied by Salkowski,^ Mörner/ and Eichholz,^ and called by them " ovo-mucoid." As a " cell-protein," this substance seemed to offer good material for our experiments. A pure product was prepared by Mörner's''' well-known process. Preparation of ovo-mucoid. (From eggs.) With increasing purity, precipitation with alcohol became correspondingly difficult. Alcohol-ether did not remove this difficulty but the addition of a few drops of sodium chlorid Solution brought about precipitation in every case. The final water-solution was freed from chlorid by dialysis in a parchment bag in the presence of toluene. The Solu- tion, which then was acid to litmus, was evaporated to dryness at 40° C, yielding yellow flakes which were ground to a white powder. Properties of the ovo-mucoid product. This ovo-mucoid was readily soluble in water and gave a good biuret test. The water- solution frothed on shaking, but was not viscid. Phosphotungstic acid, 0.2 per cent. hydrochloric acid, 4 per cent. acetic acid and tannic acid Solutions precipitated the aqueous Solution, which was acid to litmus. (From shad roe.) The roe was ground in a mortar with sand and this mixture poured into boiling, slightly acidulated, water. The remaining Steps were identical with those for the preparation of ovo-mucoid from eggs and the product responded to the same tests. These two products were used in the following studies. ' Neumeister : Zeitschrift für Biologie, 1890, xxvii, p. 331, 'Salkowski: Centralhlatt f. d. med. Wissensch., 1893, xxxi, pp. 513 and 706. ^ Mörner : Zeitschr. f. physiol. Chem., 1893, xviii, p. 525. ' Eichholz : Journ. Physiol., 1898, xxiii, p. 163. igi2] Walter H. Eddy 117 H. Strychnin ovo-mucoid (egg). The method of prepara- tion followed the lines of the morphin-mucoid process (page 112) with the following abbreviation : After dialysis the Solution was at once precipitated with absolute alcohol. No other methods of puri- fication were used. The filtrate f rom the original mixture of ovo-mucoid and strych- nin was turbid and acid to litmus, but became neutral on standing, in the presence of toluene. On dialysis, and consequent dilution with water, the Solution clarified. The dialysate on the other band became turbid but failed to give a protein or strychnin test, When the dialyzed liquid was treated with absolute alcohol, in excess, a mixed, cheesy and gelatinous precipitate was produced. The alcoholic filtrate from this precipitate was acid and gave a strychnin test, suggesting dissociation. The precipitate dissolved readily in water and the Solution was then filtered. It was nozv acid in reaction and gave no strychnin test. Precipitated again with alcohol, the solid product failed to give the strychnin test, was acid and resembled in every way the original ovo-mucoid. A portion of this precipitate was dissolved in water and the So- lution evaporated to dryness at 40° C. A new trituration with strychnin was made with this product. The results were the same as with the first preparation, viz., a turbin Solution that cleared on dilution with water by dialysis and gave in this condition both strychnin and protein tests. Alcohol again dissociated it into strychnin and ovo-mucoid (?). From the above results it was deemed desirable to make a care- ful study of the reactions of the product and a second preparation was conducted for this purpose. The turbid filtrate obtained from the initial mixture of strychnin and ovo-mucoid was found to be actually amphoteric to litmus, though acid to Phenolphthalein. Its alkalinity to litmus was not increased by retriturating it with strych- nin. Dilution with water resulted again in a clear Solution, giving both strychnin and protein tests. On standing for a considerable time in a parchment bag, in the presence of toluene, the amphoteric reac- tion gradually disappeared and the Solution became distinctly acid to litmus. It also finally yielded a precipitate in the bag and lost its power to respond to the strychnin test. The turbid dialysate grad- ii8 Sonic Protein Compounds [Sept. ually acquired protein material, but the frequent renewals of water and large voliime made it impossible to determine the presence of strychnin. Apparently complete dissociation resulted, but neither the character of the dissociation products nor the manner in which the strychnin separated was determined. Strychnin ovo-niucoid (roe). The results with ovo-mucoid from shad roe were identical with those in the case of tgg ovo-mu- coid except that the disappearance of the strychnin on dialysis was much slower. It was ten days before the contents of the bag failed to give the strychnin test. Concentration of the dialysates in this case before applying the strychnin test failed to make its detection possible. Evidence of the Compound natnre of the ovo-mucoid products. The clear amphoteric Solution, with its response to strychnin and protein tests, indicates a chemical combination, especially in view of the water-insolubility of strychnin. The dissociability in alcohol of the shad roe product, and the results of dialysis, indicate that it is more stable than the strychnin product with &gg ovo-mucoid. Again the question of whether we are here dealing with a true chemical Compound or with an adsorption product remains open for further investigation. 2. Protein-protein Compounds. The foregoing experiments were preliminary to attempts to bring about combinations between acid-reacting and basic-reacting proteins, such as protamins and histons. /. HiSTON MucoiD. Preparation of histon hydrochlorid. His- ton was prepared by the method of Huiskamp.^ Thymus glands from freshly killed calves were freed from fat with a knife and minced in a meat chopper. The hash was then placed in a large bottle and extracted in an ordinary ice box for 24-48 hours with distilled water. About 300 c.c. of water were used with each 100 grams of thymus. The extract was filtered through wet fluted filter papers. Nucleohiston was precipitated from the filtrate with 5 c.c. of IG per Cent, calcium chlorid Solution per 100 c.c. of extract. The precipitate was then filtered off and redissolved in water to which a little ammonia had . been added. This Solution was filtered and reprecipitated with calcium chlorid Solution in the usual way. The * Huiskamp : Zeitscjir. f. physiol. Chemie, 1901, xxxii, p. 145. igi2] Walter H. Eddy 119 precipitate was then extracted with 0.8 per cent. hydrochloric acid for the production of the hydrochlorid. This extract of histon hydrochlorid was finally dialyzed in a parchment bag against dis- tilled water until neutral to litmus. This Solution of histon hydro- chlorid was used for the preparation described below. Preparation of potassium mucoid. Acid-free mucoid was dis- solved in 0.3 per cent. potassium hydroxid Solution and the liquid filtered. The filtrate was then dialyzed in a parchment bag against distilled water (in the presence of toluene) until neutral to litmus. The product in this neutral Solution was presumably potassium mucoid. Preparation of histon mucoid. Histon hydrochlorid Solution was added drop by drop to the potassium mucoid Solution. A pre- cipitate formed immediately, and sedimented quickly beneath the clear supernatant liquid. Excess of the histon hydrochlorid Solu- tion dissolved the precipitate. The product was then filtered off and washed with water until the washings no longer gave precipi- tates with ten per cent. ammonium hydroxid or 0.2 per cent. hydro- chloric acid Solution. Evidence of the Compound nature of the histon mucoid product. A portion of the precipitate was triturated with 0.05 per cent. so- dium carbonate Solution. A colloidal Solution was obtained. Its filtrate gave a heavy precipitate with 0.2 per cent. hydrochloric acid Solution and a distinct precipitate with ammonium hydroxid Solution. These results did not determine whether the sodium carbonate merely dissolved the histon mucoid, or dissociated it into a histon So- lution and a sodium mucoid Solution. To ascertain these points the following tests were made : (a) A portion of the sodium carbonate Solution was poured into 95 per cent. alcohol. It failed to precipitate at once or on Standing. (b) A portion of the sodium carbonate Solution was poured into 95 per cent. alcohol, to which one drop of 10 per cent. sodium chlorid Solution had been added. A precipitate appeared on Standing. (c) Alcohol-ether failed to precipitate the Solution but with the addition of a drop of salt Solution a precipitate appeared. 120 Some Protein Compounds [Sept. {d) The precipitates obtained in {h) and (c) failed, in this first set of tests, to dissolve in water and the washings gave no hydro- chloric acid or ammonia precipitate. The precipitates dissolved in 0.05 per Cent, sodium carbonate Solution and the filtrates gave both the ammonia and hydrochloric acid tests. {e) Histon hydrochlorid Solution was not precipitated by alcohol even when sodium chlorid was present. Alcohol also failed to pre- cipitate potassium mucoid Solution but did so in the presence of a trace of sodium chlorid. These results Warrant the inference that the sodium carbonate acted as a solvent rather than as a dissociant. They also indicate that precipitation by alcohol in the presence of salt served to dif- ferentiate the histon mucoid from the histon hydrochlorid, and that our product was a Compound and not a mixture. A Solution of histon hydrochlorid was tested with an excess of alcohol in the absence of salt. A similar Solution of potassium mucoid was made. When these two clear Solutions were mixed a precipitate of histon mucoid formed at once. This histon mucoid was then dissolved in 0.05 per cent. sodium carbonate Solution and the filtered Solution precipitated with alcohol in the presence of a little salt. This precipitate, unlike that above (d), dissolved readily in zvater. The water-solution gave both the ammonia precipitate and the hydrochloric acid precipitate. This and similar results indi- cated the formation of a soluble histon mucoid Compound. Finally a new histon mucoid product w^as made by the original method. This product was washed free from excesses of both his- ton hydrochlorid and potassium mucoid, as before, and then treated as f ollows : A portion was macerated in a mortar with o.i per cent. hydro- chloric acid Solution and a second portion in another mortar with 0.1 per cent. potassium hydroxid Solution. These liquids were fil- tered. The acid filtrate gave a precipitate with ammonia but not with hydrochloric acid. The alkali filtrate gave a heavy precipitate with hydrochloric acid but none with ammonia. These results suggest that mixtures of (a) histon hydrochlorid and potassium mucoid Solutions yield a precipitate of histon mucoid; (&) pure histon mucoid and o.i per cent. hydrochloric acid I9I2] Walter H. Eddy 121 Solutions yield histon hydrochlorid and insoluble mucoid; (c) pure histon mucoid and o.i per cent. potassium hydroxid Solutions yield a potassium mucoid histon complex. The failure to get a histon precipitate with ammonia in the potassium hydroxid extract may have been due to the small amount of resultant histon mucoid or to the formation of an insoluble form o£ histon, such as ammonia produces. Whatever the explanation of this failure, there seemed to be no doubt of the power of histon to combine with mucoid to form a Compound different in proper- ties from either histon hydrochlorid or potassium mucoid. /. Histon nucleoprotein (yeast). Neutral potassium nu- cleoprotein (obtained by dissolving yeast nucleoprotein in o.i per cent. potassium hydroxid Solution and dialyzing free from hydroxyl ions) combines with histon hydrochlorid in the same way as potas- sium mucoid (page 119). Much more of the Solution of histon is necessary for the production of the salt. The product was similar to histon mucoid in being insoluble in water; in dissolving readily in 0.05 per cent. sodium carbonate Solution but incompletely in 0.5 per cent. sodium carbonate Solution ; and in forming, with sodium carbonate, a water-soluble sodium-histon nucleoprotein complex. K. Histon ovo-mucoid. Preparation. The ovo-mucoid {tgg) was purified to such a degree as to be practically soluble in salt-free alcohol (page 116). A similarly pure Solution of histon hydro- chlorid was used. When the water Solutions of these two substances were com- bined, a precipitate formed slowly. A slight excess of the histon Solution dissolved the precipitate. The precipitate dissolved to a turbid Solution in 0.05 per cent. sodium carbonate Solution. This turbid fluid was filtered and divided into two portions. One por- tion was poured into 95 per cent. alcohol to which 3 drops of 10 per cent. sodium chlorid Solution had been added. The other por- tion was saturated with ammonium sulfate. Both portions gave heavy precipitates, which were soluble in water; the Solutions were precipitated in part by ammonia. When purification of the ammo- nium sulfate precipitate by dialysis was attempted, the Compound broke down. The "alcohol precipitate" was hydrolyzed with hydrochloric acid. The resultant liquid, neutralized with potassium 122 Some Protein Compounds [Sept. hydroxid, rediiced the Fehling-Benedict reagent. This result, with the precipitation by ammonia, seemed to show the presence of both glucoprotein and histon in the precipitate. When alcohol Solutions of ovo-mucoid and histon hydrochlorid were poured together, a precipitate formed at once that gave both the ammonia test and the reduction test. The latter process is the simplest and quiekest method of obtaining this product. The results of these researches have shown that the methods for the preparation of histon, as outlined in the literature, are in serious need of revision. In fact, the results suggest that so-called histon is a protein salt rather than a simple protein. In a future paper will be presented the findings in regard to histon preparation. EFFECTS OF INTRAPERITONEAL INJECTIONS OF EPINEPHRIN ON THE PARTITION OF NITRO- GEN IN URINE FROM A DOG JACOB ROSENBLOOM and WILLIAM WEINBERGER (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) I. INTRODUCTION The action of epinephrin on nitrogenous metabolism has been the object of investigation by several authors. The experimental results of Kraus and Hirsch/ and Quest,^ indicate that intravenous or subcutaneous injections of epinephrin exert very Httle influenae on the nitrogenous metaboHsm of healthy dogs, the insignificant increase of eliminated nitrogen being caused both by the glycosuria and (after subcutaneous injections) by skin necrosis. Fasting ani- mals seem to be differently affected. Falta and Rudinger,^ and Underhill and Closson* were able to show an accelerating influ- enae, on protein metaboHsm, of subcutaneous and intravenous injec- tions of epinephrin. Underhill and Closson have shown that the subcutaneous injec- tion of " adrenalin chlorid " Solutions into dogs is not attended by any significant change in the proportions of the urea-, ammonia- and creatinin-nitrogen of the urine, in partial disagreement with Paton,^ who also found that, although on a sufficient diet, the catabolism of proteins is not interfered with, there is a markedly increased produc- tion of ammonia. In all the above mentioned experiments the epinephrin was in- jected into veins or into subcutaneous tissues. The intraperi- toneal way has not been utilized by previous observers in this con- ^ Kraus and Hirsch. Cited by Kraus and Friedenthal : Berl. klin. Woch., 1908, xlv, p. 1709. ^Quest: Zeit. f. exp. Path., 1908, v, p. 43. ' Falta and Rudinger : Central, f. klin. Med., 1908, Ixvi, p. i. * Underhill and Closson : Anier. Journ. Physiol., 1906, xvii, p. 42. ' Paten : English Journ. of Physiol., 1903, xxix, p. 286 ; 1904, xxxii, p. 59. 123 124 Epinephrin Effects on Metabolism [Sept. nection, althongh it is possible that this mode of administering epinephrin has a different effect on nitrogenous metabolism, as is the case in carbohydrate metabolism (Löwi).® IL DESCRIPTION OF THE EXPERIMENTS This investigation consisted of two metabolism experiments. One animal was used for both experiments. An intermediate pe- riod served to allow the animal to recuperate from the effects of the first experiment before the second one was begun. The metabolism work was conducted by the general methods in use in this labo- ratory.'^ We determined the nitrogen content in the several ingredients of the food. Urinary nitrogen, in the leading forms, was deter- mined as f ollows : ammonia, each day ; total, urea, creatin and Crea- tinin (as Creatinin), every third day; purins, at the end of each period. The urine was preserved with thymol. Total nitrogen was determined by the Kjeldahl process ; ammonia and Creatinin by the Polin methods f iirea^ by Benedict's method ;^^ purin nitrogen by a combination of the Arnstein" and Salkowski^^ methods. The authors used two specimens of the colorless "adrenalin Chlorid" (i:i,ooo) of Parke, Davis and Co. They were pur- chased in the open market. Each was tested for its pressor action at the conclusion of the corresponding injection experiments and was then found to be practically as active as ever. Varying amounts and concentrations of "adrenalin chlorid" were injected into the peritoneal cavity ; in the first experiment the concentration was 1 : 10,000 — in the second, i : i,ooo. In one injection period of *Löwi: Von Noorden's Metabolism and practica! medicine, 1907, iii, p. 1181. ' Mead and Gies : Anier. Journ. Physiol., 1901, v, p. 106 ; also Gies and collab- orators: Biochemical Researches, 1903, i, Reprint No. 21; Gies: Amer. Journ. of Physiol., 1905, xiv, p. 403; Gies: Amer. Journ. of Physiol., 1901, v, p. 235; also Gies and collaborators : Biochemical Researches, 1903, i, Reprint No. i ; Gies : Proc. Amer. Physiol. Soc, Amer. Journ. of Physiol., 1904, x, p. 22; Hawk and Gies: Amer. Journ. of Physiol., 1904, xi, p. 177. * Polin : Amer. Journ. of Physiol., 190S, xiii, p. 45. " No glycosuria occurred. Examinations were made repeatedly. ^^ Benedict : Journ. of Biol. Chem., 1910, viii, p. 405. " Arnstein : Zeit. f. physiol. Chem., 1897, xxiii, p. 417. " Salkowski : Salkowski's Manual of physiol. chem. and path., 1904; Arch. d. ges. Physiol., 1897-98, Ixix, p. 268. I9I2] Jacoh Rosenhloom and William Weinherger 125 eighteen days, a total of 62 c.c. o£ i : 10,000 Solution was given intraperitoneally ; in another injection period of six days, a total of 29 c.c. of 1 : 1000 Solution was administered. The accompanying tables contain the metabolic data obtained in this study. TABLE I. FIRST METABOLISM EXPERIMENT (jUNE I2-JULY l8, I9I2) A. Daily Records I. Fore Period. Normal Condition Number of the day I 2 3 4 5 6 7 8 9 10 Body weight (kilos) 6.3 270 1,017 6.33 210 1.025 6.3 270 1,020 6.3 190 1,024 6.31 192 1,024 6.3 212 1,021 6.28 275 1,020 6.27 240 1,018 6.3 188 1,021 6.33 Urine, volume (c.c.) 170 1,026 TJrine. so. er II. Dosage Period. Intraperitoneal Injections Number of the day Body weight (kilos) . , Urine, volume (c.c.) . , Urine, sp.gr , Adrenalin Solution (i 10,000) c.c. 6.34 135 1,027 S 6.27 157 1,031 5 6.28 80 1,042 5 6.26 100 1,040 5 6.35 165 1.030 8 6.25 224 1,020 6.33 160 1.030 5 8 6.26 245 1,020 6.31 180 I,02S 5 Number of the day Body weight (kilos) . . Urine, volume (c.c.) . . Urine, sp. gr Adrenalin Solution (i 10,000) c.c. 10 II 12 13 14 IS 16 17 6.26 6.33 6.41 6.40 6.42 6.37 6.35 6.34 200 105 153 175 255 230 205 270 1.023 1.037 1.025 1,023 1,019 1,017 1,020 1,020 2 3 3 — 4 — 4 4 18 6.43 167 1,023 4 III. After Period. Normal Conditions Number of the day I 2 3 4 5 6 7 8 Body weight (kilos) Urine, volume (c.c.) Urine. so. sx 6.34 135 1,027 6.40 145 1,020 6.44 200 1,022 6.40 245 1,020 6.44 220 1,018 6.41 185 1,022 6.40 214 1,019 6.46 214 1,020 B. Analytical Totais and Daily Averages of Urinary Data for Each Period 0 3 > - 4) IS Nitrogen UreaN Ammonia N Creatin and Creatinin N Purin N Period 2S H So Daily average, grams 2 S E-i dt Daily average, grams H bo Daily average, grams Total, grams Daily average, gram 15 H Daily average, gram Fore (10 days). Dosage (18 days). After (18 days). 2,217 3.206 1.558 221 178 195 41-594 74.225 33-077 4-159 4.123 4-134 36.85 63-58 28.63 3.685 3-532 3.578 I.5II 2.945 I-I95 0.1511 0.1636 0.1493 1.046 I.714 I.IOI 0.1046 0.0952 0.1376 0.067 0.147 0.081 0.0067 0.0081 O.OIOI 126 Epinephrin Effects on Metaholism [Sept, TABLE 2. SECOND METABOLISM EXPERIMENT (jULY 2I-AUGUST 9,. I912) A. Daily Records I. Fore Period. Normal Conditions Number of the day Body weight (kilos) Urine, volume (c.c.) Urine, sp.gr 6.52 190 1,020 6.53 190 1,023 6.53 230 1,019 6.54 200 1,020 6.57 193 1,021 IL Dosage Period. Intraperitoneal Injections Number of the day I 2 3 4 5 6 Body weight (kilos) . . . Urine, volume (c.c). . . Urine. so. er 6.54 ISO 1.028 4 6.69 134 1,028 5 6.68 215 1,019 5 6.63 275 1,018 5 6.70 185 1,020 5 6.71 240 1,016 Adrenalin Solution (i : 1,000) c.c 5 III. After Period. Normal Conditions Number of the day Body weight (kilos) Urine, volume (c.c.) Urine, sp.gr I 2 3 4 5 6 7 8 6.68 245 1,019 6.72 232 1,018 6.72 230 1,018 6.76 200 1,019 6.78 202 1,017 6.77 240 1,018 6.78 200 1,019 6.83 223 1,017 6.80 23s 1,019 B. Analytical Totais and Daily Averages of Urinary Data for Each Period 0 E 0 3 "o > >> 0 > > " Nitrogen UreaN Ammonia N Creatin and Creatinin N Purin N Period — B E — « 2 E 0 rt r: n E ü E V := t« E S E 0 « H So CS 0 u cd >" f^ Fore (5 days) . . . Dosage (6 days) After (9 days) . . 1,003 1,199 2,007 201 199 223 20.997 24.678 34-937 4.199 4-II3 3.882 18.22 21.46 29-39 3-643 3-5766 3.266 0.767 0.967 1.499 0.1533 0.1611 0.1664 0.671 0.7314 1.266 0.1342 O.I2I9 0.1407 0.039 0.039 0.059 0.0076 0.006s 0.0066 TABLE 3. PARTITION OF THE URINARY NITROGEN Period I. Fore period Injection period .... Post injection period II. Fore period Injection period .... Post injection period Urea N, per Cent. 88.5 85-7 86.5 86.7 87.0 84.2 Ammonia N, per Cent. 3-64 3-75 3-61 3-65 3-92 4.29 Creatin and Creatinin N, per Cent. 2.51 2.50 3-33 3-20 3-73 3-62 Purin N, per Cent. 0.16 0.20 0.24 0.19 0.16 0.17 Undeter- mined N, per Cent. 5-19 7.86 6.32 6.26 5-19 7.72 I9I2] Jacob Rosenhloom and William Weinherger 127 III. CONCLUSIONS The results of these experiments show conclusively that intra- peritoneal injections of "adrenalin chlorid" Solutions were without appreciable effect on the proportions of nitrogen (in the forms of Urea, ammonia, creatin and Creatinin, purins, and undetermined sub- stances) in the urine of a healthy dog. /THE BIOCHEMICAL SOCIETY, ENGLAND In a previous note, which appeared in the Biochemical Bul- letin (i: 484), it was stated that the recently founded Bio- chemical Club would probably develop into a society with a Journal of its own. This is now an accomplished fact, and the Biochemical Society of England has been launched into being. It has been instituted for the purpose of facilitating intercourse between those biologists and chemists who are interested in problems common to both, such as the chemical questions connected with agriculture, brewing, animal and vegetable physiology and pathology, etc. Meetings are held at different centers throughout the country for the communication of papers and demonstrations. The Honorary Secretary is Dr. R. H. A. Flimmer, University College, London, W. C, from whom further Information can be obtained. The Bio-Chemical Journal, which has hitherto been under the editorship of Professor Moore, F.R.S., of Liverpool, will in the fu- ture be conducted by the Biochemical Society, and will be issued by the Cambridge University Press, Fetter Lane, London, E. C. The editors are Professor W. M. Bayliss, F.R.S., University College, London, W. C, and Professor A. Harden, F.R.S., Lister Institute, Chelsea Gardens, London, S. W. The first issue of the Journal under these editors is expected in January next. The price is £1.1.0 ($5) per volume. One f€els sure that our American confreres will heartily support the new enterprise. W. D. Halliburton King's College, London. 128 MEETINGS OF THE SECTION (II) ON DIETETIC HYGIENE AND HYGIENIC PHYSIOLOGY OF THE FIFTEENTH INTERNATIONAL CONGRESS ON HYGIENE AND DEMOGRAPHY, WITH AB- STRACTS OF SOME OF THE PAPERS Proceedings reported by THE Secretary, LAFAYETTE B. MENDEL The meeting o£ the Congress was noteworthy for the unusual opportunity which it afforded to American men of science to meet some of their foreign colleagues, particularly those from the Con- tinent, in a personal way. It can scarcely be said that the proceed- ings of the Section on Dietetic Hygiene and Hygienic Physiology were unique in any way; nor could they be expected to attract the Chief interest where so many important disciplines and conflicting or overlapping scientific fields were involved. The Symposium on the specific dynamic action of foodstuffs deserves special comment, however, both on account of the new views which were forcefully presented there for the first time, and the preeminent part played by all of the referees in the development of this field of study. I. OFFICIAL LIST OF PRESIDENTS AND VICE-PRESIDENTS OF THE SECTION Honorary Presidents : Dr. Max Rubner, Professor of Phys- iology and Director of the Physiological Institute, Berlin, Germany ; Dr. Artur Schattenfroh, Professor of Hygiene in the Univer- sity of Vienna, Austria ; Dr. Axel Holst, Professor of Hygiene, University of Christiania, Norway ; Dr. A. B. Macallum, Profes- sor of Biochemistry, University of Toronto, Canada. President: Dr. Russell H. Chittenden, Professor of Phys- iological Chemistry, Sheffield Scientific School of Yale University, New Haven, Conn. 129 130 Biochcmical Proceedings, Hygienic Congrcss [Sept. Vice-presidents : Dr. Graham Lusk, Professor of Physiology, Cornell University Medical College, New York City; Dr. David L. Edsall, Professor of Clinical Medicine, Harvard Medical School, Boston, Mass. II. OFFICIAL PROGRAMM All the meetings of the section were held on Sept. 23 to Sept. 27, inclusive, in Washington, D. C, at the new National Museum, Room 376. 1. Monday afternoon, September 23. The physiological SIGNIFICANCE OF SOME SUBSTANCES USED IN THE PRESERVATION OF FOOD : Dr. John H. Long, professor of chemistry, Northwestern Uni- versity Medical School, Chicago, 111. (page 132) ; Dr. Artur Schat- tenfroh, professor of hygiene in the University of Vienna, Austria. 2. Tuesday morning, September 24. The specific dynamic ACTiON OF FOODSTUFFS : Dr. Max Ruhner, professor of physiology and director of the Physiological Institute, Berlin, Germany. {A) The work of digestion and specific dynamic action : Dr. N. Zuntz, Direktor des tierphysiologischen Laboratoriums der landwirtschaft- lichen Hochschule, Berlin, Germany {presented by Prof. F. G. Bene- dict).— (5) The influence of the Ingestion of food upon metabolism: Dr. Francis G. Benedict, director of the Nütrition Laboratory of the Carnegie Institution of Washington, Boston, Mass. (page 134). — (C) The influence of foodstuffs and their cleavage products upon heat production : Dr. Graham Lusk, professor of physiology, Cornell University Medical College, New York City (page 135). 3. Tuesday afternoon, September 24. Nutrition and GROWTH. (A) An anatomical analysis of growth : Dr. Henry H. Donaldson, The Wistar Institute of Anatomy, Philadelphia, Pa. — (B) Nutrition of the embryo: Dr. John R. Murlin, assistant pro- fessor of physiology, Cornell University Medical College, New York City. — (C) The nutrition and growth of bone: Dr. Francis H. Mc- Crudden, chemist at the Hospital of the Rockefeiler Institute for Medical Research, New York City (page 137). — {D) The role of proteins in growth : Dr. Lofayette B. Mendel, professor of physi- ological chemistry, Shefiield Scientific School of Yale University, New Haven, Conn. (page 138). — (E) The influence of the quantity * Abstracts of the papers appear on the pages indicated by the numerals in parenthesis. I9I2] Lafayette B. Mendel 131 and quality of food upon the growing organism : Dr. Hans Aron, director of the scientific laboratory of the University Children's Clinic, Breslau, Germany {presented by Prof. Lafayette B. Men- del).— (F) Direct calorimetry of infants, with a comparison of the results obtained by this and other methods: Dr. John Howland, Professor of pediatrics, Johns Hopkins University, Baltimore, Md. (page 139). 4. Wednesday morning, September 25. The röle of in- ORGANIC SUBSTANCES IN THE NUTRITION OF MAN. {A) The antag- onistic action of salts : Dr. Jacques Loeb, head of the department of experimental biology, Rocke feller Institute for Medical Research, New York City. — (B) The distribution of soluble salts in living cells and the forces Controlling it: Dr. Archibald B. Macalliim, pro- fessor of biochemistry, University of Toronto, Canada (page 140). — (C) The röle which common salt and water assume in the nutri- tion of man : Dr. Hermann Strauss, professor of clinical medicine, University of Berlin, Germany (page 141). 5. Thursday morning, September 26. Practical dietetics, {A) Cost and nutritive value of f oods : Dr. C. F. Langworthy, ex- pert in charge of nutrition investigations, U. S. Department of Agriculture, Washington, D. C. — {B) The influence of the prepara- tion of food on its nutritive value : Dr. Max Riibner, professor of physiology and director of the Physiological Institute, Berlin, Germany. — (C) The choice of foods, with regard to disease: Dr. Carl von Noorden, professor of internal medicine and director of the First Medical Clinic, Vienna, Austria (page 143). — {D) Diet in relation to disease, chiefly in relation to some forms of partial un- derfeeding (beriberi and scurvy) : Dr. Axel Holst, professor of hy- giene, University of Christiania, Norway. — {E) Diet and metabo- lism in fever : Dr. Warren Coleman, Cornell University Medical College, New York City (page 145). 6. Thursday afternoon, September 26. Ventilation in its HYGiENic ASPECTS, {A) Organic matter in the expired air: Dr. Milton J. Rosenmi, professor of preventive medicine, Harvard Med- ical School, Boston, Mass. — (B) A consideration of the unknown fac- tors in the ill-effects of bad Ventilation: Dr. Yandell Henderson, professor of physiology, Yale Medical School, New Haven, Conn. (page 146). — (C) The hygienic physiology of work in compressed 132 Biochcmical Proceedings, Hygienic Congress [Sept. air: Dr. J. J. R. Macleod, professor of physiology, Western Re- serve Medical School, Cleveland, Ohio (page 147). 7. Friday morning, September 27. The hygienic physiol- ogy OF EXERCISE. {A) The influence of exercise on the nervous System : Dr. Leon Asher, a. o. professor of physiology, Bern, Swit- zerland {presented by Prof. L. B. Mendel). — {B) The influence of exercise on the heart : Dr. R. Taif McKenzie, director of physical education, University of Pennsylvania, Philadelphia, Pa. — (C) Certain aspects of the influence of muscular exercise upon the respi- ratory System : Dr. Theodore Hough, professor of physiology, Uni- versity of Virginia, Charlottesville, Va. (page 148). — {D) Physical training in the United States Naval Service: Dr. J. A. Murphy, surgeon, U. S. N., U. S. Naval Academy, Annapolis, Md. Additional papers. The prevention of arteriosclerosis and heart disease in otherwise healthy individuals past middle life: Dr. Louis F. Bishop, New York City, — Tuberculosis and metabolism: Dr. Diesing, chief physician, Recreation and Convalescent Home, Gross-Hansdorf, Hamburg, Germany. — On the nature and impor- tance of the diet as the most important factor of causal therapy in severe diseases of the stomach and intestines, in nervous and mental diseases, and in disorders of the circulation and of the metabolism : Dr. W. Plönies, Hanover, Germany. — Public baths: Dr. Simon Baruch, president, American Association for Promoting Hygiene and Public Baths, New York City. — The significance of hydrother- apy for hygiene, therapeutics and medical Instruction: Prof. Dr. L. Brieger, Hydrotherapeutische Universitäts-Anstalt, Berlin, Ger- many.— The importance of the nutritive salts for healthy and sick people : Dr. R. Peters, Hanover, Germany. III. ABSTRACTS OF SOME OF THE PAPERS = The physiological significance of some substances used in the preservation of food JOHN H. long This paper dealt with the action on the human organism of a number of substances employed as food preservatives, or otherwise, in the preparation of food. * Reprinted f rom the official pamphlet containing " abstracts of papers to be read at the congress," Sept. 23-28, 1912 (pp. 11-28). igi2] Lafayette B. Mendel 133 Something of the history of food preservatives was recited, and it was shown that a considerable number of substances are added to food largely because of their preservative properties, rather than because of flavors they may impart. Some of the so-called " na- tural " preservatives come under this head. Modern conditions of living and modern scientific advances have called for the introduc- tion of more efficient substances, the so-called " chemical " or " arti- ficial " preservatives. Many of these substances have been con- demned, and perhaps properly, but frequently the condemnation is solely on the ground of their origin. This basis of condemnation has no justification in fact, as all preservatives are as truly chem- ical as are those of recent introduction made by industrial processes. The active principles in cloves, cinnamon, allspice, etc., are true chemical Compounds, and in their action on the body and final dis- position are much like benzoic acid, now made largely by laboratory processes. A number of important investigations on the physiological action of sodium benzoate have been carried out in the last few years, and the results of these were discussed. The effects of large and small amounts of benzoic acid are known, and it has been clearly shown that the use of the small quantities employed in the ordinary pro- tection of the condimental foods is quite unobjectionable. Such small amounts are normally disposed of in the human body without ill effects. The use of copper salts in coloring vegetables was next discussed. There is an enormous literature on the subject, especially from France and Germany, where copper has long been used in the can- ning Industries. Several commissions have pronounced in favor of permitting the use of copper salts, although others have opposed it. But all authorities have come to agree that the toxicity of these salts is much less than was at one time assumed. This toxicity depends somewhat on the combinations in which the salts are in- gested. The effects of copper as used in young peas or string beans are far less marked than are those of its inorganic salts. It is, there- fore, not quite justifiable to draw conclusions as to the behavior of copper from experiments with copper sulfate alone. If only very young and fresh vegetables, with plenty of chlorophyl, were treated 134 Biochemical Proccedings, Hygienic Congress [Sept. with copper, and if the amount were strictly limited, there might be but little fault foiind. But with older vegetables the combination is far less stable and the effects approach those of the inorganic salts. The amounts of copper taken up by the liver and other Organs from inorganic salts may be considerable, and such absorp- tion cannot be held free from danger. The use of these salts serves no real good purpose and should be condemned. The paper touched also on the employment of sulphurous oxide and sulphites in certain food Industries. The influence of the ingestion of food upon metabolism FRANCIS G. BENEDICT Three interpretations of the increase in metabolism following the ingestion of food are current : first, the theory in which the mechanical work of the digestive processes plays the most prom- inent röle; second, the less sharply defined theory in which the con- ception of the development of free heat unavailable to the cells is the dominant note, and, finally, the opinion expressed by Friedrich Müller, that there is absorbed out of the food certain substances w'hich are carried by the blood to the cells and there stimulate the cells to a greater metabolic activity. The evidence used for the evaluation of these views in this paper is based almost exclusively upon experiments made upon men in our laboratory. It was found that although the ingestion of sodium sulfate produced a powerful peristalsis, no measurable increase in the me- tabolism as measured by the oxygen consumption was noticed. Similarly, the ingestion of large amounts of agar-agar produced very voluminous, bulky stools, but did not increase metabolism measurably. As subsidiary evidence, in unpublished experiments on dogs with deficient pancreatic secretion, it was found that al- though the decreased assimilation of protein and fat resulted in large, bulky, fatty stools, there was not an increase in the carbon dioxide production. The evidence points strongly to the fact that the ingestion of meat by depancreatized dogs, accompanied as it is with large, voluminous, bulky stools, results in absolutely a smaller increase in metabolism than is experienced with the ingestion of meat by normal dogs. I9I2] Lafayette B. Mendel 135 Both of these pieces of evidence, therefore, can be taken as strongly contrary to the work of digestion as an explanation of any considerable proportion of the increased metabolism generally noted after the ingestion of food. Experiments both on dogs and on men show that foUowing the ingestion of food there is an increased muscle tonus as indicated by the pulse rate, and frequently by the respiration rate, showing that the animal is living on a higher metaboHc plane than formerly. The increased heat is thus a product of cell action, and the question as to its economic value acquires a new significance. A man asleep, with lowest heat production, is of little value to the world ; awake, with no external muscular activity, he has increased internal activity and is capable of intellectnal life. The ingestion of protein alone stimulates metabolism with the possibility of some differences in the kinds of protein. Carbohy- drates show rapid effects, not so great as protein, and different car- bohydrates give different results. The metabolism 12 hours after the last meal of a carbohydrate-free, fat-rich diet, with moderate amounts of protein, is much greater than the metabolism under sim- ilar time-conditions after a mixed diet with the same amount of protein. Diabetics with varying degrees of intensity of the disease show marked differences in the total metabolism 12 hours after the last meal. A high acidosis is coincident with a high metabolism. A carbohydrate-free, fat-rich diet, eaten by a normal individual, is accompanied by the presence of an acidosis and an increased metabolism. The evidence suggests that coincidental with what is commonly termed a " State of acidosis " there is present in the blood a substance or substances, probably of an acid nature, that stimulate the cells to a greater metabolism. ö' The influenae of foodstuffs and their cleavage products upon heat production GRAHAM LUSK If meat in large quantity be given to a dog, the heat production rises in the second hour almost to its maximum, reaches its maxi- mum in the third hour and continues at this level through the tenth 13Ö Biochemical Proceedings, Hygienic Congress [Sept. hour when it beglns to fall. In one instance the heat production during a morning hour was 22.3 calories, and after the ingestion of 1,200 grams of meat it had risen in the second hour to 36 calories, reaching 40 calories in the third hour at which level it remained through the tenth hour, after which it gradually feil to 25 calories in the twenty-first hour. During the second hour the nitrogen elim- ination was one third the maximal nitrogen Output as evenly main- tained between the third and tenth hours. The second hour also showed that the calculated non-protein respiratory quotient ranged between 90 and 99, which indicated that that part of the metabolism which was not due to protein, as calculated from urinary nitrogen, originated largely from carbohydrate. During the later hours, the increased heat production is proportional to the nitrogen in the urine. During a period of 15 hours, protein carbon was retained in the or- ganism and when the oxygen absorption as computed on the basis of such retention in the form of dextrose is compared with the actual oxygen absorption, the two agree within 0.9 per cent., whereas computed on the basis of carbon retained as fat, there is a discrepancy of 10 per cent. between the calculated and actual value. Administration of 50 grams of dextrose in 150 c.c. of water to a dog causes a rise of heat production from 16.2 to 20 calories, at which level it is maintained during the second, third and fourth hours, falling nearly to the basal level in the fifth hour. The skin temperature rises to a greater extent than the rectal temperature. The absorption from the intestine is completed in the fourth hour. The urine is scanty until the fourth hour when 100 c.c. are suddenly eliminated. The sugar content of the blood in per cent. rises in the first hour but becomes normal after that. After the first hour the percentage of hemoglobin in the blood falls but returns to nor- mal subsequent to the fourth hour. Hence, after sugar ingestion, osmotic phenomena cause an increased volume of blood. When the absorption is complete, the glycogenic function removes the dex- trose from the blood, and the blood returns to its normal composi- tion through the elimination of water by the kidney. Water alone or a Solution of salt or of urea have no effect on the metabolism, hence the increase in metabolism is probably due to the increased number of molecules of dextrose carried to the cells and not to I9I2] Lafayette B. Mendel 137 changes due to osmosis. Liebig's extract of beef is without influ- ence on the metabolism. Fifty grams of olive oil cause a consider- able increase in heat production. GlycocoU causes a very great in- crease in heat production, alanin also acts powerfully, leucin and ty- rosin less so and glutamic acid not at all. It is concluded that the heat production may be increased by increasing the quantity of sugar and fat reaching the cells, or it may be increased through the direct Stimulation of the cells by amino- acids, notably glycocoll and alanin. Nutrition and bone growth FRANCIS H. MCCRUDDEN The question of the nature of bone metabolism in health and disease is one that until recently can hardly be said to have been attacked experimentally. The pathologists, with one or two excep- tions, have generally considered bone as a dead tissue not undergo- ing metabolism, once it is laid down. This opinion has, of course, colored their views regarding the nature of the process in various bone diseases. In osteomalacia, for example, a disease in which there is a decrease in the mineral content of the bone, it has been supposed that the process is due to the action of an acid which dis- solves out the mineral constituents. Numerous investigations, chemical, histological and clinical, dur- ing the last few years have shown that these views regarding the nature of the process in osteomalacia and the nature of normal bone metabolism cannot be correct. Bone, like the other tissues, under- goes metabolism throughout life. Old bone is continuously being resorbed and new bone laid down. If the new bone laid down is not qualitatively of the right composition, the result may be rickets, osteomalacia, osteoporosis, or Osteitis deformans, depending on the age of the patient and other factors. The bones act as a störe of lime salts to be called on in time of need just as the subcutaneous tissue acts as a störe of fat and the liver as a störe of glycogen; and a flux of calcium from the bones started by a growing fetus, a hardening callus, metastatic bone formation, etc., may, under cer- tain circumstances, lead to decalcification enough to result in osteo- 138 Biochemical Proceedings, Hygienic Congress [Sept. malacia and similar conditions. An important factor is the degree to which overproduction takes place, a factor involved also in im- munity and, in fact, in all tissue repair. Other disturbances which may be said to involve quantitative dis- turbance in bone growth, — the rate of growth, — rather than dis- turbances in the qualitative character of the bone produced, are the various types of dwarfism. In some of these the failure to grow seems to depend on an absence of the " growing tendency " on the part of the bones; in others, some disturbance in the supply of lime salts available for bone growth seems to be at fault. The role of proteins in growth LAFAYETTE B. MENDEL Some of the views held in the past regarding the interrelation of the food supply and growth are no longer tenable. Growth has often been associated in a causal way with the relative abundance of protein in diet. The parallelism between the protein content of the milk of various species and rate of growth may, in the familiär cases be an example of correlation rather than of causation. Recent in- vestigations have shown that the assumed association of growth with high protein intake is not confirmed by the evidence at band. Growth is a function of the cells. This inherent capacity ap- parently cannot be exaggerated by feeding ; but growth can be held in abeyance by various conditions. These include inadequacy of the food supply in respect to both quantity and quality of the nutrients. Attention must be directed to the chemical as well as the energetic aspects of the problems involved. In the past physiologists have largely disregarded the relative values of the individual members of different groups of food substances in nutrition, owing to an igno- rance of the chemical characteristics of the individuals. In considering the uses of protein in the organism, the distinc- tion between the requirement for maintenance and that for growth must be clearly kept in mind. The development of a successful method of investigation by Osborne and Mendel has made it easy to approach some of the problems experimentally. The method was explained in detail. Normal rate of growth has been induced in I9I2] Lafayette B. Mendel i39 rats with dietaries containing various Single purified proteins. But not all proteins suffice to promote growth under otherwise favorable conditions. Some suffice for maintenance without growth, whereby a prolonged period of stunting, or suppression of growth, can be in- duced ; still other proteins are alone insufficient for the maintenance requirement. The capacity to grow is not lost even after comparatively long periods of dwarfing and a subsequent normal unimpaired rate of growth may be attained with a suitable protein dietary. Aside from the apparent nutritive inequalities of the different proteins, other incidental findings, such as the synthetic features in growth, and diverse questions raised thereby, present a multitude of view- points which may serve to direct further research in this field. Direct calorimetry of infants, with a comparison of the results obtained by this and other methods JOHN HOWLAND A discussion of the various methods for determining the ex- change of energy in infants with their advantages and disadvantages and their limitations. The results of direct calorimetry obtained with four children by means of a modified Atwater-Rosa-Benedict calorimeter. The carbon dioxide excretion, oxygen consumption and heat production of two essentially normal children. The effect of the Ingestion of food, and especially of an excess of protein. The effect of eighteen hours' fasting. The heat production and respira- tory exchange of two extremely emaciated children. Difficulty of comparing results with those obtained by other methods on account of the different conditions under which the experiments have been conducted, the different Information that has been supplied and also on account of the unsatis facto ry formulas for determining the surface areas of infants, which give errors of 20 per Cent, and more. A new, more accurate and simple formula for determining this. 140 Biochcmical Proceedings, Hygienic Congress [Sept. The distribution of soluble salts in living cells and the force^ Controlling it ARCHIBALD B, MACALLUM 1. The distribution of salts in living matter is held to be due to the forces that make the distribution of salts uniform in an ordinary Solution. These forces are the same as those which determine the distribution of the molecules of a gas in an enclosed space. Conse- quently, in a living cell the salts are supposed to be uniformly dis- tributed throughout the fluid of the cytoplasm, that is, the osmotic force, or the pressure exercised by the molecules and ions through- out the fluid, is due to this uniform distribution of the solute throughout the System. The quantity of a soluble salt present in living matter is, therefore, a measure of the osmotic pressure therein and hence exchange between the salts within and those without would, in all cases, if the cell membrane were permeable, develop so as to ad just the pressure equally within and without the cells. 2. This conception leaves wholly out of account the action of surface tension. Every particle of the colloid of which living mat- ter is composed presents to the fluid in which it is suspended an In- terface where the surface tension of the fluid is lower than such a fluid has at its free surface. In consequence the Gibbs-Thomson principle comes into Operation and there results a condensation of the molecules and ions of the solutes on the Interfaces of all par- ticles. As the united interfacial surfaces must, in relation to the total volume of the Solution or fluid, present a very great area, a very large proportion of each of the solutes must be so Condensed, and the general concentration is, accordingly, greatly reduced or brought to the vanishing point. This would reduce the osmotic pressure due to such solutes to a very low value or even to nil. 3. The degree to which concentration on surfaces or on In- terfaces obtains depends on the degree of diminution of the tension of the fluid at an Interface, but it also depends on the nature of the solute, for the concentration in case of certain salts greatly exceeds the value demanded by the Gibbs equation, while in other salts the ascertained value approximates the theoretical value. 4. Such surface condensation of the salts of living matter can. I9I2] Lafayette B. Mendel 141 in a number of cases, be demonstrated microchemically. In the case of potassium salts this Is especially feasible. They are in this way found Condensed in interfaces inside of living cells, and also on sur- faces in tissues, i. e., on the external surfaces of nerve cells, of renal, pancreatic and salivary tubules, and thereby relations are established which determine the processes of excretion and secretion of these salts. In such cases the potassium salts in the tissue fluids elsewhere than at the surfaces or interfaces are scarcely detectible microchem- ically. 5. Surface tension is, therefore, an all-important factor in de- termining the distribution of salts in living matter. The role which common salt and water assume in the nutrition of man HERMANN STRAUSS Common salt plays an important part as a regulator of the osmotic processes in the human organism, whereby the latter with greatest tenacity holds fast the percentage concentration of its fluids. Man can get along on relatively small quantities ( jE^ gm. ) of " salt required by the tissues." But the majority of civilized men con- sume much greater quantities of common salt, and the principal quantity taken in food plays the part of a " seasoning salt." There- fore, the reduction, in the diet, of common salt has its limits, since dis- turbances may ensue f rom too great a reduction. Where the supply is too abundant, the excess is excreted. As a result of reduction of ingested common salt, a diminution in the secretion of gastric juice has been noted in dogs. In diseases of the stomach in men, it has been proposed, in the case of lack of hydrochloric acid in the gastric juice, to introduce copious quantities of common salt ; in the case of increase in the secretion of gastric juice, to decrease the quantity of common salt in the food. But, in practice, with such a proce- dure, it has been possible to obtain only inconstant results. I myself have pointed out, that an excretory insufficiency of the renal function may be traced to a retention of common salt. Through the retention of water, this condition favors the develop- ment of dropsy, since the principal amount of the retained salt finds 142 Biochemical Proceedings, Hygienic Congress [Sept. lodgement in the organism in the form of a " seroretention," while only a small part is deposited in the form of a " historetention." In consideration of these established opinions, for a decade, I have recommended a limitation of the supply of salt in the food, and a medicinal Stimulation of salt-elimination, in the prevention and treatment of hydronephrosis. The Situation, with regard to uncomplicated diseases of the heart (as well as incipient compensation disturbances in subjects of heart disease) is different from that in cases of parenchymatous nephritis. Also, in inflammatory discharges, and in ascites resulting from cir- rhosis of the liver, the circumstances are otherwise. In these con- ditions, the results of a deprivation of chlorin are very inconstant. For alimentation in diabetes insipidus, there have been established certain correct requirements similar to those laid down for parenchy- matous nephritics with an inclination toward dropsy. The signifi- cance of a limitation of salt as a means of lessening the thirst, in all cases in which there is a question of a decrease of fluid in the aliment, is now more highly appreciated than formerly. The question of dry retention of chlorin is now not wholly clear. At present, exact investigations as to the salt-content of the skin are lacking. Also, the relation of salt-retention to the development of uremia has not yet been fully explained. I should be inclined at this time to State only that, ccoteris parihus, uremia occurs more readily in the nephritic organism which is poor in water, than in one where water is abundant, and I may also State that, in the vom- ited matter of uremics, an extraordinary quantity of common salt is found. Through recent researches, a marked relationship between bro- min and chlorin has also been brought to light. Bromid poisoning may be success fully treated by means of an abundant supply of common salt. In complete deprivation of salt, and, likewise, in thorough lim- itation of fluids, an increase in the disintegration of protein may be noted. On the other band, an increase in the combustion of fat cannot be shown. As a rule, salt-equilibrium is restored in 24-48 hours. On the contrary, following a previously sharp decrease in the supply of salt, it requires several days for the restoration of salt- 1912] Lafayette B. Mendel 143 equilibrium ; and, in extreme retention of salt, increased elimination may be checked for many days. It cannot be denied that many healthy persons consume too great quantities of common salt. Moderate amounts are not injurious. A certain quantity of salt, as seasoning, is permissible for civilized individuals accustomed to substances which stimulate the sense of taste. The choice of foods, with regard to disease CARL VON NOORDEN The discussion pertained to the lessons dietetlc therapy holds for US in its connection with various diseases and disease groups, and to the foods that are serviceable or a hindrance to the attainment of the end desired. Only the major groups of food substances, such as proteins, fats, carbohydrates, spices and salts were considered. 1. Obesity. Principle : Decrease in the caloric value of the food. This is best attained through a decrease or total exclusion of the supply of fat. Carbohydrates, where relatively plentiful in the diet, should be barred out. In anti-fat treatments, the amount of con- tained protein should, where possible, amount to not less than 100 grams. The supply of water must be curtailed only if the obesity is accompanied by disturbances of the circulation. 2. Forced alimentation. Principle : Increase of the caloric sup- ply over the diet for maintenance. Theoretically, it is all the same, whether the center of gravity rests upon a large supply of carbohy- drates or fat. In reality, 250 gm. of carbohydrate is seldom ex- ceeded, because most carbohydrate foods possess a very great vol- ume. The supply of protein may not ordinarily be increased be- yond 100-120 gm. By means of these, approximately 1,300 calo- ries, no satisfactory alimentative results may be obtained. The practical results depend always upon the increase in the supply of fat. In most cases, the latter may be increased to 250 or 300 gm. daily, and then increases in weight of about 2 kilos per week may be gained. 3. Gout and uric acid diatheses. Principle : Decrease in animal foods; eventually total exclusion of the same. It is useful, in the 144 Biochemical Proceedings, Hygienic Congress [Sept. case of every gouty patient, to undertake a separate "test of tolera- tion," and, from the result of this test, to establish the patient's diet 4. Diabetes mellitus. Principle : Avoidance of such foods as in- cite the organ of sugar-production, the liver cells, to increased for- mation of sugar. Every undue Stimulation of the sugar-forming organ has, as its result, not only an immediate lavish production of sugar, but also increases f or the f uture its morbid excitability ; while systematic care of the organ renders its recovery possible. There- fore, decrease, and, under some circumstances, total exclusion of the carbohydrates. Moreover, decrease of protein substances. It is essential, in every case of diabetes, to exactly determine under what dietetic regime and manner of living the least amount of superfluous sugar is formed. That order of diet is best under which the patient continues free from superfluous sugar. 5. Feverish diseases and morbus Basedowii. Principle: In both these diseased conditions there occurs an abnormal increase in caloric production. Simultaneously ensues a heightened sensibility in re- lation to the specific dynamic influence of proteins. In order to limit as far as possible the caloric production and the loss in weight, practical empiricism and theory likewise call for a scanty protein supply, while weight is gained by an ample provision of carbohy- drates. 6. Diseases of the digestive organs. Principle : A food supply which is sufficiently nourishing, while imposing as little tax as pos- sible upon the diseased organs. A discussion of the injurious effect of certain combinations of foods. Report upon enterotoxic neuritis. Attack by means of protracted pure milk diet. 7. Kidney diseases. Principle : As much rest as possible for the kidneys. The amount of the intake of those nutrient media whose products of metabolism leave the body through the kidneys, should be reduced. The proteins conie first in this regard. But this limitation should not be carried too far, since patients with chronic kidney diseases become anemic and weak if strict curtail- ment of the proteins is too long continued. Many spices irritate the kidneys, and indulgence therein must be limited ; the same is the case with regard to alcohol. Common salt and water severely tax the kidneys. I9I2] Lafayette B. Mendel i45 Final words : Warning against schematic employment of dietary precepts. An effort must be made, on the one band, to hold fast to the basic rules of nutrition-therapy, but, on the other, to duly take into account the individuality of the patient. Diet and metabolism in fever WARREN COLEMAN Empiricism has been a signal failure as a basis for the fever diet. Through studies of metabolism the Solution of this problem appears to be at band. With diets containing large amounts of carbohy- drate it is possible to bring typhoid fever patients into nitrogen equilibrium, or nearly so. The apparently excessive quantities of food required for the purpose are almost completely absorbed. The heat-production in typhoid fever, as determined by indirect calorimetry, averages about 35 calories per kilogram at absolute rest. Diets furnishing only sufficient energy to cover the heat-pro- duction do not Protect the body against nitrogen or weight loss. The explanation of this discrepancy has not yet been found. Respiratory quotients in typhoid fever below 0.65 to 0.70 appear to be due to errors of technique. The lowest quotient we obtained in the fasting state, during the febrile period, was 0.70. During the same period, patients on a füll diet gave quotients varying from 0.75 to 0.95 at short intervals after food. The quotient rises dur- ing the later stages of the fever and reaches i.o to 1.15 early in con- valescence. During a relapse, a quotient of 1.04 was obtained while the patient had a temperature of 102° F. (37.7° C). The oxygen consumption during the febrile stage varies between 4 and 6 c.c. per kilogram a minute. Compared with the amount used in the fasting stage, the oxygen consumption is not greatly increased by the quantity of food administered. The body burns carbohydrate by preference during fever as long as it is available. As indicated by a falling quotient, from 100 to 120 grams of lactose is, for the most part, consumed, or deposited in the glycogen depöts, after 4 to 5 hours. The optimum amount of carbohydrate in the fever must be determined for each patient in- dividually, but is always large. The optimum amounts of fat and protein have not yet been determined. 146 Biochcmical Proceedings, Hygienic Congress [Sept. A consideration of the unknown factors in the ill-effects of bad Ventilation YANDELL HENDERSON The facts regarding Ventilation present an extraordinary contra- diction. Fresh air, sunlight and dry cool climates exert a decidedly beneficial effect upon health. Ill-ventilated dwellings decrease vitality. In some persons under certain conditions even a few min- utes in a crowded room may produce acute ill-effects. As to how these effects are produced physiology has up to the present time afforded no satisfactory explanation. The evidence is almost en- tirely negative. The ill-effects of bad Ventilation can not be due to lack of oxygen. It is probable that they are not due in any con- siderable degree to excess of CO2. The idea that they are due to some poisonous substance contained in the expired air has in recent years been regarded as untenable. Recently this conception has been revived in a novel form by the brilliant work of Rosenau. Even Rosenau's investigations do not appear, however, to afford the Solu- tion of this problem. The recent investigation of Hill in England and of Flügge and his pupils in Germany makes it highly probable that the effects of fresh or vitiated air are brought about not by a direct action upon the lungs but indirectly through the skin. It appears probable that the temperature and moisture of the air sur- rounding the body are the essential Clements. According to the explanation to be suggested in this paper the condition of the skin exerts a potent influence upon the lungs. This may be in part a vaso-motor reflex acting upon the pulmonary circu- lation. More probably it is a chemical or hormone influence upon certain pulmonary processes. The evidence accumulated during recent years indicates that the lungs are not mere passive organs through which gases diffuse as through non-living membranes. The investigations of Bohr, of Haidane and his co-workers and of the recent Pikes Peak expedition all tend to indicate that the lungs are the seat of vital activities of great importance to health. Thus under certain conditions the lungs secrete oxygen into the blood, and it appears that considerable oxidation may take place in the blood during its passage through the pulmonary vessels. The evidence I9I2] Lafayette B. Mendel 147 available, although still far from complete, suggests that these pul- monary activities are indirectly but powerfiilly influenced through conditions affecting the skin, and that it is in this manner that Venti- lation influences health. The hygienic physiology of work in compressed air J. J. R. MACLEOD Although it is now a well-established fact that the Symptoms of Caisson disease and diver's palsy are due to the sudden liberation o£ bubbles of nitrogen in the blood and tissue fluids, on account of too sudden decompression, there are several peculiarities regarding the conditions which influence the safety of decompression about which there is still a certain degree of uncertainty. This is the case more particularly with regard to : ( i ) Whether the decompression should be uniform or in stages; (2) how long it should take in proportion to the time of the shift and the pressure employed; (3) the degree to which the breathing of oxygen increases the safety of decompres- sion. Although, as insisted on by Haidane and others, it is no doubt the case that " the absolute air pressure can always be reduced to half the absolute pressure at which the tissues are saturated without risk " yet, in practice, it has not been found that the method is in any way superior to that of gradual decompression. The time that should be taken in decompression depends on the length of the shift in the caisson, because the Saturation of the re- moter parts of the body with nitrogen continues for a long time after this has been attained in the blood and the more accessible tis- sues. Tables indicating what time should be allowed have been prepared by Haidane and by Japp. The advantages of breathing oxygen are not only that it accel- erates the diffusion of nitrogen out of the lungs and, therefore, out of the blood; but, if Symptoms have already appeared, it supplies enough oxygen to keep life going when the circulation is danger- ously obstructed by nitrogen bubbles. In using oxygen at higher pressures, its toxic action must however be kept in mind. Recompression, either by placing the caisson worker in a pres- sure Chamber or by having the diver descend again to a certain depth 148 Biochemical Proceedings, Hygienic Congress [Sept. whenever the first Symptoms appear, is by far the most efficient treat- ment, as both experiment and the experience of engineers testify. On account of the heat and the high relative humidity of com- pressed air, the worker in a caisson is under conditions which tend to lower his efficiency. Not only this, but his appetite is likely to suf- fer and his general condition after some time to deteriorate so that he becomes liable to infections if not to caisson disease itself. The Caissons should therefore be well ventilated and the wet bulb ther- mometer kept as low as possible. Means for doing this were dis- ciissed. In the choice of men for caisson work attention should be paid to age, body weight and f atness and while engaged in the work the men should be kept in good training. Certain aspects of the influenae of muscular exercise upon the respiratory System THEODORE HOUGH Muscular activity increases the respiratory exchange from three- to tenfold, thus making demands on the System comparable only with those of the more severe forms of dyspnea. In meeting the respiratory needs of the tissues there are secondary effects of hy- gienic importance, such as the increased aspiration of the thora'x upon the return of venous blood to the heart and also upon the flow of lymph in the larger lymphatics ; this increased lymph flow is feit in the interstitial Spaces of every organ in the body, thus favorably influencing the environment of every cell. The introduction with more vigorous exercise of physiological strain makes it important to inquire into the exact condition of the organism revealed by the accompanying respiratory phenomena. Of the conditions known to increase the work of the respiratory Center Geppert and Zuntz have excluded, as exciting causes of the in- creased breathing movements of muscular activity, afferent Impulses from the working muscles and deficiency of oxygen in the arterial blood; their work also shows a decrease of the total {i. c, free and combined) carbon dioxid of the arterial blood; it does not establish a diminished tension of this gas in the respiratory center, and it is possible (Haidane) that there may be increased tension of this gas in igi2] Lafayette B. Mendel I49 the Center along with a fall of the total amount in the blood. No direct determinations of the condition of the blood in this respect have been made. Determinations of the alveolar tensions of oxygen and carbon dioxid during and at varying periods after muscular activity show that with increasing intensity of work there is first a rise of CO2 tension, then a fall to and below normal. In the latter case the CO2 tension sinks still further after the cessation of the exercise and may remain subnormal for over half an hour or even an hour. Review of the evidence as a whole leads to the conclusion that during more vigorous exercise the main cause of the increase of breathing movements is some catabolite (other than CO2) of the working muscle. During moderate exercise the increase of CO2 tension of the blood is probably an adequate explanation ; the respi- ratory condition of the organism would thus differ not only in degree but also in kind with moderate and with more vigorous exercise. Theory that the distress which is relieved by " second wind " is due to excessive CO2 tension not well established by the evidence at band, but worthy of further study. Bearing upon this question is the effect of previous Inhalation of oxygen in lessening the distress of maximal effort. Should not the measurement of respiratory power in physical examination be extended so as to include not only the anatomic features of ehest expansion and vital capacity, but also the ability of the respiratory System to meet successfully the conditions of the more vigorous forms of muscular activity? Yale University, New Haven, Conn. MEETINGS OF THE SECTION ON BIOCHEM- ISTRY, INCLUDING PHARMACOLOGY (VIII, D), OF THE EIGHTH INTERNATIONAL CON- GRESS OF APPLIED CHEMISTRY Proceedings reported by THE Secretary, JOHN A. MANDEL I. LIST OF OFFICERS OF THE BIOCHEMICAL SECTION President, John J. Abel; Vice President, William J. Gies; Secre- tary, John A. Mandel; Executive Committee: Reid Hunt, Thomas B. Osborne and the officers. IL SECTIONAL PROGRAM^ The meetings of the Section were held on September 6 to Sep- tember 12, inclusive, at Columbia University, in Room 301 of Havemeyer Hall. The morning sessions were opened at 10 o'clock and the afternoon sessions at i o'clock. Friday morning, September 6. In the chair: The Vice President. Julius Stoklasa: Ueber die photochemische Synthese der Kohlenhydrate unter Einwirkung der ultravioletten Strahlen. — L. Marchlezvski: The present State of our knowledge of the relation- ship of the chemistry of the blood coloring matter and Chlorophyll. — L. Marchlezvski and C. A. Jacobson: On the quality of Chlorophyll and the variable ratio of the two constituents, and on methods for determining this ratio. — *Guido M. Piccinini: II manganese del punto di vista delle funzioni enzimatiche. — ^ Jules Wolff: Sur la re- sistance de la Peroxydase ä l'ammoniaque et sur son activation par contact avec l'alcali. — *Jides Wolff: Sur une nouvelle fonction du catalyseur dit " Peroxydase" et sur le transformation biochemique de l'orcine en orceine (page 53). — Walter Jones: Some new phases *The asterisks indicate the papers which were actually presented. Some of the titles were received after the official program had been printed and are included here informally. Abstracts of most of the papers were published in Volume 19 of the preliminary report of the proceedings of the Congress. 150 1912] ■ John A. Mandel 151 of the nuclein fermentation. — Carl Voegtlin: Further studies in biologic oxidations. — "^Walter R. Bloor: Fatty acid esters of glucose. — C. C. Guthrie: A comparative study of the action of Solu- tions on the preservation of the vitality of tissues. — *R. Delaimay and O. Bailly: Les pepsines fluides etude du sediment qui se produit dans certaines d'entre elles. — William J. Gies: Modified collodion membranes, with demonstrations. Saturday morning, September 7. In the chair: Prof. Mauthner, of Vienna. ^Gabriel Bertrand and F. Medigreceanu: Sur la presence normale du manganese chez les animaux. — *M, Lin- det: Sur les elements mineraux de la caseine du lait. — *P. Malvezin: La question de l'acide sulfureux dans les vins blancs. — *Z. Mimu- roto: Ueber das Vorkommen von Adenin und Asparaginsäure in Maulbeerblättern. — U. Suzuki and 5". Matsunaga: Ueber das Vor- kommen von Nikotinsäure (m-Pyridinkarbonsäure) in der Reis- kleie.— Zozo Sakaguchi: Ueber den Fettgehalt des normalen und pathologischen Harns. — W. N. Berg: Effect of sodium chlorid and cold storage upon the activities of proteolytic enzymes. — *Thonias B. Aldrich: The iodine content of the small, the medium and the large thyroid glands of beef, sheep and hogs. — *Lezvis W. Fetzer: The chemical changes taking place in milk under pathological con- ditions. — *Ma.Y Kahn: A study of the chemistry of renal calculi, Monday morning, September 9. In the chair : The Pres- ident. *M. Nicloux: Moyen de caracteriser de petites quantites d'alcool methylique dans le sang et dans les tissus. — Zennoshin Hatta: Zur Kritik der Zuckerbestimmungsmethode nach Ivar Bang. — Munemichi Taniura: Zur Prüfung der Kumagawa-Sutoschen Fett- bestimmungsmethode in Bezug auf die Oxydation der fettsäuren und unverseif baren Substanzen im Verlaufe des Verfahrens. — Yuji Sueyoski: Eine neue approximative Eiweissbestimmungsmethode bei Albuminurie. — "^ Franz Herles: Schnelles Verfahren zur Bestim- mung der Harnsäure im Harn. — W. Worth Haie and Atherton Seidell: The comparative estimation of epinephrin in suprarenal glands and in its Solutions, physiologically and by color tests. — Lyman B. Stookey: The Cammidge reaction. — ^Herbert H. Bunzel: Oxidase determinations. — /. P. Atkinson: On the Separation of cer- tain alkaloids from nerve tissue. — *W. H. Schidtz and Atherton 152 Biochcmical Procccdings, Chemical Congrcss [Sept. Scidcll: The determination of thymol in dog feces. — "^Shiro Tashiro: A new apparatus for the detection and estimation of exceedingly minute quantities of carbon dioxide in biological materials. — "^Shiro Tashiro: Carbon dioxide production in the nerve fibre during an excitation. Its apphcation for detection of life in protoplasm. — *F. Klein: Die selenige Säure — ihr Verhalten gegen Eiweiss und tierische Haut. — ^Gabriel Bertrand and H. Agulhon: Sur la presence normale du bore chez les animaux. Monday afternoon, September 9. In the chair : The Pres- ident. "^ Felix Ehrlich: Ueber einige chemische Reaktionen der Mikroorganismen und ihre Bedeutung für chemische und biologische Probleme. — ^Gilbert T. Morgan and E. Ashley Cooper: The influ- ence of the chemical Constitution of certain organic hydroxyl and aminic derivatives on their germicidal power. — Takaoki Sasaki: Ueber den Abbau einiger Polypeptide durch Bakterien. II. Unter- suchungen mit nicht verflüssigenden Bakterien. — Naganiichi Shi- bata: Zur Frage der Fettzersetzung durch einige Saprophyten. — *A. Trillat: Influence des impuretes gazeuses de l'air sur la vitalite des microbes. — *M. Javillier: Influence exercee par le zinc sur Vas- pergilliis niger au point de vue de l'utilisative par la plante. — *Car/ L. Aisberg and O. F. Black: Biochemical and toxicological studies upon Penicillium stolonifernm. Tuesday morning, September 10. In the chair : The Pres- ident. *M. Mane: Relations de la plante avec les Clements fertili- sants; loi du minimum et loi des rapports physiologiques. — *M. Gerber: Etüde comparee des pressures des Vamanite phalloide et de Vamadoiivier. — *i?. Dubois: Sur l'atmolyse et sur l'atmolyseur. — */?. Dubois: Recherches sur les vacuolides de la purpurase. — *i?. Dubois: La biophotogenese reduite a une action zymasique. — Oszvald Schreiner: The physiological röle of organic constituents in plant metabolism. — *C F. Langworthy : The study of problems of vegetable physiology by means of the respiration calorimeter; a progress report. — ^Hozvard S. Reed: The enzyme activities in- volved in certain plant diseases. — ^Ernest D. Clark: Origin and sig- nificance of starch. — William J. Gies: Studies of diffusion, with demonstrations. Tuesday afternoon, September 10. In the chair.- The jgi2] John A. Mandel 153 President. *P. Carles: Les phosphates et le son de froment dans ralimentation animale. — *P. Carles: Entretien du tissu dentaire par une alimentation appropriee. — Minoru Maeda: Versuche über die Ausnutzung von " Konnyak " (einer japanischen Speise). — *PanlE. Hozve and Philip B. Hawk: The utilization of various protein foods by man after repeated fasting. — *L. F. F oster and Philip B. Hawk: A study of the utilization of ingested food when undermas- ticated ("bolted") and overmasticated ("fletcherized"). — S. P. Beebe: The influence of the thyroid on the excretion of ammonia. — ^Andrew Hunter and Maurice H. Givens: Purin metaboHsm in the monkey. — William Salant and /. B. Rieger: The influence of alcohol on protein metabolism. — Jacob Rosenbloom: Chemical and pharma- cological studies of human duodenal contents. Wednesday morning, September 11. In the chair: The President. *M, Sauton: Nutrition minerale du bacille tubercu- leux. — * Walter J. Dilling: Charts of spectra representing visible and invisible bands of various hemoglobin derivatives, with explana- tory booklet. — *G. O. Higley: Some notes on the form of the curve of carbon dioxide excretion resulting from muscular work follow- ing forced breathing. — *G. 0. Higley: The influence of barometric pressure on the carbon dioxide excretion in man. — *Joseph L. Miller and Dean D. I.ezvis: Physiological action of the various anatomical components of the hypophysis. — Isaac Levin: Immunity and specific therapy in experimental Cancer. — Lafayette B. Mendel: The physio- logical behavior of lipoid-soluble dyes. — *5. B. Crohn: Experiences with duodenal and stool ferments in health and disease. — Hertnan M. Adler: Experimental production of lesions resembling pellagra. — William J. des: Studies of edema, with demonstrations. Wednesday afternoon, September 11. In the chair: The President. George W. Crile: Neuro-cytological changes resulting from the administration of certain drugs. — *G. A. Menge: Some new Compounds of the cholin type. — Reid Hunt: Physiological action of some new Compounds of the cholin type. — Arthur S. Loev- enhart: Further observations on the action of oxidizing substances. — W. H. Schultz: Pharmacological action of proteins and some of their derivatives. — */^. H. Schultz and Atherton Seidell: Subcu- taneous absorption of thymol from oils. 154 Biochemical Proceedings, Chemical Congress [Sept. Thursday morning, September 12. In the chair: The President. */. M. Fortesciie-Brickdale: The arylarsonates : their pharmacology considered from the experimental and practical stand- points. — *//. A. D. Jowett, V. F. L. Pyman and V. F. G. P. Remfry: The relation between chemical Constitution and physiological action, as exempHfied by the glyoxahnes, isoquinolines and acid amides. — Walther Straub: Pharmakologische Bedeutung der Zellmembranen. — Charles Baskerville: Inhalation anesthetics. — "^Thomas B. Al- drich: On feeding young white rats the anterior and posterior parts of the pituitary gland. — /. A. E. Eyster: The relation of calcium to the inhibitory mechanism of the heart. — Clyde Brooks: On the action of alcohol on the circulation. Thursday afternoon, September 12. In the chair: The President. Giovanni Bufalini: Reazioni caratteristiche del veleno del rospo (Bufo vulgaris.) — Giovanni Bufalini: Meccanismo dell' azione narcotici del chloridrine. — *E. Fonrneau and V. K. Ochslin: Chlorure de l'acide dichloroarsinobenzoique ; ethers des acides benz- arsineux et benzarsinique. — *C R. Marshall: The pharmacological action of brom-strychnins. — *C R. Marshall: The influence of hy- droxyl and carboxyl groups on the pharmacological action of nitric esters. — Isaac Adler: Studies on chronic adrenalin, lead and nicotin intoxications. — *Ivo Novi: II calcio e il magnesio del cervello in varie condizioni fiziologiche e farmacologiche. — *L. Launoy: Action de quelques amines, en particulier du chlorure et de l'hydrate de tetra- methylammonium sur la secretion pancreatique. — *R. Delaunay and O. Bailly: Examen critique des conditions d'essai des pancreatines medicinales. III. ATTENDANCE Among the many in attendance at one or more sectional meet- ings, the Secretary noted the presence of the colleagues named below : John J. Abel, T. B. Aldrich, C. L. Aisberg, J. P. Atkinson, W. N. Berg, G. Bertrand (Paris), Samuel Bookman, Harold C. Bradley, H. H. Bunzel, Ernest D. Clark, F. C. Cook, F. Ehrlich (Breslau), Frank R. Eider, B. G. Feinberg, Lewis W. Fetzer, M. S. Fine, Harry L. Fisher, A. O. Gettler, Wm. J. Gies, A. J. Goldfarb, R. A. Gortner, Isidor Greenwald, M. L. Hamlin, G. A. Hanford, 1912] John A. Mandel i55 Robert A. Hatcher, Philip B. Hawk, G. O. Higley, B. Horowitz, E. M. Houghton, Paul E. Howe, Reid Hunt, Max Kahn, F. Klein, P. A. Kober, W. M. Kraus, P. A. Levene, Isaac Levin, Alfred P. Loth- rop, Wm. G. Lyle, John A. Mandel, Samuel Matthews, T. Mauthner (Vienna), F. Medigreceanu, G. M. Meyer, Jas. L, Miller, G. T. Morgan (Dublin), Max Morse, Victor C. Myers, W. A. Pearson, F. B. Power (London), Howard S. Reed, A. I. Ringer, C. J. Rob- inson, Anton R. Rose, Jacob Rosenbloom, William Salant, Emily C. Seaman, Atherton Seidell, B. Setlik (Prague), H. C. Sherman, Torald Sollman, Matthew Steel, M. X. Sullivan, Shiro Tashiro, Rodney H. True, H, Vieth (Ludwigshavn), Charles Weisman, Louis E. Wise. University and Bellevue Hospital Medical College, New York City. SIXTH SCIENTIFIC MEETING OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION, AT THE COLLEGE OF PHYSICIANS AND SUR- GEONS, NEW YORK, JUNE 3, 1912 PrOCEEDINGS RePORTED BY THE Secretary, ALFRED P. LOTHROP The sixth scientific session (third "annual" meeting) of the Columbia University Biochemical Association was held at the Co- lumbia Medical School on the evening of June 3, 191 2. The execu- tive proceedings of this session were published on pages 570-573 of Volume I of the Biochemical Bulletin (June number). The scientific proceedings consisted of research Communications by members of the Association. Abstracts of the papers are pre- sented here (pages 158-187) in two groups: (I) Abstracts of papers on research by non-resident members^ and (II) abstracts of papers from the Columbia Biochemical Department and affiliated laboratories. The appended summary will faciliate reference tp the abstracts (1-44). A SUMMARY OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE SUCCEEDING ABSTRACTS I Allan C. Eustis. On the physio- WiLLiAM N. Berg. The physico- logical action of some of the amins Chemical basis of striated-muscle produced by intestinal putrefaction. contraction. (i) (s) William N. Berg, with L. A. Rogers, Allan C. Eustis. Solubilities and C. R. Potteiger and B. J. Davis. action of ^-imidazolylethylamin and Factors influencing the flavors of the relation to asthma and anaphy- storage butter. (2) laxis. (6) Isabel Bevier, for Anna W. Wil- A. J. Goldfarb. On the production of liams. A study of ropy bread. (3) grafted multiple embryos. (7) Allan C. Eustis. On the toxicity of Max Morse. Non-toxicity of inor- guinea pig urine and its relation to ganic colloid Solutions upon protozoa. anaphylaxis. .(4) (8) ^ Members of the Association who were not officially connected with the Columbia biochemical department when the research was conducted. 156 I9I2] Alfred P. Lothrop 157 Max Morse, for L. B. Ripley. Larvae of Lepidoptera obtained with sul- furic acid. (9) Anton Richard Rose. A study of the metabolism and physiological effects of certain phosphorus Com- pounds in milk cows. (10) II David Alperin. Contribution to the knowledge of nucleoprotein metab- olism, with special reference to uri- colysis and to the properties of uricase. (11) George D. Beal and George A. Geiger. The comparative diffusibility of vari- ous pigments in different solvents. (12) Stanley R. Benedict. The occur- rence and estimation of Creatinin in urine. (13) Louis E. Bisch. An endeavor to pre- pare Phrenosin from protagon. (14) Louis E. Bisch. Mucoid-silver prod- ucts. (15) Sidney Born. Protein-copper prod- ucts. (16) J. J. Bronfenbrenner and Hideyo NoGUCHi. A biochemical study of the phenomena known as comple- ment Splitting. (17) Ernest D. Clark. Notes on the chemical natura of Lloyd's " tannin mass." (18) Walter H. Eddy. A study of some protein Compounds. (19) Walter H. Eddy. The preparation of thymus histon. (20) Frank R. Elder and William J. Gies. The influence of proteases on the swelling of collagen and fibrin par- ticles in alkalin and acid media con- taining a biological electrolyte. (21) William J. Gies. A convenient form of apparatus for demonstrations of osmotic pressure exerted by lipins. (22) William J. Gies. Some interesting properties of thymol. (23) William J. Gies. A convenient method of preparing starch that swells rapidly in water. (24) R. f. Hare. A study of the carbo- hydrates of the prickly pear and its fruits. (25) Henry H. Janeway and William H. Welker. The relation of acapnia to shock. (26) Max Kahn. Biochemical studies of sulfocyanate. (27) Max Kahn. The chemical Constitu- tion of renal calculi. (28) Max Kahn and Jacob Rosenbloom. The colloidal nitrogen in urine from a dog with a tumor of the breast. (29) Max Kahn and Frederic G. Good- ridge. A non-protein, colloidal, ni- trogenous substance in milk. (30) John L. Kantor. A biochemical test for free acid, with a review of the methods for estimating the various factors in gastric acidity. (31) Marguerite T. Lee. A study of modi- fications of the biuret reagent. (32) Alfred P. Lothrop. A chemical study of salivary mucin. (33) C. A. Mathewson. A study of some of the more important biochemical tests. (34) Jacob Rosenbloom. A quantitative study of the lipins of bile obtained from a patient with a biliary fistula. (35) Jacob Rosenbloom and William Weinberger. Effects of intraperi- toneal injectionsof epinephrin on the partition of nitrogen in urine from a dog. (36) Oscar M. Schloss. A case of allergy to common foods. (37) Carl A. Schwarze. The comparative enzyme content of green and varie- gated leaves of Tradescantia. (38) 158 Proceedings CGlumhia Biochcmical Association [Sept. Emily C. SE.^MAN. Biochemical studies of beryllium sulfate. (39) Clayton S. Smith. Chemical changes in fish during long periods of cold storage (40) William Weinberger. An attempt to sharpen the end point in Benedict's method for the quantitative deter- mination of sugar in urine. (41) William H. Welker. Diffusibility of protein through rubber merhbranes, with a note on the disintegration of collodion membranes by common ethyl ether and other solvents. (42) Charles Weisman. A further study of the Bardach test for protein. (43) Harold E. Woodward. A study of the surface tension of dog blood-serum by the drop-weight method. (44) I. ABSTRACTS OF PAPERS ON RESEARCH BY NON- RESIDENT MEMBERS^ 1. The physico-chemical basis of striated-muscle contrac- tion. William N. Berg. {Washington, D. C). Part I was published in the June issue of the Biochemical Bulletin; part II is presentcd in this issue} 2. Factors influencing the flavors of storage butter. Wil- liam N. Berg, with L. A. Rogers, C. R. Potteiger, and B, J. Davis. {Dairy Division Research Laboratories, Bureau of Aninial Industry, Washington, D. C.) The official government bulletin on this subject is in press. 3. A study of ropy bread. Isabel Bevier, for Anna W. Williams. {Research Laboratory, Department of Household Sci- ence, University of Illinois, Urbana, III.) Published in fidl in the June issue'^ of the Biochemical Bulletin. 4. On the toxicity of guinea pig urine and its relation to anaphylaxis. Allan C. Eustis. {Laboratory of Clinical Med- iane, Department of Nutrition, Tidane University, New Orleans, La.) The nrine of guinea pigs, fed on Kohlrabi or cabbage, con- tains a great excess of indican, which readily oxidizes to indigo. Such urine also contains excess of putrefactive amins. Tests for /8-imidazolylethylamin, as well as efforts to isolate it, have been negative. Experiments on fifteen guinea pigs weighing 300 grams each, with different specimens of guinea pig urine, indicate that 1.5 c.c. constitutes a lethal dose when injected intravenously. In these ^ Members of the Association who were not officially connected with the Columbia biochemical department when the research was conducted. ^Berg: Biochemical Bulletin, 1912, i, pp. 535-7; ii, pp. loi-io. * Williams : Biochemical Bulletin, 1912, i, pp. 529-534. I9I2] Alfred P. Lothrop i59 animals, the Symptoms were identical with those observed after in- jections of /8-imidazolylethylamin. There was no delay in coagu- lation of the blood, but there was marked lowering of blood pres- sure and lowering of body temperature. After intravenous injections of filtered giiinea pig iirine into three dogs, Symptoms resembUng those of anaphylactic shock were exhibited, but the fall in blood pressure was not constant as it is after anaphylactic shock, and there was no delay in the coagulation of the blood. There is evidently some relation between the occur- rence of putrefactive amins and anaphylactic shock, but the writer's results do not bear out Pfeiffer's opinion regarding that relation. 5. On the physiological action of some of the amins produced by intestinal putref action. Allan C. Eustis. (Laboratory of Clinical Medicine, Department of Nutrition, Tulane University, New Orleans, La.) Putrescin (tetramethylendiamin) and cada- verin (pentamethylendiamin), in doses as small as o.i mg., are in- stantly fatal when injected intravenously into guinea pigs. Non- fatal doses produce marked lowering of blood pressure, dyspnea from edem.a of the lungs, salivation and prostration. The pulse is quickened. Phenylethylamin is immediately fatal to a guinea pig weighing 300 gm. when 0.05 gram is injected intravenously; 0.03 gram was fatal in two minutes when injected intravenously into a 300 gram guinea pig, with immediate prostration and paralysis of the respira- tory center ; 0.02 gm. produced a distinct chill in a 300 gram guinea pig, followed by prostration but with ultimate recovery. ß-imidasolylethylamin, in doses of o.oi gram intravenously, caused death in three minutes with typical anaphylactic Symptoms, the animals dying in attacks of forcible inspiratory effort, the heart continuing to beat after the respiration had ceased. Parahydroxyethylamin, as well as isoamylamin, produced marked rise in blood pressure. 6. Solubilities and action of /?-imidazolylethylamin and the relation to asthma and anaphylaxis. Allan C. Eustis. (Lab- oratory of Clinical Medicine, Department of Nutrition, Tiüane University, New Orleans, La.) I. A specimen of chemically pure )S-imidazolylethylamin, obtained through the courtesy of Dr. Dale i6o Proccedings Columbia Biochemical Association [Sept of the research laboratory of Burroughs, Welcome & Co., was in- soliible in cold Chloroform, benzene, tolnene, amyl alcohol, but slightly soluble in xylol, easily soluble in methyl alcohol, and soluble in cold carbon disulfide and hot amyl alcohol. Aqneous Solutions were tested with several reagents, to dis- cover if possible some means of detecting the presence of /?-imida- zolylethylamin in the tissties or blood, as follows: Bromine water, no precipitate, no coloration; copper sulfate, negative; potassium ferrocyanid, negative; Paidy's reagent, cherry red coloration; pic- ric acid, yellow precipitate insoluble in water, alcohol, ether, xylol and toluene, but which gave the positive Pauly reaction; phospho- tungstic acid, gray-blue precipitate, soluble in barium chloride Solu- tion, and in barium hydroxid Solution, which gave a positive Pauly reaction; sodiiim nitrite, negative; magnesium sulfate, negative; niercuric chlorid, negative ; gold chlorid, negative. Efforts to detect, by microchemical means, the presence of ß-\m\- dazolylethylamin in the bronchioles of guinea pigs dying from ana- phylactic shock, were without results. IL Tests of the physiological action of /J-imidazolylethylamin were conducted upon rabbits, guinea pigs and dogs by intravenous, subcutaneous and intraperitoneal injections. Intravenous injections of 0.5 mg. in guinea pigs caused immediate respiratory embarräss- ment, lowered blood pressure and diminished body heat, the animal dying in six minutes from suffocation due to complete occlusion of the bronchioles ; it being impossible to either f orce air into the lungs or to withdraw air, after the contraction had become complete. The Symptoms were typical of anaphylactic shock, and the post- mortem examination revealed the presence of enormous emphysema, the heart continuing to beat long after respiration had ceased. In dogs and rabbits there was also a lowering of the blood pressure and some respiratory embarrassment, but the occlusion of the bron- chioles was not as complete as in guinea pigs. Stibctttaneons and intraperitoneal injections were much less toxic and, in some instances, were entirely negative, suggesting that the tissues are able to utilize /?-imidazolylethylamin. The writer has seen many cases of asthma relieved entirely along dietetic lines by a " low protein " diet, and empirically has I9I2] 'Alfred P. Lothrop i6i found that red meats predispose to asthmatic attacks. )8-imidazo- lylethylamin is produced in the putrefactioii of histidin, and hemo- globin yields a large percentage of histidin on decomposition. It is possible, therefore, that /3-imidazolylethylamin causes asthma. Un- like clinical asthma, however, experimental asthma produced by iß-imidazolylethylamin is not relieved by injections of epinephrin ("adrenaHn chlorid"). 7. On the production of grafted multiple embryos. A. J, GoLDFARB. (Marine Biological Lahoratory, Woods Hole, Mass., and the Department of Natural History, College of the City of New York.) Grafted multiple embryos were first successfully produced in considerable numbers by Driesch, with the eggs of either of two genera of echinoderms, namely, Echiniis and Sphaere- chinus. Though several investigators have endeavored to repeat these experiments with American echinoderms they have failed completely. By slightly modifying the Herbst-Driesch method as described below, an unusually large number of grafted multiple embryos and larvae were produced from the eggs of Arbacia punc- tidata. After removing the fertilization membranes, the eggs were placed either directly into a sodium hydroxid Solution, or first placed in calcium-free sea water, then in an alkaline liquid of the following composition: 4 to 20 drops of 0.5 per cent. sodium hy- droxid Solution in 200 c.c. of sea water. This treatment sufficed in Driesch's experiments with Echinus and Sphaerechinus, giving rise to about 4 per cent. of agglutinated and fused embryos. For Arbacia eggs it was necessary to Supplement this treat- ment by centrifuging the eggs in tubes with very narrow bores, so that the eggs whose outer surfaces had previously been gelatinized were compressed against one another. These eggs gave rise to about 40 per cent. of agglutinated and fused embryos and larvae. The multiple embryos of Arbacia, so produced, were of the same general character as those described by Driesch, such as true twins, incomplete fusions, and complete fusions of the respective embryos. 8. Non-toxicity of inorganic colloid Solutions upon pro- tozoa. Max Morse. (Boardman Laboratories, Trinity College, Hartford, Conn.) Colloidal platinum prepared by the Bredig 102 Proceedings Columbia Biochemical Association [Sept. method, in which the house current of iio volts was reduced to 70 volts by lamps in parallel and passed through glass-distilled water by means of platinum electrodes, was used as a medium in which cultures of Paramecium and other protozoa were permitted to rest. Drop-ctilture slides were also made of these cultures in hanging drops of the platinum black. In all cases there was no augmenta- tion of division-frequence or size of the organism, nor any evi- dence of toxicity. Attempts with a Solution of mastic in ether and alcohol, which gave beautiful pictures under the Dunkelf eldtheleuch- tung of Zeiss, were not clear in their results. The colloidal Solu- tion was dialyzed for seven days in a fish-bladder, which freed it from the ether and alcohol, leaving a colloidal mass with excellent brownian movement. However, there is good reason to believe that this is not toxic in any way on protozoa. No attempt was made to " ultra-filter " the colloidal Solutions, in order to study the effects of small and larger colloidal particles upon protozoa, because of the apparent indifference of the organisms to the mixed Solution. 9. Larvae of Lepidoptera obtained with sulfuric acid. Max Morse, für L. B. Ripley. (Boardjnan Laboratories, Trinity Col- lege, Hartford, Conn.) Larvae were obtained from unfertilized eggs of the moth, Cecropia, by painting them with Baker's conc. sulfuric acid (sp. g. 1.84) for from 3 to 6 seconds and immediafely washing in pure water until entirely free from the acid. They were then left to dry and to develop. Checks were made by treating one- half of the batch from a given female with the acid and leaving the other half untouched. The females had been raised and isolated, from cocoons. The typical blueing of the developing eggs could be observed in the early stages of the eggs treated with acid while the control eggs remained white. The larvae emerged sev- eral days later in the case of the artificially fertilized eggs than in those normally fertilized. The percentage of errors was low. The larvae after emerging from the eggs were fed upon wild cherry, but thus far they have not been carried to the adult stage. This is now being tried. Petrunkevitch, Tichomorow and others have succeeded in obtaining larvae from silk-worm eggs by artificial means, but Cecropia has thus far failed to yield larvae under arti- ficial conditions. Short exposure and thorough washing may be the key to the success obtained in the present case. I9I2] 'Alfred P. Lothrop 163 10. A study of the metabolism and physiological effects o£ certain phosphorus Compounds in milk cows. Anton Richard Rose. (New York Agricultural Experiment Station, Geneva, N. Y.) The phosphorus requirement of a cow, aside from the milk phosphorus, would seem from the results of this experiment to be about 26 mg. per kilo of body weight. When the phosphorus supply is less than this amount, the physiological functions are con- tinued at the expense of the phosphorus previously stored in the tissues. Storage takes place when a greater amount than that indi- cated above is ingested. When the ingested insoluble phosphorus did not exceed 14 grams per day, there was approximate regularity in the phosphorus elimination in the feces independent of the In- gestion, suggesting that all the forms of phosphorus were digested, with liberation of phosphates; also that the fixed phosphorus of the feces was entirely due to the cellular matter from the mucosa and the intestinal flora. The soluble organic phosphorus in the feces was relatively slight in quantity, even in the periods when " phytin " was fed in liberal amounts. The calcium phytate added to the washed-bran ration was not utilized as economically as the " phytin " of the whole bran, and the "phytin" of the partially washed bran also gave a lower digestion coefficient. The addition of "phytin" to the "low phosphorus" ration in- creased the potassium Output in both feces and urine. The fecal potassium dropped in quantity when the "phytin" was withheld, but the urinary potassium did not. The amount of fecal mag- nesium was constant through the several periods except in the fourth, when it seems to have been influenced by the increased in- take of calcium phytate. At the beginning of the experiment the magnesium in the urine was equal to half that in the feces, but con- tinually decreased until the mobile magnesium of the body had been largely eliminated. The calcium in the urine increased remarkably when the phosphorus intake decreased. In the calcium phytate period, the calcium increase in the feces was approximately equiva- lent to the calcium increase in the rations. In all cases the addition of organic phosphorus to the " low phos- phorus " ration was followed by a decrease in the milk flow, and the withdrawal of this phosphorus from the ration was followed by a 164 Proceedings Coliunbia Biochemical Association [Sept. larger yield of milk. The percentage of fat in the milk fluctuated regularly with the changing amount of phosphorus ingested. The response was immediate, but the quantities of milk-fat bear no con- stant ratio to the amount of phosphorus in the rations. Aside from those pertaining to the fat, there were practically no changes in the composition of the milk, not even in the percentage of phosphorus in the fat-free solid matter. The moisture relations in the problem seem significant, though the intake and outgo of water could not be accurately measured in this experiment. The margin, after allowing for the influence of temperature, leads one to suspect a large retention of water in the last two periods. Up to the sixtieth day there was no outward sign of any physio- logical disturbance, but about that time the appetite began to wane. On the seventy-seventh day the milk-flow declined rapidly and serious trouble developed. A few days later the cow was placed in a box-stall and fed alfalfa, silage and vvheat bran, which caused all signs of malnutrition to disappear in the course of a week and also increased the milk-flow. IL ABSTRACTS OF PAPERS FROM THE COLUMBIA BIOCHEMICAL DEPARTMENT AND AFFILIATED LABORATORIES 11. Contribution to the knowledge of nucleoprotein metab- olism, with special reference to uricolysis and to the properties of uricase.^ David Alperin. The author studied the relative efficiency of the Wiener, Rosell, Croftan, Wiener and Wiechowski, and Galeotti methods for the preparation of uricase, and indicated the properties of the products. Wiener and Wiechowski have sug- gested that the subcutaneous or intravenous administration of uricase preparations is an effective procedure for the eure of gout and allied diseases. The author concludes that "practical demon- stration of the efficiency of this method of treatment has not been made." 12. The comparative diffusibility of various pigments in different solvents. George D. Beal and George A. Geiger. (Piiblishcd in füll in this issue of the Biochemical Bulletin.)^ "Alperin: Dissertation, Columbia University, 1912. 'Beal and Geiger: Biochemical Bulletin, 1912, ii, pp. ydr-^. 1912] Alfred P. Lothrop 165 13. The occurrence and estimation of Creatinin in urine/ Stanley R. Benedict, l'he work contemplates a thoroiigh inves- tigation of the question as to whether the Jaffe reaction in urine is due entirely, as is usually assumed, to the form of Creatinin which is ordinarily isolated from urine, or whether other substances may not be partially responsible for the reaction. The results indicate that there are two (or more) forms of Creatinin in urine, both of which yield the Jaffe reaction and also a zinc chlorid Compound, but which differ from each other in certain specific properties. A change in the ratio between these two forms of Creatinin in the urine has been observed in certain abnormal conditions. The most marked change was noted in inanition. There is probably a third substance contributing to the Creatinin reaction of urine which is in no wise related to Creatinin, but appears to be a weak acid. The study is in progress. 14. An endeavor to prepare phrenosin from protagon.^ Louis E. Bisch. Thudichum's method^ of isolating phrenosin has apparently never been reviewed. It was assumed that this method could be applied with success directly to protagon. The author was unable to do so, however. Repetitions of each of the numerous Steps in the process, with as much as 1450 grams of protagon at a time (in faithful accord with Thudichum's description), failed to yield sufficient material with which to complete the directions. It is possible that losses, which seem to have occurred at all stages of the process, totally consumed any phrenosin that existed in the orig- inal protagon. It is Dr. Gies' Intention to study this possibility further. 15. Mucoid-silver products.^^ Louis E. Bisch. Mo'ist, a-cid- free tendomucoid, triturated with a moderate amount of moist, alkali-free silver oxid, yields a brown to black mixture which be- comes very viscid when a small volume of ammonium hydroxid So- lution is stirred into it. A mechanical excess of 10 per cent. ammonium hydroxid Solution converts the viscid mass into a brown ^ Under the auspices of the George Crocker Special Research Fund. * Bisch : Dissertation (Part I), Columbia University, 1912. * Thudichum : A treatise on the chemical Constitution of the hrain, 1884, pp. 136-8. "Bisch: Dissertation (Part II), Columbia University, 1912. i66 Proceedings Columbia Biochemical Association [Sept. to black Solution, from which free alkali and free silver can be removed by dialysis. The neutral Solution thus prepared appears to contain argent-ammonium-mucoid, which may be obtained by precipitation with alcohol or by direct desiccation. The aqueous Solutions of these products are similar to those of argyrol in many respects yet appear to keep indefinitely. The purified material is antiseptic, and retards the growth of plants, but is seemingly non- irritant to the Cornea or other animal tissues. Fairly large quan- tities fail to induce toxic effects when injected subcutaneously or intravenously into dogs. The product in aqueous Solution is decom- posed by acidification. The purified material yields about i6 per cent. of ash. The silver content will be given special attention in the near future. i6. Protein-copper products. Sidney Born. Concentrated aqueous Solutions of various indiffusible proteins, when rendered slightly alkalin with sodium hydroxid Solution and treated with a moderate quantity of copper sulfate Solution, exhibit the typical biuret reaction in marked degree, but the excesses of alkali and copper may be removed by dialysis and, as the process continues (although no color may appear in the diffusate), the deep "biuret color" slowly changes until finally a blue or green persists. The resultant protein-copper product was isolated by precipitation of such a Solution with alcohol or by its direct desiccation. The Pro- portion of copper in six products made from edestin, gelatin, and serum protein ranged from 4.2 to 6.3 per cent. Injected subcuta- neously into frogs, the edestin and gelatin products (1.3 c.c. of concentrated aqueous Solution in each case) caused death in three hours. The properties of the dialyzed Solutions and the products therefrom will be described in some detail later. 17. A biochemical study of the phenomena known as com- plement Splitting. J. J. Bronfenbrenner and Hideyo Nogu- CHi.^^ A. It is generally accepted that complement may be split into a mid-piece and an end-piece. The mid-piece is thought to be in the globulin fraction, and the end-piece in the albumin fraction. 11 Bronfenbrenner: Dissertation, Columbia, 1912; Bronfenbrenner and Noguchi : Journal of Experimental Mediane, 1912, xv, 598-643. Most of the work was conducted at the Rockefeller Institute for Medical Research. I9I2] 'Alfred P. Lothrop 167 The restoration of complement activity by putting together the albu- min and globulin fractions does not prove, however, that each fraction contained a part of the complement, for the albumin fraction can be reactivated in the absence of the globulin fraction. Complement-splitting as brought about by hydrochloric acid, carbon dioxid, and dialysis, is really an inactivation of the whole complement by certain acids or alkalis, either added in the free State to the serum, or liberated as a result of the dissociation of certain electrolytes. That the whole complement, and not a part only, is present in the albumin fraction of the serum can be demonstrated by the re- moval of the inhibitory action of the acid or alkali. This can be effected by the addition, not only of alkali or acid, but also of any amphoteric substance. When hydrochloric acid, carbon dioxid, or dialysis are employed to produce the phenomenon known as com- plement Splitting, the complement is merely inactivated, not split. B. Thus far, most investigators have made but little distinction between the Splitting phenomenon obtained by chemical interference and that which takes place in the biological phenomenon known as complement fixation. In this study we have shown that these two sets of phenomena exhibit certain fundamental differences and that the so-called complement Splitting by physical conditions leading to chemical interaction, or directly by chemical means, is not a real Splitting of the complement, but an inactivation of the active prin- ciple of complement through an alteration in the reaction of the medium caused by an excess of either anions or cations. The modi- fication of the reaction of the medium may cause a more or less definite combination of the complement with the free ions, but the latter can readily be removed by an appropriate number of opposite ions, and render the complement active once more. The fluids that have hitherto been regarded as containing the end-piece of com- plement, contain, as a matter of fact, the whole complement tem- porarily deprived of its activity by certain ions derived either from the Salt constituents of the serum itself under a modified physical condition (dialysis against water or dilution with water) or intro- duced in the form of dissociable electrolytes. On the other hand, the Splitting of complement in the fixation i68 Proccedings Columbia Biochemical Association [Sept. reaction seems far more complicated than that caused by the phys- ical or chemical procedures. The supernatant fluid from the fixa- tion test differs from all the other end-pieces prepared by chemical methods in being active upoii persensitized sheep corpuscles only (not upon human corpuscles). The addition of various mid-pieces, obtained by different methods, to sensitized sheep corpuscles does not render the Wassermann supernatant fluid active. It is quite remarkable that the persensitized sheep corpuscles are, on the other hand, easily attacked, not only by the supernatant fluids of fixation tests, but also equally well by the other end-pieces. It is not at all improbable that in the fixation reaction, where so many factors come into play, there is a most complicated physical as well as chemical interaction leading to such an entangled mixture of factors that a substance carrying one set of ions alone cannot reverse the activity of complement, and hence the reversion takes place only when cer- tain electrolytes with both ions are employed. At all events there seems to be no doubt that the inactivation of complement is far more complicated in the Wassermann reaction or the Bordet-Gen- gou phenomenon than in the inactivation by physical or chemical means. Nevertheless, no one has as yet proved conclusively that the supernatant fluid of a fixation test necessarily contains the end- piece of complement. i8. Notes on the chemical nature of Lloyd's " tannin mass." Ernest D. Clark. Chemical studies were made upon "tannin masses" prepared by Lloyd from the fruit of the persim- mon. The original material dissolved in alkalies to form a purple jelly-like Solution. In dilute mineral acid Solutions the "masses" turned bright red in color and no swelling was observed. Upon hydrolysis with 0.2 per cent. and 2.0 per cent. hydrochloric acid Solutions, cherry-red colorations were obtained. Such Solutions contained both tannin and phloroglucin in considerable proportions. The presence of phenolic substances like vanillin was also indicated. An insoluble gelatinous substance was removed, by filtration, from the hydrolyzed acid mixture and seemed to be cellulose or a related material. Hydrolysis with 0.5 per cent. and 5.0 per cent. sodium hydroxid Solutions gave thick, dark-colored liquids and large amounts of insoluble gelatinous residue. Alkaline hydrolysis pro- igi2] 'Alfred P. Lothrop 169 duced the same kinds of materials as those that resulted from acid hydrolysis. The "tannin masses" seem to be combinations of tannin and phloroglucin associated with cellulose-like substances. With ferric chlorid, phloroglucin gives a dark blue product but not the blackish precipitate characteristic of the tannin-ferric chlorid reaction. Theref ore, " iron reagents " do not detect tannin in the presence of phloroglucin. 19. A study of some protein Compounds. Walter H. Eddy. (Published in füll in this issue of the Biochemical Bulletin. )^2 20. The preparation of thymus histon. Walter H. Eddy. As outlined by Bang, the properties of histon may be summarized as follows : Water-soluble, non-coagulable by heat, precipitated by am- monia in the presence of salts, precipitated from neutral Solution by "alkaloidal reagents," produces precipitates of several soluble pro- teins from their aqueous Solutions. The current method of preparing thymus histon, as recom- mended in Standard handbooks such as Abderhalden's and Oppen- heimer's, may be summarized as follows : Extraction of the minced glands with water. Precipitation of the water extract by acid or calcium chlorid, and extraction of this precipitate with 0.8 per cent. hydrochloric acid Solution. Precipitation of the hydrochloric acid extract with ammonium hydroxid Solution, either before or after removing free hydrochloric acid by dialysis. Washing the "am- monia precipitate " free from ammonia with alcohol and ether. Kossei, who discovered histon in goose blood, obtained it by saturating the hydrochloric acid extract with sodium chlorid. He alone calls attention to the anomaly noted in our experiments, viz., that treatment with ammonium hydroxid Solution results invariably in the precipitation of a substance that is practically insoluble in water. In a series of many preparations, extending in time over a period of two years and involving materials obtained from many calves, we have come to the conclusion that the " ammonia-pre- cipitation" of a hydrochloric acid Solution of thymus histon results invariably in a water-insoluble product. Furthermore, our experiments show that of two fractions of the same hydrochloric acid Solution, the fraction saturated with sodium chlorid invariably ^^Eddy: Biochemical Bulletin, 1912, ii, p. 111-22. I/o Proceedings Columbia Biochemical Association [Sept. yields a prodtict which (when free from sodium chlorid) is water- soluble, gives all the qualitative histon tests, and contains less nitrogen than the "ammonia precipitate" from the other fraction; the " ammonia precipitate " being water-insohtble and appar- ently a very different stibstance. Finally, when the " sodium chlorid precipitate " of histon is dissolved in vvater, and the aqueous Solution is treated with a few drops of ammonium hydroxid Solu- tion, a precipitate is produced which is insoluble in water. Quanti- tative studies now under way show marked differences in the nitro- gen content of the two products. The results suggest that " histon " as commonly prepared is an adsorption product or a salt, rather than a simple protein. The following method is suggested as a means of obtaining water-soluble histon from thymus : Mince f resh thymus glands and extract the hash with distilled water for 24 hours (best in the cold), Precipitate the aqueous extract with acetic acid Solution and ex- tract the precipitate with 0.8 per cent. hydrochloric acid Solution (after Lilienfeld) ; or add sufficient calcium chlorid to the aqueous extract to make its content of that substance 0.2 per cent. and extract the precipitate with 0.8 per cent. hydrochloric acid Solution (after Huiskamp) ; or add sufficient hydrochloric acid to the aqueous extract to make its content of the acid 0.8 per cent. and let stand 24 hours (after Kossei and Kutscher). Filter off the hydrochloric acid extract, and either remove the free acid or precipitate the histon directly by Saturation with sodium chlorid. Remove ad- mixed sodium chlorid by dialysis. Filter the resultant salt-free Solution and evaporate it to dryness at 45° C. This material, ground to a powder, may be heated to 105° C. without loss of water-solubility. 21. The influenae of proteases on the swelling of Collagen and fibrin particles in alkalin and acid media containing a bio- logical electrolyte. Frank R. Elder and William J. Gies. (Published in full in the June issue of the Biochemical Bulletin. )^^ 22. A convenient form of apparatus for demonstrations of osmotic pressure exerted by lipins. William J. Gies. The " Eider and Gies : Biochemical Bulletin, 1912, i, pp. 540-545. I9I2] 'Alfred P. Lothrop 171 writer repeated the demonstration described on page 59.^^ Instead, however, of using a thin rubber bag in a muslin sheath, he employed a i2-inch section of ordinary bunsen-burner tubing. The rubber tube had been swollen to its maximum extension by immersion in ether for about an hour previous to its use. It was then closed at one end by the Insertion of a short, tightly fitting, section of a thick glass rod, which was fastened by a ligature. After the swollen tube had been filled with olive oil and a narrow glass tube about 10 feet in length (in two sections) had been tied into the open end and held upright, the rubber-oil portion of the vertical tubulär appa- ratus was completely immersed in ether in a tall, narrow cylinder. The oil began to rise in the tube almost immediately, and rapidly proceeded upward until the liquid emerged from the open top. 23. Some interesting properties of thymol. William J. GiES. During the course of recent experiments on enzymes as pos- sible factors in the development of edema/^ we had occasion to study the effect of trypsin on elastin in ammonium hydroxid Solu- tions containing a biological electrolyte (NaCl). To our surprise we not only failed to obtain the swelling results which we had pre- viously observed under similar conditions/^ but the elastin particles in use gradually became green, ultimately blue. With repeated shaking, the elastin particles were more deeply colored, and the supernatant liquid slowly became green; finally, bluish green. The color of the particles slowly diminished in intensity as the pigment accumulated in the liquid. Unlike the elastin used in the previous experiments, this product had been prepared about 10 years before. The fresh-ligament hash had been put in water and preserved there with considerable alcoholic thymol Solution; later, had been put in alcohol; ultimately, had been dried and bottled. The main supply of the dry elastin smelled strongly of thymol. Some of the above-mentioned green and blue ammoniacal liquids, when shaken with ether or toluene, were quickly transformed into purplish, then reddish mixtures. The ether layer on the quiescent liquid was bright red — all green and blue had disappeared from the " Gies : Biochemical Bulletin, 1912, ii, p. 55. " Eider and Gies : Ibid., 1912, i, p. 540. *' Tracy and Gies : Ibid., 1912, i, p. 472. 1/2 Proceedings Columbia Biochemical Association [Sept. alkalin liquid imderneath, which was colorless. By spontaneous evaporation, the ether extract yielded a purplish-red oily product, vvith a pronounced thymol odor. When a small quantity of thymol (Kahlbaum) was mixed with lO per Cent, ammonium hydroxid Solution, the liquid became green- ish in about 2 hours ; then gradually turned blue. Alcohol appeared to accelerate the transformation. Shaken with ether, the blue was wholly removed and a beautiful, red, ether-layer obtained. Such ether extracts yielded, by spontaneous evaporation, a purplish-red oily product, which dissolved readily in ether, toluene and alcohol, the Solutions being bright red. In some cases the oily product be- came crystalline, due apparently to the presence of unchanged thy- mol ( ?). The red alcoholic Solution was turned deeply bluish by a drop of n/io sodium hydroxid Solution; the red was restored by a drop of w/io hydrochloric acid Solution. These transformations could be elicited repeatedly in the same Solution. The changes were so sharp that the material may prove to be a valuable indicator for use in the titration of alcoholic liquids. Concentrated alcoholic Solu- tions yielded reddish white precipitates when they were diluted with water — a ready means of isolating the substance. The reddish white precipitate dissolved promptly in alcohol, ether and toluene, and formed a red Solution in each case. An excess of thymol, added to a green or blue ammoniacal Solu- tion in its original condition, completely changed the green or blue to red, and wholly dissolved the red material, behaving, in this respect, like toluene and ether. These phenomena did not appear to be due to impurities in the thymol. A general survey of thymol literature has not revealed the explanation of these results, although certain inferences are sug- gested by several color reactions of thymol. The chemical nature of the colored substances derived from thymol in these preliminary experiments, the possible Utility of the products — their probable antiseptic, pharmacologic and other rela- tionships, suggest numerous interesting biochemical inquiries which will be undertaken in the near future. 24. A convenient method of preparing starch that swells rapidly in water. William J. Gies. For the purpose of study- I9I2] ^Alfred P. Lothrop 173 ing the effects of amylases on the power of starch to imbibe water (prior to hydrolytic cleavage), the writer prepared markedly hydro- phylic starch in the following way: A very thick starch paste was speedily prepared by rapidly pouring a thicl<: potato-starch Suspen- sion through musHn into boiling water while the latter was being vigorously stirred. The vessel containing the paste was inimersed in ice water immediately after the last portion of starch Suspension had been added. By constant stirring of both liquids, and by the maintenance of a low external temperature, the paste was speedily cooled,^'^ when it was poured into, and thoroughly stirred in, a large excess of 95 per cent. alcohol. After the Sedimentation of the prod- uct, and the decantation of the alcoholic liquid, the snow white ma- terial was treated with fresh portions of alcohol until its viscidity disappeared and it became firmly granulär. After several washings with ether, to remove alcohol, the product was rapidly freed from ether in a current of air from an electric fan. Although somewhat hygroscopic, the material formed hard, snow-white masses which could be granulated easily in an ordinary pulverizer. Placed in water, the particles swell very rapidly into bloated glassy forms. " Starch paste " may be made almost instantly from the product. The powder can easily be freed from its soluble car- bohydrate impurities by dialysis. The material promises to be of special Service in many connections. Mr. Nathan Rosenthal has undertaken a study of the effects of amylases on the swelling of material of this kind in various anti-hydrophylic media, such as dilute alcohol. 25. A study of the carbohydrates of the prickly pear and its fruits. R. F. Hare.^^ The difficulties encountered in the practical laboratory Separation of the sugars from the mineral matter, muci- lages, gums and dextrinoid substances have been numerous, and the Operations time-consuming. Many attempts to obtain the sugars free and in crystalline form have usually resulted unsuccessfully; so that it became necessary to make the individual tests not on the sugar crystals, but on the syrups previously purified as much as pos- sible by different methods. " The Operations were conducted rapidly in order to prevent undue hydrol- ysis. It is probable that satisfactory results can be obtained by pouring the hot paste directly into alcohol. "Hare: Dissertation, Columbia University, 191 1. 174 Proceedings Columbia Biochemical Association [Sept. The Juice of the ripe fruit contains 1.57 per cent. of pentosans and only traces of galacfan. After precipitation with lead acetate, the Juice gave the anihne acetate reaction for pentose, but none for galactose. The presence of fructose and gliicose in considerable amounts was quite definitely estabhshed by several reactions char- acteristic of these sugars. The dried mucilage of the prickly pear, when separated by pre- cipitation with alcohol from a two per cent. Solution, contained 15 per cent. of galactan, 31 per cent. of pentosan and 12 per cent. of ash. The mucilage in the aqueous extracts could not be separated completely from cell fragments, starch, crystals of calcium Oxalate and other solid particles that caused opalescence and turbidity. A dilute Solution containing 1.5 per cent. of solid matter, rendered fairly clear by repeated filtration through silk, had no effect on polarized light. This was true of all the Solutions of mucilage ob- tained in this work, both before and after subjecting them to acid hydrolysis. Harley^^ reports having found a specific rotation of + 38° for Opuntia mucilage, but places little confidence in his own results, since the reading was made on a very dilute opalescent Solu- tion and calculated from an observed rotation of + 6 minutes. Hydrolysis of the mucilage by digestion for several hours with 1,25 per cent. sulfuric acid Solution produced a sugar that had properties similar to arabinose. When its osazone was formed, oily globules rose to the surface. The precipitate was darker than glucosazone, readily soluble in hot water and melted at about 160° C. A 95 per cent. alcoholic extract of the dried stems, previously treated with ether, contained a sugar with specific rotations made on three separate Solutions of — 6.6°, — 8.25°, and — 7.1°. The osazone produced from this sugar had properties similar to those of glucosazone. These results indicate the presence of glucose and fructose^ in this extract. A 60 per cent. alcoholic extract of the dried stems contained a suhstance apparently intermediate in character hetween mucilage and sugars. It did not reduce Fehling Solution before hydrolysis, but was very readily hydrolyzed by dilute acid Solutions. Alcohol stronger than 60 per cent. reprecipitated this material as a flocculent " Harley : Journal de Pharmacie, iii, pp. 6-193. I9I2] 'Alfred P. Lothrop 17 S mass, quite different in appearance and properties from the precipi- tate of the mucilage obtained with alcohol. The precipitate was readily soluble in water, but its Solution was not mucilaginous. When hydrolyzed, it gave a plus rotation to polarized Hght. The coloring matter can be concentrated and made into a mar- ketable product, of value for coloring certain foods, by first remov- ing mucilages and gums with alcohol, and precipitating the pigment from the filtrate with acetone. The pigment is evidently a gluco- side. When separated from the juice with alcohol and acetone, and then precipitated with lead acetate, the coloring matter liberated by sulfuric acid gave a glucose-like sugar on hydrolysis. The lead salt produced by precipitating the purified pigment with lead acetate con- tains 61.42 per cent. of lead. 26. The relation of acapnia to shock.^*^ Henry H. Janeway AND William H. Welker. Henderson has published a number of papers on the relation of acapnia to shock. He maintains that a diminution of the normal amount of carbon dioxide in the blood to a sufficient degree, and maintained for a sufficient length of time, produces an irreparable disturbance of the normal balance of osmotic forces between the blood and the cytoplasm of the body cells, and that this disturbance leads to tissue asphyxia, acidosis, and fatal oligemia, accompanied by Symptoms indistinguishable from shock. He be- lieves that the essential cause of shock is acapnia. He supports this theory, not only by very thorough work on the relation of acapnia to shock from several different Standpoints, but also by furnishing control experiments, as it were, in which shock is prevented by con- servation of the animal's störe of carbon dioxide and also by suc- cessful treatment of animals, already in a condition of shock, with injections of Ringer Solution containing carbon dioxide. Whether this theory fails to stand in whole or in part, its originator deserves the greatest credit for calling attention to the possibility that härm may arise from neglect to conserve the body 's störe of carbon di- oxide, the important functions of which, in the body, have long oxide, the important functions of which, in the organism, have long been appreciated by physiologists. This theory has been of the greatest interest to one of us because of the relation of acapnia to ^ Some of the work was done in the Surgical Research Laboratory of the College of Physicians and Surgeons. 176 Procccdings Columbia Biochemical Association [Sept. artificial respiration, and to the production of shock in connection with intrathoracic surgery. It has prompted us to investigate the degree of acapnia and the associated shock produced by excessive artificial respiration. We soon found that the diminution of carbon dioxide in the blood in ordinary intrathoracic insufflation was neghgible. On the other hand, it has been quite an easy matter for us to reduce the amount of carbon dioxide in the blood to from one-third to one-half the normal amount by forced rapid inflation and deflation of the lungs. The artificial respiration was performed 45 to 90 times a minute and was continued for periods varying from 30 minutes to 3 hours. These experiments have differed from those of Hender- son in that the animals were allowed to recover. The trachea was not divided but respiration was performed by inserting a large, rather tightly fitting, tube through the larynx into the uninjured respiratory tract. The blood pressures in our experiments were not accurately measured, the animals being left as nearly normal as possible after the Operations. The degree of shock was estimated entirely from the condition of the animals after the Operation and the manner in which they recovered from it. Judged in this manner there was nothing about these animals to indicate a serious degree of shock or any greater disturbance than could be accounted for. by three other factors to which we desire to call attention in connection with these experiments and which, unless guarded against, can alone cause considerable depression and even death. ( I ) In all experiments in which excessive artificial respiration is employed there is a great reduction in the animal's body heat. The temperature can easily fall to 85° F. (2) There is a very evident possibility (which we believe to be a fact) that the rapid and complete filling of the lungs exercises a definite interference with the return of the blood to the heart. The fall of the blood pressure, as estimated with the finger, and the rapidity of the heart's actioncoin- cide closely with the pressures used to inflate the lungs; indeed, a scarcely perceptible pulse may be immediately improved by slightly lowering the latter pressures. (3) The duration of the apnea fol- lowing these experiments depends as much upon the amount of mor- phin and ether administered as upon any other factor. We do not I9I2] 'Alfred P. Lothrop i77 believe that it is possible to produce death by apnea, caused in turn by acapnia, without the assistance of the toxic effects of morphin and ether. The toxic effects of these drugs must be included as factors contributing to the shock, This report deals with only one of the phases of the relation of acapnia to shock, namely the relation of acapnia, produced by ex- cessive artificial respiration, to shock ; and as it is only a preliminary report, it is not intended as an answer to Henderson's contention. Its purpose is mainly to record two general f acts : (A) That we have reduced the amount of carbon dioxide in the blood to nearly 40 per Cent, of the normal amount, and have maintained this reduc- tion for a period of 3 hours, without producing Symptoms of shock; and (B) that there are other factors than depletion of the störe of carbon dioxid, which, unless properly guarded against, can in them- selves cause the death of the animal under experimentation. 27. Biochemical studies of sulfocyanate.^^ Max Kahn. A. The ferric chlorid colorimetric test for sulfocyanate in saliva is inexact and unreliable. A negative result by the Bunting suction method is no evidence of the absence of sulfocyanate but a positive result is suggestive of the presence of a comparatively large amount. The pink color spontaneously disappears from the ethereal layer in positive tests by the Bunting suction method. Various medicinal substances, and also certain Compounds that result from biological transformations of proteins and carbohydrates, if excreted in the saliva, give a very marked red coloration in the ferric chlorid test, similar to that produced by sulfocyanate. B. Sulfocyanate occurs in the saliva and salivary glands of man, in the salivary glands of oxen, but apparently not in the salivary glands of dogs. It occurs in the blood, but the spieen, the pancreas, the thymus, the thyroid and the testicles of dogs do not contain it. The liver seems to be the gland in the body that contains most sulfocyanate, which is also present in bile and in the small intestines. The stomach contents of dogs on an ordinary diet were free from sulfocyanate. When, however, sodium sulfid was given, the gastric mixture contained sulfocyanate. '^Kahn: Dissertation, Columbia University, 1912. Conducted under the auspices of the Dental Society of the State of New York. lyS Proceedings Columbia Biochemical Association [Sept. C. Siilfocyanate is excreted in the urine and feces. Its elimina- tion in the urine is not dependent upon the amount in the saliva. Althoiigh dog saliva is apparently always free from sulfocyanate, dog urine invariably contains it. The ingestion of amino acids (alanin) and of nitriles (acetonitrile) increases the amount of sul- focyanate in the body, as well as in the excreta. Sulfocyanate seems to be produced in the body from protein. Results with a fasting dog harmonize with this conclusion. The ingestion of sulfur, so- dium Sulfid, thioacetic acid, thiourea and taurin did not increase the Output of sulfocyanate. D. Potassium sulfocyanate is toxic to both plants and animals. Its toxicity is so marked that indiscriminate dispensation of the sub- stance to people is dangerous. The growth of molds is enhanced by potassium sulfocyanate. Yeast fermentation is not affected or is stimulated by moderate proportions of potassium sulfocyanate. Biological proportions of potassium sulfocyanate have no inhibiting influence on the growth of bacteria. The souring of milk is inhib- ited by large proportions of sulfocyanate. 28. The chemical Constitution of renal calculi. Max Kahn. Sixteen stones of nephric origin were analysed according to the method of Mackarell, Moore and Thomas. ^^ Most of the stones were composed mainly of salts of calcium. All of the stones -con- tained uric acid or urates in varying amounts, but no stone was wholly composed of urates. The shape, color and consistency of a stone are not criteria of its chemical composition. Three gouty tophi were examined by the murexid test for urates. A negative response was obtained in each case, showing that not all gouty deposits are composed of uric acid salts. 29. The colloidal nitrogen in urine from a dog with a tumor of the breast. Max Kahn and Jacob Rosenbloom. (Published in füll in this issue of the Biochemical Bulletin ).2^ 30. A non-protein, colloidal, nitrogenous substance in milk. Max Kahn and Frederic G. Goodridge. Since the figure ob- tained for " total " nitrogen in milk exceeds the sum of the values for the known nitrogenous constituents, unknown nitrogenous sub- ^ Mackarell, Moore and Thomas : Bio-Chemical Journal, 1910, iv, p. 179. ^ Kahn and Rosenbloom : Biochemical Bulletin, 1912, ii, p. 87. 1912] "Alfred P. Lothrop 179 stance must be present. The urines of man and dog contain col- loidal nitrogenous material.^* It was thought probable that such material is present in all the secretions. After a careful process, including the removal of protein without hydrolysis, substance was obtained f rom milk which is white, amor- phoiis, odorless and tasteless; insoluble in the lipin solvents, but forms in water an opalescent Solution which falls to flocculate on boiling. This material does not respond to any of the protein "color tests." It contained about 5.3 per cent. nitrogen; also car- bon, hydrogen, oxygen, and sulfur, but no loosely combined am- monia radicals. 31. A biochemical test for free acid, with a review of the methods for estimating the various factors in gastric acidity.^^ John L. Kantor. The author presented details along the lines of our original publication on this subject.^^ The test is a microscopic one and depends upon the immediate expansion of moist collagen fibrils when they are immersed in aqueous Solutions containing free organic or mineral acids of the kinds that ordinarily appear in gas- tric Contents. " Combined " acid^^ and acid salts fail to induce such effects. The test may be satisfactorily conducted with a drop of liquid and a single collagen fibril. Comparative observations indicate that for free mineral acid (HCl) the collagen-fibril test is equal in delicacy to the Töpfer and Günzberg tests, but that for free organic acid (lactic), or for mix- tures of free mineral and organic acids, it is more delicate than the latter tests. Comparative studies of common factors of interfer- ence with the several tests indicate that the collagen-fibril test ex- hibits the greater delicacy. The color of the Solution under exam- ination had no effect on the test. Further details from the clinical Standpoint, and an abstract of the historical discussion, will be pub- lished at an early date. 32. A study of modifications of the biuret reagent. Mar- GUERiTE T. Lee. This investigation was made in the endeavor to ^ Kahn and Rosenbloom : Biochemical Bulletin, 1912, ii, p. 87. ^ Kantor : Dissertation, Columbia University, 1912. ^ Kantor and Gies : Proceedings of the American Society of Biological Chemists, 1911, ii, p. 20; Journal of Biological Chemistry, 1911, ix, p. xxvi. " Goodridge and Gies : Proceedings of the Society for Experimental Biology and Mediane, 1911, viii, p. 107. i8o Proceedings Columbia Biochemical Association [Sept. discover, if possible, a more effective alkali for the biuret reagent than the Standard sodium hydroxid — or a combination of alkalies that might be better. Fairly strong Solutions o£ the following alkalies, when substi- tuted for sodium hydroxid in the biuret reagent,^^ yield Solutions that give the biuret test when they are added to dilute Solutions of Witte peptone : potassium hydroxid, ammonium hydroxid, calcium hydroxid, sodium carbonate, conin, piperidin, ethylene di-amin, tri- methyl amin, piperazin, and tetra-ethyl ammonium hydroxid. Sodium hydroxid, potassium hydroxid, ammonium hydroxid, tri- methyl amin, and tetra-ethyl ammonium hydroxid are excellent as alkalies in the biuret reagent. Tri-methyl amin appears to be more effective than sodium hydroxid. Tetra-ethyl ammonium hydroxid is seemingly as effective as sodium hydroxid when the reagent is fresh, but the efficiency of the Solution decreases on standing. Piperazin and tetra-ethyl ammonium hydroxid give most satisfac- tory tests when an excess of copper is present. There is apparently an Optimum amount of copper (sulfate) for each alkali. The study is in progress. 33. A chemical study of salivary mucin. Alfred P. Loth- ROP, Salivary mucin from the submaxillary glands of oxen.was prepared by the Hammarsten-Levene method. It is a white powder, insoluble in water, acid in reaction and readily soluble in dilute alkalin Solutions. The sodium salt can be prepared by dissolving mucin in nine parts of 0.5 per cent. sodium bicarbonate Solution plus one part of 0.5 per cent. sodium carbonate Solution. The thick Solution is then dialysed until it no longer reacts alkalin to phenolthalein but is still alkalin to litmus. (Prolonged dialysis completely hydrolyses the salt and precipitates the mucin. ) The dialysed Solution may be pre- cipitated by the addition of about six volumes of alcohol, although electrolyte (NaCl) must be present for complete flocculation. The product, washed with alcohol and ether, dries to a fine powder. ^ Gies : Proceedings of the American Society of Biological Chemists, 1910, i. P- ^7Z'> Journal of Biological Chemistry, 1910, vii, p. Ix. Also, Kantor and Gies : Biochemical Bulletin, 1912, i, p. 264. I9I2] ^Alfred P. Lothrop i8i The Salt, having an ash content of 2.7-3.3 per cent., is completely soluble in water. A 0.2 per cent. Solution is very much like a rela- tively thick natural saliva. The Solution is faintly alkalin to litmus, gives all the usual protein tests, including the Molisch test for the carbohydrate group, and is precipitated in stringy masses by acetic acid. Quantitative determinations of nitrogen and ash in mucin prep- aration III and its sodium salt gave the f ollowing typical results : Ask Nitrogen Per Cent. Found Per Cent. Calculated (Ash Free) Per Cent. Preparation III Sodium Salt III 0.28 3.27 12.49 12.20 12.53 12.61 The potassium salt was prepared in the same manner, Fre- quent reprecipitations by alcohol render the salts decreasingly solu- ble in water. These products have been made preparatory to experiments on the possible relation of salivary mucin to dental caries, in contin- uance of our studies under the auspices of the Section on Stoma- tology and Research, of the First District Dental Society, State of New York. 34. A study of some of the more important biochemical tests.^^ C. A. Mathewson. Representative substances from the following groups were studied in their influence on the tests named below: neutral inorganic salts, neutral organic Compounds, acids, acid salts, bases, basic salts, biological mixtures and miscellaneous materials. Over seventy-five substances or products were used in each case. It was found that the ten tests under examination could be arranged in the following sequence according to the percentage of factors causing interference with them: Sudan III, o; xantho- proteic, 4; Hopkins-Cole, 4; Seliwanoff, 5; MoHsch, 6.5; iodine, (for starch) 6.5 ; Fehling-Benedict, 10; biuret, 13; Millon, 22; Bar- foed, 60. The acid salts were the most potent interfering substances, the neutral organic Compounds the least potent. Of the salts, ferric chlorid was the most active agent of interference. An extension of the study is in progress. ''Mathewson: Dissertation, Columbia University, 1912. i82 Procccdings Columbia Biochemical Association [Sept. 35. A quantitative study of the lipins o£ bile obtained from a patient with a biliary fistula. Jacob Rosenbloom. Through the kindness of Dr. William Weinberger, of the Lebanon Hospital, there was placed at my disposal 3180 c.c. of human bile obtained from a patient with a biliary fistula. The fluid had the appearance of typical human bile. Its specific gravity was 1.020. The follow- ing data were obtained in a quantitative lipin analysis, the results being expressed in parts per thousand: Water, 970.2; total solids, 29.8; cholesterol, 2.61; lecithans, 6.42; fat, 6.85; fatty acids, 1.2; soaps, 2.6. Total lipins, 19.68 (1.97 per cent.). 36. Effects of intraperitoneal injections of epinephrin on the partition of nitrogen in urine from a dog. Jacob Rosenbloom AND William Weinberger. (Published in füll in this issue of the Biochemical Bulletin. )^^ 37. A case of allergy to common foods.^^ Oscar M. ScHLOSs. In a boy now 8 years old marked urticarial lesions were caused by the Ingestion of eggs, almonds and oatmeal. The idiosyn- crasy to egg was not congenital but was acquired at some time be- tween the ages of 10 days and 14 months. Symptoms due to the ingestion of oats appeared some time after the child had first eaten oatmeal when he was 22 months old. As far as can be ascertained, the idiosyncrasy to almonds was manifested the first time this food was eaten. It was found that cutaneous inoculation of these and certaln related food substances produced an urticarial wheal at the site of inoculation. The cutaneous reaction was produced only by the protein constituents of eggs, almonds and oats. Different proteins from the same source varied in activity, some being incapable of causing a reaction. Some of the active proteins caused Urticaria by mere contact with the unbroken skin. It was possible passively to sensitize guinea-pigs to ovo-mucoid (one of the active proteins from eggs) by intraperitoneal injections of the patient's blood-serum. By feeding ovo-mucoid, in gradually increasing doses, the patient became immune to egg. At the same time immunity to oatmeal and an apparently decreased susceptibility to almonds occurred. ^"Rosenbloom and Weinberger: Biochemical Bulletin, 1912, ii, p. 123. ^ Schloss : American Journal of Diseases of Children, 1912, iii, p. 341. I9I2] 'Alfred P. Lothrop 183 38. The comparative enzyme content of green and varie- gated leaves of Tradescantia.^^ Carl A. Schwarze. The re- sults of the experiments made to determine the relative enzyme con- tent of green and variegated leaves of Tradescantia show that there is a marked difference between Juices expressed from them. Etio- lated leaves are yellow in their rudimentary stage ; that is, an entirely yellow leaf presents this condition when first formed. The etio- lated leaves are free from chloroplasts and therefore possess no starch. The juice extracted from yellow leaves gives a negative Fehling test; that from green portions, a positive test. When yel- low leaves are ground in a mortar, and the juice is expressed through cheese cloth, a dark brown liquid results. Green leaves similarly treated yield a dark green liquid. Alcoholic extracts of crushed green and yellow leaves, when filtered, assume a brown color. The filtrate from yellow leaves is at first pink but the liquid gradually assumes a brown color. The filtrate from the green leaves comes through brown immediately. The juice of yellow and green leaves, when filtered, gives in both cases a brown filtrate, that from the yellow leaves being a reddish brown, When unfiltered green juice desiccates, a glossy dark green residue is deposited, at the periphery of which a few needle-shaped crystals are seen. The juice from the yellow leaves, upon desiccation, deposits a brown crystallin mass, the long crystals of which make a figure which resembles a polyaster seen in plant cells. Extracts in alcohol (80 per Cent.) deposit the greatest amount of crystals. The crys- tals from yellow leaves are darker than those from green leaves. Such reagents as guaiac and trikresol show the presence of oxidase and peroxidase in yellow and green Juices. The yellow juice seems to be richer in oxidase and peroxidase. When green juice was heated to 72° C, and tested the following day, oxidase proved to be present, that temperature having failed to destroy it. (Subjecting green juice to high temperatures results in the produc- tion of a flocculent precipitate, which Sediments promptly under a clear supernatant liquid.) Juice from yellow leaves was injected into the nodes and inter- ** Conducted in the Botanical Laboratory under Dr. Gies' guidance. 184 Proceedings Columbia Biochemical Association [Sept. nodes of healthy green Tradescantia stems. No discoloration or yellowing of the injected stem coiild be detected. 39. Biochemical studies of beryllium sulfate.^^ Emily C. Seaman. The experiments vvith beryllium sulfate have shown very conclusively that the substance has a marked effect on biochemical processes. When administered with the food it produced in dogs decided nutritive disturbances, which manifested themselves in loss of body weight, total inorganic matter, nitrogen, sulfur and phos- phorus. When large doses were administered per os the substance caused vomiting before a sufficient amount was absorbed to pro- duce any other obvious toxic Symptoms. When the calculated lethal dose was administered by a single siihcutaneous injection, the substance produced edema and necrosis of the tissue extending over a large area. No other decided Symp- toms were produced by this method. Very gradual intravenous injections of the salt produced decided toxic effect. The action of the heart became irregulär — unusually rapid and very weak: the respiration also became irregulär and shallow. During the course of the injection, there was decided tremor but this disappeared soon after the Operation. As a direct effect of the injection the temperature increased, sometimes to 105° F., but about 24 hours before the death of the animal the tem- perature began to decrease and steadily feil. After intravenous injections there was increased elimination of urine followed by re- tention. The feces became diarrheal and bloody. Vomiting began about the time the dog refused food or water. Beryllium sulfate had a decided inhibitory effect on the action of ptyalin, pepsin, and trypsin. It also retarded the action of sucrase but not to so great an extent. Solutions of the salt (i per cent. or less) did not precipitate proteins from neutral or acid Solutions. Below the concentration of M/512 Solution, beryllium sulfate did not inhibit the growth of lupin or timothy seedlings, but more con- centrated Solutions prevented growth. When present in propor- tions less than 0.5 per cent., beryllium sulfate had very little, if any, bactericidal action. 40. Chemical changes in fish during long periods of cold storage. Clayton S. Smith. Fresh fish were delivered directly ^Seaman: Dissertation, Columbia University, 1912. 1912] 'Alfred P. Lothrop 185 frort! the boat. Specimens of the same catch were immediately placed in storage and delivered to us, at intervals, in a frozen State, when they were thawed under uniform conditions and promptly subjected to analysis. Comparative data were obtained regarding moisture, organic matter, inorganic matter, and total solids; ammonia nitrogen, solu- ble nitrogen, insoluble nitrogen, coagulable nitrogen, non-coagulable nitrogen and total nitrogen; "proteose" nitrogen, both before and af ter autolysis ; f at content and f atty-acid number ; and the reducing power of the aqueous protein- free extract, as determined by the Benedict method. The flesh of fish which had been refrigerated not less than four months and not more than six months was unaltered in composition. After a period of nine months in cold storage there was a sHght, almost imperceptihle, increase in the content of ammonia nitrogen, but no other change was noted. The work is in progress. 41. An attempt to sharpen the end point in Benedict's method for the quantitative determination of sugar in urine. William Weinberger. In Benedict's modification of FehHng's sugar titration method "instead of the reduced copper being pre- cipitated as the red sub-oxid, which of its own color obscures the end point of the reaction, the copper is precipitated as cuprous sulfo- cyanate, a snow white Compound, which is rather an aid than a hin- drance to accurate Observation of the disappearance of the last trace of blue color," However, in applying Benedict's method to urine of low sugar content (below 0.5 per cent., as it frequently occurs in cases of glycosuria), one is Struck by the fact that the blue color of the mixture does not persist until the reaction is ended, for the Contents of the porcelain dish assume a dirty brownish-green hue that gradually merges into brown. This renders the correct estima- tion of the end point very difficult if not impossible. Clarifying the urine by the addition of lead acetate previous to the titration might overcome the difficulty, but this procedure would require additional manipulations and calculations ; and there is also the danger of a chemical change in the copper Solution. None of these objections apply to the simple method proposed by the author. It consists in the addition, just before heating, of approximately 10 i86 Proceedings Columbia Biochemical Association [Sept. grams (two heaping teaspoonsful) of powdered calcium carbonate to the Contents of the porcelain dish (25 c.c. of Benedict's Solution, 5-10 grams of anhydrous sodium carbonate, and a small amount of powdered pumice). The titration is then made in the usual manner. The snow white calcium carbonate, insoluble and suspended in the alkalin Solution, appears to act like the copper sulfocyanate in that it efTectively obliterates all colors except the blue color of the copper Solution. The end point obtained is sharp, the blue color being visible up to the addition of the last two drops of urine that are necessary for complete reduction. A sufficient amount of cal- cium carbonate (10 grams) must be added, otherwise the precipitate will be gray and the end point less distinct. In order to prevent sudden ebullition of the concentrated Solution, it is advisable to dilute the latter with a little distilled water, Experiments have shown that the addition of the calcium carbonate does not introduce any noticeable error. The author demonstrated these facts. 42. Diffusibility of protein through rubber membranes, with a note on the disintegration of collodion membranes by common ethyl ether and other solvents. William H. Welker. {Puh- lished in füll in this issue of the Biochemical Bulletin, )2* 43. A further study of the Bardach test for protein. Charles Weisman. {Piihlished in fiill in the June issue of the Biochemical Bulletin ).2^ 44. A study of the surface tension of dog blood-serum by the drop-weight method.^^ Harold E, Woodward. These ex- periments, about twenty in number, were planned to answer the question whether ordinary variations in the blood supply and nutri- tive condition of an individual affect the surface tension of the blood.^'^ Serum could be handled better than blood and serum from ** Welker: Biochemical Bulletin, 1912, ii, p. 70. ** Weisman : Ibid., 1912, i, p. 538. ^The animals were fed and controlled, and the blood was withdrawn and the serum coUected, by Dr. Gies and Mr. Chris Seifert. The drop-weights were made by the author in the laboratory of physical chemistry under the direction of Prof. J. L. R. Morgan. ^ These experiments were a logical preliminary to the work described else- where: Woodward, Dissertation, Columbia University, 1912. I9I2] 'Alfred P. Lothrop 1S7 clotted blood was more satisfactory than serum obtained by centri- fuging defibrinated blood. The normal surface tension of dog serum (five dogs), from the blood of animals on the usual diet in metabolism experiments in this laboratory, is about 45.5 dynes per centimeter. A daily hemorrhage of 3 per Cent, or more of the body weight, on two successive days, was without material effect on the surface tension. ^^ Small addi- tions of salt to the food raised, whereas additions of sugar lowered, somewhat the surface tension. The Ingestion of extra quantities of meat, several hours before blood was withdrawn, caused a decrease of about 1.5 per cent. in the surface tension. Fasting (1-2 days) raised the surface tension about i per cent. Copious water drink- ing (2 hours before withdrawal of blood) and the administration of magnesium sulfate, with resultant marked diarrhea (a short time prior to removal of blood from another dog), were without appre- ciable effect on the surface tension of the serum. These results suggest that the nutritive State of a given individual must be defi- nitely established before accurate conclusions can be drawn regard- ing the significance of data for surface tension of the subject's blood (or serum). [The December issue of the Biochemical Bulletin will pre- sent abstracts of the scientific Communications at the meeting of the Biochemical Association to be held on December 6, at the Columbia Medical School.] Biochemical Laboratory of Columbia University, College of Physicians and Surgeons, New York. '^When the second bleeding occurred in much less than 24 hours after the first, the surface tension was above normal. BIOCHEMICAL NEWS, NOTES AND COMMENT Contents. I. General: Necrology, i88; in memoriam, i88; anniversary celebrations, 189; honors, 190; retirements, resignations and appointments, 190; prizes, grants, endowments and funds, 193; meetings of congresses and societies, 194; buildings and general equipment, 195 ; acts of congress, 196; miscellaneous, 197. IL Columbia University Biochemical Association: General notes, 200; pro- ceedings, 201 ; biochemical department, 201. I. General Necrology. Dr. W. W. Daniels, emeritus professor of chemis- try at the University of Wisconsin. — Thomas Doliber, president of Mellin's Food Co., and one of the best known manufacturing drug- gists in America. — Dr. Morris Loeb, professor of chemistry at New York University and president of the Chemists' Club. — Dr. Her- mann Munk, formerly professor of physiology at the veterinary Col- lege in Berlin. — Dr. E. A. Holmström, Sweden's foremost pharma- cist. — Dr. Edmund von Neusser, professor of internal medicine at Vienna. — Prof. Melville Amasa Scovell, director of the Kentucky Agricultural Experiment Station and dean of the College of Agri- culture of the Kentucky State University. — Dr. Henry Adam Weber, professor of agricultural chemistry, Ohio State University. — Dr. Thomas Winter, professor of agriculture in University Col- lege of North Wales, Bangor. In memoriam. Lord Lister. A memorial to Lord Lister will be established at University College Hospital. It was in 1843 that Joseph Lister entered the College as an arts Student and graduated bachelor of arts in 1847. He then became a Student of medicine and entered the hospital to complete his studies. A special commit- tee has been formed under the presidency of the Duke of Bedford, President of the hospital. The exact nature of the tribute will be largely decided by the amount of the subscriptions received, but it has been suggested that either a bust or a tablet should be placed in both the hospital and the College. It is understood that the memo- rial will be entirely local in character, and only those who have been 188 I9I2] General 189 in some way connected with University College or the hospital are being asked to subscribe. The presidents of the Royal Society and the Royal College of Surgeons some weeks ago took the necessary steps for the forma- tion of a large and representative committee for the purpose of es- tablishing a memorial to the late Lord Lister. A meeting of the committee, which was largely attended, was held on July 22 at the rooms of the Royal Society, under the chairmanship of Sir Archi- bald Geikie. The following were appointed an executive commit- tee to recommend to a future meeting of the general committee a scheme for the memorial to Lord Lister and to organize an appeal for subscriptions : The Archbishop of Canterbury, the Lord Chan- cellor, Lords Iveagh, Rayleigh, Rothschild and Alverstone, the dean of Westminster, the Lord Mayor, the Lord Provosts of Edinburgh and Glasgow, the Master of the Rolls, Mr. Lewis Harcourt, M.P., Sir T. Barlow, Sir W. W. Cheyne, Sir R. J. Godlee, Sir H. Morris, Sir A. Geikie, Sir D. MacAlister, the Hon. Sir C. Parsons, Sir W. Turner, Sir J. Wolfe-Barry, Sir J. R. Bradford, Sir A. P. Gould, Sir A. Kempe, the Hon. W. F. D. Smith, Mr. F. M. Fry and Mr. Edmund Owen. Lord Rothschild and Sir W. W. Cheyne were appointed treasurers and Sir J. R. Bradford was appointed secretary of the Lister Memorial Committee. Dr. Paul C. Freer. The Bureau of Science of the Philippine Government has adopted resolutions in memory of Dr. Paul C. Freer, director of scientific work in the bureau, who died last April. The resolutions express the sense of his associates that " the Bureau of Science has suffered a very great loss and that the cause of sci- ence in the Philippine Islands has been deprived of one of its most zealous and conscientious advocates." Anniversary celebrations. June 30: Professor Gad, formerly director of the Physiological Institute at Graz, a pupil of Du Bois- Reymond, celebrated his seventieth birthday. — July i: Prof. Carl Binz, formerly director of the Pharmacological Institute at Bonn, celebrated his eightieth birthday. — August ß: Professor Bernstein, formerly director of the Institute of Physiology at Halle, celebrated the fiftieth anniversary of his doctorate. — September 14: Prof. W. 190 Biocltemical News, Notes and Comment [Sept. O. von Leube, the distinguished clinician, celebrated his seventieth birthday. Professor Leubc has been living at Stuttgart since last year, when he resigned his directorship of the Würzburg medical clinic. Honors. 'Awards of prizes. Dr. Alexis Carrel has been awarded the Nobel prine in medicine, in recognition of his achieve- ments in the suture of blood-vessels and the transplantation of Organs. — The Vienna Academy of Sciences has conferred its Liehen prise for 191 2 on Dr. Oswald Richter for his work on the food of algse. Honorary degree. The University of St. Andrews, Dundee, Scotland, has conferred the degree of LL.D. on Dr. S. J. Meltzer. Foreign associates. Sir William Ramsay and J. Reverdin have recently been elected foreign associates of the Paris Academie de Medecine. Retirements, resignations and appointments. Retirements. Col. Martin V. Calvin, for the past six years director of the Georgia Agricultural Experiment Station. — Prof. H. J. Wheeler, former act- ing-president of the Rhode Island State College, at Kingston, R. I., and, during the past eleven years, director of the government agri- cultural experiment Station at that Institution. Leave of ahsence. Dr. W. P. Bradley, professor of chemistry at Wesleyan University, has been granted leave of absence for the year 1912-13, to organize a department of research for the United States Rubber Goods Company. — Dr. A. F. Blakeslee has a year's leave of absence from the Connecticut Agricultural College. He has a tem- porary appointment on the staff of the Carnegie Station for Experi- mental Evolution at Cold Spring Harbor, L. L, where he will study lower fungi. Appointments have lately been announced, as follows '} Bryn Mawr College: Dr. Don R. Joseph (associate in physiology and pharmacology at the Rockef eller Institute), associate professor of physiology. Carnegie Institution, Boston Nutrition Laboratory: Mr. Joseph C. Bock (instructor in chemistry at Michigan Agricultural College), chemist. ^In the appended summary, institutions from which resignations occurred are named in parenthesis. I9I2] General 191 College of Agriculture and Mechanic Arts (Mayaguez, P. R.) : Dr. B. E. Ray (N. C. Experiment Station and College of Agriculture), Professor of chemistry. Columbia University: Mr. Ernest L. Scott (University of Kansas), instructor in physiology; Dr. Otto von Huffman (Cincinnati Hospital and Ohio Miami Medical College), instructor in clinical pathology. Commission for the study and prevention of malaria in the South : Dr. William S. Thayer, member. English Government Laboratory, London : Mr. E. Grant Hooper, deputy-government chemist (promoted), vice Mr. H. W. Davis, retired. Hamburg Botanical Institute: Dr. Hans Winkler (associate Pro- fessor of botany at Tübingen), director. Harvard University: Dr. Geo. R. Lyman (assistant professor of botany in Dartmouth College) will take the work of Professor Roland Thaxter during a sabbatical leave of absence. Institute for experimental research on Cancer, established by the Kaiser Wilhelm Society for the promotion of science: Prof. A. von Wassermann, director. Margaret Morrison School for Women of the Carnegie Institute (Pittsburgh) : Miss Mary D. MacKenzie (professor of biology at Western College, Oxford, Ohio), head of the department of biology. McGill University (Montreal) : Prof. Francis E. Lloyd (professor of botany in the Alabama Polytechnic Institute and plant physiologist to the Alabama Experiment Station), MacDonald professor of botany; Dr. F. R. Miller, lecturer in physiology. Medico-Chirurgical College (Philadelphia) : Dr. H. Lowenherg, assistant professor of infantile dietetics and also pediatrist to Mount Sinai Hospital, succeeding the late Dr. Edwin Rosenthal. Municli medical clinics : Prof. Friedrich Müller, instead of retain- ing the second clinic, has taken the first, left vacant by the death of Professor Bauer; Prof. E. v. Romberg (Tübingen) succeeds Professor Müller. N. Y. State Food Laboratory (Ithaca) : Mr. /. T. Ciisick (assistant in nutrition investigations, N. Y. Agricultural Experiment Station), analyst. N. Y. State School of Agriculture (Alfred University at Alfred) : Prof. W. J. Wright (Pennsylvania State College), director. N. C. Agricultural Experiment Station (West Raleigh) : Dr. Joseph F. Brewster, chemist. Ohio State University: Dr. W. G. Stover (Oklahoma Agricultural Experiment Station), assistant professor of botany. 192 Biochemical Nezvs, Notes and Comment [Sept. Ontario Agricultural College : Mr. R. E. Stone, lecturer in the botan- ical department. Pennsylvania Chestnut-Tree Blight Commission: Dr. F. D. Heald (professor of botany in the University of Texas), pathologist; Miss Caroline Rumbold (Missouri Botanical Garden), physiologist in charge of tree medication ; Mr. Joseph Shrawder, chemist. Reed College (Portland, Oregon) : Dr. Harry Beal Torrey (asso- ciate professor of zoology in the University of California), professor of biology. Skin and Cancer Hospital of Maryland : Mr. /. M. Codd, chemist. State University of Oregon Medical College (Portland) : John M. Co;mo//y, Ph.D., M.D. (Harvard Medical School), professor of physio- logical chemistry. U. S. Bureau of Animal Industry : Dr. Frederick J. Birchard (as- sistant in chemistry at the Rockefeiler Institute), research chemist in the Dairy Division. U. S. Bureau of Mines (Pittsburgh) : Dr. /. K. Phelps (U. S. Bu- reau of Chemistry, Washington, D. C), chemist. U. S. Bureau of Plant Industry : Dr. R. Kent Beattie (professor of botany in the State College of Washington), expert in the office of forest pathology; Dr. Neil E. Stevens (assistant pathologist in Kansas Experiment Station), forest pathologist. University College, Reading: Dr. vS". M. T. Auld (lecturer in the chemical department of the Southeastern Agricultural College atWye), professor of agricultural chemistry; Mr. John Goding (Midland Agri- cultural College), research chemist in dairy ing. University of Bonn: Professor Johannes Fitting (director of the State Botanical Institute at Hamburg), successor of Professor Stras- burger. University of Illinois: Dr. /. Howard Beard, instructor in physi- ology (promotion). University of Maryland : Dr. Jsaac M. Macks, pathologist. University of Minnesota : Dr. Robert B. Gihson, assistant professor of physiological chemistry (promotion) ; Dr. Rodney M. West, assist- ant professor of agricultural chemistry (promotion). University of South Dakota : Mr. Herbert Otto Lussky (assistant in physiology at the University of Chicago), director of the department of physiology in the College of arts and sciences and the College of medicine. University of Vienna: Prof. Wilhelm Türk, temporary successor to Prof. E. von Neusser in the medical school (page 188). I9I2] General 193 Washburn College : Dr. Edith M. Twtss, head of the department of botany ; Mr. James P. Poole, instructor in botany. Washington State College (Pullman) : Dr.IraD. Cardiff (professor of botany in Washburn College), professor of plant physiology. Prizes, grants, endowments and funds. Prizes. The Col- lege of Physicians, Philadelphia, announces that the next award of the Alvarenga prise, amounting to about $180, will be made July 14, 191 3. Essays may be devoted to any subject in medicine but must not have been published, and shonld be received by May i, 1913, by the secretary of the College, Dr. Thomas R. Neilson, 1937 Chestnut Street, who will furnish particulars, on request. — Madame Dieula- foy, widow of the late clinician, has given to the Academy of Medi- cine, of Paris, in memory of her husband, the sum necessary to found the Dieulafoy prise of $400, which will be awarded every two years to the author of the best work on the subject of internal pathology. — The Riberi prise, amounting to $4,000, will be awarded by the University of Turin, after the close of the year 1916, for the work which is adjudged to have most advanced the science of medicine. Grants. Grants for research, at the recent meeting of the Brit- ish Association: Mr. A. D. Hall, plant enzymes, £30; Prof. E. A. Schäfer, the ductless glands, £40; Prof. E. H. Starling, oxy-hemo- globin, £15 ; Prof. F. Gotch, mammalian heart, £20; Sir W. Ramsay, for the International Commission on Physical and Chemical Con- stants, £40. Endowments and funds. The London School of Tropical Med- icine is making an appeal for $500,000 to provide for the equipment and more efficient conduct of its work. — The late Dr. J. E. Robinson, first governor of Kansas, bequeathed $100,000 to the University of Kansas. The gift will be used for the medical school. — Mr. James B. Brady, of New York, has given the sum of $220,000 to the Johns Hopkins Hospital, for the establishment of a ward for the treatment of diseases of the kidney. — The late Mr. Allan Octavian Hume, well known as an ornithologist and botanist, lately bequeathed about £14,000 to the South London Botanical Institute, to which in 1907 he gave £10,000. — Under the will of the late Augustus W. Open- hym, Columbia University will receive a third of a trust fund of 194 Biochemical News, Notes and Comment [Sept. $275,000 for the endowment of research into the cause, prevention and eure of Cancer. Mr. Openhym's will stipulates that if at any time further investigation of Cancer is not required, the income of the fund may be used for research in any brauch of medicine or sur- gery. The endowment under Mr. Openhym's will is to be known as the Openhym Research Fund, and the terms of the gift are sub- stantially the same as those of the Crocker Research Fund, which amounts to $1,440,777.13. Meetings of congresses and societies. The Fifteenth Inter- national Congress on Hygiene and Demography was officially opened in the Continental Memorial Hall on September 23 and continued until September 27. President Taft delivered an address at the opening exercises. The delegates numbered about 3,000, represent- ing 33 foreign governments, every American State and territory, over 300 American cities, and leading Colleges and universities and many scientific, medical and social institutions throughout the world. The congress was divided into eleven sections and four general ses- sions were held. President Taft was honorary president, Dr. Henry P. Walcott, of Massachusetts, was president, and Dr. John S. Fulton, of Maryland, was secretary-general, of the congress. A füll account of the proceedings is given in the Journal of the Amer- ican Medical Association, beginning at page 1207 (September 28). The proceedings of the biochemical section — " dietetic hygiene ; hy- gienic physiology" — are reported at page 129 of this issue of the Biochemical Bulletin. The Eighth International Congress of Applied Chemis'try was officially opened at Continental Memorial Hall, in Washington, on September 4, and continued in New York from September 6-13, in- clusive, where the work was centralized at Columbia University and the College of the City of New York. About 2,500 members were in attendance. Dr. Edward W. Morely was honorary president, Prof. William H. Nichols was president, and Dr. Bernhard G. Hesse was secretary, of the congress. The scientific work of the congress was organized in twenty-four sections. Among the gen- eral addresses was one by Prof. Gabriel Bertrand on " The part played by infinitely small quantities of chemicals in biological chemistry." igi2] General i95 Professor W. H. Perkin delivered a lecture on " The polymeriza- tion of butadiene and isoprene," before the Sections on Organic Chemistry and India Rubber. Prof. Perkin outlined bis original method of making synthetic rubber,^ and then described the follow- ing new method : Take ethyl alcohol, which may be easily oxidized to acetaldehyde. This is Condensed by means of potassium carbon- ate to aldol and the aldol can be quantitatively converted into butyl- idine gycol. All the yields of these reactions are practically quan- titative. The butylidine glycol is then converted into a chlorid and passed over soda-lime, when practically the same product is pro- duced as the isoprene from isoamyl chlorid and, when treated with sodium, gives even better rubber than isoprene. Professor Perkin exhibited samples of what he called the first synthetic rubber ever made (the product of Tilden). A general review of the proceedings of the Congress will appear in the October issue of the Journal of Industrial and Engineering Chemistry (pages 706-719). The proceedings of the biochemical section are reported at page 150 of this issue of the Biochemical Bulletin. The eighty-second annual meeting of the British Association for the Advancement of Science, which opened at Dundee on Septem- ber 4, had a registration of 2,504 members, which is considerably larger than the average. At the opening session the President, Prof. E. A. Schäfer, delivered a notable address on the "Nature, origin and maintenance of life," which has been published in Nature (90: 7-19) and Science (36: 289-312). It was announced that Dr. J. K. Caird, of Dundee, had given £10,000 to the funds of the association. A general account of each sectional meeting will ap- pear in Science (36 : 446-452). The Royal Society recently celebrated its 25oth anniversary. The I4th meeting of the Australasian Association for the Ad- vancement of Science will be held in Melbourne in January, 19 13. Buildings and general equipment. The work of the Herriman Dispensary of the Brooklyn Hospital was inaugurated on July 17. The dispensary will be open daily. It is a two and one-half story * Biochemical Bulletin, 1912, i, p. 566. 196 Biochemical News, Notes and Comment [Sept. brick and marble structure and was given by Mr. William H. Herri- man in memory of his wife. Mr. Herriman donated $100,000 for this purpose, $25,000 of which will be used as an endowment fund. — Messrs. Jacob H. Schiff, Sei. R. Guggenheim, Ferdinand Sulz- berger and Samuel Sach have each given $50,000 to a fund for the construction of a private hospital for persons suffering f rom chronic diseases, to be built by the Montefiore Home, in the Bronx, New York City. — The Medical Faculty of the University of Utah is re- questing the Regents of the University to ask the Legislature for a special appropriation of $25,000 for the medical school. It is not generally known that the State of Utah is doing better by its Uni- versity, proportionately, than any other State, in that this Institution receives 28 per cent. of the state's income in taxes. The State of Utah contains about 400,000 inhabitants. Acts of Congress. Public Health Service. The following is the text of the act of congress concerning the Public Health Ser- vice : Be it enacted by the Senate and House of Representatives of the United States of America in Congress assembled. That the Public Health and Marine-Hospital Service of the United States shall hereafter be known and designated as the Public Health Serv- ice, and all laws pertaining to the Public Health and Marine-Hos- pital Service of the United States shall hereafter apply to the Public Health Service, and all regulations now in force, made in accord- ance with law for the Public Health and Marine-Hospital Service of the United States, shall apply to and remain in force as regula- tions of and for the Public Health Service until changed or rescinded. The Public Health Service may study and investigate the diseases of men and conditions influencing the propagation and spread thereof, including sanitation and sewage and the pollution either directly or indirectly of the navigable streams and lakes of the United States, and it may from time to time issue Information in the form of publications for the use of the public. 'Amendment to the food and drug act. Congress, before ad- journment, passed an amendment to the food and drug act which the President has signed, making it illegal " if its package or label shall bear or contain any Statement, design, or device regarding the curative or therapeutic effects of such article, or any of the ingredi- I9I2] General 197 ents or substances contained therein, which is false and fraudulent." It will be remembered that the act of 1906 declared that a drug is misbranded "the package or label of which shall bear any Statement . . . which shall be false or misleading in any particular . . . " ; but the supreme court, by a majority of five to three, decided that this did not refer to false Statements regarding the curative effect of a drug. Miscellaneous items. Proposed State medical service in Eng- land. During the recent meeting of the British Medical Associa- tion at Liverpool, a State Medical Service Association was formed under the inspiration of Dr. B. Moore, professor of biochemistry at the University of Liverpool. Prof. Moore lately produced a book entitled " The dawn of the health age," in order to demonstrate the necessity for entirely remodeling the present System of medical prac- tice in the interests of the whole Community. The object of the new association is to advocate a State medical service on the follow- ing basis : (i) the whole profession to be organized on the lines of the other State Services now in existence; (2) entry to the profes- sion to be by one state examination; (3) each member of the serv- ice to be paid an adequate salary, increasing gradually according to the length of service and position in the service, and to be entitled to a Pension after a specified number of years or in case of perma- nent disablement; (4) members of the public to have, as far as possible, free choice of physicians, but no physician to be called on to have charge of more than a specified number of patients; (5) one of the primary objects of the State service to be to unite preventive and curative medicine ; all hospitals to be nationalized and used for the purpose of consultative, operative and therapeutic work at the request of and in conjunction with the patient's own physician; (6) the Services of the state physicians to be open to €very one, rieh or poor; (7) the state medical service to be administered by a board of health under a minister of public health with cabinet rank, assisted by expert medical advisers. This movement was started before the insurance act was passed and is quite independent of the present impasse. It is intended that the work of the association shall form a brauch of sociologic science, and membership will be open to all prominent sociologists, whether lay or medical. 198 Biochemical News, Notes and Comnient [Sept. (London correspondent, Journal of the American Medical Associa- tion, 19 12, lix, p. 663 : August 10). Detection of formaldehyde in foods. In view of the introduc- tion of a mixture of nitrite and formaldehyde with the object of masking the reactions of the latter when used as a food preserva- tive, the following experiments may be of interest. A sample of f resh mixture was divided into four portions and treated as follovvs : (i) A small amount of commercial formaldehyde Solution was added; (2) small amounts of formaldehyde and sodium nitrite were added; (3) a small amount of sodium nitrite was added; (4) no addition was made. Portions of each of these were tested with Rimini's test (Phenylhydrazin hydrochlorid, sodium nitroprussid and sodium hydroxid). Prompt reactions for formaldehyde were obtained in i and 2; negative results in 3 and 4. Other portions of the samples were tested with the well-known test for nitrite (sul- fanilic acid and alphanaphthylamin) . The responses of 2 and 3 were prompt and distinct. No color was produced in i and 4. The original mixtures were allowed to stand 24 hours at room tempera- ture and the tests repeated with the same results as obtained at first. It seems easy, therefore, to unmask nitrite and formaldehyde in the presence of each other. Henry Leffmann. {Journal of Industriell and Engineering Chemistry, 1912, iv, p. 626: August.) Joiirnalistic. With the September number Prof. A. R. Cushny, of University College, London, becomes Joint editor with Prof. John J. Abel, of Johns Hopkins University, Baltimore, of the Jour- nal of Pharmacology and Experimental Therapeutics. At the same time, Sir T. Lauder Brunton, of London, Professors J. T. Cash, of Aberdeen, W. E. Dixon, of Cambridge, J. A. Gunn, of Oxford, Sir Thomas R. Fräser, of Edinburgh, J. N. Langley, of Cambridge, C. R. Marshall, of the University of St. Andrews, R. Stockman, of Glasgow, F. Ransom, of London and Dr. H. H. Dale, of London, join the board of associate editors. By this arrangement the ablest representatives of phannacology in Great Britain unite with the American and Canadian colleagues in the conduct of the Journal and the publishers feel confident that it will henceforth serve as the medium of publication for the best pharmacological researches of the I9I2] General 1 99 english-speaking countries. (Publisher's announcement, Septem- ber number, vol. iv, no. i.) Visiting agriadturalists. Mr. Paul Korchoof, agricultural ex- pert, department of the Russian ministry of agriculture, and Mr. Vaseelie Yurieff, assistant director, Kharkow Central Agricultural Experiment Station, have been visiting the agricultural Colleges and stations in this country. — Dr. E. B. Copeland, dean of the College of Agriculture, Los Bafios, P. I., who has been visiting the United States, recently returned to the Philippines. Parsons in Washington. Dr. Charles L. Parsons, secretary of the American Chemical Society, moved from Durham, N. H., to Wash- ington on September i. The headquarters of the American Chem- ical Society may now be addressed, Box 505, Washington, D. C, Remsen to remain at Hopkins. Owing to the difficulty of find- ing a suitable occupant for the post, Dr. Ira Remsen will remain at the head of Johns Hopkins University for the ensuing session, or part of it at least. Petroleum production in the United States, in 191 1, surpassed its own record (made in 1910) by an increase of nearly 11,000,000 barreis. In 1910 the Output was 209,557,248 barreis. The total production of the world also surpassed all previous records, amount- ing to over 345,000,000 barreis. Johns Hopkins limits enrolment. The dean of Johns Hopkins Medical School announces that it has become necessary to limit the number of students owing to the restricted space and facilities in the various laboratories. The present enrolment is 355, the largest in the history of the school, and fifty other students were refused ad- mission prior to the beginning of the session. Standard rations for nutrition experiments. A Conference was held at the Graduate School of Agriculture, Lansing, Mich., on July 24, to discuss the formulation of Standard rations for experi- mental work in determining the comparative value of feed stuffs. Mr. B. H. Rawl, chief of the dairy division, U. S. Department of Agriculture, President H. J. Waters, of Kansas Agricultural Col- lege, Prof. C. H. Eckles, of Missouri Experiment Station, and other leading workers in this field were present and led the discussion. 200 Biochemical News, Notes and Comment [Sept. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION I. General notes Miscellaneous items. Dr. Carl L. Alsherg was one of the dis- tinguished non-resident scientists to participate, by invitation, in a series of lectures, during the late summer at Fordham University Medical College, in New York, on nervous and mental diseases. — The following members of the Association conducted investigations at Woods Hole, Mass., during the summer: Cora J. Beckwith, H. B. Goodrich, Louise H. Gregory, Charles Packard, Alwin W. Pap- penheimer, Henry J. Spencer, Charles R. Stockard, Isabel Wheeler, and L. L. Woodruff. — Dr. 'A. Richard Bliss is editor-in-chief of TheMask, the official national organ of the Kappa Psi Fraternity. — Prof. R. Burton-Opitz is now in Europe, where he is spending a half-year leave of absence. Officers of societies. Section (V) on Control of Infectious Diseases of the I5th International Congress on Hygiene and Demog- raphy (page 194) : Dr. Charles F. Boldiian, secretary. — New York Post-Graduate Medical School and Hospital : Dr. Arthur F. Chace, secretary (reelected). — Section (IV) on Organic Chemistry of the 8th International Congress of Applied Chemistry (page 194) Dr. Harry L. Bisher, secretary. — N. Y. Entomological Society: Prof. Raymond C. Oshiirn, president. — American Association for the Study and Prevention of Infant Mortality: Dr. Philip Van Ingen, secretary. Appointments. Jefferson Medical College (Philadelphia) : Dr. Philip B. Hawk (professor of physiological chemistry, University of Illinois), professor of physiological chemistry and toxicology. — Rockefeiler Institute for Medical Research: Dr. Michael Heidel- berger (recently returned from Zürich), fellow in chemistry. — Johns Hopkins University: Dr. John Howland (professor of pediatrics in Washington University, St. Louis), director of the Harriet Lane Home for Invalid Children, professor of pediatrics, and physician in charge of the pediatric department of Johns Hopkins Hospital. — ^Cornell University Medical College, Loomis Laboratory : Miss Jessie A. Moore (assistant at the Rockefeiler Insti- tute for Medical Research), chemical assistant. — N. J. Agricultural Experiment Station: Mr. Carl A. Schwarze, assistant plant pathol- ^, Ol*jJJ_ "X-^Wv^n^, 1912] Columbia University Biochemical Association 201 ogist. — Long Island Medical College: Dr. Matthew Steel (assist- ant Professor of physiological chemistry, University of Missouri), as- sistant professor of physiological chemistry and pharmacology. — At a recent annual meeting of the Imperial Cancer Research Fund, in London, Dr. William H. Woglom was appointed first assistant in New York, a position maintained under the auspices of the Crocker Fund for the investigation of Cancer. Dr. Woglom has returned from London, where he had been pursuing a course of study under Dr. Bashford, director of the Imperial Cancer Research Fund. 2. Proceedings of the Association. Abstracts of the scientific proceedings of the third annual meet- ing (June) are pubHshed on pages 156-187 of this issue. 3. Columbia Biochemical Department. The new Assistant Professor, Dr. Paul E. Howe, B.S., A.M., PhD.^ Memorandum which was presented to the Faculty OF Medicine with Dr. Howe's nomination to the Assistant PROFESSORSHIP IN BIOLOGICAL CHEMISTRY. Paul Edward Howe was born in Chicago, Illinois, on July 29, 1885. His early education was received in the public schools of Chicago, Champaign and Urbana, Illinois (1890-1901). He at- tended the Urbana High School (1899-1901) and spent a year (i90i-'02) in the Preparatory School of the University of Illinois. At the end of a four-year course at the University of Illinois he received the degree of B.S. in Chemistry in 1906. Since 1906 he has been a graduate Student and officer at the Uni- versity of Illinois, passing by promotion through the grades of Scholar in chemistry in the graduate school (i9o6-'o7), assistant chemist in the laboratory of physiological chemistry (1907-08), assistant in physiological chemistry (1908-10), and instructor in physiological chemistry (i9io-'i2). In 1907 he received the degree of M.A. ; in 1910, the degree of Ph.D. His major subject for the Ph.D. degree was physiological chemistry, with Professor P. B. Hawk; his minor subjects were physical chemistry, physiology and histology. * Biochemical Bulletin: 1911-12, i, pp. 136, 570, 573 and 574. 202 Biochcmical N'czvs, Notes and Comment [Sept. Dr. Howe is a member of the American Society of Biological Chemists, American Chemical Society, American Society of Animal Nutrition, American Association for the Advancement of Science, Illinois Academy of Science, Sigma Xi, Phi Lambda Upsilon, and the Gamma Alpha Graduate Scientific Fraternity. Dr. Howe's publications. 1907. The electrolytic corrosion of brasses (with A. T. Lincoln and David Klein) ; Journal of Physical Chemistry, 11, 501. 1908. Comparative tests of Spiro's and Folin's methods for the determination of ammonia and urea (with P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, i, 104; Journal of Biological Chemistry, 4, p. x. 1909. Comparative tests of Spiro's and Folin's methods for the determination of ammonia and urea (with P. B. Hawk) ; Journal of Biological Chemistry, 5, 477. — On the preservation of feces (with T. A. Rutherford and P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, i, 196; Journal of Biological Chemistry, 6, p. xlix, 1910. On the preservation of feces (with T.A. Rutherford and P. B. Hawk) ; Journal of the American Chemical Society, 32, 1683. — A study in repeated fasting (with P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, i, 259; Journal of Biological Chemistry, 7, p. xlvi. — Fasting studies on men and dogs (with H. A. Mattill and P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, i, 260; Journal of Biological Chemistry, 7, p. xlvii. — Nitrogen partition in repeated fasting; Dissertation (pp. 42), pre- sented in partial fulfilment of the requirements for the degree of Doctor of Philosophy (University of Illinois). 191 1. On the differential leucocyte count during prolonged fasting (with P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, 2, 15; Journal of Biological Chemistry, 9, p. xxi. — Fasting studies : I. Nitrogen partition and physiological resistance as influenced by repeated fasting (with P. B. Hawk) ; Journal of the American Chemical Society, 33, 215. — Fasting studies: III. Nitrogen partition of two men through two seven-day fasts following the prolonged Ingestion of a low-protein diet : Supplemented by comparative data from the sub- sequent feeding period (with H. A. Mattill and P. B. Hawk) ; Journal of the American Chemical Society, 33, 568. — Fasting studies: V (Studies on water drinking: XI). Influenae of an excessive water Ingestion of a dog after a prolonged fast (with H. A. Mattill and P. B. Hawk) ; Journal of Biological Chemistry, 10, 417. I9I2] Columbia University Biochemical Association 203 1912. A metaboHsm study on a fasting man (with P. B. Hawk) ; Proceedings of the American Society of Biological Cheniists, 2, 65 ; Journal of Biological Chemistry, 11, p. xxxi. — Hydrogen-ion concen- tration of fecal extracts (with P. B. Hawk) ; Proceedings of the American Society of Biological Chemists, 2, 66; Journal of Biolog- ical Chemistry, 11, p. xxxii. — Studies on water drinking: XIII (Fast- ing studies: VIII). Hydrogen-ion concentration in feces ; Journal of Biological Chemistry, 11, 129. — A comparison of the data from two fasts each exceeding one hundred days in length and upon the same subject (with P. B. Hawk) ; Proceedings of the American Physiological Society, American Journal of Physiology, 29, p. xiv. — On the differ- ential leucocyte count during prolonged fasting (with P, B. Hawk) ; American Journal of Physiology, 30, 174. — Fasting studies: VI. Dis- tribution of nitrogen during a fast of 117 days (with H. A. Mattill and P. B. Hawk) ; Journal of Biological Chemistry, 11, 103. — The gen- eral aspects of fasting: Address before the Columbia University Bio- chemical Association, May i, 1912; Biochemical Bulletin, 2, 90. — The distribution of urinary nitrogen as influenced by the Ingestion of moderate and copious quantities of distilled water at meal time (with D. W. Wilson and P. B. Hawk) ; (in press) Journal of the American Chemical Society, 34, Proceedings, p. ^^i- — Addendum. The utiliza- tion of individual proteins by man as influenced by repeated fasting (with P. B. Hawk) ; Proceedings of the Eighth International Congress of Applied Chemistry (preliminary edition), 19, 145. (William J. Gies, Secretary of the Faculty of Medicine. ) Resignations and appointments. The following changes in the staff for the year 1912-13 were officially authorized prior to October i, IQ12: Dr. Paul E. Howe, assistant professor, vice Prof. Wm. H. Welker, resigned; Dr. Clayton S. Smith, instructor (pro- moted), vice Dr. Ernest D. Clark, appointed instructor in chemistry at the Cornell University Medical College; Dr. Frederic G. Good- ridge, assistant, vice Reuben Ottenberg, resigned ; Messrs. E. G. Mil- ler, Jr., and Arthur Knudson, assistants, vice Dr. C. S. Smith (pro- moted) and Mr. A. R. Rose, resigned; and Misses Ethel Wickwire and Tula L. Harkey, assistants (at Teachers College), vice Mr. E. G. Miller, Jr. (promoted) and Miss Blanche Harris, resigned. Summer session. Courses. Professor Gies kept the labora- tory open daily throughout the summer and conducted courses (July 5-August 15) in nutrition at the College of Physicians and 204 Biochemical News, Notes and Comment [Sept. Surgeons with Dr. C. S. Smith's assistance, and at Teachers' Col- lege with the aid of Dr. Emily C. Seaman and Miss B. E. Shaffer. Investigators. The workers named below conducted research, in the biochemical laboratory at the College of Physicians and Sur- geons, during all or part of the summer vacation : Louis Berman, R. J. Cook, Edward Cussler, F. R. Eider, N. B. Foster, Wm. J. Gies, Samuel Gitlow, Isidor Greenwald, W. M. Kraus, Alfred P. Lothrop, H. A. Mattill, H. O. Mosenthal, Jacob Rosenbloom, Emily C. Seaman, C. S. Smith, William Weinberger, Charles Weisman, Wm. H. Welker, Harry Wessler. Miscellaneous notes. Professor Gies was vice president of the Section on Biochemistry including Pharmacology of the 8th Inter- national Congress of Applied Chemistry (page 150). — Dr. H. 0. Mosenthal recently returned from Tübingen, where he had been working in the medical clinic under the direction of Prof. E. von Romberg. — Dr. Jacob Rosenbloom has resigned the affiliated Posi- tion of assistant pathologist at the German Hospital. — Mr. A. R. Rose has lately completed the requirements for the Ph.D. degree and will be publicly examined in October. He has begun a special study of amylase with Prof. H. C. Sherman. — Mr. Joseph Hepburn- has begun work as " university fellow " in biological chemistry. EDITORIALS Ernst Schulze was one of the great pioneers in biological chem- istry. He worked in an inspired way along the zone between the old zoöchemistry and the ancient phytochemistry, and achieved the distinction of removing the barriers between rnst c uze these two fields and uniting them in one great open biochemical territory. He brought Hght and understanding into large domains where darkness and doubt prevailed. His ex- ample in industry, patience, perseverance, devotion, enthusiasm, abil- ity and productiveness has been an inspiration to biochemical work- ers the world over. Schulze's classical achievements and Service will be forever linked with the history of fundamental developments in a great formative scientific era. His name and Service will be justly remembered, as his memory will be venerated, for very many generations. As the methods of chemical analysis become more delicate and refined there appears ever increasing evidence that the maintenance of health and nutrition depends not alone on the caloric values of T . ^ ^1. u food-stuffs and the relativ^e proportions of nitro- Important though ^ . unknown factors in gen and carbon in the diet, but quite as much on nutrition other factors which we are beginning fully to appreciate. Scurvy has long been one of the indications that there are certain unappreciated factors in a normal diet, and the antiscor- butic action of vegetables and vegetable Juices is strong emphasis on this point. The researches of Hart, McCollum, Steenbock and Humphrey/ on cattle, and of Osborne and Mendel,^ on rats, are among the many recent studies that reveal the importance of such unknown though influential factors in their broad bearing. The disease beriberi is a concrete example of the disturbance of such subtle influences. For a considerable time physicians in the ^ Hart, McCollum, Steenbock and Humphrey : University of Wisconsin Agricultural Experiment Station Research Bulletin, No. 17 (June, 1911). * Osborne and Mendel: Carnegie Institution, Puhlication 156. 205 2o6 Important Factors in Nutrition [Sept. Orient have believed that certain foods were responsible for this form of Polyneuritis. Miura believed the noxious agent to be contained in a certain fish, which is much eaten raw ; but more re- cently the blame has fallen on rice. It has been asserted that in the prisons of Java, beriberi occurs in one out of every forty prisoners when shelled rice is eaten; in one out of ten thousand, if the un- shelled grain is used. The classical studies of Schaumann were sug- gested by observations of this kind. Schaumann believed that since polished rice is poor in phosphorus, beriberi is due to a deficiency of certain organic phosphorus Compounds. This hypothesis had some Support in the fact that materials which relieve the pain of neuritis, such as bran, are rieh in phosphorus, but the later investigations of Wieland^ cast doubt on the accuracy of these deductions, since it could not be shown that the total body-phosphorus was much in- fluenced by feeding mice on polished rice. In this connection the researches of Fingerling,^ and of McCollum and Halpin,^ are sug- gestive, for they have shown a synthesis of organic phosphorus Com- pounds from inorganic phosphates. The latest contributions to the study of beriberi were made by Chamberlain and Vedder,^ by whom it has been shown that extracts of rice-bran are effective as therapeutic agents and that these ex- tracts contain mere traces of phosphorus. The active substance in the bran has not yet been identified, but the interesting feature dis- closed by the present evidence as to the etiology of kakki is that a food stuff may contain an ingredient which is essential in order to prevent injury to the tissues by other components of such food ma- terial. Rice grain is harmless when eaten with the pericarp but, if the latter is removed by " polishing," a malady ensues which may be cured by extracts made from the pericarp. These facts present a new face to the idea of " balanced rations " and also remind us of the broad biological significance of Loeb's "balanced Solutions." N. B. F. ' Wieland : Archiv für experimentelle Pathologie und Pharmakologie, 1912, Ixix, p. 293. * Fingerling: Biochemische Zeitschrift, 1912, xxxviii, p. 448; xxxix, p. 239. "McCollum and Halpin : Journal of Biological Chemistry, 1912, xi (Pro- ceedings of the American Society of Biological Chemists, p. xiii). * Chamberlain and Vedder: Philippine Journal of Science, 1912, vi, p. 251. 1912] Editoriais 207 In a circular with this title, Director Russell of the Wisconsin A'gricultural Experiment Station^ has recently given an interesting summary of the perfection of the Babcock quantitative test for milk- rru ~- t ^ fat and the influence which it has exerted on The Coming 01 age of the dairy science and practice throughout the world. Babcock test Milk and its numerous products play so impor- tant a röle in the economy of the home and in the dietary of the sick that the significance of Professor Babcock's contribution cannot re- main unnoticed in the annals of the medical world. The simple, yet highly accurate Babcock method of estimating the fat content of milk and cream finds daily application not only in dozens of analytic laboratories, but likewise in hundreds of creameries, in milk establishments, and even in the office of the busy practitioner of medicine, where a few inexpensive devices enable him to gauge the richness of a breast-milk or a modified milk mixture with facility. Every pediatrist appreciates what the Babcock test means for the exigencies of practice and successful feeding. Today, twenty-two years after the introduction of this procedure which, as Ex-Gover- nor Hoard remarked, has made dairymen more honest than the Bible because it has removed all opportunity for them to profit by any deceit, it is interesting to note that no change has been made in the essential features of the test during all this period. The tech- nic of the Operation remains the same as when the details were pub- lished by Dr. Babcock in 1890. The Stimulus which it has given to scientific dairying, to the standardization and improvement of our milk-supplies, to the possibilities of rational infant-feeding, and to what these in turn involve in the direction of the public health, is scarcely appreciated by the medical profession. Director Russell has written that the Babcock test frees the dairy farmer from the fetters of past traditions, and removes him from the category of "mossbacks." The influences here referred to have in fact been even more far-reaching. An additional feature deserves mention: No patent was taken out on either the method or the apparatus required to carry out the Babcock test. There zuas no copyrighting of a name — no commer- ' University of Wisconsin Agriculitiral Experiment Station, Circular of Information, No. 2^, 1912. 2o8 Babcock Test [Sept. cialism. In accord with a code of ethics now more generally recog- nized than at any time, the discoverer, becaiise of his connection with the State experiment Station, gave his invention freely to the world. We may gladly join in acknowledging our Obligation to the man whom the grate ful State of Wisconsin has presented a medal in recognition of " his iinselfish dedication of these inventions to the public Service." (Editorial : Journal of the American Medical As- sociation, 191 2, lix, p, 544.) The discovery and investigation of the specific secretions of the so-called ductless glands and of other organs make one of the most interesting chapters in physiology. Much has been learned con- cerning these secretions and their röle in the rgano- er py j^^Q^jy These extracts, theoretically, should be of great value in the treatment of diseases in which a certain gland or glands are deficient or entirely lacking in function. But actual ex- perience has been disappointing as a rule, for two reasons : ( i ) The diagnosis of insufficiency of secretion on the part of a certain gland or Organ is usually most difficult; (2) and even when a correct diagnosis is made, it is rarely possible to administer the gland sub- stances in such a way as to develop their specific activity. A notable exception to this experience is the successful use of thyroid extract in thyroid insufficiency or myxedema. Suprarenal substance has also proved highly use ful as a circulatory stimulant and hemostatic, but not for the treatment of Addison's disease. It can safely be said that the administration of gland substance from the thymus, hypophysis, ovaries, pancreas, testicles, etc., for dis- eases of these organs, has hitherto met with failure. Only härm can come from their promiscuous use before careful experimentation fully determines their value. A wholesome skepticism concerning the efficiency of preparations of the digestive enzymes is likewise commended. After years of usage many of our best clinical observers believe that pepsin, " pan- creatin " and the amylases are of little or no value. The use of se- cretin more recently has been similarly disappointing. The con- tinued routine use of these preparations is due chiefly to the claims of manufacturers. I9I2] Editorids 209 We congratulate our English confreres on the successful con- summation of their plans for the formation of a biochemical society and the publication of a biochemical Journal^ under their associated Biochemical So- control. In this coimtry we have long derived ciety, England great benefit from the meetings and activity of the American Society of Biological Chemists and are confident our English colleagues will have a similar experience. We felicitate the biochemical profession at large on this further evidence of the rapid growth in usefulness, and the prominent place of Service, of biochem- ical art and science. The Bio-Chemical Journal has been highly esteemed in America, and we wish it long life and distinguished Service under its new management. " Science is essentially mutual- istic and the success of one Organization is the gratification of all — the triumphs and discoveries of one are shared with the many, and the feeling of pride in the progress of the one may he shared. zvithout loss by sister organizations. As the discovery made in one brauch of science may be the necessary foundation for the Solution of some problem in another, so the contribution from one society may be the stepping stone to advancement in another. It is all hail then, greetings and felicitation — and Godspeed in the accomplish- ments of your future destiny." The name of the writer of this note might suggest a strong par- tiality on his part for the incorporation into biochemical discussions, in English, of such words as " Baustein." He believes, neverthe- " Baustein " or ^^^^' ^^^^ English phrases of equal f orce thoug'h " construction Unit** of more abstract significance, such as "construc- tion Unit" for "Baustein," are more acceptable, especially to stu- dents receiving their introduction, in English, to the subject of pro- tein synthesis and similar processes. The foregoing remarks recall the common use, in English, of " Splitting product " or " split product " as equivalents for " Spal- tungsprodukt," when the substance referred to is neither " Splitting Splitting productsor "^^ "split," but has resulted from cleavage. cleavage products Why not term such substances "cleavage prod- ucts" in conformity with good usage in analogous relationships ? "Halliburton: Biochemical Bulletin, 1912; ii, p. 128. 2IO X-Rays fSept. We received recently, with very great pleasure, a foreign money Order for twelve dollars instead of tzvelve Shillings in payment of Volume I of the Biochemical Bulletin. In view of the fact that this overpayment did not excite a desire to dis- A rare complimen ^^ontinue the subscription, we have proceeded with more enthusiasm than ever with our editorial work, in the hope that future volumes of the Bulletin may be much more deserving of such a compliment. The doing that makes commerce is born of the thinking that makes scholars. — Ruskin. Perhaps the most valuable result of all education is the ability _. _ to make yourself do the thing you have to do, when it ought to be done, whether you like it or not. — Huxley. The fabric of medical progress — indeed, of all progress — is woven from legitimate dreams to a greater extent than the " practi- cal" man is wont to realize or willing to admit. Editorial: Journal of the American Medical Association, 1912, lix, p. 1195. Who is it that, when years are gone by, we remember with the purest gratitude and pleasure? Not the learned or clever. But those who have had the force of character to prefer the future to the present, the good of others to their own pleasure. — Stanley. A fig for yesterday's convictions ! They were the cocksure beliefs of children lost in the dark. This is another day, and we've grown overnight. Do you plead the dignity of fixed opinion? It is enough for us to say : " We believed it when we affirmed it ; we have learned and changed our minds." — Ana Phylactic. The successful man, whether in business, in the professions and trades, or in politics, enjoys the game for its own sake. He is not a conscript in life's battles, but a volunteer, The way interests him as much as the goal. Not only the result, but the exercise of powers necessary to achieve it, gives him satisfaction. — AI I. Phatic. Speaking mentalwise, overfed conceit equals the blind staggers. The easiest kind of intoxication is that which feeds upon the poisons distilled by a self-caressing imagination. Open the floodgates of self-approval and soon you won't know whether you are making good or not, for you won't be able to present an intelligent compari- son of your own achievements with those of others. — Jaun Dice. BOOKS RECEIVED The BiocHEMicAL Bulletin will promptly acknowledge, under this heading, the receipt of all publications that may be presented to it. From time to time, selections will be made for review on pages of the volume to be appropriately indicated here. Reviews will be matter-of-fact Statements of the nature and Contents of the publications under consideration, and will be intended solely to guide possihle purchasers. The wishes or expectations of publishers or donors of volumes will be disregarded, when they are incompatible with our convictions regarding the interests of our colleagues. The size of the printed pages, in inches, is indicated in the appended notices. Practical physiological chemistry. A book designed for use in courses in practica! physiological chemistry in schools of medicine and of science. By Philip B. Hawk, professor of physiological chemistry and toxicology in the Jefferson Medical College of Philadelphia. Fourth edition, revised and en- larged. Pp. 475 — 4J^X8; $2.50 net. P. Blakiston's Sons & Co.. Philadelphia, 1912. The protein element in nutrition. (One of the International Medical Mono- graphs.) By Major D. McCay, professor of physiology, Medical College, Cal- cutta. Pp. 216 — 4X7, with 8 füll page portraits of human subjects; $2.00 net. Longmans, Green and Co., New York; Edward Arnold, London, 1912. Oxidations and reductions in the animal body. (One of the Monographs on Bio chemistry.) By H, D. Dakin, The Herter Laboratory, New York. Pp. 135 — 41^X8; $1.40 net. Longmans, Green and Co., 1912. Researches on cellulose. III (1905-1910). By C. F. Gross and E. J. Bevan. Pp. 173 — 3> Average Control ( average ) II. Extract of Fibroma of Uterus A 8 6 8 6 7 8 12 12 10 8 10.5 10 6 8 8 8 7-5 8 10 B. 10 C. 12 £> 16 Average 12 Control (average) II III (0). Extract of a Carcinoma of the Breast A 18 12 15 15 17 9 7 8 8 8 7 12 12 9 15 12 16 8 B 6 c. 8 D 8 Average 7.S 8 Control (average) III (b). Extract of Normal Breast Tissue Near the Cancer^^ A 20 18 24 22 21 8 6 8 6 7 26 17 24 25 23 6 B 8 C 8- D 7 Average 7.3 III. (c). Extract 0 f Pectoral M uscle Removed at Operation^^ A 14 26 14 24 19.5 7 6 II 8 8 26 18 26 24 23-S 9 9 12 B C D 10 Averaee 10 Preparation of Cancer extracts. Fresh cancerous tissue, direct f rom the operating room, was minced, then triturated with saiid and water, and the thin mixture frequently shaken for about an hour. ""Control" figures are given in section III (a). 232 Infliicnce of Cancer Extracts on Lupin Seedlings [Jan. The liquid was strained through gauze, then filtered. Portions of this filtered extract (boiled or unboiled) were used in the manner indicated above. Data pertaining to grozvth. The summary on page 231 pre- sents the results of this study. General conclusion. The extracts failed to inhibit growth of the seedlings. The observed acceleration of growth was probably due to inorganic salts in the extracts. It is possible, of course, that deleterious action by Cancer toxins was neutralized or overcome by the stimulating power of associated nutrient substances. This par- ticular poinit requires special investigation. THE BIOCHEMISTRY OF THE FEMALE GENITALIA^ 3. A quantitative study of certain enzymes of the ovary, Uterus, and bladder, of pregnant and non-pregnant sheep THUISCO A. ERPF-LEFKOVICS' and JACOB ROSENBLOOM (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) Introduction. In this study we used the pregnant and non- pregnant ovaries and uteri, and also the bladder, in order to com- pare our genital results with those for an organ with a supposedly non-dynamic function. We desire to express our thanks to Dr. Robert T. Frank for his interest, and for his kindness in placing at our disposal the genital material employed. Methods. i. Preparation of extracts. {A) Five grams of finely divided fresh material, washed free from blood and thor- oughly triturated with sand, were treated with 100 c.c. of water and allowed to stand for 24 hours, under toluene, with frequent shakings. At the end of this time the extract was filtered through muslin, made up to 100 c.c, and aliquot portions used for the en- zyme tests. {B) Glycerol extracts were made in the same manner. 2. EsTiMATiON OF ENZYMES. In cach casc, a control test was made with boiled extract. Lipase. A mixture of 10 c.c. of the extract, 0.5 c.c. of neutral ethyl butyrate and i c.c. of toluene was placed in a bottle and allowed to digest at 40° C. After 24 hours n/20 sodium hydroxide Solution was used to determine the acidity, with Phenolphthalein as the indicator. From this amount was subtracted the "control" acidity (10 c.c. of extract and i c.c. of toluene). Amylase. To 10 c.c. of i per cent. freshly prepared starch *The first paper in this series (a general review of the subject) has been accepted for publication in a later issue of the Biochemical Bulletin. The second paper appeared in the January issue of the Journal of Biological Chetn- istry, 1913, xiii, p. 511. See also Biochemical Bulletin, 1911, i, p. 115. ^ Mr. Lefkovics died shortly after the completion of this work. See Bio- chemical Bulletin, 1912, i, p. 573. 233 234 Biochcmistry of the Female Genitalia [Jan. paste were added lo c.c. of extract and i c.c. of toluene, and the mixture allowed to digest at 40° C, until duplicates no longer be- came blue with iodin Solution. At this point the bottles contain- ing the digestive mixtures were placed in boiling water to stop the digestions simultaneously. The contents of each bottle were then made up to 50 c.c. and run from a burette into boiling Fehling Solu- tion, acetic acid and potassium ferrocyanid being used to determine the end point. When the amount of sugar in the digestive mix- ture was less than that required completely to reduce the copper, a Standard glucose Solution was employed for that purpose. Acid- and alkali-proteases. Ten grams of gelatin were dis- solved in 100 c.c. of warm i per cent. Solution of sodium fluorid col- ored with methyl violet. This Solution was drawn into glass tubes I mm. in diameter and the filled tubes quickly placed in cold water to congeal the gelatin. The tubes were then cut into lengths of 2-3 cm. Ten c.c. of extract were placed in a small bottle closed with a perforated cork through which the gelatin tubes could be inserted; i c.c. of toluene was added and the digestions kept at room temperature for 48 hours. In the estimation of acid-protease (pep- sin) the mixture was made acid with 0.2 per cent. hydrochloric acid Solution and for alkali-protease (trypsin) they were rendered alka- line with 0.5 per cent. sodium carbonate Solution. Table showing enzyme values of pregnant and non-pregnant ovary, uterus and bladder of sheep A. Non-pregnant condition Aqueous extract Glycerol extract Organ Lipase, c.c. Amy- lase, mg. Acid- pro- tease, mm. Alkali- pro- tease, mm. Lipase, c.c. Amy- lase, mg. Acid- pro- tease, mm. Alkali- pro- tease, mm. Ovary 0.65 305 1-3 5 10 10 2 I 1-5 2 I 0 0.65 1-95 1-35 6 5 IS 6 3 3 0.25 4 0 Uterina mucosa Bladder mucosa B. Pregnant condition Ovary Uterina mucosa . Bladder mucosa. 1-3 10 2 7 2.0 7 7 8.35 25-5 I 7 8.3 12 3 i.i 7.5 1-5 0 1-35 6 4 0.5 5 o The accompanying table presents the results obtained in this Study. The lipase values are given in terms of the amount of ji/20 1913] Thuisco A. Erpf-Lefkomcs and Jacob Roscnhloom 235 sodium hydroxid Solution necessary to neutralize the acidity devel- oped by i gram of tissue. The amylase values are given in terms of the amount of maitose in mg. formed per gram of tissue. The acid- and alkali-protease values are given in terms of the number of millimeters of gelatin digested in a tube by i gram of tissue. The results show that lipase and amylase were most abundant in both the ovaries and uterine mucosae of pregnant animals. Preg- nancy had no quantitative effect on the acid-protease (pepsin), but alkali-protease was increased in both the ovary and uterine mucosa. The bladder extracts contained lipase, amylase, and acid-protease (pepsin), but no alkali-protease (trypsin). THE BIOCHEMISTRY OF THE FEMALE GENITALIA^ 4. On the absence of certain enzymes from the human chorion^ JACOB ROSENBLOOM (Laboratory of Biochemistry, University of Pittshurgh, Pittsburgh, Pa.) During pregnancy the chorion frondosum unites with the de- cidua serotina to form the placenta. The enzymes of the placenta^ have often been studied, but I am unable to find any record of a study of the enzymes of a human chorion. Through the kindness of Dr. Robert T. Frank, of New York, the vvriter received a fresh human chorion for such an investigation. The available chorion, which weighed 10 grams after it had been washed free from all blood by means of a small amount of water, was finely minced and two portions, 5 grams each, were taken for the preparation of extracts, which were made as follows : ( i ) The material was triturated with sand, 200 c.c. of water added, and the mixture allowed to stand with frequent shakings for 24 hours under toluene. At the end of that time, the extract was filtered through muslin, and the filtrate used in the tests for various en- zymes. (2) A glycerol extract was made in the same way. In testing for enzymes, control portions were always taken, which were boiled before their addition to the Solutions or suspensions of Substrate.'' The accompanying table presents the data obtained in this study. The data show that both glycerol and aqueous extracts of a human * See the first footnote of the preceding paper in this issue of the Biochem- ICAL Bulletin. " The analytic work was done in the Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York. ^ Frank: Surgery, Gynecology and Obstetrics, 1912, xv, p. 561. ' See the description of methods in the paper preceding this one. 236 I9I3] Jacob Rosenhloom 237 chorion were free from amylase, sucrase, maltase, lactase, lipase. peptidase, ereptase, acid-protease and alkali-protease. Table showing results of ensyme tests in glycerol and aqueous extracts of a human chorion Enzyme Substrate Glycerol extract Aqueous extract Amylase Sucrase Starch Absent Absent Sucrose Absent Absent Lactase Lactose Absent Absent A'Ialtasp Maltose Absent Absent Lipase Ethyl butyrate Glycyltryptophan Witte Peptone Absent Absent Ppntida'sp Absent Absent Absent Absent Acid-protease Alkali-protease Gelatin and fibrin^ Gelatin and fibrin^ Absent Absent Absent Absent It would seem, from the results of this study, either that the enzymes of the placenta are formed at a comparatively late period or that the decidua serotina furnishes the enzymes which subse- quently appear in the placenta. Possibly the presence of blood in the placenta accounts for the occurrence of certain enzymes that have been detected in the placenta. ° Extract rendered acid with 0.2 per cent. hydrochloric acid Solution. * Extract rendered alkaline with 0.5 per cent. sodium carbonate Solution. A DEPARTMENT OF BIOCHEMICAL RESEARCH AT VINELAND, NEW JERSEY^ AMOS W. PETERS To the Training School at Vineland, N. J., belongs the credit for the first establishment anywhere in the world of a biochemical laboratory as one means of investigation of the problem of feeble- mindedness in children. To the writer of this article has fallen the honor as well as the heavy duty of testing what are the possibilities of biochemical research in the field of feeble-mindedness. The large problem which this unfortunate affliction of a considerable portion of humanity presents to organized society is becoming daily more evident, as its economic bürden and its social consequences force themselves on public attention. Research on the problem is a crying need not simply f rom the humanitarian Standpoint, but also as an economic necessity. The care and treatment of these cases and the governmental management of this problem, including its amelior- ment and prevention, will in the future rest on the basis of data ob- tained by scientific research. At present we are proceeding on a very small amount of such data and we are just discovering, after some preliminary efforts made in the psychological direction, how extensive and manysided this problem is. What assurance have we that our present method of dealing with the problem is in rational accord with the nature and origin of the condition? Our proce- dures are in the stage of costly empiricism and in the very infancy of scientific investigation. It is therefore an important step for- ward when this Institution ventures to add to its present psycho- logical method of investigation that of the rapidly growing and fundamental science of biochemistry. The need for this additional method of attack, and the tendency of expert thought toward it, is well illustrated by the following quotation from the words of a leader in the study of problems of psychopathology, Dr. Southard : ^Reprinted from The Training School, 1912, ix, p. 70 (Sep.). 238 1913] Arnos W. Peters 239 The majority of cases of mental diseases are, I am convinced by special studies, characterized by the occurrence of obvious brain lesions, i. e., even in the present stage of science they possess a structural pa- thology. Do they therefore possess no functional pathology? Their possession of the two aspects is a truism. Should we not study both aspects ? Furthermore, suppose we learn that, whereas three quarters of our cases of mental disease exhibit obvious irrecoverable brain lesions, another quarter falls to shovv these. Suppose the methods of micro- scopic research should still fall to show in many cases essential or irreversible brain lesions, should we not stultify ourselves if we did not abandon jor the research campaign both that psychopathology which has taught us the main course of our disease and the neuro- pathology which has proved usefully negative? Should we not repair at once to the chemistry of metabolism, the physiology of internal secretions, and the entire point of view of psychopathology? Dis- coveries in the latter fields, concrete and pertinent facts, would carry US back to the tissues and back to the processes of the nervous System, to neuropathology, structural and functional and to psychopathology, and enlighten many dark corners therein. He who adheres to the clas- sical Problems as they He within the teaching divisions of any science is not apt to change the face of that science.^ It is the method of science to develop the ultimate truth with its numerous and involved qualifications, which are due to the infi- nite complexity of natura itself, by means of hypotheses. These are repeatedly set up and repeatedly confirmed or refuted and re- placed by others of better construction in view of previous expe- rience. Whether the hypothesis was exactly correct or not — ulti- mately tenable or untenable — ^becomes a matter of no practical sig- nificance. The testing of hypotheses develops facts, and facts, dem- onstrated and adequately qualified truths, are the precious heritage of the race f rom previous human endeavor. Now, then, the hypoth- esis which underlies the use of the biochemical method in this problem is that which postulates, simply, a relation between patho- ^ Southard, E. E. : " Psychopathology and Neuropathology : The Problems of Teaching and Research Contrasted," Amer. Jour. of Psychol., 23 : 230-235, 1912. Read by invitation in a Symposium at a meeting of the American Psycho- logical Association, December 28, 191 1, at the Hospital for the Insane, Wash- ington, D. C. 240 Biochcmical Research at Vineland, N. J . [Jan. logical mental action on the one band, and the physical condition of the brain and body on the other. We will not discuss this propo- sition — no, this hypothesis — with our readers. It is not worth while. We only wish gently to call their attention to it and to prevent them from shying at this subject on theoretical grounds. This, then, is our generalized hypothesis, and it is clear that finally our logical efforts will be directed toward the correlation of data, psy- chological and biological, taken in their widest sense. This part of our effort will be small, however, compared with the requirement for painstaking and persistent experimental determination of facts which are the real values we are seeking. In this connection it should be noticed that the present literature of chemical biology contains numerous concrete examples of investigations which have an evident relation to the problems of psychopathology viewed from the broad Standpoint of Southard, as above quoted. In future numbers of the Biochemical Bulletin we shall, from time to time, present our readers with notes and criticisms on this literature. It is important that the general aim of this biochemical effort should not be misunderstood, nor its results misinterpreted. The primary and only initial object is to contribute toward the elncida- tion of the conditions of psychopathological action by means of the biochemical method. The curing of tuberculosis was an entirely premature and abortive expenditure of effort before the elucidation of the cause and conditions of that disease. When once these con- ditions have been adequately determined, valuable applications of the new knowledge always follow, and sometimes with astonish- ing results. But now we are only in the beginning of the period of strenuous seeking after much needed information. We wish also to emphasize that we regard the biochemical as only one, but after the psychological the next in importance, of the methods that are available for determining conditions of abnormal mental action. We picture our final understanding of these conditions to be a com- posite and correlated result obtained by different methods, none of which alone would have ever yielded adequate knowledge. Now we are asked just what, concretely, is the field of application of biochemistry to the problems of feeble-mindedness. This question could be best answered by illustrations from the 1913] Arnos W. Peters 241 literature of investigation along biochemical lines; but, as above stated, this we shall continue to present in future numbers of this Bulletin. At present, before we have actually begun oiir own experimental work, we can give only an outline of the topics we plan to pursue to such extent as workers and material resources per- mit. The field is so rieh as to tax the judgment in the selection of the first attacks, and we are well aware that we are outlining more than our present resources permit to be done in the near future. Publicity and hearty Cooperation with other individuals and in- stitutions is, of course, our policy. In the present article, however, and at the very beginning of our work, we are describing only the nature of the work to be done without specific detail regarding par- ticular problems or methods. Our primary line of effort to which the others are logically re- lated is the study of the conditions of metabolism presented by the feeble minded of this institution. Very few studies of this nature have been made, and the material for them is here presented under favorable conditions for investigation. Promiscuous examinations or experiments will not be made. But at first typical and psycho- logically well-known and defined cases will be selected. For orien- tation they will at first be studied in their undisturbed condition before the experimental factor is introduced. By metabolism we understand, of course, the sum total of the chemical changes which a living organism continually performs within its tissues and upon the substances which it utilizes. The progress of biological science has made the term practically synonymous with the processes of life in so far as they are non-psychical. Under this head we intend to subject the idea of intoxication, whether endogenous (autointoxi- cation) or exogenous, to a rather thorough testing, especially in its relation to psychopathological phenomena. Two other related topics with which we will be compelled to deal in this connection pertain to the subject of glandulär secretions and that of lipoid or phosphorus metabolism. It is well known that the method of glan- dulär feeding is extensively practiced in psychopathological cases and institutions. It appears that this is usually done in a promiscu- ous way with but little of the Clements of control experiments or of adequate therapeutic indications. In our future notes and 242 Biochemical Research at Vineland, N. J. [Jan. criticisms on the literature, we shall treat this subject more fully. It seems a pity, f rom both the scientific and the humanitarian stand- point, that such potentially valuable experiments on human subjects should pass without an examination of their most important factor — that of the metaboHsm of the physiologically much afifected subject. Our second line of effort will be that of lipoid and brain chem- istry. It will not be pursued extensively until we have obtained, from the observations of metabolism and the third line of effort described below, some indications of the directions in this large and inherently difficult field that it would be best to pursue. Contrary to the common Impression, the present literature already shows the important and practical bearing of this little developed field of chem- istry on the psychopathological problem. A third kind of work which in the near future will become practically inevitable is the study of heredity, growth and develop- ment from the particitlar angle of view of the psychopathologist. It is well known how strongly the scientific and the public attention is now fixed upon the hereditary and congenital (if not hereditary) factors involved in the conditions of abnormal mental action. With- out going into detail, we wish to emphasize the fact that the hered- itary factor in this problem by no means removes it from the field of biochemical study, nor makes the pathological conditions any less amenable to elucidation by that method. In fact, the only real hope for the elucidation of the processes of reproduction and heredity seems, in the light of experiments already made, to lie in the direc- tion of an intimate knowledge of the chemistry and physics of the protoplasmic basis of Hfe. Biochemical Laboratory, Training School, Vineland, New Jersey. BIOCHEMISTRY IN NEW YORK TWENTY YEARS AGO^ E. E. SMITH The Status of biochemistry in New York City, in 1891, is well depicted by an incident that occurred in that year at the library of the New York Academy of Medicine. The writer had just come to the city and was seeking, for reference, a copy of Maly's Jahres- bericht für Thier-Chemie. In reply to an inquiry, he was informed that such a work was not in the Academy library and would most likely be found at the Veterinary College. Chemistry was, indeed, well established at that time in the curriculum of the medical schools of the city, but it consisted largely of descriptive organic and in- organic chemistry, and found relatively scant application in physiol- ogy and pathology.^ To the younger graduates of Yale, the pioneer work of Chitten- den was known, but it had not been implanted here. It did lead, however, to the Inspiration of Dr. C. A. Herter, then beginning to specialize in neurology ; and when he came to realize, as he soon did, how closely related was this field to the pathology of nutrition and determined to establish a laboratory for the investigation of this sub- ject, he naturally turned to Prof. Chittenden for someone with tech- nical training to undertake this work. ^For previous special contributions to the history of biological chemistry in New York see the Biochemical Bulletin, 191 i, i, p. 245, and 1912, i, p. 377. *"To appreciate the significance of all this, it should be remembered that, with the exception of the work in the pathological laboratories of the Colleges, the work of the Board of Health, and the work done by Dr. S. J. Meltzer, there was practically no scientific investigation in medicine worthy of the name in New York City at that time (when the 'Laboratory of C. A. Herter' was created). What was true of New York was essentially true of the country at large. . . . Dr. Herter found the study of the nervous System so abounding in confusion that he soon turned his attention to chemical problems, especially those connected with pathological conditions. Among those intimately associated with him in this work have been E. E. Smith, A. J. Wakeman and, of late, H. D. Dakin." Lusk: Science, 191 1, xxxiii, p. 846. [Ed.] 243 244 Biochemistry in New York Twcnty Ycars Ago [Jan. How little Dr. Herter appreciated the equipment that would be required is indicated by bis Suggestion that the work be conducted in the art studio of his brother, then absent in Europe. It did not re- quire many months, however, to reveal to him something of the technical scope of the field to which he was to devote the two decades permitted him for the completion of his hfe work, At the outset, he desired adequate equipment ; and when he returned f rom his sum- mer rest, in 1891, he was enthusiastically appreciative of the well- equipped laboratory awaiting him in the basement of his residence. This, three years later, was transferred to his newly built home where the entire upper floor, 50 X 100 feet, was devoted to this special work. It was no unusual, though an unique, experience to house in his animal room, rabbits, dogs, monkeys, füll grown hogs, and other animals in an array that would have astounded the unin- formed passerby in this district of elegant homes. It was not, however, the equipment that invites attention to Herter's early work nor was it the display he made of his devotion to this new field. Both were modest. What has lingered and al- ways will remain in my memory of twenty years ago is the serious- ness with which the work was undertaken. In later years, when his life was so filled with the success and magnitude of his work, it was to be expected that he would throw all that was in him into it ; but that he should have devoted himself so largely to it when its value was uncertain, or at least not demonstrated, indicates the profound purpose that was leading him to undertake it. Not in- frequently, when night had come and the day's work was done, we forgot ourselves in both discussing what we had attempted and planning what we hoped to do; finally awakening to a realization that we were neglecting the proper demands of our respective family circles. Only a fraction of the work done at that period was ever pub- lished. The first paper, " Uric acid elimination in health and dis- ease," was a record of investigations inspired by the extravagant Claims of the English physician, Haig. We differed with him in many important conclusions. We did not find that uric acid forma- tion was always constant and that elimination was determined by the degree of alkalinity of the blood, but found it to vary with the diet 1913] -E- E. Smith 245 in health and with conditions unknown to us in disease. Moreover, we did not recognize it as a causative factor in the many diseases to which this role was assigned by Haig, but rather regarded its in- creased elimination as a result of the morbid condition. Horbac- zewsky's work did not come to our attention tili after the publication of this first paper. The study of epileptics led to the conclusion that, in some cases of so-called idiopathic epilepsy, the onset of the seizures was deter- mined not by a uric acid accumulation, as claimed by Haig, but by a toxemia of gastro-intestinal origin. Indican, which had received scant attention from clinicians up to this time, was found to be a valuable index to the condition; as was also the elimination of phenol and ethereal sulphates. The occurrence of these products in undue quantity seemed to bear a direct relation to the onset of the seizures. As was natural, there followed an elaborate study of the gastro- intestinal conditions in other diseases, especially those with marked neurotic manifestations ; and the conclusion was reached that the neu- rotic exacerbations in many conditions were due to a gastro-intes- tinal toxemia. An entirely different line of study was the presence of lead and its distribution in cases of chronic lead poisoning. The results of these analyses were never published. Investigations to which was devoted a very great amount of work and which covered a very wide scope, as well, were the studies of the causes of uremic intoxication. The many theories which had been elaborated to explain this condition were each in turn subjected to investigation, involving extensive animal experimentation as well as intricate chemical research. The work covered several years and the results were of very great interest to us, and certainly influenced Herter's later work, but they were never published. During this period, there was a striking lack of activity in re- search in chemical pathology in New York; indeed, this was only the time of the awakening of general interest in the most active Cen- ters of medical science. Aside from Herter's work, only a single paper presented at the Academy of Medicine in that period comes to my mind ; and that was so glaringly f aulty that one hesitates to con- sider its sincerity. Von Noorden's Pathologie des Stoffwechsels, which appeared 246 Biochemistry in New York Twenty Years Ago [Jan. at this time, was as a beacon light on a dark night. I received my copy before Dr. Herter's attention was called to the work. He saw it on my table and, borrowing it, informed me shortly afterwards that if I really wanted a copy I had better send for another. True to bis word, I never saw this first copy again and I doubt not that it rests now in his library well worn with eager study, which it received at that time. My second copy served a similar purpose in my own hands. My personal relation with Dr. Herter was interrupted by the decision to study medicine. The modest beginnings of his work, which hardly interested more than a narrow circle of personal friends and admirers, grew to a proportion that brought him into national and, indeed, international prominence. A man of unusual personal charm and sincere purpose, he demonstrated how personal opportunity could find unselfish application to the benefit of his fellowmen in the field of applied medical science. Laboratory, 50 East Forty-first Street, New York City. IMMUNITY IN SOME OF ITS BIOCHEMICAL ASPECTS^ CHARLES FREDERICK BOLDUAN (Department of Health, New York City) (WITH PLATE 2) Contents. — Infection, 247, Immunity: natural, 248; acquired, 248; speci- ficity, 249; additional defenses, 249; Behring's discovery of antitoxin, 250; bac- teriolysins, hemolysins (cytolysins), 250; complement and immune body, 251; agglutinins, 251; Opsonins, 252; precipitins, 252; anti-antibodies, 252. Immunity from the Standpoint of cell niitrition, 253; Ehrlich's " side chain " theory, 253; receptors, 254; Weigert's " overproduction " theory, 254; natural immunity, 256; anaphylaxis, 256; results of enteral and parenteral introduction of protein, 257; significance of period of incubation, 258, and bearing on intoxication by infection (endotoxins), 258. Modern chemotherapy according to Ehrlich, 259. Chemical nature of antibodies, 260. Infection. One of the most interesting problems to all of us is that presented by disease, especially by what we call " infectious " disease. Under this term we mean disease produced by living organ- isms or their products. Among the organisms producing disease in man are bacteria, molds, yeasts, and protozoa, and we may con- veniently speak of these collectively as germs. The manner in which the various germs produce disease in man, their mode of entrance into the body, the part of the body attacked — all these differ considerably with the different germs. Some like the bacillus of diphtheria and the bacillus of tetanus (lockjaw) secrete very powerful poisons, and while the germs themselves do not penetrate deeply into the body tissues, their poison is absorbed and gives rise to severe Symptoms. In the case of other germs, for example the tubercle bacillus, the organisms penetrate deeply into the body tissues and there multiply. In their growth they destroy the cells in which they lodge and, by their poisons, affect the entire body. ^ Lecture delivered, by invitation, under the auspices of the Columbia Univer- sity Biochemical Association, at the College of Physicians and Surgeons, Novem- ber 16, 1912. 247 248 Immnnity in Some of its Biochemical Aspects [Jan. Most germs, for some obscure reason, affect by preference cer- taiii parts of the body. The typhoid bacillus usually lodges in the wall of the small intestine; the meningococcus prefers the lining membranes of the brain and spinal cord; the gonococcus is very prone to attack the mucons membrane of the genital organs and of the eye; the pneumococcus affects chiefly the respiratory organs; the diphtheria bacillus lodges in the throat and nasal passages ; the malaria parasite lodges only in the red blood cells; and certain molds affect only the skin. Immunity. Natural immunity. It is very well known, however, that certain infectious diseases occur naturally only among some of the lower animals and do not affect man, while conversely, others appear to attack only man. Among the latter may be men- tioned typhoid fever, syphilis, gonorrhea. In speaking of the re- sistance evidently possessed by certain species we make use of the term natural immunity. Thus chickens and frogs possess a natural immunity against tetanus (lockjaw) ; dogs, a natural immunity against anthrax; goats, a natural immunity against tuberculosis ; and man, a natural immunity against certain diseases of cattle. This natural immunity, however, is not always absolute. Chickens, for example, can be infected with tetanus if their bodies are chilled, and frogs can be made susceptible to tetanus by keeping them un- duly warm. Acquired immunity. Another form of immunity is that ob- served in individuals who have had one attack of a particular in- fection; thereafter they are practically safe from a second attack. These individuals are said to possess an acquired immunity. This form of immunity is well illustrated in scarlet fever, measles, small- pox, yellow fever. Often this immunity lasts throughout the life- time of the individual though there are exceptions. In studying this form of immunity, Pasteur conceived the idea of artificially producing an attack of a given infection in order to Protect the individual against another attack. He realized that it was necessary, however, to so control matters that the original attack should run a very mild course and not endanger the life of the individual. After considerable experimental work, Pasteur found that this could be accomplished by artificially weakening the 1913] Charles Frederick Boldnan 249 bacteria with which the original attack was produced. Subse- quently Salmon and Smith, in this country, showed that it was not necessary to produce even a mild attack of the disease by injecting living germs, but that the injection of dead germs would produce an immunity against that particiliar infection. Specificity of acquired immunity. Acquired immunity, whether caused by a previous natural attack of the disease, or artificially by the inoculation of living or dead germs, is always strictly specific; that is, the protection extends only to the particular disease which has previously occurred or against germs of the kind previously injected. An attack of scarlet fever protects only against scarlet fever but not against measles. Inoculating an individual with ty- phoid bacilli protects him only against typhoid fever, but not against dysentery, plague or cholera. This acquired immunity is often transmitted from mother to offspring, transmission being effected mainly, according to Famulener, through the Colostrum. Additional natural defenses against DISEASE. Beforc ex- amining into the nature of specific acquired immunity, let me call attention to certain important means by which the body is protected against infectious diseases in general. Many of these means are so commonplace that their significance is often overlooked. The protection afforded by the unbroken skin is undoubtedly one of the most important means of defense. A similar protection, though less effective, is afforded by intact and healthy mucous mem- branes. The acid gastric juice undoubtedly destroys large numbers of swallowed germs. It has been found that fresh blood serum is able to kill a considerable number of germs, and this is therefore another mode of defense. The white blood cells (leucocytes) appear to be designed especially to destroy invading micröorganisms. These cells take hold of, or rather engulf, the germs and digest them. Still another mode of defense is seen in what takes place in abscesses. When these are examined, it is found that the body has built a wall of cells around the infected area, thus shutting off the germs and their poisonous products from the rest of the body. Finally, mention may be made of the collection of fluid, i. e., of serum, as perhaps a means designed to dilute irritant poisons (pleu- risy, Peritonitis). 250 hnmunity in Some of its Biochemicol Aspects [Jan. The means of protection we have just recited are all general in their action, that is, not directed specifically against only one partic- ular infection, Let us now return to a consideration of the specific acquired immunity already mentioned. Behring's DISCOVERY OF ANTITOXIN, Most of our knowlcdgc concerning specific acquired immunity dates from Behring's discov- ery of the antitoxins of diphtheria and tetanus, in 1890. Behring found that when an animal is injected with gradually increasing doses of toxin, e. g., with diphtheria toxin, it is able, after a time, to withstand doses of the poison sufficient to kill hundreds of animals not so treated. He found that the blood serum of the treated animals contained something which neutralized the diph- theria poison, and rendered it harmless. This something he called an antitoxin. Investigation showed that the antitoxin was strictly specific, the antitoxin for diphtheria neutralized only the toxin of diphtheria, the antitoxin for tetanus, only that of tetanus. Bacteriolysins and hemolysins (cytolysins). Another important advance was made in 1894 when Pfeiffer showed that, just as an animal injected with gradually increasing doses of toxin produces an antitoxin in its blood, so also, when injected with bac- teria (cholera bacilli), it produces substances which kill and dissolve the injected micröorganisms. We have already said that fresh blood serum is able to kill a considerable number of bacteria, and that this probably constitutes one of the defenses of the body against bacterial Invasion. When the animal is injected with gradually in- creasing amounts of bacteria, however, this destructive power in- creases very greatly, but only for the particular kind of bacterium used for injection. In other words, the action is strictly specific. If an animal is injected with cholera bacilli, the serum will, after a time, even in very small doses kill enormous numbers of cholera bacilli ; tested against typhoid bacilli, or on other bacteria, its de- structive effect is merely that of normal serum from an untreated animal. When the action of the serum is studied under the micro- scope, it is seen that the bacteria are actually broken up and dis- solved. Hence such a serum is spoken of as a " bactcriolysin." Since the bacteria are also killed by this action, we also use the term " bactericidal " in speaking of such a serum. 1913] Charles Frederick Bolduan 251 It has been found that this action may be developed against cells other than bacteria. When red blood cells are used for the injec- tions, the serum acquires dissolving properties for these ; and here again the action is strictly specific, so that when blood cells from a chicken are injected into an animal, the serum of the injected animal acquires increased solvent powers only for chicken blood cells, not for blood cells of other animals. Sera directed against blood cells are usually spoken of as hemolysins. The term cytolysin is used to embrace all these cell-dissolving sera. Complement and immune body. Investigation has shown that the mode of action of these dissolving sera is somewhat complex, and consists of the Joint action of two substances. It may be re- called that this dissolving action was observed in fresli serum. Serum which had stood for several days no longer possessed this property. The researches of Metchnikoff and Bordet showed that the füll solvent power could be restored by the addition of a little fresh serum, even from a normal, untreated animal. Evidently, then, of the two substances concerned in this dissolving action, one is quite stable, and the other highly labile. The labile substance, derived from a normal untreated animal, is spoken of as the com- plement; it is not specific. The stable substance, present only in the serum of the treated animal, is called the immune body; it is highly specific. When an animal is repeatedly injected with grad- ually increasing doses of bacteria, or other cells, it responds by man- ufacturing large quantities of this "immune body" directed spe- cifically against the injected cells. The complement is not increased in the process. Agglutinins. When the serum of an animal which has been repeatedly injected with gradually increasing doses of bacteria is brought into contact with some of the bacteria, careful Observation under the microscope reveals a very interesting series of changes. Thus, if typhoid bacilli are mixed with a specific antityphoid serum (obtained, let us say, from a rabbit previously injected with typhoid bacilli), one notices, first, that the motility of the bacilli becomes markedly diminished. This is followed by the gradual collection of the bacilli into clumps. At the end of an hour or two, in place of countless bacteria moving quickly through the field, one sees 252 Iinmiinity in Some of its Biochemical Aspccts [Jan. merely several groups of absolutely immobile bacilli. If the reac- tion is feeble, the clumps are small, and one finds comparatively many isolated and, perhaps, also moving bacteria. This phenome- non is spoken of as aggliitination, and the substance in the serum which brings this about is called agglutinin. The clumping thus broiight about does not kill the bacteria ; moreover, it makes no dif- ference whether the serum is freshly drawn or has been kept for some time — it will agglutinate equally well ; and it does not require the addition of fresh serum as do the bacteriolysins. Like the an- titoxins and the bacteriolysins, the agglutinins are strictly specific, so that serum from an animal previously injected with typhoid bacilli will agglutinate only typhoid bacilli; one from an animal injected with dysentery bacilli, only such bacilli, etc. Opsonins. We have already said that the white blood corpus- cles (leucocytes) take up bacteria and destroy them. Wright, of England, showed that certain substances in blood serum have the power of increasing the appetite, as it were, of the leucocytes, and furthermore, that the amount of these substances can be increased by properly graduated injections of the appropriate bacteria. These substances he called Opsonins. They are specific, just as are the antitoxins, the bacteriolysins, and the agglutinins; that is to say, when typhoid bacilli are injected into the body, only the Opsonin for typhoid bacilli is affected; when staphylococci are employed, only the Opsonin for such organisms is affected, etc. Precipitins. If, instead of injecting bacteria or other cells, we inject an animal with Solutions of albuminous material ; for example, if we inject a rabbit with chicken-egg albumin, we find that the rabbit serum acquires the power to produce a precipitate when mixed with chicken-egg albumin. This action, too, is highly specific, so that if the serum is tested against the albumin from any other animal, e. g., from a duck tgg, no precipitate will be produced. If a rabbit is treated with human blood, the rabbit serum will produce a pre- cipitate when mixed with human blood, but not when mixed with any other blood. The substance in the treated animal's serum is spoken of as a precipitin. This test, as you probably know, is used in criminal cases to determine whether or not certain stains are those of human blood or otherwise. 1913] Charles Frederick Bolduan 253 Anti-antibodies. But even this list does not exhaust the list of "antibodies" which it is possible to produce. When enzymes are injected into an animal, the latter responds by producing anti- enzymes, and when certain " antibodies " are injected, anti-anti- bodies are produced. Immunity regarded from the Standpoint o£ cell nutrition. The whole subject of infection and immunity, and particularly the production of the antibodies just discussed, is best appreciated when regarded from the Standpoint of nutrition; for what, after all, is this apparent conflict between bacteria and the animal body but the mutual attempt of each to use the other for food. Let it be noted that production of the various antibodies takes place only when the bacteria or other allen cells are introduced parenterally, i. e., by ways other than the gastrointestinal tract. We may ex- plain this by saying that when introduced by the gastrointestinal tract the molecules of the food stufifs (organic) are split up and rebuilt in such a way that the material requires no further extensive altera- tion in order to serve as food for the various cells of the body. In the animal body this breaking down and building up is delegated to certain specialized cells ; in the primitive organisms, however, we must believe that each cell was required to break down and build up its own food. When parenterally situated cells of the higher animal are thus presented with the unprepared food which the parenteral introduction brings them, it may be assumed that they behave as does the primitive cell, and proceed to lay hold of and attempt to assimilate the injected material. With this introduction, we may pass at once to a consideration of Ehrlich's "side chain theory," which still offers the best explanation for the formation of the various antibodies. It is essentially a theory of cell nutrition. Ehrliches "side chain" theory. According to Ehrlich's conception, every cell is armed with a large number of chemical groups whose function is to lay hold of nutriment and anchor this in the cell. These groups he calls receptors or side chains. Only such substances can serve as nutriment which can thus be bound chemically to the cell protoplasm. He believes that the receptors are of at least three different kinds, and speaks of receptors of the " first Order," "second order" and " third order." These are best de- scribed with the aid of a diagram such as the accompanying one. 254 Immunity in Some of its Biochemical Aspccts [Jan. Rcceptors. In view of what has been said it is obvious that the simplest mechanism by which the cell can lay hold on food par- ticles is a receptor which merely anchors food, leaving the digestion entirely to the cell proper. It may be assumed that this type of re- ceptor suffices for comparatively small food molecules. When a larger and more complex food molecule presents itself, it may be as- sumed that a receptor would be reqiiired which not merely anchors but also acts on the food molecule to make it more readily assimi- lable. These two types are shown in A and B respectively (Plate 2). It will be noted that the receptor in B possesses an anchoring group {h) and an active group (Z) which acts on the food molecule. It is conceivable that an economy in structure could be effected in B, if, in place of the active group (Z), there were merely provision for the anchoring of an enzyme. The active group (Z) could then be dispensed with and the enzyme called upon only when a food mole- cule had been anchored by the receptor. Such an arrangement is shown in C (Plate 2). Weigert's " over prodtiction " theory. At this point you may very properly inquire why we assume the existence of receptors of these types. To explain this, let us go back to the productiön of an- titoxin in response to injections of toxin. It will be recalled that the toxin can be neutralized by the antitoxin. Moreover, and this is the important point, this action is strictly specific, so that, for example, diphtheria antitoxin neutralizes only diphtheria toxin; against any other toxin it is absolutely without effect. Since it can be satisfactorily shown that the antitoxin is not altered toxin, it is necessary to explain the productiön of antitoxin by the body cells. We have said above that only such substances can serve as nutriment for the cell which can be tied chemically to the cell pro- toplasm. Expressing this in terms of receptors, we would say that only such substances as possess groups fitting the receptors of the cell can be anchored to the cell. In thinking of these groups and the way in which they fit together, we must have stereochemical rela- tions in mind. Ehrlich cites with approval a simile used by Emil Fischer, saying that the relation of the two groups must be that of lock and key. Granted, now, that certain food molecules have been anchored by fitting cell receptors, what follows? To explain this. CM < H ■X < c (U H tu 3 CO _l < o UJ I Ü O CO 1 M- ^ - Q c ß3 1913] Charles Frederick Bolduan 255 Ehrlich makes use of an hypothesis advanced by Weigert in con- nection with regeneration. According to this, physiological func- tion and structure depend upon an equilibrium of the tissues that is maintained by virtue of mutual restraint between their component cells. Destruction of a single integer or group of integers of a tissue or a cell removes a corresponding amount of restraint at the point injured, and therefore destroys equilibrium. This permits of the abnormal exhibition of bioplastic energies on the part of the remaining uninjured components, which activity may be viewed as a compensating hyperplasia. When such bioplastic activity is called into play there is always hypercompensation ; that is, there is always more plastic material generated than is necessary to compensate for the loss. Thus far Weigert. Ehrlich, in line with Weigert's over production theory, points out that, owing to the combination of toxin with receptors of the cell, the receptors are practically lost (at least temporarily) to the cell; that the cell or its fellows now produces new receptors to re- place this loss ; but that this production always goes so far as to make a surplus of receptors ; that these receptors are thrown off by the cell, as unnecessary bailast so to speak, and then circulate in the blood as antitoxin. The same substance therefore, which, when part of the cell, combines with the anchoring group of the toxin, enabling this to act on the cell, when circulating free in the blood combines with and satisfies this anchoring group of the toxin, and prevents the poison from combining with and damaging the cells of the organism. It is obvious that this affords a complete explanation of specificity. If we now go back to our diagrams (plate 2) we shall see that all of the antibodies discussed above fit readily into this scheme. So far as the antitoxins are concerned, these would be merely receptors of the first order, thrust off from the cell and circulating in the serum. Agglutinins and precipitins would belong to the second Order; they have the active group as an integral part of the receptor. The hemolysins and bacteriolysins would be in the third order; fresh serum is active because it contains complement, but since the complement is very labile, the serum after a while contains only the immune body, i. e., that part of the receptor which anchors the food molecule on the one band and the ferment substance on the other. 256 Imniunity in Some of its Biochemical Aspccts [Jan. Explanat'wn of natural immimity. Ehrlich's views concerning the necessity for fitting receptors in order that a microörganism may attack the body cells afford a satisfactory explanation of the im- munity possessed by certain animals against particular infections. Thus, it is obvious that the entire absence of receptors fitting a cer- tain microörganism renders the body immune against infection by that microörganism. Moreover, the location of the receptors may be responsible for the relative immunity of an animal under natural conditions and its susceptibility when these conditions are changed. Thus, if receptors for a particular poison are present both in a vital tissue, like the brain, and in an indifferent tissue, like the muscles, it is clear that while an intracerebral injection of the poison might prove fatal, an intramuscular one might be almost without effect. Anaphylaxis. Most of you are probably familiär with the work of Vaughan and Wheeler concerning the cleavage products of proteins, and recall that some of their products were highly poison- ous. Certain observations of the past few years indicate that, in the parenteral digestion of proteins, similar cleavage products are pro- duced. Historically this aspect of immunity may be said to date from 1906, from the studies undertaken by Ehrlich's pupil, Otto, and from experiments made about the same time in the U. S. Hy- gienic Laboratory by Rosenau and Anderson. In the course of the standardization of diphtheria antitoxin, it had been noted that guinea pigs, which had previously been injected with toxin-antitoxin mixtures, were often killed by a subsequent injection of horse serum. When the subject was studied it was found that, when an animal is injected with an allen protein, there develops after a time a specific hypersusceptibility for this protein. After a definite interval, if the animal is given a second injection of the same pro- tein, violent Symptoms occur, which may end fatally. The reac- tion is specific, so that animals sensitized, for example, to horse serum, manifest little or no hypersusceptibility to other sera. It is possible, however, to sensitize an animal to several proteins simul- taneously. The sensitizing dose may be very small — even as little as one-millionth of a cubic centimeter of horse serum has sufficed to render a guinea-pig sensitive. A varying length of time must elapse after the sensitizing injection before the animal becomes fully 1913] Charles Frederick Bolduan 257 sensitive. In guinea-pigs treated with small doses of horse serum, from 12 to 14 days suffice; with large doses, the time required is longer, and may extend over weeks or even months. In any case, in Order to produce severe Symptoms, it is important that the second injection be large, say 5 to 10 c.c. in a guinea-pig. This phenom- enon, spoken of as anaphylaxis, has come to occupy an important place in the theory of infection and immunity. What is the ex- planation of the phenomenon? Enteral and parenteral introdiictions of protein contrasted. We know that the subcutaneous, intraperitoneal, or intravenous intro- duction of alien protein is followed by the formation of antibodies ; at the same time it can readily be shown that no antibodies develop after the oral introduction of milk, eggs, or even of raw meat. In other words there is a marked contrast in the behavior of the body toward enteral and parenteral introductions of protein. In the former the protein is acted on by specialized cells, which, through their pepsin, trypsin and enterokinase, and erepsin, break down the protein molecule so that it loses its species identity. After this, absorption takes place, and with it there is a synthesis or rearrange- ment of the molecule whereby it is built up into protein specific to the body. Under normal conditions it is impossible to produce antibodies by feeding alien protein, though precipitins have been produced by overfeeding animals with large amounts of alien pro- tein. When protein is introduced parenterally it gives rise to the formation of specific antibodies. In the case of the sensitized ani- mals described above, the first injection causes the production of specific antibodies, among them specific cytolysins acting on the alien protein molecule. When the second injection comes, the alien pro- tein is at once laid hold of by this antibody, protein cleavage results, and with it the liberation of poisonous cleavage products. These cleavage products cause the severe Symptoms and even death that characterize anaphylaxis in guinea-pigs. That similar Symptoms do not arise in the enteral digestion of protein would then be explained by saying that, in the specialized digestive apparatus, provision has been made either for preventing the formation of such poisonous cleavage products or for neutralizing them (conjugations?) before they can cause injury. 2S8 Imnmnity in Some of its Biochemical Aspects [Jan. Significance of the pcriod of inciibation in anaphylaxis. An in- teresting result of these studies on anaphylaxis is the light they shed on the significance of the period of incubation, and also on the poi- sonous Symptoms produced by bacteria f rom which no very poisonous siibstance can be extracted. Taking up first the latter point : It has been held that the various pathogenic bacteria, like the diphtheria and tetanus bacilli, either secrete toxins or, at least, contain such toxins bound np in their protoplasm. In the latter case, it was believed that these endotoxins, as they were termed, were set free during the destruction of bacteria in the body. From his studies on anaphylaxis, Friedberger concludes that it is entirely unneces- sary to assume the existence of specific endotoxins in bacteria to account for the various Symptoms seen in bacterial infections. By repeatedly injecting sensitized animals with minute doses of sheep or horse serum, he found it possible to produce all manner of fever curves at will, merely by varying the size of the dose and the inter- val between the injections. From this he concludes that the diver- sity of clinical Symptoms of various infectious diseases can readily be explained, even on the assumption of but a single poison. He speaks of this as anaphylatoxin, and regards it as a cleavage product of protein of whatever origin introduced parenterally. Just as in enteral digestion, uniform cleavage products are formed from most diverse proteins, so, he believes, that in the parenteral protein de- composition leading to the formation of anaphylatoxin, a certain poison is uniformly produced. Whether or not, in addition to anaphylatoxin, there are other specific poisons for the various in- fectious diseases is immaterial; their existence (except in certain diseases) has not been proved, and the assumption of their existence is unnecessary. According to Friedberger, the assumption of a common anaphylatoxin is only apparently in contradiction to the well known law of specificity of infectious diseases. In the infec- tious diseases it is not the poison which is specific, but only the mode of its production. The production of anaphylatoxin requires the action of antibodies; the mere Solution or disintegration of bacteria by other means does not suffice. In other words, a particular cleavage of the protein molecule is necessary. Bearing of the period of incubation on intoxication by bacterial 1913] Charles Frederick Boldiian 259 infection in general. With this conception of the effects of paren- teral protein cleavage, it is a simple matter to explain the significance of the period of incubation. For those of you who are not medical students, I will say that every infectious disease manifests itself only in a certain period of time after infection has taken place. More- over, this interval is fairly constant. Thus, after a person has con- tracted typhoid fever, some ten to fifteen days elapse before Symp- toms develop. In measles, the incubation is regularly fifteen to eighteen days; in scarlet fever, regularly from three to five days, etc. Formerly this period was explained as the time necessary for the development of germs in sufficient number to produce Symptoms. This explanation was unsatisfactory, because, in artificial in- fections, no matter how large the dose, it was never possible to shorten the incubation period below a certain minimum, and this minimum could not be explained. If, however, we regard in- fecting bacteria as protein introduced parenterally, we shall have no difficulty in explaining the incubation period as the time necessary for the body to develop antibodies which shall act on the bacteria and produce poisonous cleavage products. Even if we do not accept Friedberger's assumption of but a single anaphylatoxin, the same explanation holds for the liberation of endotoxins. In this connec- tion, I ought to say that bacteria invading the body through the in- testinal tract, e. g., the typhoid bacilli, may still be regarded as intro- duced parenterally, because they pass the intestinal barrier and gain access to the other tissues of the body. Modern chemotherapy according to Ehrlich. Before leaving the subject of infection and immunity, I should like to say a few words about the chemistry of the cell in relation to chemotherapy. I have already pointed out that Ehrlich holds that the action of a chemical substance on a given cell denotes the existence of definite chemical affinities between the substance and the cell. Applying this conception to the germicidal action of chemicals, he maintains that the latter must have a certain chemical affinity for the parasites in Order to kill them. Substances having such affinities he terms parasitotropic. It is clear, however, that substances which can de- stroy parasites will also be poisonous for the animal body, i. e., they will have chemical affinity for the tissues of the host. They are 200 Immunity in So ine of iis Biochemical As pect s [Jan. therefore also Organotropie. In the employment of chemical sub- stances to combat infectious diseases, it follows that success can only be attained if the affinity of the chemical substances for the infecting parasite bears certain relations to their affinity for the infected body. Ehrlich's studies in this direction have, therefore, aimed to find poisonous substances whose parasitotropic affinity should be great in comparison to their Organotropie affinity. In his studies on Syph- ilis, he tested a very large number of substances, many of them com- binations of arsenic. As each substance was tested it received a serial laboratory number, and finally, in " 606," a substance was found which fitted the requirements to a high degree. This sub- stance, salvarsan, has produced really marvelous results in the treat- ment of syphilis. This line of work appears very promising. Chemical natura of antibodies. A closing word concerning the chemical nature of antibodies. Most of the studies have been made on diphtheria antitoxin, and although little is known concern- ing the Constitution of this substance, it seems proable that it is protein in character. Certain it is that the antitoxin is associated with the globulins of the serum, and highly concentrated Solutions of antitoxin have been prepared, by Gibson, by precipitating and then redissolving these globulins. Moreover, as Atkinson showed, the globulins increase markedly in the serum of immunized horses as the antitoxic strength of the serum increases. The influence which the development of the field of immunity has had on biochemistry has been tremendous, for it has contributed not only new view points, but also entirely novel methods. Much has been learned about substances which no one had ever yet seen and which we know only through their action. That all this has been achieved is due mostly to one master mind, Paul Ehrlich. May he long continue to lead us ! A PLAN FOR THE ORGANIZATION OF THE AMER- ICAN BIOLOGICAL SOCIETY^ ALBERT P. MATHEWS The present condition of the biological interests of the country may be called chaotic. There is no general Organization and little Cooperation between various subdivisions of the science; there are a multitude of small societies and a large number of Journals, few with any permanent support. This condition renders the science as a whole less effective in the Community than it ought to be, and is expensive both of time and money. The time has come to effect some kind of Cooperation of all biologists to secure the advantages which come f rom Cooperation. These advantages could be obtained by the formation of a general society, to be called the American Biological Society, along the lines of the American Chemical Society. (This Society might act as the Biological Section of the American Association for the Advancement of Science.) Objects of the society: (i) To unite the biological interests of the country for purposes of education; mutual support; increased Cooperation, defense and encouragement of scientific investigation; and to increase the influence of biological knowledge in the country; (2) to Start and support a Biological Abstract Journal; (3) to pro- vide for the permanent support of the biological Journals of the country and to provide for new ones as necessity arises ; (4) to ^ This plan was proposed by Professor Mathews in 1908, in multigraphed circular form, to the members of the American Physiological Society. The plan was formally laid before the Council of the Physiological Society in December, 1908, in the hope that the Physiological Society would endorse the essential fea- tures of the Suggestion. The author was appointed a committee of one to agitate the matter. Nothing further was done, however. Theunsuccessful eflfort in December, 191 1, to bring about an Organization of a greater American Physio- logical Society, and the recent formation of the Federation of American Societies for Experimental Biology, give new interest to Professor Mathews' plan, which is published here in its original form, at our reqiiest, and with the permission of the author. See pages 269 and 271 of this issue of the Biochemical Bulletin. [Ed.] 261 202 Organization of the American Biological Society [Jan. diminish the cost, to the members of the society, of dues to societies and siibscriptions for these Journals. Details of Organization. Membership. All members of the present biological societies should be eligible for membership with- out further action and should become members on payment of the dues. Such societies are those of Anatomy, Physiology, Zoology, Botany, Experimental Medicine, Pharmacology when organized, Psychology, Biochemistry, Bacteriology, and so on. All persons sufficiently interested in the progress of biology to pay the dues of the society should be eligible for membership. LocAL SECTiONS. The Constitution should provide for the for- mation of local sections in different cities, a certain per cent. of the dues of the members of such a local section to be repaid to the section for local expenses. Affiliation of present societies. The present societies should ultimately organize as sections of the Biological Society, thus saving extra dues. Membership in these sections might be determined by the sections themselves. Dues. Dues should be sufficient to provide that each member should receive the Biological Abstract Journal, and some or all of the other biological Journals. How this may be arranged is shown beyond (page 265 ) . The cost of the Journals should be much lower to the members of the society than to Outsiders. Explanation of the proposed plan. The plan presented in the foregoing Statements is virtually that adopted with such great success by the chemists of the country. A few years ago the chem- ists were in the position of the biologists today. There were sev- eral small societies; there was nominally a general Organization dragging out an unprofitable existence. There were several Jour- nals badly supported. The American Chemical Society was orga- nized and ultimately the smaller societies became convinced of the advantage of Cooperation. Now, most of them have become sec- tions of the general society. The growth of this society has been very rapid ; and it has grown in vigor as well as in size. Two years ago^ the society started a chemical abstract Journal and it is not too much to say that this has done more for the chemical interests *The reader is reminded that this was written in 1908. [Ed.] 1913] Albert P. Mathews 263 of the country than any other step taken. Chemical Abstracts has welded the various divisions of the science together, and so great has its value proved to be, that the membership in the society has almost doubled since it was started.^ The society publishes three Journals, Chemical Abstracts (the abstract Journal), the Journal of the American Chemical Society, and the Journal of Industrial Chemistry, which are distributed to all members of the society for the dues, $10 a year. Chemical Abstracts appears every two weeks ; the other two are monthly Journals. Relation of the society to the naturalists. Two possibilities are open to us in forming the Biological Society: we could make use of the American Society of Naturalists, reorganize that and change it into a new society with new aims; or we might Start a new society, leaving the " Naturalists " to f ulfill some other usef ul function (such as that adopted in their recent reorganization). The name of the " Naturalists " is badly chosen for a general biological society, such as that proposed; and, since its partial resuscitation along its old lines might weaken our efforts (if the two societies should Cover in any way the same field), it appears wiser to me to organize a new society, and to allow the " Naturalists " to have its aim changed to the one sketched in the plan of reorganization. Discussion of the objects of the proposed biological society. The importance of a biological abstract journal. ( i ) The objects in paragraph i, page 261, are so desirable as not to need dis- cussion. (2) The desirability of starting a Biological Abstract Journal, in English, has long been apparent. Funds alone have been lacking in the past to accomplish this object. The Organization of this society would make it possible to issue such a Journal. This would do more to unify and stimiilate biology than any move we could make. (3) How the ends sought in objects 2, 3, and 4 (page 261) could be attained, will now be shown. List of the present biological Journals and their esti- MATED cosT AND PRiCE OF SUBSCRIPTION.^ The figures submittcd in this list are approximate only and are based on estimates supplied by various firms and individuals. The subscription list is a rough * The reader is reminded that this was written in 1908. [Ed.] 264 Organization of the American Biological Society [Jan. estimate only. The cost is estimated on an edition of 500 copies. Estimated cost of Journals, containing tables and cuts, and printed on good paper with press work included; edition of 500 copies: 12-point (a good sized body type) $1.40 a page; lo-point (used for reviews) $1.80 a page; 8-point (bibliography) $2.11 a page; 8-point (tables) $3.40 a page. Blank pages, and pages made up wholly of figures, $0.90. For half-tones, in the text, there is an extra charge of $1.00 each for makeready. For process-plates on coated paper: single plates, $4.00 ; double plates, $8.00. An additional 1,000 copies would increase the cost only for inserts, the press work and the paper, and may be estimated at about $500 or $600 a year, on a Journal of say 1,000 pages. The cost of a Journal is thus seen to be almost wholly the cost of putting it on the press, or the cost of its first 500 copies. Name of Journal. Subscription price per year Estimated on basis of present issues.- cost of 500 copies. American Journal of Physiology $15 $7,000 American Journal of Anatomy 5 3,000 Journal of Comparative Neurology 4 2,000 Journal of Morphology 9 3,000 Journal of Infectious Diseases 5 3,000 Journal of Experimental Medicine 5 2,500 Journal of Medical Research 8 4,000 Biological Bulletin 6 2,500 Journal of Biological Chemistry 8 3^500 Journal of Experimental Zoology 5 2,500 Anatomical Record 3 2,000 PsychologicalReview{hu.\\etmand'mdQx) 5 3,000 Botanical Gazette 7 4,000 Total number, 13 $85 $42,000 Biological Abstract Journal 4,000 American Journal of Psychology 5 2,000 There are several other Journals which might be added to this list and there are a few Journals on the list which might be taken ' The reader is reminded that this was written in 1908. [Ed.] 1913] Albert P. Mathews ^ 265 care of by special Institutes.^ It is obvious, however, that the biol- ogists have before them the problem of putting on a permanent foun- dation Journals costing about $50,000 a year. This can best be done by making each Journal seif supporting, and this is only possible by increasing the number of subscribers. HOW TG INCREASE THE NUMBER OF SUBSCRIBERS FOR THE JOUR- NALS. At present it costs, let us say, $28 a year to subscribe for the Journal of Physiology, the Journal of Biologkai Chemistry, and the Journal of Infectious Diseases. Each of these Journals probably has on the average a paid subscription list of something under 400.^ Membership in the corresponding societies costs, in addition, about $2 a year for each society, or a total yearly expense of $34. Now, if we could make a society of 2,000 members and charge each member $25 a year for all dues or, to be more liberal, let us say $30 a year, the society would have an annual income of $60,000 ; and for this sum, it could publish and supply to its members not three but thirteen Journals without further cost. Moreover, each of these Journals would have a large circulation, beneficial alike to the man who published in it and to the Journal itself. Furthermore, the amount of the individual society expenses would be greatly re- duced since, by proper Organization, one or two paid secretaries would look after notices of meetings; bills for postage, announce- ments, programs, etc., would be less; and a saving would be effected all along the line, with a great gain in efficiency. The income of the Journals would also be augmented beyond the dues by the constant sale of back numbers, the sale of extra re- prints and, in some cases, by legitimate advertising. Moreover, by keeping the present prices in effect for all non-members, nearly everyone would hasten to join the society ; thus increasing our num- bers and increasing the number of those among whom the expense would be divided, and making it possible, from time to time, to Start new special Journals with little increase in expense to the members. Furthermore, by maintaining the present prices to libraries and for- eign subscribers, a considerable sum would be added to the treasury, For example, the present cost of these Journals to subscribers is $83 a year. If there were a hundred subscribers at this price, it would add $8,300 to our income. Of course the total may easily be less * The reader is reminded that this was written in 1908. [Ed.] 266 Organisation of the American Biological Society [Jan. than this, but it will certainly amount to half that sum, since there must be fifty libraries subscribing at the old rate. How to get the necessary 2000 members. As a nucleus of the Society all members of the biological societies would probably join. There are 1,500 names in Cattell's American Men of Science vvho would be eligible and upon whose support we might confidently count. In addition probably 300 have joined the ranks of biology since that publication was issued or whose names were omitted through oversight. Let us say, at a liberal estimate, 1,800 all told. There probably could be found in addition 500 intelligent and public spirited physicians, and others sufficiently interested in biology, to join such a society with such great advantages in the matter of Jour- nals. These figures are maximum figures but they suffice to show that we could count on perhaps a thousand members at the start; and there is no doubt that the numbers would increase rapidly, just as they have done in the Chemical Society. We might also soon Start a Journal of Biological Industries, or in other ways increase Cooperation between the practical applications of biology and the science itself. Other arguments might be presented, but these suffice to show the great advantages of Cooperation and to make it evident that, in this way, we could attain these desirable objects: increase the in- fluence of biology, increase Cooperation; knit the science together, strengthen its practical applications; start a Biological Abstract Journal; provide for the support, and enlarge the usefulness, of our present Journals, and provide for new ones as the need arises; and diminish the cost, to each one of us, of subscriptions and dues. We should also accomplish more than this, for, by such an Organ- ization, we should be providing for the future, and organizing with the object of attaining certain well defined ideals. Whatever Organ- ization is attempted at this time should have in view the practical attainment of these ideals and should not be a mere repetition of what we have, with no definite plan and without foresight. In view of the foregoing facts I move the adoption of the fol- lowing: That the American Physiological Society expresses its ap- proval of the objects sought in the plan presented for the forma- tion of the American Biological Society; and it recommends. 1913] Albert P. Mathews 267 further, that the Society transmit to the other societies copies of this plan with the request that the plan be presented to the members of the societies; that each society appoint one delegate to meet mem- bers appointed by the other societies to act as a committee of Or- ganization of the American Biological Society; and that such committee shall carefully examine into the feasibility of such an Organization and, if possible, draft a Constitution and report to the societies at their next annual meeting. Suggestions for carrying out, practically, the Journal part of the plan. ( i ) It will possibly be f ound that $30 or $20 a year is more than the majority of the Society feel able or willing to pay. Arrangements could be made whereby at a somewhat larger relative cost such members could subscribe to two, three, or half a dozen of the Journals as they desired. Arrangements could be made with the Journals whereby copies would be sent to the members of the Society at a reduced price, if a certain number of subscribers was received in this way. For example, the Society might offer the Biological Abstract Journal, and any two others, for $10 a year ; the Biological Abstract Journal, and five others, for $20 a year; and the whole number, say, for $30 a year. In this way there would always be an incentive for the members, who could not at the Start pay the füll sum, to increase their subscriptions and thereby enable everyone to get his subscription at a reduced cost. It would not, however, be possible on this basis to give so much to the members as if all subscribed to all the Journals, but still a great reduction of cost could be obtained. The object aimed at should be to increase as rapidly as possible the numbers of those taking the whole number of Journals. (2) The relation of the Society to the management of the Jour- nals would, of course, have to be worked out gradually. Several courses are open to the society. One is to leave the Journals as they are under their present control and for the Society to make such arrangements with the Journals as would be most advantageous to the members. This is the club-rate principle, the society buying so many copies at a reduced rate to distribute to its members. This. arrangement might do as a temporary makeshift, to get started, but would probably be unsatisfactory in the long run, since it would not be permanent enough. 268 Organization of the American Biological Society [Jan. The Society might take over the financial responsibihty of such Journals as the Council of the Society deemed best; beginning, for example, with one or two with the largest circulation, adding the Biological Abstract Journal, and Publishing the three for $io or $12 a year, and distributing them to all its members. Then, as the Journals wished and the Council and the Society decided, one after another of the other Journals could be added until the whole list was included. This scheme would be feasible if we had a thousand members at the start. In any such arrangement the editorial boards of the Journals would retain entire charge of the editorial manage- ment, so that the independence of the Journals would be secured. (3) If such an arrangement could be made with the Wistar In- stitute of Anatomy, it might become the Publishing house for the Society, taking over the financial responsibihty for additional Jour- nals, as the Institute has already done for several, and thus greatly extending the usefulness of the Institute. In this way the Society would aid the Institute in getting the Journals on a firm basis by uniting in its support the biological interests of the country. At the Start this plan might involve an increased outlay by the Wistar Institute, but, in the long run, the dues of the Society should suffice to maintain the Journals. This plan would aid the Wistar Institute in doing the work it has undertaken. (4) Provision could be made for the starting of new Journals at any time, or for the support by the Society of those established by Outsiders. University of Chicago, Chicago, Illinois. ORGANIZATION OF THE FEDERATION OF AMER- ICAN SOCIETIES FOR EXPERIMENTAL BIOLOGY^ Comprising the American Physiological Society, the American Society of Biological Chemists, and the American Society for Pharmacology and Experimental Therapeutics JOHN AUER Among the most enjoyable features of the recent meetings at Cleveland (pages 271, 275, and 279) were the subscription dinners and smokers, held on the evenings of December 30 and 31, at the Colonial Hotel. These informal dinners were attended by the pharmacologists, physiologists and biochemists, and a pleasant flavoring of naturalists, zoologists and anatomists. At the last of these dinners perhaps the most important develop- ment of the Cleveland sessions, so far as the pharmacological, physiological and biochemical societies are concerned, took place. At this dinner, delegates f rom the three societies, empowered to act, met in Conference on the formation of an alliance which should more closely knit together the three societies zvhile yet jealoiisly preserving the individtiality of each compotient Organisation. The delegates f rom the Physiological Society were Drs. Meltzer, Lee and Cannon ; from the Biochemical Society, Drs. Lusk and Wells ;2 from the Pharmacological Society, Drs. Sollmann, Loevenhart and Auer. Dr. Meltzer was elected temporary chairman and Dr. Cannon temporary secretary. The outcome of the proceedings of this Con- ference committee can best be shown by a transcript of its minutes. The f ollowing motions were voted unanimously : That a Federation of the three societies be hereby established. ^ This account was presented, originally, as a part of Dr. Auer's report of the proceedings of the Society for Pharmacology and Experimental Therapeutics, page 279. [Ed.] ^Dr, Gies, the third delegate from the Biochemical Society (page 278), was unable to attend the Cleveland meetings because of the illness of his eldest son. 269 270 Fcderation of American Biological Societics [Jan. That the presidents and the secretaries of the constituent societies form the executive committee of the Federation. That the chairmanship of the executive committee be held in turn by the presidents of the constituent societies who shall succeed one another annually in the order of seniority of the constituent societies (Physiological, Biochemical, Pharmacological). That the secretary of the society whose president is chairman shall be the secretary of the executive committee. That the secretaries of the three societies shall consult in pre- paring the programs of the annual meeting, and that, so far as prac- ticable, and with the authors' consent, papers be so distributed as to be read to the society in which they properly belong. That the programs of the three societies be published by the secretary of the Federation under one cover and that the expense of publication be shared pro rata by the societies according to the number of members. That the official title of the new Organization be the "Federa- tion of American Societies for Experimental Biology : Comprising the American Physiological Society, the American Society of Bio- logical Chemists, and the American Society for Pharmacology and Experimental Therapeutics." That a common meeting place of the Federation with the socie- ties of Anatomists, Zoologists and Naturalists is desirable but not mandatory. That, in the name of the Federation, the International Physio- logical Congress be invited to meet in the United States in 191 6. That the present Conference committee delegate all its powers to the executive committee of the Federation. The first meeting of the new Federation will be held in Decem- ber, 191 3, in Philadelphia. Rockefeller Institute for Medical Research, New York City. ANNUAL MEETINGS OF THE ORGANIZATIONS COM- PRISING THE FEDERATION OF AMERICAN SOCIETIES FOR EXPERIMENTAL BIOLOGYi Proceedings reported by THE Secretaries, JOSEPH ERLANGER, A. N. RICHARDS, and JOHN AUER I. THE AMERICAN PHYSIOLOGICAL SOCIETY Joseph Erlanger^ The Society held its twenty-fifth annual meeting in the Medical Building of Western Reserve University, Cleveland, Ohio, Decem- ber 29, 1912 to January i, 1913. Sixty-nine members were in at- tendance. Two executive sessions and six scientific sessions were held, two of the latter being Joint sessions, one each with the Amer- ican Society of Biological Chemists and Section K of the American Association for the Advancement of Science. The Joint session with the American Society of Biological Chemists was opened with exercises in memory of the late Waldemar Koch. After the mem- bers of the Society had arisen as a token of respect to the memory of Doctor Koch, Prof. A. P. Mathews delivered a memorial address. Papers and demonstrations. The titles of the papers and demonstrations, fifty-two in all, which were read and discussed, with the names of the authors, are appended : S. Simpson: The rate of growth in the dog. — G. N. Stezvart: Further observations on the blood-flow in man. — /. A. E. Eyster and W. J. Meek: Experiments on the sinus region of the mammalian heart. — G. C. Robinson (by invitation) and /. Aner: Cardiac anaphylaxis as shown by the string galvanometer. — W. T. Porter: The functional relations of cells in nerve centers. — R. S. Lillie: Correlation between the anti-stimulating action and the anti-cyto- ^ See report on the recent Organization of the Federation, page 269. ''Acting Secretary, vice Dr. A. J. Carlson, unavoidably absent. 271 2/2 Anmial Meetings of Federated Socicties [Jan. lytic action of anesthetics. — E. B. Meigs: Studies inthe general phys- iology of smooth muscle. — W. P. Lombard: The tickle sense. — O. FoUn, W. B. Cannon, and W. Denis (by invitation) : A new colori- metric method for the determination of epinephrin. — /. Aiier and 5. /. Meltser: The splanchnic as a depressor nerve. — F. R. Miller: The saHvary secretion centers in the medulla. — M. Dresbach (by invitation); A bloodless method of recording blood pressure in animals. — W. T. Porter: A new electrica! clock. — S. P. Beebe: A new form of apparatus for artificial respiration. — A. D. Hirsch- felder: Some new apparatus. — R. S. Hoslins: Relation of fatigue metabolites to epinephrin efficiency. — D. R. Hooker: Perfusion of the respiratory center in f rogs ; the influence of calcium and potas- sium on the respiratory rhythm. — A. Hunter: The nitrogen excretion of normal and of thyroidectomized sheep. — A. L. Tatum (by in- vitation) : Studies in experimental cretinism with suggestions as to a biological test for thyroid secretion. — R. Gesell (by invitation) : The relation of pulse pressure to renal secretion. — C. Brooks and A. B. Lnckhardt: The arterial blood pressure during vomiting. — T. Sollmann and /. D. Pilcher (by invitation) : The effects of aortic compression on the circulation. — E. G. Grey (by invitation) and A. D. Hirschfelder: Clinical observations upon the carbon dioxide per- centage of alveolar air. — C. W. Greene and W. Y. Skaer (by invi- tation) : On the fat contents of the mammalian gastric glands in re- lation to the stages of digestion. — 5". Toshiro (by invitation).- The chemical change in nervous tissue during excitation. — /. F. Zucker (by invitation) : The pressor property of shed blood. — H. Cushing, L. H. Weed (by invitation) and C. Jacobsen: Further studies on the role of the pituitary gland in carbohydrate metabolism, with special reference to the autonomic control of the posterior lobe secre- tion.— S. A. Matthews and D. D. Lewis (by invitation) : The pars intermedia; its place in Diabetes insipidus. — Lydia M. Degner (by invitation) and A. E. Livingston (by invitation) : Effects ofthyroid- ectomy and castration, respectively, on the pituitary in the rabbit. — P. W. Cobb and L. R. Geisler (by invitation) : The influence on foveal vision of the brightness of surroundings. — D. E. Jackson: Some observations on the peripheral action of certain drugs. — G. L. Kite (by invitation) : The relative permeability of the surface and 1913] Joseph Erlanger 273 the interior portions of the cytoplasm of animal and plant cells. — — J. D. Pilcher (by invitation) : The excretion of nitrogen subse* quent to ligation of successive branches of the renal arteries. — W. E. Biirge: The uniform rate of destruction of ptyalin and pepsin by the electric current. — G. H. Whipple: Hematogenous jaundice and its relation to the liver. — 5". /. Meltzer: Is the pulsation of the anterior lymph hearts responsible for the action of some drugs in cardiec- tomized frogs? — H. McGnigan: The Synergie action of morphin and strychnin. Joint programs. With Section K (Physiology and Experi- mental Medicine) of the American Association for the Advancement of Science : page 277 ; with the American Society of Biological Chemists : page 275. The following ten papers were read by title: — C. D. Snyder: The influence of temperature on the mammalian heart. — A. J. Carlson: Some observations on the physiology of the empty stom- ach and esophagus in man and dog. — H. C. Bradley: The problem of enzyme synthesis. — G. W. Crile: The relation between the phys- ical State of the brain cells and brain functions ; experimental and elinical. — F. Henderson and C. T. Flynn (by invitation) : Oligemia in acute disease. — H. McGuigan: The secondary depression by epinephrin; the rate of destruction of the pressor and the hypergly- cemic actions of epinephrin. — W. B. Wherry (by invitation) : On the transformation of amoebae into flagellates and vice versa. — P. E. Howe (by invitation) and P. B. Hazvk: The influence of fasting on the creatine content of muscle. — C. D. Snyder: A study of the electromyograms. — A. J. Carlson: The correlation of the physiolog- ical States of the thyroid of the fetus and of the mother. New members: G. C. Robinson, Rockefeiler Institute for Med- ical Research. — /. D. Pilcher, P. J. Hanzlik, R. S. Pearce, Western Reserve Medical School. — E. C. Schneider, Colorado College. — A. H. Ryan, University of Pittsburgh. — M. Dresbach, Cornell Uni- versity. — G. Bachmann, Atlanta, Ga. — H. G. Barbour, Yale Medical School. — W. DeB. MacNider, University of North Carolina. — A R. Moore, University of California. — H. B. Williams, Columbia University — V. H. K. Moorhonse, Washington University. The federation. At this meeting considerable progress was 274 Anmial Meetings of Federated Societies [Jan. made toward the formation of a close federation of the American Physiological Society, the American Society of Biological Chem- ists and the American Society for Pharmacology and Experimental Therapeutics. The Society expressed its desire to enter into such a federation, and a committee was appointed to confer with similar committees of the sister societies with a view to bringing about such a federation. The committee was granted power to make the ar- rangements for the next annual meeting. This committee was also directed to confer with a similar committee of the American Society of Naturalists to consider the advisability of establishing closer rela- tions with that society (page 278). Future programs. With regard to the measures of remedying the threatening congestion of programs that were referred to the Council at the last annual meeting, it was decided that should the federation of the three societies be accomplished (page 269), the secretaries of the federated societies be empowered to attempt the equalization of the programs of the three societies by placing papers on the program of the society to which its subject is most closely related. It was also decided to place at the end of the program papers presented by non-members, and, in the event of congestion of the program, to read these by title. Officers-elect. The following officers were elected for the year 1913: President — S. J. Meltzer; Secretary — A. J. Carlson; Treas- URER — Joseph Erlanger. Additional memeers of the COUNCIL — W. B. Camion and F. S. Lee. Editorial committee on the publication of the American Journal of Physiology for 1913 — W. T. Porter, A. J. Carlson, Joseph Erlanger, W. H. Howell, F. S. Lee, Graham Lusk, S. J. Meltser. (Appointed by the president.) Local entertainment. The local Committee on Enterta%iment, following the plan that was first tried last year at Baltimore by the members and friends of the Society, again agreed to dispense with all private entertainment, and to Substitute for it informal subscription dinners followed by smokers each evening while the Society was in session. These functions were open to all members 1913] Alfred N. Richards 275 and guests of the Societies of the Experimental Biological Sciences. It was again demonstrated that this method of entertainment, by bringing all of the members together under conditions permitting informal discussion and exchange of ideas, adds greatly to the pleasure and value of the meeting. Abstracts of the papers. The abstracts of the papers will be published in the February number of the "American Journal of Physiology. Washington University Medical School, IL THE AMERICAN SOCIETY OF BIOLOGICAL CHEMISTS Alfred N. Richards The sessions of the seventh annual meeting of the American Society of Biological Chemists were held in the medical buildings of Western Reserv^e University, Cleveland, Ohio, December 30, 1912-January I, 191 3. The scientific programs, which were of ex- ceptional interest, are appended. First Session. December 30, 9 a. m. Presiding officer: President A. B. Macallum. A. B. Macallum: Presidential address, on The energy of muscu- lar contraction; thermodynamic or chemodynamic ? — M. H. Givens and A. H. Hunt er: The excretion of pure catabolites in sundry types of mammalia. — O. Polin and W. Denis: The occurrence of uric acid in blood. — L. J. Henderson and W. W. Palmer: Studies of the ex- cretion of acid. — A. E. Taylor and A. I. Ringer: On the utilization of ammonia nitrogen in the protein metabolism. — W. McK. Mar- riott: The determination of acetone substances in blood and tissues by micro methods. — *jF/. McGuigan and F. C. Becht: The com- pression of the lung by inert gases. — /. Rosenhloom: A new method for drying tissues and fluids. — W. N. Berg: Surface tension in muscle contraction. Second session. December 31, g.oo a. m. Joint Session wiTH THE American Physiological Society. Presiding offi- CERS : President A. B. Macallum and President S. J. Meltzer. * Read by title. 276 Anmial Meetings of Federated Societies [Jan. A. P. Mathews: Memorial address on Waldemar Koch. — L. Loeb: The infliience of pregnancy on the cydic changes in the Uterus. — G. Ltisk: Metabolism of a dwarf. — H. S. Gasser (by in- vitation) and A. S. Loevcnhart: The mechanism of Stimulation by oxygen want. — T. B. Osborne and L. B. Mendel: Feeding experi- ments relating to the nutritive value of the proteins of maize. — A. I. Ringer: The fate of fatty acids in diabetic organisms. — A. B. Macallum and W. R. Campbell: On the secretion of pure acid by the kidney (with demonstration). — D. Marine: Hypertrophy and hyperplasia of the parathyroid in birds. — G. H. Whipple: Intestinal obstruction; study of a toxic substance present in the intestinal mucosa. — E. V. McCollum: The influence of the plane of protein intake on nitrogenous retention in the pig. Third session. December 31, 2.00 p. m. Presiding offi- cer: President A. B. Macallum. *W. Salant and /. B. Rieger: Further observations on the influ- ence of cafTein on creatin and Creatinin metabolism. — *//. H. Bunuel: Quantitative oxidase measurements. — *H. S. Reed: The regulating function of amylase in the fungus, Glomerella. — A. P. Mathews: A new method of determining valence based on molecular cohesion. — H. G. Wells: The entrance of chemical substances into diseased tissues. — H. C. Bradley: The problem of enzyme synthesis. — R. T. Woody att (by invitation) : Certain 3-carbon atom complexes in metabolism. — *£. G. Hastings and E. B. Hart: The presence of a lactic acid producing enzyme in Bact. lactici acidi. — E. V. Mc- Collum and Marguerite Davis: The influence of the composition and amount of the mineral content of the ration on growth. — H. C. Bradley: Connective tissues of Limulus. — S. Tashiro (by invita- tion) : A new method for the detection of minute amounts of carbon dioxid. — *L. H. Davis and A. D. Emmett: A study of the chemical changes in meats during the process of drying by the vacuum method. — *H. T. Leo and P. E. Howe: Muscle creatin; dialysis of creatin f rom dog muscle. — *N. Stadtmüller, M. Kahn and /. Rosen- bloom: Studies on sulfur metabolism; the urinary sulfur partition in various diseases. — *£. V. McCollum and H. Steenbock: The meta- bolic end-products of the lipoid nitrogen of tgg yolk. * Read by title. 1913] Alfred N. Richards 277 Fourth Session. January i, 2.30 p. m. Joint Session with Section K of THE American Association for the Advance- MENT OF Science, and the American Physiological Society. Presiding officer : Professor J. J. R. Macleod. Symposium. Some recent applications of physical chemistry in biology — {A) A. B. Macallum: Surface tension; {B) L. J. Hen- derson: The control of neutrality in the animal body; (C) A. S. Loevenhart: The physical chemistry of enzyme action. Memorial addresses. In the presidential address at the open- ing of the first Session, President Macallum made extended refer- ence to the life, character, and achievements of the late Waldemar Koch, a charter member of the Society. At the opening of the Joint Session with the American Physiological Society, Prof. A. P. Mathews delivered a memorial address on Professor Koch. New members: Louis Baumann, University Hospital, Iowa City; F, J. Birchard, U. S. Department of Agriculture; Samuel Bookman, Mt. Sinai Hospital, New York; E. D. Clark, Cornell University Medical College; H. J. Corper, University of Chicago; A. A. Epstein, Mt. Sinai Hospital, New York; M. S. Eine, N. Y. Post Graduate Medical School; Isidor Greenwald, Montefiore Home, New York ; W. H. Howell, Johns Hopkins Medical School ; N. W. Janney, The Herter Laboratory, New York; /. S. Kleiner, Rockefeller Institute for Medical Research ; P. A. Kober, Roosevelt Hospital, New York; E. C. Koch, University of Chicago; Leo Kristeller, Berlin, Germany ; E. B. La Borge, Rockefeller Institute for Medical Research; A. P. Lothrop, Columbia University; W. DeB. MacNider, University of North Carolina; A. L Ringer, Uni- versity of Pennsylvania; W. C. Rose, University of Pennsylvania; E. C. Schneider, Colorado College ; Harry Steenbock, University of Wisconsin; R. T. Woodyatt, University of Chicago. Officers-elect. The following officers were elected for the year 1913: President — A. B. Macallum; Vice-president — Graham Lusk; Secretary — Philip A. Shaffer; Treasurer — Donald D. Van Slyke. Additional members of the COUNCIL — H. P. Armsby, Lafa- yette B. Mendel, H. Gideon Wells. Nominating committee — Carl L. Aisberg, H. D. Dakin, P. B. 2/8 Annual Meetings of Federated Societies [Jan. Hau'k, Reid Hunt, Walter Jones, T. B. Oshorne, A. N. Richards, H. C. Shernian, F. P. Underhill. Special committees. President Macallum appointed William J. Gies, Graham Lusk and H. Gideon Wells a committee to confer with similar committees from the American Physiological Society and the American Pharmacological Society concerning the forma- tion of a federation of the three societies having for its object "the establishment of a stable connection between the three societies, for the purpose of fixing the time and place of the annual meetings, the arranging of Joint sessions whenever possible, and in general to establish, officially, closer scientific and social affiliations between the sister societies, while retaining their individual independence." This committee was further empowered to act upon such matters connected with the proposed federation as should require decision before the next annual meeting of the Society, and was also author- ized to confer with representatives of the American Society of Naturalists concerning closer affiliation with that Society (p. 274). The Committee appointed at the sixth annual meeting to prepare a report concerning the nomenclature of the lipoids reported prog- ress. Professor Leathes, a former member of the committee, was appointed chairman to succeed the late Professor Koch, Dr. E. K. Dunham was appointed to the vacancy created by Professor Koch's death, and Dr. P. A. Levene to the vacancy created by Dr. Jacques Loeb's resignation. The members of the reconstructed committee are J. B. Leathes, chairman, H. D. Dakin, E. K. Dunham, WiUiam J. Gies and P, A. Levene. Vote of thanks. A unanimous vote of thanks was extended by the Society to Professors Macleod, Sollmann, Stewart and Pearce, and to the members of the "Local Committee," for the hospitality which the Society enjoyed. Attendance. The following members were present at one or more of the sessions : J. J. Abel, Samuel Amberg, S. P. Beebe, W. N. Berg, W. R. Bloor, H. C. Bradley, H. J. Corper, Otto Polin, W. E. Garrey, H. D. Haskins, Shinkishi Hatai, R. A. Hatcher, P. B. Hawk, L. J. Henderson, A. H. Hunter, J. B. Leathes, A. S. Loevenhart, Graham Lusk, A. B. Macallum, J. J. R. Macleod, W. DeB. MacNider, W. McK. Marriott, A. P. Mathews, H. A. 1913] John Aller 279 Mattill, E. V. McCollum, F. H. McCrudden, L. B. Mendel, V. C. Myers, H. S. Raper, A. N. Richards, A. I. Ringer, E. W. Rock- wood, Jacob Rosenbloom, L. G. Rowntree, Torald Sollmann, H. C. Wells, R. T. Woodyatt. Abstracts of the papers. Abstracts of the papers will be pub- lished in the March number of the Journal of Biological Chemistry. University of Pennsylvania. III. THE AMERICAN SOCIETY FOR PHARMACOLOGY AND EXPERI- MENTAL THERAPEUTICS. John Auer The fourth annual meeting of the Society was held in the Med- ical Building of Western Reserve University, Cleveland, Ohio, on December 30 and 31, 1912. The scientific programs are appended: First Session. December 30, 9.00 a. m. — *W. Salant: The influence of temperature on the toxicity of caffein. — *fF. Salant: Further observations on the influence of cafTein on the circulation. — S. P. Beebe and Eleanor Van Alstyne: The effect of high protein diet on the growth of transplantable tumors of the white rat. — L. B. Mendel and R. L. Kahn: The physiological action of some methyl purins. — /. A. E. Eyster and W. J. Meek: The action of certain drugs on the electrocardiogram. — P. J. Handik (by invitation) : The intestinal absorption of alcohol. — P. J. Handik (by invitation) : The "toxic dose" of salicylates according to clinical statistics. — W. H. Brown and A. S. Loevenhart: The effect of hematin upon the circulation and respiration. — W. DeB. MacNider: The effect of anesthetics on the Output of urine in uranium nephritis. — G. B. Roth: The physiological assay of aconitin. Second session. December 30, 2.00 p. m. L. G. Rozvntree and R. Fitz: Renal function in experimental passive congestion. — R. Fits and L. G. Rowntree: The effect of temporary occlusion of renal circulation on renal function. — W. W. Ford: Observations on three poisonous fungi not previously described. — /. D. Pilcher: The protective action of lipoids against hemolysis. — H. G. Barboiir (by invitation) : The action of histamin upon surviving arteries. — G. W. * Read by title. 28o Annual Meetings of Federated Societies [Jan. Crile and /. B. Austin: Nitrous oxide sleep compared with normal sleep; brain cell studies. — W. T. Porter and /. H. Pratt: The action of diphtheria toxin on the vasomotor centre. — H. Nogttchi and /. Bronfenbrenner: The effects of certain disinfectants and therapeu- tic preparations upon the cultivated spirochetes. — F. M. Surface (by invitation) : The effect of surplus cow serum on complement fixation with infectious abortion. — */. Adler and C. L. Aisberg: Studies lipon the long continued administration of adrenalin and nicotin. — *C L. Aisberg: The hemolytic power of various plants. Third session. December 31, 9.00 a. m. — *F. H ender son: Demonstration of a carbonator for quantitative carbon-dioxide therapy. — P. Lcivis: Further observations on the relations of vital stains to the tubercle. — T. S. Githens and 6". /. Meltser: On the course of the toxic effects of ether and Chloroform under intra- tracheal insufflation. — T. S. Githens: On the influence of decerebra- tion upon morphin tetanus in frogs. — /, 5*. Kleiner (by invitation) : On the effect of sodium bicarbonate and sodium chlorid upon the convulsions produced by heroin and strychnin. — /. Aiier and 5. /. Meltzer: The influence of pituitrin upon the depressor action of the vagus nerve in cats. — B. T. Terry: The influence of heat upön the toxicity for trypanosomes of blood containing transformed atoxyl. — B. T. Terry: Variations in the toxicity of transformed atoxyl for trypanosomes, caused by altering the number of organisms. Officers-elect. The following officers were elected for 1913 : President — Torald Sollmann; Secretary — JohnAuer; Treas- URER — A. S. Loevenhart. New members of the Council — /. 7. Abel and Wm. DeB. MacNider. Membership committee — C. W. Edmunds was reelected to serve three years, and the place made vacant by Dr. Sollmann's elec- tion to the presidency was filled by the election of Reid Hunt. New members. Among the candidates for membership under investigation by the Membership Committee, the following were favorably reported to the Council, recommended for election, and elected by the Society : H. G. Barboiir, Yale University ; Clyde Brooks, University of Pittsburgh ; Gary Eggleston, Cornell Uni- * Read by title. 1913] John Auer 281 versity Medical College; P. J. Hanslik, Western Reserve Univer- sity; D. E. Jackson, Washington University; /. 5'. Kleiner, Rocke- feiler Institute for Medical Research; O. H. Plant, University of Pennsylvania; A. H. Ryan, University of Pittsburgh; F. P. Under- hill, Yale University. Attendance. The following members were present at one or more sessions of this meeting: J. J. Abel, Samuel Amberg, John Auer, S. P. Beebe, E. D. Brown, G. W. Crile, J. A. E. Eyster, W. W. Ford, T. S. Githens, C. W. Greene, Worth Haie, R. A. Hatcher, V. E. Henderson, A. W. Hewlett, A. D. Hirschfelder, D. R. Hooker, D. R. Joseph, P. A. Lewis, A. S. Loevenhart, W. DeB. MacNider, S. J. Meltzer, L. B. Mendel, J. D. Pilcher, J. H. Pratt, A. N. Richards, G. B. Roth, L. G. Rowntree, Torald Soll- mann, G. N. Stewart, B. R. Terry. Vote of thanks. At the last meeting the Society passed a vote of thanks to the Western Reserve University for the hospitality ex- tended and to the " Local Committee," Drs. Macleod, Sollmann and Pearce, for their thorough arrangement of all the details which made the Cleveland meeting so pleasant. Abstracts of the papers. Abstracts of the papers will be pub- lished in the March number of the Journal of Pharmacology and Experimental Therapeiitics. [The report on the Organization of the Federation of Amer- ican SociETiES FOR ExPERiMENTAL BiOLOGY, which appears on pages 269-70, was taken bodily from Dr. Auer's account of the pro- ceedings of the Pharmacological Society. (Ed.)] Rockefeiler Institute for Medical Research. MEETING OF THE AMERICAN SOCIETY OF ANIMAL NUTRITION (American Society o£ Animal Production) PROCEEDINGS REPORTED BY LEWIS W. FETZER The annual meeting of the American Society o£ Animal Nutri- tion was held at Chicago, 111., on November 30, 191 2. The ad- dress of the President, H. J. Waters, of the Kansas State Agricul- tural College, at Manhattan, Kansas, dealt with a report on a " Study of the effects of different proteins and ash constituents on the growth of pigs." Professor E. W. Morse, of the Office of Experiment Stations, U. S. Department of Agriculture, read a paper entitled " Sugges- tions concerning the planning and reporting of feeding trials," in which he pointed out some needed improvements in planning feed- ing tests so that the results obtained could be interpreted by modern biometrical and Statistical methods, and that the work as a whole could be so systematized and coördinated that the results of each investigation could be compared with those obtained by others. At present the work in this direction is so uncorrelated that a compila- tion of results on any uniform basis is out of the question. The Standing committee on methods of reporting results of feeding experiments made a special report, which contained recom- mendations urging uniform methods of reporting data obtained in such experiments. The recommendations include a summary of the opinions expressed by a large nuniber of investigators in response to questions previously propounded to members of the Society. The recommendations were adopted. It was voted to enlarge the scope of the work of this society, to include all animal husbandry interests — problems connected with the breeding, judging and management of live stock. This is to be in addition to investigations in regard to nutritive value of feeds 282 1913] Lewis W. Fetzer 283 and other problems pertaining to animal nutrition. The name of the Society was accordingly changed to "The American Society of Animal Production." The officers elected for the coming year are as f ollows : Pres- ident, C. F. Ciirtiss (Iowa State Agricultural College) ; Vice Pres- ident^ E. B. Forhes (Ohio State Agricultural Experiment Station) ; Secretary-Treasurer, D. H. Otis (University of Wisconsin) ; CoMMiTTEE ON Experimentation, H. J. Wüters (Kansas State Agricultural College) and E. B. Forhes (Ohio State Agricultural Experiment Station). A meeting of the Society will be held at the time of the Panama- Pacific International Exposition, at San Francisco in 19 15. Office of Experiment Stations, U. S. Department of Agriculture. EIGHTH SCIENTIFIC MEETING OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION AT THE COLLEGE OF PHYSICIANS AND SUR- GEONS, NEW YORK, DEC. 6, 1912* Proceedings reported by THE Secretary, ALFRED P. LOTHROP The eighth scientific scssion of the Columbia University Bio- chemical Association was held at the Columbia Medical School, at 4 p. m., on December 6, 1912.^ Abstracts of the papers are pre- sented here (pages 285-96) in two groups : (I) Abstracts of papers on research by non-resident members^ and (II) abstracts of papers from the Columbia Biochemical Department and affiliated labora- tories. The appended summary f acilitates ref erence to the abstracts (45-62).^ A SUMMARY OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE SUCCEEDING ABSTRACTS Allan C. Eustis. Biochemical rea- sons why free purgation is necessary in combating acidosis of diabetes ; results of clinico-chemical observa- tions. (45) A. J. GoLDFARB. Studies on the effects of salinity changes upon regenera- tion. (46) I. Greenwald. The procedure of Salomon and Saxl as a diagnostic test for Carcinoma. (47) J. Arthur Harris and Ross Aiken GoRTNER. On the relationship be- tvreen the weight of the sugar beet and the composition of its juice. (48) Paul E. Howe (with H. C. Biddle), Fasting Studies. XL Note on the composition of the muscle of fast- ing dogs. (49) Max Morse. The role of phagocytes in the involuting tail of amphibian larvae. (50) * Scientific meetings are held regularly on the first Fridays of December, February and April, and on the first Monday in June. ^ Proceedings of the sixth meeting were published in the last number of the Biochemical Bulletin, 1912, ii, p. 156. Proceedings of the seyenth meeting are published on page 322 of this issue. * Members of the Association who were not officially connected with the Columbia Biochemical Department when the research was conducted. 'For abstracts 1-44 see Biochemical Bulletin, 1912, ii, p. 156. 284 I9I3] Alfred P. Lothrop 285 Max Morse. Laboratory hints. (51) Jacob Rosenbloom. The diflfusion o£ iodo-eosin from ether through rub- ber into ether. (52) Jacob Rosenbloom. The absence of certain enzymes from the human chorion, (53) II Walter H. Eddy. The preparation of histon from flounder sperm. (54) Thuisco A. Erpf-Lefkovics and Jacob Rosenbloom. A quantitative study of certain enzymes of the ovary, Uterus and bladder, of preg- nant and non-pregnant sheep. (55) Nellis B. Foster. Pathological devia- tions in the chemistry of uremic blood. (56) Nellis B. Foster. Effect of phlor- hizin on a dog with Eck fistula. (57) Frederic G. Goodridge and Nellis B. Foster. The relation of uricolysis to sub-oxidation. (58) William J. Gies. A demonstration of some of the tinctorial properties of pigments produced from thymol by ammonium hydroxid. (59) B. Horowitz. Experiments on pig- ments produced from thymol by the action of ammonia. (60) W. M. Kraus. Influence of uranium nephritis on the excretion of Crea- tinin, uric acid and chlorids, and the effect of Creatinin injections during uranium nephritis. (61) Charles Weisman. On the question of protein in expired air. (62) I. ABSTRACTS of PAPERS on RESEARCH BY NON-RESIDENT MEMBERS* 45. Biochemical reasons why free purgation is necessary in combating acidosis of diabetes; results of clinico-chemical ob- servations. Allan C. Eustis. (Laboratory of Clinical Medicine, Medical Department, Tulane University, New Orleans, La.) Ob- servations were made upon twelve patients in the von Noorden clinic in Vienna and lipon four patients in the private practice of the writer during the past year. Tests for indican were made by Salkowski's method, the same amount of urine being used for each test, and the grade of the color imparted by the indigo was recorded +, ++, + + +J etc. The ammonia determinations were made according to Folin's method. In each case there was a marked degree of acetonuria, also a large percentage of ammonia nitrogen in the urine, and a high indican index. According to the experiments of Dale, when />-hydroxyphenyl * Members of the Association who were not officially connected with the Columbia Biochemical Department when the research was conducted. 286 Proceedings Columbia Biochemical Association [Jan. ethylamin is perfused through the liver, it yields phenyl acetic acid. As the amin is a product of intestinal putrefaction of tyrosin, and assuming that some such cleavage takes place with other aromatic amins of intestinal putrefaction, their formation should be markedly hindered by free purgation, with consequent reduction of acidemia. In the cases observed there was a drop in the proportions of am- monia, acetone, and indican when free purgation was instituted in conjunction with a low protein diet. In two cases of fatal acidemia it was impossible to obtain free purgation owing to intestinal paresis, while in those cases in which free purgation was obtained, prompt relief from acidemia was noted. 46. Studies on the eff ects of salinity changes upon regenera- tion. A. J. GoLDFARB. (Marine Biological Laboratory of the Carnegie Institution^ Dry Tortugas, Florida, and the Biological Laboratories of the College of the City of New York.) This in- vestigation was made with the idea of ascertaining to what extent changes in density of sea water afifected an organism, Cassiopea xamanacha, normally subject to relatively great changes in density of the sea. It was furthermore intended to compare the results, on one band, with those upon the hydroid, Endendriuin, which lives in more dilute and more static densities, and on the other, with the classic results of Loeb on the Tubidaria of Messina. Much care was devoted to reducing the number of variable factors, or in ren- dering them uniform, or eliminating them altogether, such as, varia- tions in respiration due to varying volume, surface and depth of the Solutions, variations due to differences in size of the medusas, to level of amputation, to cyclical variations in density, to limited num- bers (323 arms were used), etc. The results are not easily summarized. Normal and super- normal regeneration occurred in normal sea water and in sea water diluted from 5 to 15 per cent. In the gradient series, regeneration was at first gradually, then more rapidly, reduced until a 50 per cent. Solution was reached, in which regeneration was inhibited altogether. The medusae however lived in this and in the 45 per cent. solutions. The other half of the curve was strikingly different, in that re- generation feil off very rapidly, ceasing completely in 125 or 133 per cent. solutions. The whole curve was strikingly different in character from the one described by Loeb. I9I3] Alfred P. Lothrop 287 It is particularly noteworthy that the curve for the Eudendrium is intermediate in character between Tuhnlaria and Cassiopea. It seems altogether certain that Loeb's curve can no longer serve as a type, expressive of the behavior of organisms under varying condi- tions of density of the sea water, and it is doubtful whether the phenomenon can be expressed in a simple curve based on two vari- able factors. It appears also probable that the relatively high con- centration in which Cassiopea normally lives may be associated with the high Optimum density for regeneration in this animal. It is also probable that the minimal effects of increasing dilution may be an adaptive response to the extreme dilutions to which Cassiopea is normally subject. 47. The procedure of Salomon and Saxl as a diagnostic test for Carcinoma. I. Green wald. (Chemical Laborafory of the Monte fiore Home, New York City.) The procedure of Salomon and Saxl,^ proposed as a test for Carcinoma, was tried in a number of urines. All were positive. The precipitates obtained were filtered off, ignited, and weighed, with the results shown in the appended summary. Total sulfur Was also determined. There was no apparent relation between the amount of sulfur precipitated by the Salomon-Saxl procedure, either absolute or relative to the total sulfur, and the presence or absence of Carcinoma. Number of urines Number of cases Sulfur precipitated in the test : Types of cases As BaS04 average, mg. Relation to total sulfur, per Cent. Normal 8 5 9 2 6 5 8 2 9-5 7.2 6.6 7-7 I.2I Pathological I-5S 1.23 Carcinoma Sarcoma 48. On the relationship between the weight of the sugar beet and the composition of its juice. J. Arthur Harris and Ross AiKEN Gortner, {Carnegie Institution of Washington, Sta- tion for Experimental Evolution, Cold Spring Harbor, L. /.) Al- though the literature pertaining to work on the sugar beet is very voluminous, but little attention has been paid to the relationships "Salomon and Saxl: Wiener klinische Wochenschrift, 191 1, xxiv, p. 449; Deutsche medizinische Wochenschrift, 1912, xxxviii, p. 53. 288 Procccd'mgs Columbia Biochemical Association [Jan. that may exist between the weight of the root of the beet and the chemical composition of its juice. We have compiled such data from varions federal and State bulletins, and have examined them by calculating the intensity of such relationships on the — i to + i Scale of the coefficient of correlation. We have also written the regression equations showing the absolute change in solids, sugar, or purity, associated with a unit change in the weight of the beet.° We find that composition and purity are very closely correlated with the weight of the beet; as the weight increases, total solids, purity and sucrose fall rapidly. The following is a representative summary showing the rate of fall on the relative scale of — i to + i of the coefficient of correlation, and the rate on an absolute scale by the second term of the regression equation, where w= weight, j=rz sucrose, /) = purity and ;f = total solids. Data pertaining to 475 Beets'' rwt = — 0.497 — 0.023 ^ = 20.1 19 — 0.096 w rws = — 0.576 ± 0.021 s = 17.644 — o. 122 w rwp = — 0.474 ± 0.024 p = 88.516 — 0.273 w Inasmuch as our results show the necessity of taking into account the weight of the individual beets in all studies on composition, and because of the bearing of our data on the beet sugar industry, we shall publish them in füll in the Journal of Industrial and Engineer- ing Chemistry (1913, v, p. 192). 49. Fasting studies. XI. Note on the composition of the muscle of fasting dogs. Paul E. Howe (with H. C. Biddle). (Laboratory of Physiological Chemistry, University of Illinois.) To be published in füll in the April number of the Biochemical Bulletin. 50. The role of phagocytes in the involuting tail of amphib- ian larvae. Max Morse. (Boardman Laboratories, Trinity Col- lege, Hartford, Conn.) Barfurth, Metchnikoff, Mercier, and others have sought, in the phagocytes, the principal factor in the absorption of tissues. This has been held in question mainly by Looss, who believes that a chemical dissolution is at the basis of the 'Harris: American Naturalist, 1910, xliv, p. 693. * Nevada Data. Wilson: Bull. 32, Nev. Agri. Exper. Sta., 1896. 1913] Alfred P. Lothrop 289 process and that this has no primary relation to the activity of the phagocytes. It is conceivable that if phagocytosis is the principal factor, the blood-counts (differential and with the hemacytometer) would show both an increase in the total number of leucocytes and a difference between numbers of polymorphonuclear forms, in the bloods of a larva in which the process of metamorphosis has not begun and of a transforming individual. On account of the great difficulty in identifying the various forms of leucocytes in hemacytometer preparations, this method of counting was not adopted, but the counts were made upon smears, stained with Wright's stain. Three sets of individnals were used: (i) Larvse in which the appendages had not appeared «and hence no absorption of the tail had begun; (2) individuals in which the process of absorption had progressed to some extent and (3) those in which the absorption had been com- pleted for a number of months, i. e., adult frogs. Twenty-eight specimens were used and the percentage results are as f ollows : Polymorph 9.8 8.6 18.3 Basoph Eosinoph 6.5 7.0 0.4 T,arge M Small M Individuais absorbing the tail Non-absorbing Adults 4.2 4.7 6.2 36.1 20.6 13.2 42.4 S9.0 61.2 The polymorphonuclear type runs slightly more numerous in the individuals undergoing metamorphosis than in individuals before the process has begun, but they are found in much larger quantities in adults than in either of the other two groups. As Friedsohn and Neumann have shown, however, it is impossible to distinguish young polymorphonuclear leucocytes in the blood of amphibian larvse f rom young erythrocytes and other forms of leucocytes, since all of the corpuscles originate from cells similar in appearance in all cases. Hence, the number of large nucleated forms doubtless includes young polymorphonuclear leucocytes; and if this were so, it would be expected that the number of the large ones would be smaller in adults, which is the case as seen in the above column of values for large mononuclear leucocytes. For these reasons, it is doubtful if there is any decided difference in the number of polymorphonuclear leucocytes in any stage of frog development; and therefore it is 290 Proceedings Columbia Biochemical Association [Jan. doubtful whether these bodies play an important röle in the absorp- tion of the tail of the tadpole. With reg-ard to the basophiles and eosinophiles, it will be seen that they occur in smaller numbers in individuals undergoing meta- morphosis than in young tadpoles and, so far as the basophiles are concerned, they are fewer in number in larvse than in adults. The reverse is the case with the eosinophiles, being found in small num- bers in the adult. The small mononuclear leucocytes occur in smaller numbers in the "absorbing" animals than in either of the other types. They are regarded by the investigators mentioned above as the young, indifferent, forms of the several types of leucocytes and probably, also, of the erythrocytes, but since they occur in larger numbers in the blood of the adult, this view is not borne out by the results of the present investigation. It may be concluded, then, from this set of data, that leucocytes do not play an important role in protein and other transfer in the involuting tadpole tail nor do they initiate the process. 51. Laboratory hints. Max Morse. {Boardman Labora^ tories, Trinity College, Hartford, Conn.) An ULTRA-FILTER. Colloids may be filtered to advantage by coating the surface of an alundum filter-disc, placed in a Buchner funnel, with a thin collodion Solution and applying the funnel to a pump, or the house vacuum. The first drainage through the filter will be coarse, the finer suspensions Alling up the interstices of the filter disc; afterwards, fine particles filter. A CONVENIENT HOT-WATER BOTTLE HOLDER. Sclcct 3. FloreUCC flask with a neck whose sides are as nearly parallel as possible for a distance and wrap half a yard of quarter-inch twine around the neck, spirally ; then wrap electrician's tape or surgeon's tape around the whole, again spirally, thus keeping the twine from unwinding. The heat does not soften the rubber of the tape sufficiently to cause it to leave the twine. If in place of the twine, small round leather belt- ing be used, the apparatus is perfectly satisfactory for all time. The belting may be obtained in any sewing-machine shop or in a belting supply house at small cost. 52. The diffUsion of iodo-eosin from ether through rubber into ether. Jacob Rosenbloom. (Laboratory of Biological 1913] Alfred P. Lothrop 291 Chemistry of the University of Pittsburgh.) Boas and P have shown that various cholesterol esters diffuse from ether Solution through rubber into ether; cholesterol-stearate, for example, with a molecular weight of 652.51, diffuses very readily. Recently I found that the acid form of iodo-eosin diffuses very quickly under similar conditions. This substance, with a molecular weight of 836, has the composition indicated by the appended structural formula : co<^ • >c< >0 53. The absence of certain enzymes from the human chorion. Jacob Rosenbloom. (Laboratory of Biological Chem- istry of the University of Pittsburgh.) Published in füll in this issue of the Biochemical Bulletin, page 236. See also abstract 55- IL ABSTRACTS OF PAPERS FROM THE COLUMBIA BIOCHEMICAL DEPARTMENT AND AFFILIATED LABORATORIES 54. The preparation of histon from flounder sperm. Walter H. Eddy. Testes, obtained from cold storage flounders and pre- served in alcohol, were ground and extracted with 0.8 per cent. hydrochloric acid Solution. The histon was obtained from the acid extract after the manner of Kossei and Kutscher, and purified by washing with water, re-dissolving in 0.8 per cent. hydrochloric acid Solution and reprecipitating with ammonia, several times. Like ammonia-precipitated thymus histon, this histon was insoluble in water and the hydrochloric acid Solution, after dialysis to neutrality, gave all the characteristic histon tests. The "ammonia precipi- tate," washed with water, alcohol and ether, and dried to constant weight at 105° C, contained 18.08 per cent. of nitrogen (one determination only, because of lack of pure material). Like that of thymus histon, the hydrochloric acid extraCt of the histon from flounder testes yielded, when saturated with sodium chlorid, a pre- cipitate which was soluble in water. Such a Solution gave the char- acteristic ammonia and alkaloidal tests, but was not precipitated * Rosenbloom : Biochemical Bulletin, 1912, ii, p. 67 ; Boas and Rosen- bloom: Proc. Soc. Exp. Biol. and Med., 1911, viii, p. 132. 292 Proceedings Columbia Biochcmical Association [Jan. by cold nitric acid Solution. Histon from fresh testes is in course of prqjaration for comparisons with the product from cold storage material. We lare also investigating the presence of histon in flounder roe. 55. A quantitative study of certain enzymes of the övary, Uterus, and bladder, of pregnant and non-pregnant sheep. Thuisco A. Erpf-Lefkovics and Jacob Rosenbloom. Published in füll in this issue of the Biochemical Bulletin, page 233. See abstract 53. 56. Pathological deviations in the chemistry of uremic blood.^ Nellis B. Foster. In general it was found that there is a considerable increase in the amount of non-protein nitrogen in the blood in severe cases of nephritis. However this is not an invariable rule, as very severe cases sometimes show an approximately normal figure; and a high non-protein nitrogen value has been noted in other diseases, such as valvulär heart cases and pneumonia. The results must be interpreted in the light of the whole clinical picture. When the total non-protein nitrogen is i gram or over, per liter, the prognosis is probably to be regarded as extremely grave. The percentage of non-protein nitrogen that occurs as urea is extremely variable, and seems to bear no constant relation to the total non- protein nitrogen. The data are so lacking in concordance that it must be left to further investigation, now in progress, to disclose in uremic blood chemical substances which either qualitatively or quantitatively present a constant divergence from normal. 57. Effect of phlorhizin on a dog with Eck fistula. Nellis B, Foster. It has been stated by Rosenfeld that the administration of phlorhizin to dogs with Eck fistula does not induce glucosuria. Such a result would have so much bearing upon our ideas of the mode of action of phlorhizin that the matter required further study. One gram of phlorhizin in olive oil emulsion was given to a dog with an Eck fistula. Glucose was found in the urine, in considerable quantity, for nine days subsequent to the phlorhizin administration. 58. The relation of uricolysis to suboxidation.^*' Frederic G. Goodridge and Nellis B. Foster. In order to investigate the 'Foster: Archives of Internal Mediane, 1912, x, p. 414. ^* Goodridge and Foster : Ibid., 1912, x, p. 585. I9I3] Alfred P. Lothrop 293 subject of diminished oxidation in its relation to uric acid excretion, we &tudied cases of poisoning by illuminating gas (people) and by potassium Cyanide (dogs). The results show that retardations of the oxidizing processes, either by deprivation of oxygen (gas) or by interference with cellular functions ( Cyanide), were not followed by increased excretion of uric acid. It appears improbable, there- fiore, that uric acid destruction in the body is a simple oxidizing process. 59. A demonstration of some of the tinctorial properties of pigments produced from thymol by ammonium hydroxid. William J. Gies. The phenomena described in a previous com- munication on this subject^^ were demonstrated, as an introduction to the succeeding communication by Mr. Horowitz. Ammonium hydroxid produces from thymol a blue^ pigment (or mixture of pigments?). Ether extracts the blue material from the alkalin liquid, but in ether Solution the pigment is red. After evaporation of the ether from such an extraot, a purplish oily product remains. This residue yields a purplish Solution in alcohol, which becomes hliie when it is rendered slightly alkalin. Filter paper, soaked in such a blue, alkalin, alcoholic Solution, and then dried at room tem- perature, assumes a bright red color as the alcohol disappears. Treated with alcohol, such red filter paper, particularly if slightly moist, becomes bright green. Interesting probabilities suggested by these results, and the possible relationship of these color phenomena to the pigments in the Monardas^^ and other plants, will be in- vestigated. 60. Experiments on pigments produced from thymol by the action of ammonia. Benjamin Horowitz. Professor Gies has found that thymol, in contact with ammonia, develops a blue color.^^ Under his direotion I have been making a study of this phe- nomenon. The question early arose as to whether ammonia and thymol alone are sufficient for the formation of the pigment. Certain ob- "Gies: Biochemical Bulletin, 1912, ii, p. 171. "Wakeman: Bulletin of the University of Wisconsin, No. 448; Science series, 191 1, iv, p. 81. "Gies: Biochemical Bulletin, 1912, ii, p. 171, See also the preceding abstract, above. 294 Proceedings Columbia Biochemical Association [Jan. servations by Liebermann^"* (whose work on colored Compounds produced from phenols has been of great importa'nce ) , as well as the many uses o£ oxidizing agents in the production of phenol pig- ments, suggested that 'atmospheric oxygen takes an active part in the process. This supposition was confirmed. A current of hydrogen passed throiigh an aqneous ammonia-thymol mixture inhibited the f ormation of pigment. Nascent hydrogen ( f ormed by the addition of Zn dust or sodium amalgam) acted similarly. On the other band, the addition of a few drops of hydrogen peroxid greatly accelerated the production of pigment by oxidation. A rise has been noticed in an aqueous ammonia-thymol mixture inverted over water, in- d'icating the absorption of oxygen. The addition of Zn dust to the blue Solution in an open vessel caused the color to disappear, except near the surface, where blue always remained, showing the influence of atmospheric oxygen. Ether was added to ascertain whether the reduced substance could be extracted by it and whether the chromogen thus removed by ether would yield pigment in the presence of oxygen. The bottle was now tightly stoppered and allowed to stand. Within 24 hours the red ether layer (the blue pigment yields a red Solution in ether) had become colorless. On releasing the stopper the ether Solution be~ came colored again. This was repeated many times with different samples but always with the same result. Attempts have since been made to isolate the reduced product by evaporating the ether Solu- tion in a current of hydrogen, but so f ar without success. The work is in progress. 61. Influence of uranium nephritis on the excretion of Creatinin, uric acid and chlorids, and the effect of Creatinin in- jections during uranium nephritis. W. M. Kraus. In acute uranium nephritis in dogs, Creatinin was excreted in decreased amounts ; uric acid, in increased amounts. In subacute uranium nephritis, Creatinin was eliminated in decreased amounts ; uric acid and dhlorids, in increased amounts (2 weeks). Creatinin, injected in normal dogs, appears to be excreted " in toto." Creatinin, in- jected during acute uranium nephritis, is not wholly eliminated. Such an injection causes decreased Output of endogenous Creatinin, "Liebermann: Ber. d. d. Chem. Gesell., 1874, vii, p. 247; 1875, viii, p. 1649. I9I3] Alfred P. Lothrop 295 also uric acid, chlorids and wäter ; and death may ensue. Creatinin, injected during subacute uranium nephritis, is excrelted "in toto" and apparently does not affect the excretion of endogenous cre- aitinin, uric acid, chlorids or water. Two of the animals with acute nephritis died after injection of Creatinin, in marked contrast with the apparent lack of effect of Creatinin injections in th^ dogs with subacute nephritis. In acute nephritis, where Creatinin (endogenous and injec'ted) and all the other substances named above were excreted in decreased amounts, there was apparently a partial arrest of renal function. In the subacute nephritis this did not occur. In acute nephritis there is probably not only insufficient intact tubulär epithdium to carry the additional bürden, viz., the injected Creatinin, buit this additional bürden aggravates the preexisting condition. In subacute nephritis, on the other hand, regeneration occurs, thus increasing the func- tional possibilities of the kidney, so that injected Creatinin can be excreted " in toto." Fever increases Creatinin excretion. It is also known that cer- tain infeotions, e. g., diphtheria, cholera, pneumonia and colon in- fections, cause tubulär nephritides. If, instead of adding hyper- creatininemia to a nephritis which is predominatingly tubulär (as described for the above experiments with uranium nephritis), there should be hypercreatininemia follozved by tubulär nephritis, it is probable that a similar reaction would result, namely, a uremia-like toxemia, ending perhaps in death. Creatinin has been taken only as an example of urinary sub- stances normally excreted by the kidney tubules. It seems probable that other normal substances, which are increased in fever and ex- creted by the tubules, would act in a similar way, i. e., to overtax an already overfunctioning kidney in a condition aggravated by a tubulär nephritis. This Suggestion as to the production of uremia dioes not concern the substances directly causing rt, merely the mechanism of their retention. 62. On the question of protein in expired air. Charles Weisman. Rosenau and Amoss^^ recently pu'blished a paper in which they stated that expired air contains volatile prcytein. Their "Rosenau and Amoss: Journal of Medical Research, 191 1, xxv, p. 35. 296 Procccdings Columbia Biochemical Association [Jan. conclusions were dependent 011 anaphylactic phenomena that ap- peared to be obtained with condensations from expired air. At Dr. Gies' Suggestion I am repeating their experiments with a view of applying the findings to problems in Ventilation and disease. Six repetitions of the experiments by Rosenau and Amoss have been made thus far, with negative results in each instance. We believe that Rosenau and Amoss failed to conduct adequate control experiments, both on the toxic effects of blood serum and on their anaphylactic tests. Their choice of sites (heart, brain) for the injeotions is open to criticism. Injeotions into the heart may produce lesions of the heart or lungs as well as pericardial in- jury and hemorrhage; and, after injury to the bündle of Hiss, resu'ltant Symptoms like the Stokes-Adams Syndrome, with oon- sequent difficulty of respiration, may simulate anaphylactic effects. Besides, in such injections, there is no assurance that the entire amount of injected fluid goes into the heart. As for injections into the brain, Rosenau and Amoss themselves say : " When the second injection was placed under the dura, through the optic foramen, the results were sometimes clouded by the appearance of Symptoms which were interpreted to be the result of central Irritation." In cur work, the second injection was made intravenously (externa! jugular vein) . The conclusion by Rosenau and Amoss, that expired air contains protein ( " volatile " ) , appears to be unwarranted. The experiments are in progress with the Cooperation of Drs. J. Bronfenbrenner and S. Gitlow. [The proceedings of the February and April meetings of the Biochemical Association will be published in the April number of the Biochemical Bulletin.] Biochemical Laboratory of Columbia University, College of Physicians and Surgeons, New York. FOLIA MICROBIOLOGICA Perhaps it will be of some interest for readers of the Biochem- ICAL Bulletin to learn that the recently founded Nederlandsche Vereeniging voor Microbiologie publishes a quarterly Journal enti- tled Folia Microbiologica. The editors are : Professors Beijerinck (Delft), Klein (Groningen), Poels (Rotterdam), and Sleeswijk (Delft). The subscription price of Folia Microbiologica is Five Dollars ($5.00) a year. The first volume is now complete and con- tains the following papers : Numbers i and 2 : Beijerinck, Mutation bei Mikroben ; Klein, Ueber die biologische Analyse des Kaseinantiserums ; Jacobsen, Die Kulturbedingungen von Hcematococciis pluvialis. Number 3 : Söhngen, Ueber fettspaltende Mikroben und deren Einfluss auf Mölkereiprodukte und Margarine; Ross Van Lennep, L'influence des substances fixes sur l'anaerobiose dans les milieux de culture liquides ; Fresemann Victor, Ueber die proteolytische und antiproteolytische, resp. antitryptische, Wirkung des menschlichen Blutserums; Reeser, Complement fixation of different sera prepared at the State Serum Institute, Rotterdam ; Boeseken und Waterman, Ueber die Wirkung der Borsäure und einiger anderen Verbindungen auf die Entwickelung von Penicillium glaucum und Aspergillus niger. Number 4: Eijkman, Untersuchungen über die Reaktionsge- schwindigkeit der Mikroorganismen; Beijerinck, Die durch Bakte- rien aus Rohrzucker erzeugten schleimigen Wandstoffe; VanCalcar, Ueber die Kenntnis des anaphylaktischen Zustandes des tierischen und menschlichen Organismus; Waterman, Beitrag zur Kenntnis der Kohlenstoffnahrung von Aspergillus niger; Jacobsen, Die Oxy- dation von elementarem Schwefel durch Bakterien. The editorial introductory notice concludes with these words: " If foreign authors should wish the fruits of their researches to appear in our columns, they would be most heartily welcomed. The Latin title of our Journal indicates that this new publication has not sprung from chauvinism. We entertain very earnest con- victions on the ' internationalism of science.'" C. A. Pekelharing University of Utrecht, Holland. BIOCHEMICAL BIBLIOGRAPHY AND INDEX WILLIAM J. Gl ES (Biochemical Lahoratory of Columbia University, at the College of Physicians and Surgeons, New York.) Biochemical literature is becoming so abundant and comprehen- sive, and so detailed and miscellaneous, that earnest efforts to assimi- late a considerable part of it are apt to induce severe attacks of indi- gestion. Ways and means for rapid and accurate sifting and Classification of subjects, experimental data, conclusions, and theo- ries, gain importance witli the increase of activity, and the growth of interest, in biochemistry. Appropriate year books, reviews, Zen- tralblätter, and especially the Department of Biological Chemistry in Chemical Ahstracts, acquire cumnlative usefulness as biochemical research develops in quantity, variety and extent. The writer finds it very desirable to maintain a running, quar- terly, card index of the titles of the papers in the leading biochemical Journals. This index is particularly valuable during the intervals between the publication of indices of volumes of abstracts and re- views. In the belief that it may be helpful to others, a copy of a portion of the index is presented below. Sections of the bibliog- raphy and index will be printed regularly in the Biochemical Bul- letin, if the writer's opinion on its probable general Utility proves to be correct. In the appended bibliography, titles are shortened in a free and easy manner, redundant words are ignored, common words are ab- breviated, surnames of collaborators are connected by hyphens, and punctuation marks are omitted, for the sake of condensation. Vol- ume numerals are given in Roman, and the Arabic numerals imme- diately following them, or placed at the beginning of sections, desig- nate respective issues of the volume ; numerals separated by a slanted line indicate month and day of issue. The numeral at the end of each item is that of the initial page of the corresponding paper. The numeral at the beginning of each item indicates sequence in the 298 1913] William J. Gies 299 bibliography. The latter numerals are used in the index of subjects at the end (page 305). Abbreviations of words in the index are similar to those in the bibliography. The System of notation in the index, although a space-saving device, makes it easy to refer to any title in the bibliography, The index is a compass not an encyclo- pedia; its purpose is achieved if it acts as a convenient guide to the sources of information — if it helps the reader speedily to locate and follow the main currents through the literature. Biochemical bibliography and index: 1912; fourth quarter (Sept.- Dec.) ; Journals : Biochemische Zeitschrift, Zeitschrift für physiolo- gische Chemie, Journal of Biological Chemistry, Bio-Chemical Journal, Biochemical Bulletin. Biochem. Zeit. XLIV: 1-2; 9/6. — iHäri Einfl intraven Bluttransf a Gaswechs,!. — 2Häri-Pesthy Hat Temp d Nähr Einfl a Gaswechs ?,6. — ^Rudö-Cserna Wirk intraperiton Blutinfus a Gaswechs,40. — 4Häri Einfl Kohlenhyd a Energieums,66 ; 5Wirk intraperiton Blutinfus a Energie- verbr,84. — GAlexand^r-Revess Einfl optisch Reiz a Gaswechs Ge- hirns,95. — ^Alexander Blutgaswechs Gehirns, 127. — SOrnstein Parenter Ernähr,i40. 3-4;9/9. — gQuagliariello Änd H'-konz währ Hitzekoag Prot,i57; loH'-konz Blut bei Temp'erhöh nach Wärmestich,i62. — iiRohonyi Veränd H'-konz b Pepsinwirk u Saurebind'vermög hydrol Spalt'prod Eiweis,i65. — i2Glaser Entwick'arbeit im Fundulusei,i8o. — iT^Berczeller Lipolyt Wirk Organextr,i85 ; i4Bestim Fet u Lipoid d Blutes u üb Lipol,i93. — i^Verzär Arb Pankr u Einfl a Verbren Kohl- enhyd,20i. — i6Beläk Wirk Phloriz a Gaswechs u Nierenarb,2i3. — lyTangl Respir'app für Schwein, Schaf ,235 ; iSMinim Erhalt'arbeit Schwein i Hunger,252. — igWeiser Ca, Mg, P, N-Umsatz wachs Schwein,279. — 2oZunt2 Versuchsergebn v Chauveau Mind'wert d Fet Kohl'hyd gegenüb als Energiespend b Musk'arb,290. — 2iSiegfried- Zimmermann Bericht, 292. 5-6;9/2o. — 22Pribram Diastas,293. — 2^Loeb-Beutner Verletz'strom,303. — 2^alladin Eiweissabbau u At- mung Pflanz,3i8. — 2^Forssmann-Hintze Heterol Toxiz Antiser,336. — 26Kopacze'wski Einfl Antisept a Wirk Maltase,349. — 2yTschernoruski Wirk V Nucleinsäu u nucl'spalt Ferm im tier Organis,353. — 28Fasal Colorim Meth quant Tryptophanbest u üb Tryptophangeh Horngebild u Eiweisskör,392. — 2gBierry Verdau Inulin, 402; 3oSaccharos spaltend Ferm,4i5; 3iRafiinos u Gentianos spalt Ferm,426; 32Stachyos u Man- ninotrios spalt Ferm,446. — ;^^Ohta Hitzebeständ Tryps u Peps,472; 34Verh Äpfelsäu im Tierkör,48i. — ^^N euherg-Schewket Polarim Bestim Glucosamingeh v Ovomucoid u Pseudomucin,49i ; 36Veränd 300 Biochemical Bibliography and Index [Jan. Arzneimit i Licht,495 ; 37Nachw gepaar Glucuronsäu i Harn, 502. — 2ßAltschul " Agfa "-Lecith, 505. (Pages, 508.) XLV:i-2;9/25. — ^gEmbden-Krans Milchsä'bild durchström Leber, I. — 4oSchmits Verh Glycerin künst Durchbl Leber, 18. — 41 Oppen- heimer Milchsä'bild durchström Leber,30. — 42Embden-Kalberlah-Engel Milchsä'bild Muskelpresss,45. — 4^Kondo Milchs'bild Muskelpresss,63. —44Kraske Milchs'bild B\ut,8i. —4SKondo Milchs'bild Blut,88.— 46^; Noorden Milchs'bild Blut,94. — 4yEmbden-Baldes-Schmits Milchs'bild a Traubenz i Tierkör, 108. — 480ppenheitner Einwirk verdün NaOH a Glyc'ald u Dioxyaceton,i34. — 4gMasiida Auftret aldehydart Subst b Leberdurchblut u üb Acetessigsäu'bild aus Äthylalk, 140. — ^oEmbden- Baldes Umwandl Acetaldeh i Äthylalk tier Organism, 157. 3-4;9/30. — ^lOhta Acetessigsä'bild aus Dicarbonsä mit 4C-Atom,i67. — ^2Emb- den-Schmita-Baldes Glycer'bild Tierkör, 174. — ^T^Embden-Oppenheimer Abbau Brenztraub'säu i Tierkör, 186. — $4Höber Verteil Blutzuck auf Körperch u Plasm,207. — ^^Langer Heroinaussch u -gewöhn,22i ; 56 Alkaloidaussch nach Magen unt Einfl i Mag gebracht Salz,239. — $yPescheck Einwirk NH4-salz u essig Salz a N-wechs Fleischfr,244. — ^SCytronberg Cholest'ase Blutkör,28i. — sgDavidsohn Magenlipase,284. — 6oEisenberg Formaldehydhämoly,303. — GiMaidorti Blutgiftanäm, 328. — 62Allemann Bedeut H' Milchgerin,346. 5-6;io/i2. — 6^Endler Durchtr Salz durch Protoplas,359. — 64N othmannZuckerhandl Wirk Narkot a Plasmaström,4i2. — 6$Tschermak Adaptat Ferm'bild Ver- dau'kan,4S2. — 66Bickel-Tsividis Einfl Digital'kör Kurv d Elekt'kar- diogr,462. — GyFreudenberg Fettstoffwechs,467. — 6SPreti Katal Einwirk Blei a Harnsä'bild u Harnsä'zersetz, 488. (Pages, 502.) XLVI:i-2;io/23. — 6gPorges Resp Quot Säurevergift, i. — yoBre- dig-Fiske Katal bewirk asym Synth,7. — yiChristiansen Frei u geb HCl i Mag'inh,24; y2lbid., 50; yT,Ibid., 71; y4lbid.,S2. — y^Müller Einfl Be- handl Milch a Labfähigk,94. — y6Würts Verteil P-säure Harn u Kot, 103. — yyTschernorutzky Wirk NaaCOg auf Alkaloidsalz u Farbst,ii2. — ySLöb Verbal Stärk unt Einfl stillen Entlad,i2i ; 79Pankr'diast,i25. — SoMichaelis-Davidoff Elektrom Bestim Blutalk,i3i. — SiKammann Pollentox,i5i. — 82Rohland Adsorp Tone,i70. 3-4;ii/9. — S^Galeotti Aussch H2O bei Atmung, 173. — 84Gramenitzki Blut- u Hamzuck b Adren'inf,i86. — ^^Siegfried-Zimmermann Entst Phenol a />-Kresol i Hund,2io. — 86Rosenthal Einfl Osmiumsäu a Receptor'app Erythrocy, 225. — 8yOhta Bedeut Proteol f spezif Hämol, 247. — 88 Fränkel Lipoid, 253. — 8gChristiansen Mett Meth u Acid'opt Peps'wirk,257, — goLöb- Gutmann Glykoly,288. — giHaas Schick Glyoxylsäu i Tierkör,296. — 1913] William J. Gies 301 g2Rona Schicks tierabgeb Eiweisskör i Darm,307. 5;ii/i4. — g^Bat- telli-Stern Oxyd />-Phenylendiam i Tiergevveb,3i7; g4lbid.jT,4T,. — g^Mohr-Heimann Norm u Eklamp Placent,367. — g6Rohland Ad- sorp Tone,374. — gySchuls Kieselsä'gehalt Schilddr,376. — gSvFenyvessy Natur u künst Komplem verhalt sich in " Regenerat " ident,393. — ggBattelli-Stern Nomenkl Polyphenoloxydas,395. 6;ii/25. — lOoDox- Neidig Spalt a- u ^-Methylglucosid dur Asp niger,2g7. — lOiHassel- balch Neutral'reg u Reizbark Atemzent i Wirk a COa-span Blut,403. — i02Winterstein App Mikroblutgasanal u Mikrorespirom,440. — lO^Pig- hini Chem Nerv'syst,450. — lO^Freund-Kaminer Bezieh zw Tumorzel u Blutser,47o. — lo^Lehedew Mechan alkohol Gär,483. — io6L6pezSuärez HCI-bild Magen,490. — loyBang Erwid,5oo. — loSEissler Physostigmin, 502. (Pages, 504.) XLVII:i ;ii/30. — logPalitzsch-Walbum H"-konz b trypt Gelat'ver- flüs.i. — I loKlausner Klausner Serumreak,36. — 1 1 iMichaelis-David- sohn Abhäng spezif Fäll'reak von H*konz,59. — ii2Beutner Physikal Nat bioelek Potent'dif, y^. 2;i2/g. — ii^Johannessohn Einfl org Sau a Hefegär,97. — ii4Loeb Verh Eissigsäu b künst Durchblut Leber,ii8; ii5Permeab u antag Elek'lytwirk (neu Meth),i27. — iißHirata Diast Kraft Mundspeich,i67. — iiyZaleski-Marx Carboxylas höh Pflanz, 184. — iiSStoklasa-Sebor-Zdobnicky Photochem Synth Kohlenhydr,i86. 3-4;i2/i2. — iigBerrär Chem u quant Bestim Leim, 189. — i2oScaffidi Purinstoffwechs,2i5. — i2iChristiansen Mechan Peps'verdau,226. — 122 Michaelis Isoelekt Punkt,250. — i22,Michaelis-P eckst ein Ibid., Casein, 260. — 124W olfgang-Faleki: Physikal Zustandsänd Kolloid,269. — 125 Starkenstein Ferm'wirk u Beeinfl d Neut'salz,3oo. — i26Grafe-Vouk Inulinstoffwechs Cichorium intyb L. (Zichorie), 320. — 127a/ Klercker Pentos d Guanylsäu,33i. — i28Loezvy-Gerhartz Aussch HoO b Atmung, 343. 5;i2/i8. — i2gBickel-Pawlo'W Pharm Wirk /J-Oxyphen'äth'am, 345. — i^oReale C-wechs; Labil u stab C d Harn,355. — i^iSsobolew Milchs'bild b antisep Organautol,367. — 12,2V ernon Abhäng Oxydase- wirk V Lip'd,374. — i^^Pollini Katal Wirk Eisensalz Leberautol,396. — i^4Neuberg-Kerb Zuckerfrei Hefegär,405 ; 135 Ibid,4i^. 6;i2/3i. — iTjSMoldovan Wirk Chinins,42i. — i^yMichaelis-Rona Umlag Glucos b alkal Reakt; Theor Katal,447. — i^SSasaki Abbau Polypept dur nicht verflüss Bakt,462; I39lbid dur verflüss Bakt,472. — i4oSignorelli Ver- hältn zwisch Amin-N u Ges-N Harn unt versch Beding,482. (Pages, 508.) Zeit. f. physiol. Chem. LXXX:5;8/28. — i4iOsborne-Mendel- Ferry Wachst bei Fütter'vers m isol Nahr'subst,307. — i42Rogosinski 302 Biochcmical Bibliography and Index [Jan. Methylier Clupein,37i. — i^T^Pringsheim Ferm Abbau Hemicellulos ; Trisacch Zwisch'prod d Hydrol Mannan,376. 6;io/2. — i^Wacker Cholesterin u Begleitsubst i Depotfet b Carcin,383. — i^^Trier Um Am'äth'alk (Colamin) i Cholin,409. — i46Letsche Einwirk Hydrox'am Blut färb (Methäm),4i2. — I4y Mörner Ovomucoid u Zucker Weiss d Vogelei,430. (Pages, 473). LXXXI: i-2;io/io. — i4SAbderJialdcn-Fodor Aus Glykok, d-Alan u /-Leuc darst struk'isom Tripept,i. — 14(^80 hui ze-Trier Verbreit Cholin, 53. — i^oEider-Palm Zusam u Bild Enzym; Entwick Hefe in Nahrlös, 59. — i^ilnonye Entst Kreatin, 71; i52Xan'prot'reak,8o. — is^Grabow- ski-Marchlewski Blutfarbst, 86. — 154 Abderhalden Feststel Schwan- gersch,90. — i^^Dorner Beeinfl alkoh Gärung d Zell u Zellpresss,99. — i^6Buglia-Costantino Muskelch; Formol titr Ges'am'-N d glat, querges u Herzm, 109; I57lbid, d N Ext'kt'st,i20; I581bid, frei, Formol titr Amin-N,i30; i59Chem Embry; Formol titr Ges'-am'-N d embry Mus- kel, 143 ; 160 Ibid, frei Am'-N Musk d Ochsenembr,i55. — i6iCostan- tino Muskelch; S d glat, querges u Herzm, sow Myoprot d Säuget,i63. — i62Usui Bind Thymol rot Blutz,i75. — i6^Sieber H2O2 als hydrol Prinzip, 185. — i64Rohland Tongeruch,200. — i6^Warbiirg Bestim kl i HoO-gelös C02-Meng,202. — 166J olles Nachw Glukuronsä diabet Harn, 203; i67Nachw Album Harn, 205. — i68Grafe Bericht,2o6. 3;io/22. — i6gAbderhalden-Weil Nerv'syst Aminosä; Am'sä periph Nerv u Leitungsb Rück'mark (weis Subst),207. — lyoAbderhalden- Weil Die b Isolier Monoam'sä m Hilf Est'meth entsteh Verlust ; infrei- heitsetz Est m Bleihydr,226. — lyiAbderhalden-Hanslian Verh a-Pyr- rolid'carb'sä im Organis,228. — iy2Abderhalden-Weil Dreh'gsvermög Blutpias u -ser versch Tierart versch Alt u Geschl,233. — lyT^Laquer- Brünecke Einfl Gasen (O) Tryp- u Pep'verdau,239. — iy4Siegfried- Schutt Absch Am'sä mit Hilf Carbaminoreak,26o. — ly^Kossel-Gazv- rilow Frei Amidogr d Prot,274. — iy6Grafe Antw Bemerk v W. Völtz üb N-reten u N-gleichgew b Füt NH4-salz,28o. — lyy London Aus An- lass Mit'g Folin-Lyman, Prot metab standp bl'd a tis anal; absorp f stom,283. 4; 10/30. — lySAbderhalden-Kashizuado Kern Thymus u Anaphylax'vers mit Kernsubst (Nuc'prot, Nuc'n, Nuc'nsä),285. — 179 Abderhaldeti-Kautssch Fäulnisvers rf-Glutam'sä; y^Am'but'sä,294. — — iSoAbderhalden Isol Glycyl-phen'alan a Chym d Dünnd; Stud mit Hilf versch Abbaustuf Prot u synth darges Polypep,3i5. — iSiAbder- halden-Hirsch Füt'vers Gelatin, NH^-salz, vollst abgebau Fleisch u aus all bekan Am'sä besteh Gemisch,323. — iS2Pfeiffer-Modelski Verh Am'sä u Polypep geg Neut'salz,329. — iSsPekelharing Anorg Salze auf 1913] William J. Gies 3^3 Wirk Pankreaslip,355. 5-6 ;i 1/5. — i^^London-Ritvosch-Mepissow- Stassow-Dagaew- Masijezvski- Gabrilowitsch- Krym- H olmberg - Wide- mann-Gillels-Solowjew Norm u path Verdau (Hund), 369. — iSsHen- riques-Gjaldhcsk Plasteinbildung,439. — i86Abderhalden-ValettePetti- bone Einfl physik Zust Prot a Raschh ihr Abbau dur Ferm; Bedeut Verdau Prot dur Pep-HCl für weit Abbau dur Tryp,458. — iSyAbder- halden-Lampe Schicks von in Magendarmk eingeführ Am'sä, Am'sä'- gemis, Pepton u Protein,473. (Pages, 507.) LXXXII:i-2;ii/i2. — i88Abderhalden-Hirsch Synth Fähigk tier Zell; Verwert versch N-quel im Organis,i. — iSgAbderhalden-Lampe Ibid,2i. — igoFischer-MeyerBets Porphyrinbildung,96. — igiAbderhal- den AnaphylaXjioQ. — ig2Küster Methylier Hämin,ii3. — ig^Abderhal- den-Wurm Pyrrol'carb'sä u aus ihr Aufgeb Polypep,i6o; I94a-Am'but'- rsä u Derivjiö/. — ig^Arnold Hämatoporphyr'urie b Abdom'typhus,i72. 3-4 ',11/25. — ig6Hedin Reak zw Enzym u and Subst,i75. — igyJahn- sonBlohm Einwirk kol'd Subst auf Hem d Enzymwirk, 178. — igSRinger- Schmutzer Quad'ur,209. — iggKylin Färbst d Fucoid,22i. — 20oJegoroff Verh Schim'p (A niger, P crustac) z Phytin,23i. — 20iWiener Zusam's art u ven Blut,243. — 202Fischer-Krollpfeiffer Einwirk Phthalsä'anhyd a Pyrrolderiv,266. — 20^Arnold Darst Hämatoporphyr a CO-Blut,273. — 204Fanser Einwirk HCl-gas Diastas,276. — 20^Jansen Konstit Cholsä verm Bromier,326. — 2o6Jansen Cholsä'resor Hundedarm,342. — 207 Bürker Nomenkl Blutfarbs'deriv,346. 5;ii/3o. — 2oSGrafe N-reten b Füt NH^-salz (Schwein), 347. — 2ogPanser Einwirk HCl Invertas, :^'//'. — 2ioFischer-Röse Garfarbst,39i. — 2iiMariconda Funkt Verh Darmsegm nach lang Period funkt Untätigk,4o6. — 2i2Marchlewski Bemerk, Blutfarbst,4i3. — 2iT,Sieber Bericht,4i4. 6;i2/24. — 214L0M- don-Dobroivolskaja Pfortaderbl ; fistel,4i5. — 2i^Starkenstein-H enze Nachw Glykog Meeresmol'k (spez b Cephal u Aplys),4i7. — 2i6Kasch- izvabara Einfl J Autol,425. — 2ijBuglia-Costantino Muskelch; Ext'kt'st u frei Formol titr Am'-N d Musk,439. — 2i8Küster Bilirub u Hämin, 463. — 2igRinger-Trigt Einfl Reakt auf Ptyarwirk,484. — 22oPincus- sohn Abw; Erwid an Gräfe, 502. (Pages, 502.) Jour. Biol. Chem. XII:3;9. — 221 Koch Meth guanid urin p'thy'dect anim,3i3. — 222Williams Anim calorim ; sm resp cal,3i7. — 223 Williams- Riche-Lusk Ibid., met dog aft ing meat larg quant,349. — 22äfLevene- Jacobs Guan'hexosid i hydrol thym-nucl ac,377; 225Cerebron ac,38i ; 226Cerebrosid of brain,389. — 22yVanSlyke-Meyer Am'ac-N bl'd; prot assim,399. — 228Levene-Jacobs Struc thym-nucl ac,4ii; 229Guanyl ac,42i. — 22,oJacobs Prep glucosid, 427; 23iRemov phos'tung ac fr aq 304 Biochemical Bibliography and Index [Jan. 501,429. — 232 Ringer Prot metab i exp diabet,43i. — 2T^;^Anderson Org P- ac compd wh bran,447. — 2^4Kendall-Farmer Bact metab,465 ; 235 Ibid,46g. — 2T,60sborne-Mendel-Ferry Gliad i nutr,473. — 22i7Ringer Chem gluconeogen ; quant conv propion ac i glucos,5ii. (Pages, 522.) XIII:i;io. — 2^SKober-Siigiura Cu compl'x am ac, peptd a peptn,i. — 229UnderhiIl Mech phlorh diabet,i5. — 24oLusk Anim calorim; metab aft inges dextr a fat, incl behav H2O, urea, NaCl,27. — 24iFisher- Wishart Ibid, absorp dextr a eff on comps brd,49. — 242Kendall- Farmer Bact metab,63. — 24^Mendel-Daniels Behav fat-sol dye a st-fat in anim org,7i. — 244Menge New compd cholin type; acyl deriv a- meth'ch, ")8-homoch" (/?-meth'ch), y-homoch, 97. 2;ii.—24sWhite- Thomas Tryp proteol Cynoscion regal,iii. — 246Epstein-Bookman Form glyc'ol i body,ii7. — 24yHart-Humphrey-Mornson Comp effic f growth total-N fr alfalfa hay a corn,i33. — 248Lusk-Riche Anim calorim; infl ing am'ac metab,i55 ; 249lbid, infl mix food-st metab,i85. — 2^oM cCollum-Steenbock Creatin metab grow pig,209. — 2$iMcCol- lum-H aipin-Drescher Synth lecith i hen a charac of lec produced,2i9. — 2S2Denis Metab cold-bl anim; urin fish,225. — 2^T,Osborne-Mendel Mainten w isol prot,233. — 2^4Levene-Birchard Kyrin f ract part'l hydrol prot,277. 3;i2. — 2^^Keyes-Gillespie Gas metab bact; prod ferm dextr Bcoli, Btyphosis, Bzvelchii,2gi ; 256Ibid, absor O by grow cult Bcoli, B'welchii,2,oS- — 2^7 Anderson Org P- ac cot seed meal,3ii. — 258 Robertson Bld relat'ship i compos serum prot; compar ser hors, rabb, rat, ox, norm a fast,325. — 2^gRettger-Neivell Putrefac, proteus gr,34i. — 26oLewis Behav hydant deriv in metab ; hydan a ethyl hydan,347. — 26iDakin Racemiz prot a deriv resul fr tautom ch'ge,357. — 262Folin- Macallum Colorim deter uric ac in ur,363. — 26'T,Hunter-Givens Metab endog a exog purin (monk),37i. — 264F olin-Lyman Absorp fr stom; repl London,389. (Pages, 391.) Bio-Chem. Journ. Vl:4\io.—26'^MacLean Phos'tid kidn,333; 266 Purif phos'tid, 355. — 26yCooper Relat phenol a w-cresol to prot (mech o disinfec),362. — 268Bostock Distr N in autol, spec ref deaminiz,388. — 26gCoppin Efif purin deriv a org compd grow a cell-div plant,4i6. — 2'joMelvin Glycol i brd,422. — 2'jiHarris-Creighton Reduc FeClg surv organ,429. — 2y2WatkynThomas Act opium alkaloid,433. — 2y:^Harding- Ruttan Detec aceto-acet ac b sod nit'prus a NH40H,445. — 2y4Smed- ley Fat ac butter,45i. (Pages, 129.) Biochem. Bull. II:5;9. — 2'jsWinterstein E Schulze,i. — 2y6Rose Lit inos-P- acid (phyt),2i.— 277i/arW3' Artif cell,5o.— 278^^0/^ New igi3] William J. Gies 3^5 func peroxidas, trans orcin i orcein,53. — 27gGies Diff thr rub'r ; lipin and lipin-sol subst,55. — 2SoRosenbloom Ibid,64. — 2SiWelker Ibid, Diff prot; disin'gr col'dion membr b ethtr, 70.— 282Beal-Geiger Ibid, pigm, 7S.—2SsKahn-Rosenbloom Col'd-N ur, dog tumor hr'st,8,7.—284Howe Fast,go.—2SsBerg Contr str musc; surf tens,ioi.— 286£;(/rf3; Prot compd,iii. — 28yRosenbloom-Weinberger Eff intrap inj epineph partit N uT,i2:^.—288Hallibtirton Bioch Soc, Eng,i28.—28gMendel Proc See (2) Diet hyg a hyg phys'l, 15 Int Con Hyg a Demog,i29. — 2goMandel Proc See Bioch ine Phar'col (8d) 8 Int Con Appl Ch,i50.—29iLothrop Proc Col Un Bioch As'n, 156. (Pages, 210.) Subject index of the foregoing bibliography/ Absorpi 77,206-41-64; acetaldehso; acetat57; acet-acii4; acet-acet-ac49,5 1,273; acidi 1,89,1 13; acidosög; acyl-deriv244 ; adaptat65; adsorp82,96; agei72; "agfa"-lecith38 ; (i-alani48; alb'in 167; alcoh49,50,fermi05-55; aldeh49; alfa-hay247; al-tract65,i87; alkali8o,i37; alkard56,77,272 ; am-aci69-70-<^-8i-^-7,238-48; a-am'butyr-aci 79-94; am'eth'alc 145; am-grpi7S; am-Ni40-56-5-9-6o,2i7-27; ammon57,273,saltsi77-8i,2o8; amylas 79,116; anaphyli78-9i ; anem6i ; antag-actiis; antisep26; antiser25; Aplysiid2i5; appi7,i02; A-nigerioo,2oo; assim92,227; asym-synth7o ; autoli3i-i,2i6-68. Bact i38-9,2S5-6,metah234-5-42,coli,typhos,welch2S5-6; bil-pig2io; bilirub2i8; Bioch- Soc-Eng288; blood3,5,7,io-4,39,40-/-'^-5-<5-9,S4,84,ioi-i4-72-7,20i-j-7-i2-4-27-4i- 58-7o,alk8o,corp58,i62,gasi02,serio4,transfi; brain6,7,226 ; breast283; brom20S; butter274. Cai9; calorim222-j-40-7-^-9 ; carbam-reaci74; carbohydr4, 15,20,1 18; CO203; CO2I01-65; C-metabi3o; carbox'asii7; carcini44; cardiogr66; caseini23; catal68,70, 133-7; celliS5-88-9,277,div269; Cephalop2i5; cerebron-ac22S ; cerebrosid 226; cholest'as58 ; cholest'oli44; chol-ac2os-ö ; cholin 145-9,244; chymi8o; clay82, 96, 164; clupeini42; coag9; colamini45; cordion28l ; colloidi24-97,N283 ; Col-Un- Bioch-As29i; complem98; CU238; correci68,2i3; corn-gr247; corp54; cot-seed- m'l257; creatini5i,metab25o; cresol, m267,^85 ; cult-medi5o; Cynoscton-regal245. Deamin268; developi2; dextr(gluc) ; diabet 1 66,232-^ ; di-acet-ac49,5i,273 ; diastas 22,79,204; di-carb-ac5i ; diffus63,279-8o-J-2 ; digest29,i2i-73-84-<5; digital66; di- oxyacet48; disinfec267; drug36; dye243. Eclamp95; eggi2,whi47; electr23,78,8o, 1 12-22-5; e'lytli5; embry 159-60; energ4,5,20; enzym30-7-2,65,99, 125-43-50-96-7; epid-tis28; epineph84,287 ; erythrocy86; ether28o-i; ethy-hydanto26o ; excr55-<5, 83,128; extr'tivi57,2i7. Fast258-84; fati4,20,i44,24O-j,metab67,sol-dy243; fat-ac 274; feces76; fermi 13-34-5,255; FeCls27i ; fish252; fistula2i4; food2,8,i4i,249; formald6o, 156-5-9-60,21 7; Fucoidesi99; fungiioo,200. Gas io2-73,metabi, 2,3,6,7, 16,255-0 ; gelati09-i9-8i ; gentianos3i ; gliad236; gluconeogen237 ; glucosam35; glucos47,54,84, 137,237-40-1-55; glucosid23o; glucuron-ac37,i66; d-glutam-aci79; glycer40,52; glyc-ald48; gly'col 148,246; glycog2i5; glycolys90,270 ; glyc-phen'alan 180; glyoxyl-ac9i ; growi4i,247-5o-<5-69; guanid22i ; guan-hexos224 ; guanyl-ac 127,229. H*9,io-7,62,io9-ii ; hay247; HCl7i-^-5-^,io6,204-9; heart 66,156-7-5-61 ; "heat-punc"io; hem'porph 195,203; hemicelluli43; hemin 192,2 18; hemogli46-53, 207-12; hemol6o,87; heroin55 ; homochol244; hungeriS; hydanto26o; H2O2163; hyd roll 1,1 37-43-63,254; hy'ox'ami46. Inj-curr23 ; inos-P-ac276 ; Int-Con,(8)-App ' See explanation on page 298. 3o6 Biochemiccd Bibliography and Index [Jan. Ch290;(i5)-Hyg-a-Dem289; intest92,i8o,2o6-ii ; inul29,metabi26; invert209; I216; ironi33. Kerat28; kidn 16,265; Klausnr-reaciio; kyrin254. Lact-ac39,4i-^-j-^-5- 6-7,131; Iead68,hydri70; lecith38,25i ; /-leucini48; light36; lipassg, 183; lipin 14,27g- 80; lipoid 14-88, 132; Iipoli3-^; livr39,40-/-9,l 14-33. Mgig; mainten 18,253; tnal- ac34; maltas26; manani43; maninotn32; meati8i,223; metabi,2,3,4,5,6,7,8,i5-ö-5, 24,57,67, i2o-<5-30-76-7,223-32-^-5-4(>--?-S-9-50-2-5-6-6o-5 ; methemogl 146 ; method 28,80,115-9-54-65-0-7-70-^,205-30-62-6-73; meth'ation 142-92; meth'chol244 ; meth- glucioo; meth-guan22i ; Mett-methSg; mlk75,coag62 ; morsk2i5; muscl42-5, 156-7- 5-^>-6o-i,2i7-85,contr20,285 ; myoproti6i. Ni9,i40-57,227-47-6S-83-7,foodi88-9, met57, 176,208; narcot64; nerv- sy 3103-69; neut-regioi ; nomencl99,207 ; nucleas27; nucl-ac27, 178,224-^; nucleini78; nuc-proti78; nutri8,i4i-5o-8i-5-9,208-36-53-5. Op'm-alk272; optic6,i72; orcin(ein)278; osm-ac86; ovomuc35,i47; oxidasi32; oxidat93-<^; 0173,256; /'-oxyphen'eth'ami29. Pancri5,dias79,lipi83; p'thy'dect22i ; parenter-f eed6,8 ; Pen-crustac200; pentosi27; peps 11,33,89,121-73-86; peptid238; Pepton 187,238; pernieabii5; peroxid278; phenol85,267 ; ph'endiam93-4 ; phloriziö, diabet239; phosphatid265-6 ; P-ac76,233-57 ; P19; phos'tung-ac23i ; pho'ch-syniiS; phtal-ac-anhy202 ; physostigioS; phytin200-76 ; pigment77, 199,210-82; placent95; plant24,i 17,269; plasma54; plasteini85; polemicio7-76-7,220-64 ; porn-tox8i ; poly- pep 138-9-80-^-93; polyphen'oxidas99; port-brd2l4,fist2i4; porphyri9o; potent'l 112; p'p't'niii; pregnani54; propion-ac237 ; protein9,ii,28,92,i75-8o-6-7,253-<^-5- 6i-7-8i,assim227,compd286,metab24,i77,232; proteol87,24S ; proteus gr 259; pro- toprm63,streani64 ; pseudomuc35; ptyalin2i9; purini20-57,263-9;putrefaci79,259; pyrol202; pyrorcarb-aci7i-93; pyr-tart-acS3. Quad'urigS; quini36. Racemizat 261; receptor86; rafinos3i ; reduc27i ; regen98; renin75; respi7,24,83,i02-28,calor 222,centrioi.quot69; rub'r279,8o-/- 1 77. extr 13 ; toxic25 ; toxin8i ; toleranceSS; tranfusi ; tripepti48; trisac'ari43 ; tryps33,i09-73-86,proteol245; tryptoph28; tumor283,celli04; typhusi95. Urea24o; uric-ac68,i98,262 ; urin37,76,84,i30-4O-66-7-95,22i-52-62-83-7. Water83, 128,240; ■wheat-bran233. Xanthoproteic-reaci52. Yeasti34-5-S0,fermentaii3. BIOCHEMICAL NEWS, NOTES AND COMMENT Contents. I. General: Necrology, 307; in memoriam, 307; honors, 308; re- tirements, resignations, declinations and appointments, 310; lectures, 312; endow- ments, funds and buildings, 313; societies, associations, etc., 314; officers-elect of biological organizations, 315 ; miscellaneous items, 316. II. Columbia University Biochemical Association: i. General notes, 321; 2. Proceedings of the Associa- tion, 322; 3. Columbia Biochemical Department, 324. I. GENERAL Necrology. David Axenfeld, professor of physiology at Perugia. — Carl Bins, professor of pharmacology at Bonn. — Edzvard Curtis, emeritus professor of materia medica and therapeutics at Columbia University. — Elie de Cyon, some time professor of physi- ology at the Academy of Sciences of St. Petersburg, lately of Paris. — Wilhelm Ebstein, professor of internal medicine at Göttingen. — Humphrey O. Jones, proiQssor of chemistry at Cambridge. — Oszvald Kohts, professor emeritus of diseases of children at Strassburg. — Ewen Mcintyre, for many years president of the N, Y. College of Pharmacy. — /. W. Mallett, professor emeritus of chemistry at the University of Virginia. — Hermann Miink, professor emeritus of physiology at the veterinary College in Berlin. — Clarence V. Murphy, bacteriologist and medical chemist at the Mass. State Sanatorium, Rutland. — Aime Pagnoul, formerly director of the Agricultural Station at Pas-de-Calais. — Heinrich RittJmusen, professor emeritus of agricultural chemistry at Königsberg. — Preston B. Rose, for- merly assistant professor of physiological chemistry and toxicology, and lecturer on renal diseases, at the University of Michigan. — O. T. Williams, lecturer on pharmacology and demonstrator of bio- chemistry at the University of Liverpool. In memoriam. Lord Lister. The Lister memorials com- prise a " Lister International Memorial Fund," f rom which will be drawn from time to time a Lister international award for the most notable contribution to surgery in any part of the world, and which will also Support fellowships and studentships in surgical research; a monument in London; a memorial tablet in Westminster Abbey; 307 3o8 Biochemical News, Notes and Comment [Jan. a monument in Glasgow; and the preservation of the ward in the old building which is now being torn down to make way for a new building of the Royal Infirmary. This ward will be arranged as it was in Lister's time, furnished with contemporary articles and pro- vided with exhibits showing the work that Lister did, and with articles of a personal nature associated with the man in his work. Contributions may be made to any of the memorials, and may be sent to Dr. W. W. Keen, 1729 Chestnut Street, Philadelphia. Each contributor is asked to designate the particular memorial to which he wishes his contribution to be applied (p. 189), Paul C. Freer. The July issue of the Philippine Journal of Science was a memorial to the late Dr. Paul C. Freer, director of the Bureau of Science of the Philippine Islands, dean of the College of Medicine and professor of chemistry at the University of the Philippines, founder and editor of the Philippine Journal of Science (p. 189). Honors. Nobel prizes were presented by the King of Sweden at a bancjuet in Stockholm, on December 10. Those to whom awards had been made were present, including Dr. Alexis Carrel, of the Rockefeller Institute for Medical Research (p. 190). — The Nobel prize for chemistry has been divided between Professors Grignard, of Nancy, and Sabattier, of Toulouse. — Professor Sabat- tier has given his portion of the Nobel prize in chemistry to the building fund of the Toulouse Institute of Chemistry. — A reception was given in honor of Dr. Alexis Carrel, at New York University, on November 16, when President Taft, the French ambassador, and others, delivered congratulatory addresses and Dr. Carrel responded. Order of merit. Dr. Paul Ehrlich, of Frankfort, and Dr. Emil Warburg, president of the "Reichsanstalt" at Charlottenberg, have been made members of the Bavarian-Maximilian Order, which is the highest Bavarian decoration for scientific Services. Corresponding member. Prof. F. E. Lloyd, of McGill Uni- versity, has been elected a corresponding member of the Centro de Sciencias, Letras, e Artes, Campinas, S. Paulo, Brazil, in recogni- tion of his work on the desert rubber plant, guayule. Honorary member. At its meeting on December 3 the Acad- 1913] General 309 emy of Medicine, of Paris, elected Professor Delezenne of the Pasteur Institute an honorary member of the section on anatomy and physiology. Anniversary celebrations. On October 22 Prof. A. Kossei, of Heidelberg, celebrated the twenty-fifth anniversary of his pro- fessorship. — The twenty-fifth anniversary of Prof. Charles Richefs appointment to the chair of physiology in the Faculty of Medicine, of Paris, was celebrated on December 22. Professor Chauveau presided at the celebration and presented Dr. Riebet with a Fest- schrift to which three score scholars had contributed from different countries, among them being Pavloff, Kossei, Verworn, Sherring- ton, Chauveau and Bouchard. After the presentation, addresses of congratulation were made by Professors Landouzy, Dastre, Gley, Langlois, and others. CoMPLiMENTARY DINNERS. Dr. Jacques Loeb was the guest of honor at the second annual dinner of the Columbia University Bio- chemical Association, at the Chemists' Club, N. Y., Nov. 6 (p. 322). — Prof. R. H. C hütenden will be the guest of his many pupils and friends at a dinner at Delmonico's, N. Y., on March i. AwARDs OF MEDALs. By the Royal Society: The Davy medal to Prof. Otto Wallach, of Göttingen, for his researches on the chem- istry of the essential oils and the cycloölefines; the Buchanan medal, to Col. Wm. C. Gorgas, of the U. S. Army, chief sanitary officer of the Panama Canal zone. — By the Prussian government: The gold medal for science to Dr. Walther Nernst, professor of chemistry at Berlin. — By the Swedish Medical Society: The Retsius medal to Dr. J. N. Langley, professor of physiology at Cambridge, for his work on the nervous System. Prizes. The Gedge pri::e of Cambridge University has been awarded to Mr. A. V. Hill, of Trinity College, for his essay on the heat production of amphibian muscle and of cold-blooded animals. — Prize for work on diabetes: The medical society of Carlsbad has offered $1,000 for the best work or works on "Treatment of dia- betes mellitus, with special reference to balneotherapy." Competi- tion is open to physicians of all countries, and any language may be used. All Communications should be addressed to the Vereini- gung Karlsbäder Aertze, Carlsbad, Austria. The jury consists of 3IO Biochcmical News, Notes and Comment [Jan. Professors von Jaksch of Prague, Lüthje of Kiel, Ortner of Vienna, Schmidt of Innsbruck, and Dr. Ganz. Essays must be received by Dec. 31, 1913. Retirements, resignations, declinations, appointments. Re- TiREMENTS : Dr. Frana Pf äff , professor of pharmacology and thera- peutics, Harvard Medical School. — Captain R. W. Silvester, for tvventy years president of Maryland Agricultural College. (He has been made president emeritus and librarian of the Institution.) — Dr. G. R. Kraus, professor of botany at Würzburg. Declinations. Prof. 'Andrezv Boss, in charge of the depart- ment of farm management of the department of agriculture, Uni- versity of Minnesota, has declined an offer of the position of director of the new government demonstration farms and trial gardens, at Mandan, N. D. — Prof. E. M. Freem-cresol. These were the fundamental investigations which led to the Solution of the question as to how the action of the intestinal bacteria might be checked or diminished. The lactic acid bacillus has proved to be the best for this purpose. An obstacle to the work of the lactic acid bacilli has been their need of sugar, which does not reach the rectum in a usable form. Dr. Metchnikoff and Dr. Wollman, his pupil, have overcome this difficulty by cultivating bacteria that pro- 320 Biochemical News, Notes and Comment ■ [Jan. duce sugar from starch. It is now possible to supply a diet capable of supporting the bacillus that limits the action of the intestinal flora. Of course, concludes Metchnikoff, the struggle against senility is not concluded. Whether these discoveries will actually tend toward the lengthening of human life is a question of the future, but it cannot be denied that a beginning has been made, and we have reason to hope that from these investigations mankind may derive practical benefit (Journ. Amer. Med. Assn., 1912, lix, p. 815). Electrons. Abstract of an address before the American Philo- sophical Society at Philadelphia, Nov. i, by Sir William Ramsay. The actual existence of electrons in motion has been conclusively demonstrated ; the mass of an electron is not far from one iSßoth of that of an atom of hydrogen; and as the mass of an atom of hydro- gen is now known with fair accuracy, that of an electron is nearly 0.8X10'^'^ gram. Electrons in motion are negative electricity; they constitute a form of matter, which, at present, has more claim to the term "elementary" than have most of the " Clements." In- deed, metals must be regarded as Compound substances, of which one component consists of one or more electrons; these electrons are, as a rule, not very firmly attached, as is evident from the gener- ally easy oxidation of most metals. Non-metals are also composed partly of electrons, not so easily detached. The " combination of Clements with each other" consists in the shifting of one or more electrons from the more metallic to the less metallic dement; no doubt it will some day be possible to give " structural formulae " to the Clements, showing the relationship in position, or in directed mo- tion, between the true Clements and their attached electrons. The word " electricity" has a dual meaning; it may mean first, an assem- bly of electrons, stationary or in motion; or second, waves in the ether, produced by the stopping or starting of electrons in motion. The motion of electrons constitutes one factor of electrical en- ergy; wave-motion in the ether can be used as a means of gene- erating electrical energy, by employing the waves in making elec- trons move. Progress in man's command of natural forces has been made by learning how to direct and control the motion of masses — in other words, by acquiring a knowledge of mechanics; progress in the future will consist in acquiring the power to control 1913] Columbia Biochemical Association 321 and direct the motions o£ electrons. This has already been largely achieved by electric contrivances : it is, however, only by the use of concrete ideas regarding the "material" used, viz., electricity, that the progress of invention and discovery can be hastened {Science, 191 2, xxxvi, p. 684). IL COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION I. General notes Marriages. On October 2, Miss Rowena Farmer and Dr. Oscar M. Schloss. — On October 15, Miss Ethel P. Willcox and Dr. Harold E. Woodward. Appointments. Columbia University : Dr. Russell L. Cecil, Proudfit fellow in medicine; Dr. Wm. H. Woglom, assistant pro- fessor engaged in cancer research (page 201). — Cornell University Medical College : Dr. Stanley R. Benedict, professor of chemistry (promotion). — Johns Hopkins Medical School: Dr. Edwards A. Park (Columbia University), assistant professor of pediatrics. — Massachusetts Agricultural College: Dr. H. D. Goodale (Carnegie Institution, Station for Experimental Evolution), research biologist. department of poultry husbandry. — Turck Research Laboratory (New York) : Dr. Anton R. Rose (Columbia University), chemist. — University of Texas (Austin) : Mary E. Gearing (Public Schools, Houston, Tex.), professor of domestic science; Anna E. Richardson (Agnes Scott College, Decatur, Ga.), assistant pro- fessor of domestic science. — U. S. Department of Agriculture, Bureau of Chemistry: Carl L. Aisberg, chief chemist (p. 211) ; H. E. Buchbinder, assistant chemist. (P. 324.) New members and ofEcers-elect of societies. American So- ciety of Biological Chemists : Members, Louis Baumann, Samuel Bookman, Ernest D. Clark, Isidor Greenwald, Alfred P. Lothrop; Nominating Committee, Carl L. Aisberg, P. B. Hawk and Alfred N. Richards; Lipoid nomenclature committee, Wm. J. Gies; Com- mittee on the Organization of a federation of biological societies, Wm. J. Gies (p. 277) . — American Society of Naturalists : E. Newton Harvey. — American Society of Zoologists : E. Newton Harvey. — Phi Lambda Upsilon: National president, A. D. Emmett; national secretary, H. L. Fisher; registrar, George D. Beal. — Society for 322 Biochemical News, Notes and Comment . [Jan. Clinical Serology and Hematology (New York) : Secretary, D. J. Kaliski. — Nu Sigma Nu Alumni Association (New York) : Execu- tive Committee, Wm. K. Terriberry; Nominating Committee, Ralph G. Stillman. 2. Proceedings of the Association Second annual dinner. The second annual dinner of the As- sociation (and the first meeting of the Association for the academic year I9i2-'i3) was held at the Chemists' Club, 52 East 4ist Street, on Wednesday evening, November 6.^ About 125 members and their guests were present, the attendance being so large, in fact, that the main dining room of the Club was filled to capacity and it was necessary to use a room on the floor above for the accommodation of about 20 members. The dinner was given in honor of Dr. Jacques Loeb, of the Rockefeiler Institute for Medical Research. AdeHghtfulhalf hour's infomial social gathering preceded the banquet. The president, Dr. Walter H. Eddy, was the toastmaster and appropriately introduced the Speakers of the evening. Prof. Lafayette B. Mendel, of Yale University, spoke briefly on the characteristics of scientific men. Prof. Marston T. Bogert, of Columbia University, President of the Society of Chemical Industry, brought to the Association, and its guest of honor, the greetings of that Society. Profs. C.-E. A. Wins- low, of the College of the City of New York, and Graham Lusk, of the Cornell University Medical College, also spoke informally and in a very entertaining way. The main address was delivered by Dr. Loeb, who gave a very interesting and instructive account of the results of some of his recent work on the permeability of cells. Prof. Max Morse, of Trinity College, proposed that Dr. Loeb be elected an honorary member of the Association. The motion was seconded by Dr. E. Newton Harvey, of Princeton University, and unanimously carried by a rising vote. The names of those present and the groupings at the tables are indicated on pages 323 and 324. * An account of the first annual dinner, in honor of Prof. R. H. Chittenden, was published in the Biochemical Bulletin, 191 i, i, pp. 334-339. I9I3] Columbia Biochemical Association 323 *Charles Baskerville *Marston T. Bogert Walter H. Eddy *Jacques Loeb *Graham Lusk *S. J. Meltzer *Lafayette B. Mendel *C-E. A. Winslow Ella Hazel Clark Helen Gavin *Marie L. Minor Helen G. Russell Emily C. Seaman Mary B. Stark Helen S. Watt *Mary D. Womack J. J. Bronfenbrenner tJ. G. M. Bullowa A. J. Goldfarb Benjamin Horowitz Louis Hussakof fMax Kahn fWilliam Weinberger Charles Weisman *Ch'lotte G. Bultman Mary C. de Garmo *tMary B. Kirkbride Jessie A. Moore *Fairfax T. Proudfit *Mird D. Schlesinger Tula Lake Harkey *Israel S. Kleiner Alfred P. Lothrop Anton R, Rose *Mary D. S. Rose * Guest. * James Ewing *Cyrus W. Field Nellis B. Foster F. G. Goodridge *P. A. Levene ♦William H. Park *E. E. Smith fAlexander Smith *Karl Vogel *John Auer C. Stuart Gager *Walter A. Jacobs *F. H. McCrudden H. O. Mosenthal *Edgar W. Olive *D. D. Van Slyke *Jerome Alexander *tJ. P- Atkinson *Edwin J. Banzhaf Charles F. Bolduan fWm. B. Boyd fF. T. Van Beuren Harry Wessler tH. B. Wilcox B. C. Gruenberg Max Morse Raymond C. Osburn *Louisa Bruckman *Harriet C. Jacobson Marguerite T. Lee Helen McClure Arthur Knudson ♦fAlbert Plaut Edward Plaut *Eugene Unna *E. H. Bartley ♦Walter A. Bastedo *tS. P. Beebe ♦Charles A. Doremus ♦Henry C. Sherman Matthew Steel ♦Frederick H. Sykes ♦H. T. Vulte Ula M. Dow Ada M. Field Ruth S. Finch ♦Edith C. Keefer ♦Ethel MacMillan ♦Alice E. Skinner ♦Wilhelmina Spohr Helen B. Thompson ♦Ronald M. Ferry T. Stuart Hart Paul E. Howe ♦Ewing H. Rand Anna M. Connelly ♦Mathilda L. Mayer Elizabeth Rothermel Ethel W. Wickwire Harvey B. Clough Samuel Gitlow Fred W. Hartwell C. A. Mathewson Donald Gordon John L. Kantor Daniel R. Lucas Ralph G. Stillman t Detained or obliged to leave before the conclusion of the dinner. 324 Biochcmical Ncivs, Notes and Comment' [Jan. *Orabel Chilton ♦Helen Ide Gray Alice H. McKinney *Bessie G. Pond S. R. Benedict R. A. Cooke *M. S. Fine *V. C. Myers *tM. I. Falk *R. G. Reese fWm. W. Tracey G. W. Vandegrift Will H. Chapman *Fred'k H. Morrison *Harold E. Smith Ernst Boas Ernest D. Clark Harry L. Fisher Ross A. Gortner Isidor Greenwald E. Newton Harvey Michael Heidelberger *John J. Kenny J. Buren Sidbury Louis E. Wise William J. Gies Joseph S. Hepburn Walter F. Hume Walter M. Kraus S. Kubushiro E. G. Miller, Jr. P. W. Punnett *T. B. Reed Grover Tracy Proceedings of the eighth scientific meeting. The second meeting of the Association for the academic year 1912-13 was held at the Columbia Medical School, on Friday, Dec. 6, at 4.15 p. m., instead of the regulär weekly departmental seminar. At Dr. Gies' Suggestion the Association began, with this meeting, a series of quarterly sessions for the presentation of the results of research by its members. As presiding officer at the seminars, Dr. Gies out- lined the plan and purpose of these scientific sessions and formally turned over the meeting to the Association. President Eddy then took the chair. The scientific program and abstracts of the papers are given on page 284. The remaining meetings of this series, for the year I9i2-'i3, will be held on February 7, April 4 and June 2. Abstracts of the Communications will be published in the succeeding issues of the BiocHEMicAL Bulletin, Alfred P. Lothrop, Secretary. 3. Columbia Biochemical Department Resignations and appointments. Stapf. The following further changes in the stafT, for the year I9i2-'i3, \vere officially authorized during the quarter ending Dec. 31 : Dr. Jacob Rosen- hloom, associate, resigned to accept the assistant professorship of * Guest. t Detained or obliged to leave before the conclusion of the dinner. 1913] Columbia Biochemical Association 325 biochemistry at the University of Pittsburgh. — Dr. Herman O. Mosenthal, instructor, appointed associate, vice Dr. Rosenbloom re- signed. — Dr. Max Kahn appointed instructor, vice Dr. Mosenthal promoted. — Dr. Clayton S. Smith, instructor, resigned to accept an assistantship in pharmacology in the Bureau of Chemistry, U. S. Department of Agriculture, Washington. — Dr. Louis E. Wise ap- pointed instructor, vice Dr. Smith resigned (page 203). The retirement of Drs. Rosenbloom and Smith from the depart- ment as noted above, after active and very successful terms of Serv- ice, occasioned deep regret among their associates at Columbia, whose hearty good wishes attend them in their new fields of use- fulness. ( See bibliography, below. ) Students. Arbuckle Sugar Co. (Brooklyn) : Abraham Gross, research chemist. — Harriman Research Laboratory (Roosevelt Hos- pital, N. Y.) : Marston L. Hamlin, research assistant. — Industrial School (New Bedford, Mass.) : Constanze C. Hart (Teachers Col- lege), assistant. — State Normal School (Truro, N. S.) : Blanche R. Harris (Teachers College), assistant. — Texas (North) State Nor- mal School: Blanche E. Shuffer (Teachers College), professor of home economics. — N. Y. University and Bellevue Hospital Medical College: Percy W. Punnett, assistant in chemistry. — ^University of Kentucky: Mary E. Sweeny, head of extension department. — Uni- versity of Porto Rico; L. A. Robinson, professor of psychology. — Washington State College (Pullman) : Louise McDanell, instructor in domestic science. — West High School (Rochester, N. Y.) : David F. Renshaw, instructor in chemistry. Dr. Rosenbloom's career. Jacob Rosenbloom was born in Braddock, Pa., on Feb. 25, 1884. His early education was received in the public schools and high school of North Braddock, Pa. At the end of a four-year course at the University of Pittsburgh he received the degree of B.S. in chemistry, in 1905. From 1905 to 1909 he was a Student here at the College of Physicians and Sur- geons, receiving the degrees of M.D. and Ph.D. in 1909. His ma- jor subject for the Ph.D. degree was biological chemistry, with Professor Gies. Dr. Rosenbloom was assistant in this department for the year 1909-19 10; associate (also assistant pathologist to the German 326 BiocJicmical News, Notes and Commcnt [Jan. Hospital, N. Y.), during 1910-12. The summer of 1912 was spent at Johns Hopkins University in clinical medicine. Last October he received and accepted appointment as assistant professor of bio- chemistry in the University of Pittsburgh. Dr. Rosenbloom is a Fellow of the American Association for the Advancement of Science, a member of the American Society of Bio- logical Chemists, Society for Experimental Biology and Medicine, American Chemical Society, Chemists' Club of New York, Sigma Xi, International Psychoanalytic Association, and the Society for Biological Research of the University of Pittsburgh. He married, in June, 191 1, Miss Merla Cohen of Baltimore, Md. Dr. Rosenbloom's publications. 1905. A colorimetric method for the determination of tungsten; Thesis for the degree of B.S., Univ. of Pittsburgh. 1907. Some azolitmin Compounds of mucoids, nucleoproteins and other proteins, with exhibition of products (with Wm. J. Gies) ; Proc. Amer. Soc. Biol. Chem., i, p. 48; Jour. Biol. Chem., iii, p. xxxix. 1909. A contribution to the study of the nature and origin of the Bence Jones protein, with bibliography ; Dissertation, Columbia Uni- versity. Pp. 64. 19 IG. On the effects and fate of injected connective tissue mucoid (with Wm. J. Gies) ; Proc. Amer. Soc. Biol. Chem., i, p. 271 ; Jour. Biol. Chem., vii, p. Iviii. — Is the Bence Jones protein produced from osseoalbumoid ? ; Ibid., p. 227 and p. xiv. — A study of the duodenal Contents in man (with M. Einhorn) ; Arch. Internal Med., vi, p. 666; Int. Beitr. z. Path. 11. Ther. d. Ernähr. Stoffw. u. Verd'krank., ii, p. 184. 191 1. A histological and chemical study of the fatty matter of normal and cryptorchid testes (with F. M. Hanes) ; Jour. Exp. Med., xiii, p. 355. — A study of the nitrogen metabolism in three cases of duodenal alimentation (with M. Einhorn) ; Amer. Jour. Med. Sei., cxlii, p. 7 ; Int. Beitr. z. Path. u. Ther. d. Ernähr. Stoffw. u. Verd'krank., iii, p. 5. — ^A new process for the purification of lipins, with demon- strations (with Wm. J. Gies) ; Proc. Amer. Soc. Biol. Chem., ii, p. 8; Jour. Biol. Chem., ix, p. xiv. — A demonstration of the osmotic pressure exerted by fat (with Wm. J. Gies) ; Proc. Soc. Exp. Biol. and Med., viii, p. 71. — The effects of intraperitoneal injections of epinephrin on the partition of nitrogen in the urine of dogs (with W. Weinberger) ; Ibid., p. 131. — Experiments on the diffusibility of cholesterol esters and 1913] Columbia Biochemical Association 327 of lecithan Compounds (with E. Boas) ; Ibid., p. 132. — The importance of the colloidal nitrogen in the urine in the diagnosis of Cancer (with M. Einhorn and M. Kahn) ; Anier. Jour. of Gastro-Enter., i, p. 12; Arch. f. Verdauungskrank., xvii, p. 557. — On the lipins of the heart muscle of the ox; Science, xxxiv, p. 221 ; Biochem, Bull., i, p. 114. — The effect of pregnancy on the Upins of the ovary and corpus luteum of the cow; Ibid., p. 222 and p. 115. — ^A proposed chemical Classifica- tion of lipins, with a note on the intimate relation between cholesterols and bile salts (with Wm. J. Gies) ; Ibid., p. 51. — Intracellular lipins; Ibid., p. 75. — A review of the history of Bence Jones protein and mul- tiple myeloma; Ibid., p. 161. — The older theories of edema; Ibid.. P- 275- 1912. Osseoalbumoid as a possible precursor of Bence Jones pro- tein; Arch. Internal Med., ix, p. 236. — Spontaneously precipitated Bence Jones protein in urine; Ibid., p. 255. — The glycyltryptophan and tryptophan tests for Cancer of the stomach (with C. H. San- ford) ; Ibid., p. 445. — A note on the distribution of chlorate in a woman fatally poisoned by potassium chlorate; Biochem. Bull., i, p. 483. — A study of the diffusibility of lipins from ether through rubber mem- branes into ether ; Ibid., ii, p. 64. — The colloidal nitrogen in urine from a dog with a tumor of the breast (with M. Kahn) ; Ibid., p. 87. — Effects of intraperitoneal injections of epinephrin on the partition of nitrogen in urine from a dog (with W. Weinberger) ; Ibid., p. 123. — A quanti- tative study of the lipins of bile obtained from a patient with a biliary fistula; Ibid., p. 182. — A disturbing factor in Lieben's and in Gunning's test for acetone in urine ; Jour. Amer. Med. Assn., lix, p. 445. — A report of some new chemical analyses of urinary calculi, with indications for treatment (with M. Kahn) ; Ibid., lix, p. 2252. — The diffusion of iodo- eosin from ether through rubber into ether ; Proc. See. Exp. Biol. and Med., X, p. 48. [Dr. Rosenbloom's papers in this issiie (pp. 22g, 2^^, 2^6, and 2<)Q) were submitted for publication in December 1912.] Awards of degrees at Columbia. -Mr. Anton R. Rose re- cently passed a public examination for the Ph.D. degree, thus com- pleting the requirements for that degree in biological chemistry. His dissertation is entitled Biochemical studies of phyto-phosphates. — Miss Clara W. Hasslock and D. F. Renshaw completed on Oct. 10 the requirements for the degree of A.M. Miscellaneous items. Professor Gies delivered a lecture in 328 Biochemical News, Notes and Comment [Jaa the autumn series at the New York Botanical Garden, October 26, on The chemical production of albuminous substances in plants. On October 7 he addressed the Section on Research of the First District Dental Society of the State of New York, at the Academy of Medicine, on Recent developments in the study of dental caries. Dr. Lothrop followed with a paper on the work he has been doing in this connection on salivary mucin.^ — Professor Gies was one of the Organizers of the New York Gastro-Enterological Society (P- 315)- He is Secretary of a Committee of Twenty-five, of Prof. R. H. Chittenden's pupils, in charge of a dinner to be given at Del- monico's on March i in honor of Professor Chittenden, and of a fund to be given to Yale in the name of Professor Chittenden. •Gies: Journal of the Allied (Dental) Societies, 1912, vii, pp. 397 and 478; Lothrop : Ibid., p. 410. EDITORIALS A year's experience in the conduct of the Biochemical Bulle- tin has induced us to change our plan of quarterly issue. The manuscript of future numbers will be sent to the printer on the first New plan of ^^Y ^^ ^^^^^ natural quarter; the months of issue quarterly issue of will be January, April, July and October; and the Bulletin ^^g contents of each issue will pertain to the quar- ter preceding the month of issue. Volume II will close with the July number. We congratulate President Taft and the country on the appoint- ment of Dr. Carl L. Aisberg, in succession to Dr. Harvey W. Wiley, as Chief of the Bureau of Chemistry. Dr. Alsberg's training in chemistry, in general biology and in medicine has been unusually broad and deep. His chemical knowledge, his sanitary comprehension, his scientific wisdom, and his zeal as an investigator, have had exceptional fruitage through- out his entire professional career. Admired as a gentleman by all who know him and respected by his colleagues everywhere as a sci- entist of eminent capacity, Dr. Aisberg is also universally esteemed for his habitual fidelity to duty, his moral integrity and his high professional purpose. We look forward with great confidence to a career for Dr. Aisberg which will be distinguished by a patriotism, a zeal in public Service, a personal courage, a common sense, a sci- entific exactness, an aggressiveness in the detection of violations of law, an executive capacity, that will be the delight of all his biochem- ical colleagues and the pride of his countrymen — and in this faith we tender him our felicitation and support. As we are about to close this issue of the Bulletin, we learn that the following testimonial (as proposed by Prof. Graham Lusk), which was sent a few days ago (Jan. 21) to about 275 of Dr. Alsberg's fellow workers in the American Physiological Society and 329 330 Biological Che mists in Hospitals [Jan. the American Society of Biological Chemists, has already been signed by nearly all of them : To Dr. Carl L. Aisberg: We who, like yourself, are active workers in the field of experimental biological science, congratulate the country and yourself on your appointment as Chief of the Bureau of Chemistry in the Department of Agriculture. We wish to express our confidence in your ability and integrity. We desire for you a successful admin- istration which shall promote the public welfare, shall jealously guard the public health, and shall uphold the dignity of the science which you represent. We are greatly indebted to Dr. Adler for the biographical and bibliographical facts, pertaining to Dr. Aisberg, on pages 211-216 of this issue. During the past winter a number of cases of stock poisoning due, apparently, to the feeding of spoiled or moldy silage, were brought to our attention. At that time we were unable to give the Stock poisoning niatter due consideration. During the present due to spoiled si- winter, however, we shall be in a position to make läge. Help! some preliminary studies to ascertain the cause of toxicity in silage. Our work would be greatly facilitated if those readers of the Biochemical Bulletin who know of such cases of stock poisoning would bring this matter to the attention of the owners so that samples of the silage might be forwarded to the Chemical Section of the Iowa State College. The samples should be accompanied by füll particulars regarding the apparent cause of spoilage and the Symptoms exhibited by the animals to which the silage was fed. Arthur W. Dox. Until recently the pathological work in our larger hospitals has been done by attending physicians. The growth of this special field, however, has made it impossible for any one to keep abreast with Demand for bio- ^^ ^"^ ^^ anything eise. The progress of medical logical chemists in science constantly tends toward what is most re- the hospitals fined and subtle. As the microscope revealed new fields of research, so chemistry has opened previously unim- 1913] Editoriais 33 ' agined paths for investigation. The directors of Hospitals are alive to this fact, as is shown by the increasing demand for biological chemists to cooperate with pathologists in the investigation of dis- ease. Up to now relatively few men have been adequately trained to be hospital chemists. What is required of them, and what will be demanded of them more and more, is not the making of routine analyses at the Suggestion of some attending physician, who in all probability has but a vague idea of what he wants; but rather that, unguided, they shall be able to discern in any disease process a defi- nite problem for investigation, and shall be competent to establish the conditions and conduct the details of suitable experimental re- search thereon. In order to do this with any degree of success, a hospital chemist must, in addition to his knowledge of biological chemistry, have a fairly good understanding of general pathology and bacteriology. It is not necessary that he be a physician, since technical familiarity with the clinical aspects of disease will not be reqnired of him. In the next decade, if not sooner, there will be a great demand for this type of highly trained biological chemist. The beginning of this demand is seen now in the growing number of biological chemists attached to the main hospitals of our larger cities. These chemists are on the same footing with the pathologists and the bac- teriologists. Although at present their remuneration is not what it should be, this will be remedied as soon as their value to hospitals is clearly shown. This field of work should be particularly attrac- tive to those who do not care for an academic career but who are devoted to biochemical research and averse to commercial chemistry. In the fall of 191 1, when it was seriously proposed to merge the American Society of Biological Chemists into the American Physio- logical Society, we were among the many who objected to the plan Federation of °^ ^^^ ground that such a merger would be detri- American Societies mental to biological chemistry as a science and for Experimental as a profession — sufficient reason for dissenters, 10 ogy ^^^^ •£ ^ mej-ger were ordered, to maintain the existence of an independent American Biochemical Society.^ In ^Editorial: Biochemical Bulletin, 1911, i, p. 364. 332 Electrons [Jan. presenting these objections informally to our colleagues \ve empha- sized, however, the desirability of more intimate affiliation between the leading biological societies, in harmony zvith the policy of the American Society of Biological Chemists front its estahlishment, and suggested the Organization of a " federation" of independent societies for the attainment of that purpose and other mutually advantageous objects. There was much discussion but no decision. The Organization of the Biochemical Society of England, mean- while, with all that its existence implies,^ gave added weight to the objections that were raised against the assimilation of the Amer- ican Society of Biological Chemists by the American Physiological Society. During the past year the " federation " idea has won its way into unanimous acceptance, as is indicated by the account in this issue of the Organization of the Federation of American Societies for Ex- perimental Biology (page 269). The Federation is, in effect, an embryonic American Biological Society, the independent societies being its working divisions. The plan of federation has not weak- ened the independence or impaired the autonomy of any of the con- stituent societies. We heartily commend to the attention of all our readers the Mathews plan for the Organization of the American Biological So- ciety, which is published in füll in this issue (page 261 ). We believe that the logical development of the Federation would secure all the many desirable results at which Professor Mathews' excellent and far-reaching plan is aimed, including the establishment and success- ful conduct of a Biological Abstract Journal. We hope to present the views of some of our colleagues on this and related subjects in our July issue. The knowledge of nature as it is — not as we imagine it to be — constitutes true science. — Paracelsus. „ Liability to error is the price we pay for forward Electrons -^ o- , • i. movement. — o idgwick. The secret of all who make discoveries is to look upon nothing as impossible. — von Liehig. ^Halliburton: Biochemical Bulletin, 1912, i, p. 484; ii, p. 128; 1913, ii (this issue), p. 318. BOOKS RECEIVED The BiocHEMicAL Bulletin will promptly acknowledge, under this heading, the receipt of all publications that may be presented to it. From time to time, selections will be made for review on pages of the volume to be appropriately indicated here. Reviews will be matter-of-fact Statements of the nature and Contents of the publications under consideration, and will be intended solely to guide possible ptirchasers. The wishes or expectations of publishers or donors of volumes will be disregarded, when they are incompatible with our convictions regarding the interests of our colleagues. The sises of the printed pages are indicated, in inches, in the appended notices. Glycosuria and allied conditions. By P. J. Cainmidge. Pp. 467 — 4 X 6.;4 ; $4.50 net. Longmans, Green & Co.. New York; Edward Arnold, London, 1913. The chemical Constitution of the proteins: Part II, Synthesis, etc. 26. ed. (One of the Monographs on Biochemistry.) By R. H. A. Plimmer, Univ. reader and ass't prof. of physiological ehem., University Coli., London. Pp. 107 — 4^ X 7l/i ; $1.20 net. Longmans, Green & Co., 1913. Microscopy and the microscopical examination of drugs. By Chas. E. Gabel, microscopical food and drug analyst, Iowa State Dairy and Food Commis- sion. Pp. 116 — 4X6H', $1.00. Kenyon Co., Des Moines, la., 1911. Collected papers: Laboratory of physiological chemistry, Sheffield Sci- entific School, Yale University. 1911-1912. (35 reprints.) Medical and surgical report of Bellevue and Allied Hospitals in the City of New York. By Van Home Norrie, John A. Hartwell, A. Alexander Smith and Charles E. Nammack. Vol. iv, 1909-1910. (55 reprints.) Report of the laboratories of the University of BufFalo, medical depart- ment; including the third Harrington lecture (Hektoen). No. 4. 1912. (8 reprints.) Contributions from the physiological laboratory of the Medico-Chirurgi- cal College, Phila. By Isaac Ott and John C. Scott. Part xix of Ott's con- tributions to physiology, 1912. (13 reprints.) Report of the Pellagra Commission of the State of Illinois. Pp. 250 — 4]4 X 7. Nov., 1911. Practical physiological chemistry. A book designed for use in courses in practical physiological chemistry in schools of medicinc and of science. By Philip B. Hawk, professor of physiological chemistry and toxicology in the JeflFerson Medical College of Philadelphia. Fourth edition, revised and en- larged. Pp. 475—45^X8; $2.50 net. P. Blakiston's Sons & Co., Philadelphia, 1912. The protein dement in nutrition. (One of the International Medical Mono- graphs.) By Major D. McCay, professor of physiology, Medical College, Cal- cutta. Pp. 216 — 4X7. with 8 füll page portraits of human subjects; $2.00 net. Longmans, Green and Co., New York; Edward Arnold, London, 1912. Oxidations and reductions in the animal body. (One of the Monographs on Biochemistry.) By H. D. Dakin, The Herter Laboratory, New York. Pp. 135 — 4)/^X8; $1.40 net. Longmans, Green and Co., 1912. Researches on cellulose. III (1905-1910). By C. F. Gross and E. J. Bevan. Pp- U3 — 3/^X6; $2.50 net. Longmans, Green and Co., 1912. An introduction to the study of the protozoa, with special reference to the parasitic forms. By E. A. Minchin. professor of protozoology in the Univer- sity of London. Pp. Si/— 4X7l^; $600 net. Longmans, Green and Co., New York; Edward Arnold, London, 1912. OFFICERS OF THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-1913* OFFICIAL REGISTER, DEC. 31, 1912 William J. Gies: Professor and Chairman of the Stoff; Consulting chemisl, New York Botanical Garden; Pathological chemist, Bellevue Hospital; Mem- bcr of the Faculties of N. Y. Teachers College and N. Y. College of Pharmacy. [B.S., Gettysburg College, 1893 and M.S., 1896; Ph.B., Yale University, 1894 and Ph.D., 1897. Instructor, i898-'o2; adjunct professor, 1902-05; Professor, 1905-.] Paul E. Howe: Assistant Professor. [B.S., University of Illinois, 1906; A.M., 1907 and Ph.D., 1910. Assistant professor, 1912-.] Nellis B. Foster: Associate; Associate Physician, New York Hospital ; Chemist, St. Luke's Hospital. [B.S., Amherst College, 1898; M.D., Johns Hopkins University, 1902. Instructor, i9o6-'o8; associate, 1908-.] Walter H. Eddy : Associate and Secretary of the Staff. [B.S., Amherst Col- lege, 1898; A.M., Columbia, 1908 and Ph.D., 1909. Assistant, i9o8-'io; associate, 1910-.] Alfred P. Lothrop: Associate and Departmental Registrar. [A.B., Oberlin, 1906 and A.M., 1907 ; Ph.D., Columbia, 1909. Assistant, i9o8-'o9 ; instructor, i909-'i2; associate, 1912-.] Herman O. Mosenthal : Associate; Assistant Attending Physician, Presbyterian Hospital; Assistant Physician, Vanderbilt Clinic; Instructor in medicine. [A.B., Columbia, 1899 and M.D., 1903. Assistant, i9o8-'09 ; instructor, i909-'i2 ; associate, 1912-.] Emily C. Seaman: Instructor. [B.S., Adelphi College, 1899; A.M., Columbia, 1905 and Ph.D., 1912. Tutor, i90(>-'io; instructor, 1910-.] Max Kahn: Instructor. Director of the chemical and physiological laboratories of Beth Israel Hospital. [M.D., Cornell University Medical College, 1910; A.M., Columbia, 191 1 and Ph.D.. 1912. Instructor, 1912-.] Louis E. Wise: Instructor. [A.B., Columbia, 1907 and Ph.D., 191 1. Instructor, 1912-.] Edgar G. Miller, Jr. : Assistant, 1911-. [B.S., Gettysburg College, 1911.] Frederic G. Goodridge: Assistant, 1912-. [A.B., Harvard University, 1897; M.D., Columbia, 1901.] Arthur Knudson: Assistant, 1912-. [A.B., University of Missouri, 1912.] Ethel Wickwire: Assistant, 1912-. [A.B., Tri-State College, 1909.] Tula L. Harkey: Assistant, 1912-. [A.B., Colorado College, 1909.] Benjamin Horowitz: Assistant, 1913-. [B.S., Columbia, 1911 and A.M., 1912.] Christian Seifert: Laboratory assistant, 1898-. Stella Waldeck : Recorder, 1908-. Blanche E. Shaffer: Laboratory assistant. summer Session, 1912. Joseph S. Hepburn : University fellow, 1912-13. [A.B., Central High School, Philadelphia, 1903 and A.M., 1908; B. S., University of Pennsylvania, 1907 and M.S., 1907.] *The work of the department was inaugurated in October, 1898, by Prof. R. H. Chittenden (lecturer and director), Dr. William J. Gies (instructor), Messrs. Alfred N. Richards and Allan C. Eustis (assistants), and Christian Seifert (laboratory assistant). COURSES OFFERED BY THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY. 1911-1913 Courses 51, 105 and 215 are given during the first half-year only. Course loi is given during the first half-year and is repeated (102) during the second half-year. Courses 104 and iio (52) are given only during the second half year. All other courses are conducted throughout the entirc academic year. All courses not otherwise specified are given at the College of Physicians and Surgeons. (Abbreviations: C, Conference; D, demonstration ; L, lecture; Lw, labora- tory work; R, recitation.) ORGANIC CHEMISTRY 51. Elkmentary ORGANIC CHKMisTRY. Introductory to courses loi, 102 and HO (52). {Required of first year siudents of medicine.) L, i hr. D, i hr. R, 2 hr., each section (2). Lw. 6 hr. each section (2). Profs. Gies and Howe, Drs. Wise and Goodridge, and Messrs. Miller and Knudson. NUTRITION (PHYSIOLOGICAL AND PATHOLOGICAL CHEMISTRY) 101-102. General physiological chemistry. A course in the eletnents of normal nutrition. (Teachers College, School of Practical Arts.) L, 2 hr. R. l hr., each section (2). Lw, 5 hr., each section (2). Prof. Gies, Dr. Seaman and Misses VVickwire and Harkey. (This course is designated "Chemistry 51" and "Household Arts Education 125" in the Teachers College Announcement.) This course is designated "Chemistry s 51 " in the Teachers College Division of the Summer School Announcement. The course was given last summer by Prof. Gies, Dr. Seaman and Miss Shaffer. 104. General patholooical chemistry. Lectures on nutrition in disease. (Teachers College, School of Practical Arts.) L, i hr. Prof. Gies. (This course is designated "Chemistry 52" in the Teachers College Announcement.) HO (52). General physiological chemistry. A course in the Clements of normal nutrition. (Required of first year students of medicine.) L, 2 hr. R. i hr.. each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe. Dr. Wise, and Messrs. Miller and Knudson. This course is designated "S — 104" in the Medical Division of the Summer School Announcement. It was given last summer by Prof. Gies and Dr. Smith. 209-210. Chemistry of nutrition. (School of Pharmacy. Required of candidates for the Degree of Doctor of Pharmacy.") L, 1 hr. Prof. Gies. 211-212. General biological chemistry. Specially adopted to the needs of secondary school teachers of biology. L, i hr. Lw, 4 hr. Dr. Eddy. 213-214. Advanced physiological chemistry, including methods of re- SEARCH IN nutrition. (Teachers College, School of Practical Arts.) L, i hr. Lw, 5 hr. Prof. Howe, Dr. Seaman and Mr. Horowitz. (This course is desig- nated " Household Arts Education 127" in the Teachers College Announcement.) 215. General biological chemistry. A course in the eletnents of normal nutrition. L, i hr. Lw, 7 hr. Prof. Gies, Dr. Lothrop and Messrs. Miller and Knudson. 217-218. BlOCHEMICAL methods OF RESEARCH, INCLUDING CLINICAL METHODS AND URiNARY ANALYsis IN GENERAL. L, I hr. Lw, 7 hr. Profs. Gies and Howe, Dr. Lothrop, and Messrs. Miller and Hepburn. 219-220. Nutrition in health. A laboratory course in advanced physio- logical chemistry. L, 2 hr. Lw, 14 hr. Profs. Gies and Howe, and Dr. Lothrop. Courses in Nutrition (continued) 221-222. Nutrition in Disease. A lahoratory course in advanced patholog- ical chemistry. L, 2 hr. Lw, 14 hr. Prof. Gies. 223-224. Nutrition in Disease. L, i hr. Profs. Gies and Howe, and Drs. Foster, Mosenthal, Kahn and Goodridgc. 225-226. Advanced physiological and patholocical chemistry, including ALL PHASES OF NUTRITION. Research. C, I hr. (individual students). Lw, 16 hr. Profs. Gies and Howe, and Dr. Lothrop. fm^COLOGY 231-232. Effects and detection oi«*poi.sons, including food preservatives and adulterants. Lw, 6 hr. Prof. Gies and Mr. Miller. BOTANY 235-236. Chemical piiYsiOLOGY of PLANTS. (New York Botanical Garden.) L, 1 hr. Lw, 5 hr. Prof. Gies. BACTERIOLOGY 241-242. Chemistry of microorganisms: fermentations. putrefactions AND THE behavior of enzymes. An introducHon io sauitary chemistry. L, i hr. Lw, 7 hr. Prof. Gies. SANITATION 105. Sanitary chemistry. (Teachers College, School of Practical Arts). L. I hr. Lw, 3 hr. Dr. Seaman and Miss Harkey. (This course is designated " Chemistry 57 " and " Household Arts Education 129 " in the Teachers College Announcement. ) BIOCHEMICAL SEMINAR 301-302. Biochemical Seminar, i hr. Prof. Gies. RESEARCH IN BIOLOGICAL CHEMISTRY Biochemical research may be conducted, by advanced workers, independently or under guidancc, in any of the departmcntal laboratories. LABORATORIES FOR ADVANCED WORK IN BIOCHEMISTRY The laboratories in which the advanced work of the biochemical department is conducted are situated at the College of Physicians and Surgeons, Teachers College, New York Botanical Garden and Bellevue Hospital. Fach lahoratory is well equipped for research in nutrition and all other phases of biological chemistry. BIOCHEMICAL LIBRARY Prof. Gies* library occupies a room adjoining the main biochemical lahora- tory at the College of Physicians and Surgeons and is accessible, by appoint- ment, to all past and present workers in the Department. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION The Biochemical Association holds scientific meetings regularly on tiie first Fridays in December, February and April, and on the first Monday in June. These meetings are open to all students in the University. SUMMER SCHOOL COURSES Summer Session courses are mentioned in the foregoing references to Courses 101-102 and iio (52). Prof. Gies will have charge of both courses next Summer. He will also conduct a special lecture course in nutrition. The labora- tories will be open for research throughout the summer. ANNOUNCEMENTS Professional Assistance Offered to Biological Chemists The Columbia University Biochemical Association will be glad to cooperate confidentially with all who desire the Services of biological chemists and with all who seek positions in biological chemistry. Address inquiries to William J. Gies, 457 West sgth St., New York. Joumalistic New JOURNAL. Physiological Researches. To appear at irregulär intervals. Edited by Burton E. Livingston, Manager, Johns Hopkins University ; jDamV/ T. MacDoiigal, Carnegie Institution of Washington ; Herbert M. Richards, Columbia University. " The recent rapid advance of physiological science has been accompanied by a realization of the Community of interest and uniformity of method which characterize the physiology of plants and of animals, and it has seemed highly desirable that the general physiological field thus indicated should possess an Organ of publication in which its more comprehensive and technical papers might appear. This need is emphasized by the fact that pres- ent facilities for publication in physiology are generally taxed beyond their capacity and papers are consequently subject to long delays in appearance. It has therefore been decided to inaugurate a new series of scientific papers which will embrace contributions towards the advance of fundamental physiological knowledge." " The plan of publication of the new series, for which the title of Physiological Researches has been adopted, is one in which practical ownership is vested in the contributors. It is hoped that the project will receive the interest and support of biologists of all classes. (Each volume will contain about 450 pages; each number will contain but a Single contribution ; and the numbers will be issued irregularly). Pub- lication of the first contribution ma)'^ be expected in a short time. Sub- scriptions will be received by the volume, the price being $5.00 per volume, payable in advance. Subscriptions to volume I, which are received prior to the date of publication of the first research, may be made at the reduced price of S4.00. At the date of the appearance of the first research the price will automatically become the regulär one. Remittances should be made payable to Physiological Researches, and all correspondence should be addressed to Physiological Researches, Station N, Baltimore, Maryland, U. S. A." (Editors' announcement.) Reduced subscription price of the Journal of Biological Chemistry, The directors of the Journal of Biological Chemistry have announced that "beginning with the February issue of 1913 (Vol. 14, No. i) the subscription price of the Journal to domestic subscribers will be reduced f rom $4.00 to $3.00 per volume ; to foreign subscribers, $3.25. Any one engaged in biochemical work who subscribes for the Journal at this rate (beginning with Vol. 14) may secure Volumes 1-13 for $20.00, plus cost of transportation. The price at which a complete set has hitherto been sold is $50.00. Subscribers for the Journal who wish to complete their files may secure early volumes for $1.50 each, plus cost of transportation. Address : Alfred N. Richards, Secretary, University of Pennsylvania. Meetings of Societies and Congresses A-MERicAN Chemical Society: Annual meeting (47th) at Mil- waukee, Wisconsin, March 25-28. Charles L. Parsons, Secretary, Box 505, Washington, D. C. At the last meeting of the Society the Council authorized the formation of a Div^ision of Biological Chemistry. At that meeting the details of Organization of the Division were entrusted to a committee. The committee will report in Milwaukee, the final Organization of the Division will be perfected, and officers will be elected. Tenth International Congress of Agriculture: Ghent, Bel- gium, June 8-13. Secretary-general, Dr. P. de Vuyst, 22 Avenue des Germaines, Brüssels. American committee: Dr. L. O. Howard, mem- ber of the International Commission on Agriculture and chief of the Bureau of Entomolog>'; and Dr. A. C. True, director, Mr. John Ham- ilton, specialist in farmers' institutes, Dr. C. F. Langworthy, chief of nutrition investigations and Dr. J. I. Schulte, assistant agriculturist, of the Office of Experiment Stations. American Medical Association, Annual meeting: Minneapolis, Minn., Jiine 17-20. General secretary, Geo. H. Simmons, 535 Dear- born Ave., Chicago. General meeting of the International Association of Botan- iSTs: Copenhagen, June 23. Secretary-general, J. P. Lotsy, Haarlem, Holland. Seventeenth International Congress of Medicine: London, Aug. 6-12. General secretary, Dr. W. P. Herringham, 13 Hinde St., London, W. FouRTH International Congress on School Hygiene: Buflfalo, N. Y., Aug. 25-7,0. Secretary-general, Prof. Thomas A. Storey, Col- lege of the City of New York. NiNTH International Physiological Congress: Groningen, Hol- land, Sept. 2-6. American Secretary, Prof. W. T. Porter, Harvard Medical School. Third International Congress of Refrigeration : Washington, D. C, Sept. if) (opening meeting) ; Chicago, Sept. 17-23 (business and scientific meetings). Secretary-general, Mr. J. F. Nickerson, 431 So. Dearborn St., Chicago. The Biochemical Bulletin The Biochemical Bulletin is a quarterly biochemical review. 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Vol. II April, 1913 No. 7 Biochemical Bulletin Edited, for the Columbia University Biochemical Association, by the EDITORIAL COMMITTEE: ALFRED P. LOTHROP, Chairman, PAUL E. HOWE, Secretary, WILLIAM J. GIES, Treasurer, WALTER H. EDDY, JOSEPH S. HEPBURN, H. O. MOSENTHAL. NELLIS B. FOSTER, MAX KAHN, EMILY C. SEAMAN, F. G. GOODRIDGE, ARTHUR KNUDSON, ETHEL WICKWIRE, TULA L. HARKEY, EDGAR G. MILLER, JR., LOUIS E. WISE, all of the Staff of the Biochemical Department of Columbia University. CONTENTS Heinrich Ritthausen (Portrait) 334 Appreciation. Thomas B. Osborne ; 335 BiBLiOGRAPHY. Lewis IV. Fetzer. 339 Dinner To Professor Chittenden : Testimonial by his pupils, '94S. 349 Society for Experimental Biology and Medicine : Tenth anniversary meeting and dinner. Nineteen O. Three 358 Methods for the Electrometric Determination of the Concentration of Hydrogen IONS IN BiOLOGiCAL FLUIDS. K. A. Hasselbalch 367 A Method for the Determination of Tryptophan derived from Protein. Jesse A. Sanders and Clarence E. May 373 Physical Chemistry of Muscle Plasma. Filippo Bottazzi '. 379 Fasting Studies : h. A Note on the Composition of Muscle from Fasting Dogs. H. C. Biddle and Paul E. Howe 386 Some Notes on the Form of the Curve of Carbon-dioxide Excretion Result- iNG FROM Muscular Work Following Forced Breathing. G. O. Higley... 390 The Influence of Barometric Pressure on Carbon-Dioxide Excretion in Man. G. O. Higley 393 The Relation of Acapnia to Shock, and a Consideration of the Mechanical Effects of Artificial Hyper-Respiration upon the Circulation. Henry H. Janeway and Ephraim M. Ewing 403 Cleavage of Pyromucuric Acid by Mold Enzymes. Arthur W. Dox and Ray E. Neidig. 407 Analysis of the Ash of the Castor Bean. Marston Lovell Hamlin 410 Notes on the Chemical« Nature of the " Tannin Masses " in the Fruit of the Persimmon. Ernest D. Clark 412 HiSTON AND ITS Preparation. Walter H. Eddy 419 Did von Wittich Antedate Ostwald in the Definition of Enzyme Action ? William N. Berg 441 The Biochemical Society, England 446 Scientific Proceedings of the Columbia University Biochemical Association. Alfred P. Lothrop, Secretary 452 Biochemical Bibliography and Index. William J. Gies 470 Biochemical News, Notes and Comment 476 Editorials : Including numerous quotations from letters on the Mathews plan for the Organization of an American Biological Society 487 NEW YORK Columbia University Biochemical Association. Entered as second-claas matter in the Post Office at Lancaster, Pa. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Members PROF. R. H. CHITTENDEN, First Director of the Columbia University De- partment of Biological (Physiological) Chemistry; Director of the Shef' field Scientific School of Yale University PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Switserland PROF. SAMUEL W. LAMBERT, Dean of the Columbia University School of Mediane DR. JACQUES LOEB, Member of the Rockef eller Institute for Medical Re- search; Head of the Department of Ex perimental Biology PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. LEON ASHER, University of Bern, Switserland PROF. FILIPPO BOTTAZZI, University of Naples, Italy PROF. VLADIMIR S. GULEVIC, University of Moscow, Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden, — C. Stuart Gager. College of the City of New York. — ^Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Goldfarb. Columbia University: Departments. — Anatomy: Alfred J. Brown, H. von W. Schulte; Bacteriology: James G. 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Russell Sage Institute of Pathology. — Eugene F. DuBois. TuRCK Institute. — Anton R. Rose. Vettin School. — Laura l. Mattoon. Leopold L. Falke, 5316 Thirteenth Avenue, Brooklyn ; Mahel P. Fitzgerald, 416 East 65th Street, Manhattan; Abraham Gross, c/o Arbuckle Sugar Co., Brooklyn; Alfred H. Kropff, 619 Kent Avenue, Brooklyn. Non-Resident Members Allegheny General Hospital (Pittsburgh). — James P. McKelvy. Carnegie Institution (Cold Spring Harbor, L. I.). — Ross A. Gortner. Cornell University (Ithaca). — Jean Broadhurst. Drake University Medical School (Des Moines, la.). — E. R. Posner. Forest School (Biltmore, N. C). — Homer D. House. Iowa University Hospital (Iowa City). — Louis Baumann. Isolation Hospital (San Francisco, Cal.). — L. D. Mead. Jefferson Medical College (Phila.). — P. B. Hawk, Edward A. Spitzka. Johns Hopkins University (Baltimore). — John Howland, W. M. Kraus, Burton E. Livingston, Edwards A. Park. Lehigh University (Bethlehem, Pa.). — William H. Welker. 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University of Montana (Missoula). — J. E. Kirkwood. University of Pennsylvania (Phila.). — A. N. Richards. University of Porto Rico (Las Pietras). — L. A. Robinson. University of Tennessee (Memphis). — Edwin D. Watkins. University of Texas (Austin). — Mary E. Gearing, Anna E. Richardson. University of Toronto (Canada). — Olive G. Pattcrson, University of Utah (Salt Lake City). — H. A. Mattill. University of Wisconsin (Aladison). — W. H. Petersen. Vassar College (Poughkeepsie, N. Y.). — Winifred J. Robinson. Washington State College (PuUman). — ^Joscphine T. Berry, Louise McDanell. Wesleyan University (Middletown, Conn.). — David D. Whitney. West Penn Hospital (Pittsburgh). — Jacob Rosenboom. Williams College (Williamstown, Mass.). — John S. Adriance. Yale University (New Haven, Conn.). — Lorande Loss Woodruff. Albert H. Allen, Saranac Lake, N. Y. ; Emma A. Buehler, Newark, N. J.; George A. Geiger, West Orange, N. J.; Edward G. Griffin, Albany, N. Y. ; F. C. Hinkel, Utica, N. Y.; Cavalier H. Joüet, Roselle, N. J.; A. E. Olpt>, West Hoboken, N. J.; Adeline H. Rowland, Pittsburgh, Pa. ; William A. Taltavall, Redlands, Cal. ; David C. Twichell, Saranac Lake, N. Y. EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the collaboration of the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockefeiler Institute for Medical Research PROF. WILDER D. BANCROFT, Cornell University,- Ithaca DR. CHARLES A. DOREMUS, 55 W. 53d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station, Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR. LEWIS W. FETZER, U. S. Dep't of Agriculture, Washington, D. C. PROF. MARTIN H. FISCHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. M. HARDISTY, McGill University, Montreal, Canada DR. K. A. HASSELBALCH, Finsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, A^. Y. Univ. and Bellevue Hospital Med. College PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan PROF. LAFAYETTE B. MENDEL, Yale University PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven DR. AMOS W. PETERS, The Training School, Vineland, N. J. PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon PROF. UMETARÖ SUZUKI, University of Tokyo, Japan MISS ANNA W. WILLIAMS, University of Illinois, Urbana, lll. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switzerland DR. JULES WOLFF, 26 Rue Dutot, Paris HEINRICH RITTHAUSEN. BiocHEMiCAL Bulletin Volume II APRIL, 191 3 No. 7 IN MEMORIAM HEINRICH RITTHAUSEN Born January 13, 1826 Died October 16, 1912 Bv the death of Heinrich Ritthansen in Berlin, on October 16, 1912, at the age of eighty-six, a long life spent in biochemical research was terminated. Beginning as a Student under Liebig, and inspired by this great teacher, he niade agricnltural chemistry bis life profession. His first work was as assistant to Professor Erd- mann at Leipzig. From 1854 to 1856 he was director of the scien- tific department of the experiment Station at Moeckern. He theo became director of the Station at Saaran in Schlesien. In 1858 he was appointed professor of chemistry and physics in the Royal Agricnltural Academy at ^^"aldau ; in 1867, professor of chemistry and director of the experiment Station at Poppeisdorf; and in 1873, professor of chemistry at Königsberg, where he remained until 1899. when his active career was concluded by advancing years. The latter part of his life was spent in Berlin. At the present time, when the development of agricnltural sci- ence has made piain the value of the agricnltural experiment Station not only to the f armer jjut to the entire Community, the life of one who commenced his career in the first established institution of this kind is of special interest. A\'hen Ritthausen began his work, the brilliant writings and lectures of Liebig had directed the attention of the whole world to the importance of applying the discoveries of science to the practice 't -1 - 3,3^ ^Ic'mricli l\inh(nisrn lApril of ai;ricu]lui"c. As i»nly ihe nidinicMils of a kiiowlcdge of llie cliemical and ])hysical ])r()l)lenis of ihe i^rowlh and niaintenance of plants and animals liad l)een ac(|nire(l, it seemed a sim])le matter to instrucl ihe farnier in proper melhods to l)e eniployed in raising liis crops and stock. No dou1)t remained in the minds of the early scientists \\\\t) promoted this propaganda that ihe practical retiirns of their efforts wonld soon l)e reahzed. Planis were to l)e fed with carbonic acid, nitrogen, and inorganic salts, and ihe proper (jnaniity of each essential dement of plant food was to l)e determined l)}- chemical analysis of the tissnes and ash of the plant, the fertilizer snpplied and the soil in w hich it grew. Animals were supposed to be composed of substances directly or indirecll}" assimilated from their foods. Their heat and mechanical energy was snpplied by the carljohydrates and fats ; the all)nmin, fibrin. casein and gelatin ])resent in their Ijlood, mnscle, milk, etc., \vas fnrnished !)}■ identical proteins contained in the \arions vege- table ])r()dncts \\\{h wliich they were fed. Here again chemical analysis was to fnrnish the gnide to proper practice, which shonld replace crude and wastefnl methods fonnded in ignorance of the real factors involved. I.ittle did the enthusiasts who established the first agricultural experiment Station realize what was before them nor what dis- coxeries in e\'ery Ijranch of l)ii)logical science their efforts were to lead to. Little did they dream of enzymes and liydrolyses, of toxins and antitoxins and specificity of li\-ing tissues, of optical isomers and tantomeric componnds, of nncleic acids and pyrimidines, of amino-acids and Polypeptides, of colloids and surface tension, nor of mnltitndes of other discoveries ^vhic]^ have followed chiefly from the inspiration which Liebig imparted to those who worked Avitli liim. All tliese disco\-eries had to ])e made l^efore the practical jjrob- lems of agricnltnre conld be satisfactorily dealt with by the scientist ; and it is evident that Ritthausen was one of tlie first to realize this, for we find him, after a short experience in attempting an immediate application of chemistry to the feeding of cattle, tnrning bis atten- tion to a carefnl study of the protein constituents of their vegetable foods. 1913I Thomas B. Osbonie 337 His earliest paper on this subject, which appeared in the Jonnial für praktische Chemie in 1862, described the proteins of wheat ; and for five successive years he pnblished papers on this same sub- ject. In the conrse of this work he isolated glutaminic acid from the products of hydrolysis of the gluten proteins, a discovery which ranks among the more important made by biochemists. He then extended his investigations to seeds of importance for nutrition, hardlv a year passing when he did not contribute two or more papers on the resnlts of his work. In 1872 he pnl)hshed a review of his earher work nnder the title Die Eiz^'cisskörpcr der Getreidearten. HiUseufrüeJiteii und Oelsamen. This was the first attempt made to furnish an account of wliat had been learned respecting the properties of proteins of vegetable origin. Akhongh this work contained nmch of vahie to animal physiologists, and was suggestive in man\- ways in con- nection with the problems then claiming their attention, few of them appear to have read it with the care that it deserved. Authors of text-books on physiological chemistry, for many years after, dis- covered in it nothing more than the fact that Ritthausen employed dikite alkahne Solutions in isolating his preparations, and con- sec[uently dismissed his results with the Statement that all his products were altered in their preparation and so deser\'ed little consideration on the part of physiologists. Although such a criti- cism did not apply to the proteins soluble in strong alcohol which Ritthausen had described, and which after fifty years have l)ecome of much importance in the study of problems of nutrition. these remained for man\- years unknown to nearly every [)hysiological chemist. After the pulilication of this review, Ritthausen con- tinued his work until it included most of the seeds used for feeding men and animals. When Hoppe-Seyler and \\'e}l introduced neutral saline S(j1u- tions as solvents for many of the proteins of animal and vegetable origin, Ritthausen's results were regarded with increasing disfavor by physiological chemists. Undiscouraged by the unfair treatment accorded him, Ritthausen re-examined ])y the aid of salt solutions nearly all of the seeds which lie had previously studied, and also showed that most of his earlier preparations were still soluble in 33^ llc'uiricli RittJuuiscii [April neutral sah snliuiun and were nnaltered in elcnienlar\- cumposition. Linie attention was, howex-er. paid \n this later work, although bis pa])ers were filled witli int'orniatiun that has proxed morc helpful in de\"el()i)ing" nur present knowledge of the ehemistrv uf proteins in general than has mosl of that ftn'nished l)\' bis critics. Aniong the many proteins which he acenralely described were several that could easily be obtained in well-formed crystals, a fact which. at that tinie. was of great importance in i)rotein chemistry. Thns. in 1881, he described a crystalline protein of the hemp-seed and the method for its preparation, which is essentially that now in nse. For more than twenty years this protein remained almost unknown although in recent years, under the name of edestin, it has been employed in hundreds of i)hysiological experiments in con- nection with a great variety of problems in protein chemistry. As a result of bis later w^rk he proved that wide difTerences exist between different food proteins; and he was the first to direct at- tention to this fact, and to discuss its probable bearing on their relative value in nutrition. Ritthausen's studies were not confined solely to the \-egetable proteins, as is evident from bis extensive bibliography which ap- pears at page 339. He made man}- im-estigations of other con- stituents of seeds. obtaining \icin and convicin from vetch seeds, and discovered in the cotton-seed " melitose " now known as raffinose. If we are to judge Ritthausen's work fairly we must remember that it was begun under the influence of Liebig's erroneous assump- tion that only a few forms of protein existed ; that at that time organic chemistr}- was in its infanc}-; that few methods were known by which jjroteins could be isolated from the tissues containing them, or by which the different proteins could be separated from one another and be purified : that the only means for preventing the changes caused by bacteria and enzymes were low temperatures ; and that the facilities for conducting such investigations were very limited. "Ko the writer, who has had a long experience in this same field. under the vastly more favorable conditions pre\'ailing a gen- eration later, it is astonishing that Ritthausen accomplished so much, and that the data he secured were in the main so accurate. What- 1913] Lewis W. Fcfccr 339 ever may have been the sh(jrtcomings of Ritthaiisen's work, the fact remains that he made a niost valuable contribiition to biological chemistry : and that instead of criticism, he deserves our gratitude and admiration for his patience and perseverance in one of the most difficult fields of investigation. Thomas B. Osborne, Connecticut Agricultiiral Experiment Station, N'ew Haz'cn. PUI5LICATI(JXS OF PROFESSOR HEINRICH RITTHAUSEX I. Journal für praktische Chemie Ueber die Aschenbestandtheile einiger Lycopodiumarten : Lvc. coiii- planatum, Lyc. Chaiiicrcyparissiis, Lyc. clavatuiii. sowie über die Säure von Lyc. coinplaiiatum : 1851, 53, 413. Xeue Analysen der Aschen einiger Lycopodiumarten: 1853, 58, 133. Ueber die Einwirkung des Sahniaks auf Kupfer; 1853. 59, 369. Zersetzung des Sahniaks durch Zink; 1853, 60, 473. Ueber einige Kohlenwasserstoffe des leichten Steinkohlentheeröls ; 1854. 61. 74. Chemische L'ntersuchung der Runkelrübe; 1855, 65, i. Chemische Zusammensetzung des rothen und schwedischen Klees (Tr//. pratcusc und Trif. hyhriduni ) in verschiedenen \'egetations- Perioden ; 1855, 65, 8. A'eränderungen des Heus von Rothklee durch Auswaschung von Regen : 1855, 65, 13. Ueber den Einfluss der Düngung mit Asche und Gyps auf die chemische Zusammensetzung des Klees; 1855, 65, 15. Destillations-Rückstände von der Spiritus-Fabrikation aus Kartoffeln (Schlempe) ; 1855. 66, 289. Rückstände von der Spiritus-Fabrikation aus Getreide; 1855, 66, 308. Rückstände, welche bei der Bierproduktion gewonnen werden (mit H. Scheven) ; 1855. 66, 311. Analysen der Asche von Gerstenmalz, Trebern und Malzkeimen; 1855, 66, 315. Ueber die Bestandtheile des Weizenklebers ; 1862, 85, 193. Ueber die Zusammensetzung des Pflanzenleims und das \^erhalten des- selben zu \\ asser ; 1862, 86, 257. 340 Heinrich Rillhaiiscii [April Ueber die Ziisanunensetzung des Pflanzenleims, 1863, 88. 141. Reactionen des l'flanzenleinis : 1863. 88. 142. Zur Darstellung des Pflanzenleinis ; 1803. 88, 145. Cholesterin im Fett des Weizens; 1863, 88. 145. Trimethylamin aus Weizenbrand; 1863, 88, I-17. L'eber die P>estandtheile des Weizenklebers; 1864, 91, 296. L'ntersncliungen ueber einige liestandtbeile des Roggensamens ; 1866, 99. 439- Ueber die Cilutaminsiiure ; 186O, 99, 454. Ueber die Bestandtheile des Weizenklebers ; 1866, 99, 462. Ueber einige Bestandtheile des Roggensamens : In Weingeist lösliches Gummi; Cholesterin und Palmitinsäure im Fette des Roggens; Buttersäuregährung des Roggenmehls; 1867, 102, 321. Dolomitreicher Mergel; 1867, 102, 369. Lithionhaltiger Mergel und Boden in Ostpreussen ; 1867, 102, 371. Bildung von Mvianit im Grunde einer Düngergrube; 1867, 102, 373. Blasenstein (eines Ochsen) von Kieselerde; i8C>7, 102, 374. Soda als sogenannter Mauersalpeter; 1867, 102, ;i,j=^. Portland-Cement von Powunden ; 1867, 102, ^/(). Ueber das Pflanzen-Casein oder Legumin ; 1867, 103, 65, 193, 273. Ueber die Zersetzungsprodukte des Legumins und des Prote'inkörpers der Lupinen und Mandeln beim Kochen mit Schwefelsäure; 1868, 103. 233- Ueber die (dutansäure, das Zersetzungsprodukt der ( llutaminsäure durch salpetrige Säure; 1868, 103, 239. Asparaginsäure und (Glutaminsäure, Zersetzungsprodukte des Legu- mins beim Kochen mit Schwefelsäure; 1869, 106, 445. Prote'instofife des Maissamens; 1869. 106. 471. Asparaginsäure und Glutaminsäure, Zersetzungsprodukte des Legu- mins und Conglutins beim Kochen mit Schwefelsäure; 1869, 107, 218. Ueber die Säuren der Samen der gelben Lui)inen ; 1870, 2 ( n. f.), 339. Lieber das A'orkommen von Amygdalin und eine neue dem Asparagin ähnliche Substanz in Wickensamen (mit L^. Kreussler ) ; 1870, 2 Ol. f.). 333- Leucin aus Pflanzenproteinstoffen (mit l". Kreussler) ; 187 r, 3 (n. f.), 307. Lieber die \^erbreitung der Asparaginsäure und Glutaminsäure unter den Zersetzungsprodukten der Proteinstoffe (mit R. Pott) ; 187 1, 3 (n. f-)- 3U- 1913] Lc7\.'is JJ\ Fctzer 341 A'erbindungen der Prote'instoffe mit Kupferoxyd ( Legumins, Con- glutins. GlutenL.-seins) ; 1872, 5 (n. f.), 215. Ueber das Drehungsvermögen von Glutan- und Aepfelsäure ; 1872, 5 (n. f.), 354- Untersuchungen über A'erbindungen der Eiweisskörper mit Kupfer- oxyd (mit F. Weger); 1873, 7 ( n. f.), 361. Notiz ueber die asparaginähnhche Substanz im Wickensamen; 1873, 7 (n- f-). 374- Ueber die Bestimmung des Stickstoffs der Eiweisskörper mittelst Na- tronkalk ; 1874, 8 (n. f.), IG. Neue Methode zur Analyse der ^vlilcli und ein vom Milchzucker ver- schiedenes Kohlehydrat in der Kuhmilch; 1877. 15 (n. f.), 329. Nachtrag hierzu; 1877, 16 (n. f. ), 22)7- Krystallinische Eiweisskörper aus verschiedenen Oelsamen ; 1881. 23 (n. f.). 481. Ueber \'icin und eine zweite stickstoft'reiche Substanz der Wickensa- men. Convicin : 1881, 24 (n. f.), 202. Ueber die Einwirkung von Salzlösungen auf Conglutin und Legumin ; 1881, 24 (n. f.), 221. Ueber die Eiweisskörper von Oelsamen; 1881, 24 (n. f.), 257. Ueber die A'erbreitung der ^lyronsäure in den Samen von Brassica iiapiis und ra[^a; 1881, 24 (n. f.), 2/2,. Zusammensetzung der Eiweisskörper der Hanfsamen und des krystal- lisirten Eiweisses auf Hanf- und Ricinussamen ; 1882, 25 ( n. f.), 130. Ueber die Zusammensetzung des Krystallisirten Eiweisses aus Kürbis- samen; 1882. 25 (n. f.), 137. Ueber das A'erh alten des chromsäuren Bleis bei A'erb rennungen und zu Sauerstoff; 1882. 25 (n. f.), 141- lieber das A'erhalten des Conglutins aus Lupinensamen zu Salzlösun- gen; 1882. 26 (n. f.), 422. Ueber die Eiweisskörper der Pfirsichkerne und der Pressrückstände von Sesamsamen; 1882, 26 (n. f.), 440. Ueber das A'erhalten des Legumins zu Salzlösungen; 1882, 26 (n. f.), 504- Ueber ^lelitose aus Baumwollensamen; 1884, 29 (n. f.), 351. A'orkommen von Citronensäure in verschiedenen Leguminosensamen ; 1884. 29 (n. f.), 357. \'orkommen von \'icin in Saubohnen (U/cfa /aöa) ; 1884,29 (n. f.), 359. Lieber die Löslichkeit von Pflanzenproteinkörpern in salzsäurehaltigem Wasser; 1884, 29 ( n. f.), 360. 342 Heinrich J\illliaiiscii [April Ueber Zusainiiicnsct/.uni^- der mittelst Salzlösung (laryestellten luweiss- körper der Saubohnen { ricia falhi) und weissen iUjhnen { I'hasc- olus), 1S84. 29 (n. f. ), 448. Ueber Betain aus Pressrückstiinden der lUiuniwollsanicn (mit Prcnss) ; 1884. 30 (n. f.), 32. L'eber die Fdweisskörper des Weizenklebers oder Glutens ; 1890. 59 (n. f.).474- Löslichkeit von Eiweisskörpern in Glycerin ; 189g, 59 ( n. f.), 479. Ueber die Zusammensetzung des Vicins ; 1899, 5g (n. f.), 480. Ueber Divicin ; 1899, 59 (n. f.), 482. Zusammensetzung des Convicins aus Wicken- und Saubohnensamen ; 1899, 59 (n. f.), 487- 2. Berichte der Versuchs-Station zu Möckern^ Untersuchungen des Grünfutters von dem amerikanischen Zahnmais und dem österreichischen Mais (mit E. Wolff ) : 1854, 3, i. A'ergleichende Untersuchung des schwedischen und des gewöhnlichen rothen Klees (mit E. Wolff) ; 1854, 3, 11. Chemische Untersuchung von Gras, Heu und Grummet ( mit E. Wolff) ; 1854. 3. 18. ^ . Chemische Untersuchung der Runkelrübe: (a) Einfluss des Blattens auf die Zusammensetzung; (b) Einfluss der Grösse auf die Zu- sammensetzung: (c) Einfluss der A'arietät auf die Zusammenset- zung (mit E. Wolff") : 1854, 3, 22. Beobachtungen über die ~\Iilchproduktion bei dem Uebergang von der Winterfütterung zu der Grünfütterung (mit J. G. Bahr und E. Wolff ) : 1854, 3, 38. Ueber den Einfluss des im Dampf gekochten Futters auf die Alilch- produktion (mit J. G. Bahr) : 1855. 4, i. Ueber den Einfluss der Zuckerrüben auf Milchproduktion (mit J. G. ' Bahr) : 1855. 4, 13. Düngungsversuche mit Knochenmehl, guanisirtem Knochenmehl, Blut- dünger, und Guano (mit J. G. Bahr und W. Knop) : 1855, 4, 15. \'ersuche mit Ueberdüngung von Chilsalpeter, Kochsalz und Guano bei Weizen und Roggen (von J. G. Bahr, mitgetheilt von H. Ritt- hausen) : 1855, 4- 22. ^ Compiled from Nobbe's Quellenverzeichniss der hauptsächlichsten in den Jahren 1852 liis 1877 von den Versuchs-Stationen veröffentlichten wissenschaft- lichen Arbeiten, in Ilntii'ickhi)tg 11. TJüitigkcit d. laiuln'irtscliaftlichcii l'crsuchs- Sfafioiicii. etc.. 1877, pp. 284-435 (Berlin). 1913] Lewis W. Fctzcr 343 Düngung des Roggens mit Peruanischem Guano, Chilsalpeter, gebrann- tem, reinen Knochenmehl und Polenz'schen Guano (von J. G. Bahr, mitgetheilt von H. Ritthausen) ; 1855, 4, 29. N'ersuche mit verschiedenen Sorten Guano und Knochenmehl, Raps- kuchenmehl und Stallmist zu Weizen und Kartoffeln (von J. G. Bahr, mitgetheilt von H. Ritthausen) ; 1855, 4. 31. Einfluss der Düngung des Klee's mit Asche und Gyps ; 1855, 4, 41. l'ntersuchungen des schwedischen und rothen Klees; 1855, 4, 65. \"eränderungen des Heus von Rothklee durch Auswaschung von Regen ; 1855. 4. 72>- Vergleichende Untersuchung der \\'intergerste, Annat- und Probstei- ger ste : 1855,4, 76. Ueber den Einfluss der Lupinen auf die ^lilchproduktion, ein Fütter- ungsversuch (mit J. G. Bahr) ; 1856, 5, i. Ueber die Zusammensetzung und den Nahrungswerth einiger in der Landwirtschaft als Futtermittel angewendeter Fabrikationsrück- stände (Kartoffelschlempe, Malz, Presshefe, Getreideschlempe) ; 1856, 5, 15. Ueber einige Eigenschaften von Kulturpflanzen, die in gleicher Vege- tationszeit einen verschiedenen (irad der Entwickelung zeigen (mit H. Scheven) : 1856, 5, 67. Entwicklung und Thätigkeit der land- und forstwirthschaftlichen Ver- suchs-Stationen in den ersten 25 Jahren ihres Bestehens ; Fest- schrift zur Feier des 25 jährigen Jubiläums der A'ersuchs-Station Möckern ; 1877, p. 56. (A statement, made by Professor Ritt- hausen at the request of Prof. G. Kühn, in regard to work con- ducted by or under him while Director of the Experiment Station at Möckern.) 3. Jahresberichte der Versuchs-Station Ida Marienhütte (bei Saarau) nach Breslau, gegründet A. D. 1857 (Aus den Mittheilungen des landwirtschaftlichen Centralvereins für Schlesien )- Zusammensetzung" der Kuhmilch ; i, 59. Lieber Dünger-Fabrikation; i, 60. Analysen des Bodens der Ida-Marienhütte (mit P. Bretschneider) ; I, 82. " Compiled f rom Nobbe's agricultural bibliography, 1877. See footnote. page 342. The numerals for the years of publicatinn are not given in Nobbe's bihlioQrapli>-. 344 II einrieb RiithiUiscn I April \'ersuclio über SaiiieiKlungung ; i. 85. \'ersiiche mit Ueberdüngung des Roggens ; i, 95. Düngungsversiicbe bei Rüben (Beta ) ; i. 104. Untersuchung von in gleicher A'egetationszeit, ungleich entwickelten Kulturpflanzen; i. 134. Untersuchung eines Torfes und seiner Asche; i, 145. Bestimmung der Asche in \'egetabilien ; i. 147. Bestimmung der Phosphorsäure; i, 148. Bestimmung der Kieselsäure in Pflanzenaschen; i. 149. Analysen des Bodens der Ida-Marienhütte (mit P. Bretschneider ) ; 2. 36. Untersuchung von Zuckerrüben; 2, 66. Untersuchung von Zuckerrüben ; 4. 59. Analysen des Bodens der Ida-Marienhütte (mit P. Bretschneider) ; 6. 100. 4. Sächsiches Amts- und Amzeigeblatt ^'ersuche über den Xahrungswerth der Kartofl:'elschlempe in A'ergleich zu Kartofi'eln und ]\Ialz, und süsser Maische, bei gleichen Mengen Rohmaterial (Kartoffeln und Malz) : Fütterungsversuche mit Kühen ( mit J. G. Bahr) ; 1856, p. 87. Fütterungsversuclie mit Kühen ueber den Einfluss von geschrotenem und gekochtem Getreide auf Milchproduktion (mit J. (i. Bahr) ; 1856, p. 96. Ueber die Zusammensetzung einiger Wurzelgewächse (Rüben, Kohl- rüben und Strunkkraut) und den Einfluss der Grösse und Schwere, sowie starker Düngung auf die Zusammensetzung derselben; 1857, \'ersuch über die A'erdaulichkeit der Holzfaser des Futters beim Rind (mit H. Scheven) ; 1858, p. 58. 5. Landwirtschaftlichen Versuchs-Stationen Orittheilungen aus dem Agriculturchemischen Eaboratorium der Fni- versität Königsberg i. Pr.) Untersuchungen ueber den Einfluss einer an Stickstoff' und Phosphor- säure reichen Düngung auf die Zusammensetzung der Pflanze und der Samen von Sommerweizen (mit R. Pott) ; 1873. 16, 384. Ueber die Einwirkung freier Phosphorsäure auf kohlensauren Kalk ; 1877, 20. 401. I'eber den Fettgehalt der käuflichen K]eber]irä]:)arate ; 1877, 20. 408. iyi3] Lci^'is IV. Fct::cr 345 Analysen einiger Futtermittel: 1877, 20. 409. Ueber den angeblichen (Jehalt des Roggensamens an Stearinsäure; 1877, 20. 412. Berichtigung zu der Mittheilung von ]\I. von Sivera : Ueber den Stick- stoffgehalt des Torfbodens; 1880. 25. 169. l'eber Zerstörung von Fett durch Schimmelpilze Unit H. Baumann) ; 1896, 47, 389- Ueber die Berechnung der Proteinstoft'e in den Wanzensamen aus dem gefimdenen Gehalte an Stickstoff; 1896. 47, 391. 6. Berichte der deutschen chemischen Gesellschaft Ueber Mein: Ijestandtheil der Samen von l'icia sativa : 1876, 9, 301. ^^'assergehalt und Reaktion des Alloxantins : 1896, 29. 892. Ueber Alloxantin als Spaltungsprodukt des Convicins aus Saubohnen (Ficia faba minor ) und Wicken (Jlcia satkv) : 189C). 29, 894. Ueber Galactit aus den Samen der gelben Lupine: 1896. 29. 896. Reaktionen des Alloxantins aus Convicin der Saubohnen und Wicken ; 1896, 29, 2106. Mein ein Glycosid ; 1896, 29. 2108. Ueber Leucinimid. ein Spaltungsprodukt der Eiweisskörper beim Kochen mit Säuren; 1896, 29, 2109. 7. Archiv für die gesammte Physiologie (Pf^üger) Die Eiweisskörper der Ptlanzensamen : 1877, ^S- -^V- Ueber den Stickstoffgehalt der Pflanzen-Eiweisskörper nach den ^leth- oden von Dumas und Will-\'arrentrapp (mit H. Settegast) ; 1878, 16, 293. Ueber die Zusammensetzung der Proteinsubstanz der Bertholletia- (Para-) Nüsse; 1878, 16. 301. Ueber den Stickstoft'gehalt der Pflanzen-Eiweisskörper nach den ]\Ieth- oden von Dumas und WlU-Varrentrapp : 1878. 18, 236. Ueber die Eiweisskörper der Ricinussamen. der Proteinkörper, sowie der Krystalloide dieser Samen; 1879. 19. 15. Ueber die Eiweisskörper verschiedener Oelsamen ; 1880, 21, 81. 8. Chemiker-Zeitung In Weingeist lösliches (iummi aus Roggen: Secalin : 1897, 21. 717. Zur Darstellung der Alkaloide der gelben Lupinen (Lup. Intens) ; 1897, 21, 718. 340 llciurich Ritlluuiscu [April 9. Book Die lü\vcisski"ir])er ^\Q\■ ( ictreideartcn, I liilscii fniclite und (MsaiiK'n : Beiträge zur i'hysiologie der Samen der Kullurgewäehse. der Nahrungs- und Futtermittel; pages 252. Jionn (Max Cohen und Sohn), 1872. 10. Miscellaneous publications Versuciie über Düngung von Rüben: Chemische .Ickeyniami, 1858, p. 130; abs. in Jahresber. Ayr. Cheiii.. 1858, i, 226. Die Aschen einiger Futterpflanzen; M ift/ieiliiiu/en aus Jf'aldaii, 1859, p. 91 ; abs. in Jahresber. Jgr. Cheiii.. 1850. 2. 84. "Mug," ein Dungmittel; JJ'ocJieiiblatt der Annalcii der Laiid-ccirfseliaff, 1861, p. 8; abs. in Jaliresber. Ayr. Chciii., 1861, 4, 195. Das A'erhalten der freien Ph^sphorsäure der Superphosphate ; Laiid- 7cirtsch. Zeitung für das iiordöstlich.e Deutselüand, 1875, 11; abs. in Jahresber. Agr. Chein., 1875, ^^> 5^- Verlust an DüngstofTen im PJoden einer Düngerstätte (mit Ritsch- mann ) ; . Igrieulfureh. Ceiifralbl., 1876, p. 35; abs. in Jaliresber. Agr. Chein.. 1876, ig, 40. Ueber Proteinkörner, Krystalloide und krystallisirtes Ei weiss. Schriften der Physikalisch-Ökonouiisclieu Gesellschaft .::u Königsberg ; 1881, 22, 15. Lewis W. Fetzer. Office nf E.vpcriincnf Statioits, U. S. Drparliiicnt of Agricultitre, und Ccorgcfon'ii t^iii'c'ersify, Jl'ashingtoii. D. C. DINNER TO PROFESSOR CHITTENDEN Testimonial by his pupils In December, 191 1, a nnml^er of the former pupils of Prof. Russell H. Chittenden, at a Conference in P)altimore. concluded " that the time had arrived when it would be appropriate to provide some formal expression of the esteem in which Professor Chittenden is held by those who appreciate his contributions to physiological chem- istry and education." Drs. S. W. Lambert, F. S. Meara, Holmes C. Jackson, S. P. Beebe, and William J. Gies were requested to serve as a provisional committee of five, to consider the matter further and to proceed with Organization and execution of plans, "if some step in this direction seemed appropriate, after further consideration." The pro\-isional committee of five decided to organize Professor Chittenden's pupils for the purpose indicated, and invited twenty additional former pupils of Professor Chittenden's to serve with them as a Committee of Twenty Five, as follows : John A. Hartwell, chainiiaii, T. S. Arbuthnot, S. P. Beebe, Joseph A. Blake, Harvey Cushing, H. H. Donaldson, Isadore Dyer, P. B. Hawk, Theodore C. Janeway, Elliott P. Joslin, J. H. ^[. Knox, Samuel W. Lambert, P. A. Levene, Frank S. ]Meara, Lafayette B. Mendel, Charles Norris, Thomas B. Osborne, Alfred N. Richards, E. AV. Rockwood, W. T. Sedgwick, W. Gilman Thompson, H. Gideon Wells, E. B. Wilson, Holmes C. Jackson, trcasiircr, and William J. Gies, sccrctary. The Committee of Twenty-Five rec[uested Drs. Hartwell, Beebe, Jane- way, Jackson, and Gies to serve as a subcommittee for the execu- tion of the committee's plans. The general committee has invited Professor Chittenden's pupils to coöperate in raising a Russell H. Cli'ittcndcn Fund, to be pre- sented to the Yale corporation without any other condition than that it be used for the advancement of the work of the department of physiological chemistry in the Sheffield Scientific School. The committee also authorized the secretary to invite Professor 349 350 Dinner lo Professor Chili enden [April Chittenden to be the gnest of bis pupils at a dinner in Xew York on ^larcli T. In tlic formal imitation, the secrttar\- w n itc to Professor Chittenden. in part. as follows : The above-named Committee of Twenty Five has instructed nie to invite you to be the gnest of your many pupils and friends at a dinner in yonr bonor in Xew York City on Alarcb i, 1913. It is onr desire not only to bave the pleasure of yonr Company but also to extend to you onr personal and professional greetings. anfl to evidence onr friend- ship and respect. The dinner in bonor of Professor Chittenden was held at Del- monico's. on Satnrday evening, March i, and proved to be a delight- fnl event in evcrv particnlar. Althongh man}- who expected to be present were unable to attend, and sent letters of regret, about seventy-fi\-e pnpils and a dozen in\-ite(l friends comprised the en- thusiastic Company tbat made the dinner a cordial testimonial of affection and esteem for Professor Chittenden. Among those who were unable to accept invitations to be present at the dinner. and to speak afterwards. were President Hadley, of Yale, and Prof. William H. Welch, of Johns Hopkins University. In a letter expressing regret for bis unavoidable absence, President Hadle}' wrote. in part. as follows : When }ou are having the Chittenden dinner, I shall be three thou- sand miles away. But I do not want to let the occasion go by without a word of greeting. ( )ur universities are on the lookout for men who are either discoverers or teachers or Organizers. In Chittenden Yale has a man who is all three. Professor \\'elcb sent a teleoram in which he said : -fc>' Deeply regret unavoidable absence from banquet. Affectionate greetings and heartiest congratulations to Chittenden — the man and friend, the great teacher, investigator, administrator — who has ren- dered inestimable service to science, Yale, and countr}-. ^fay many vears of health and vigorous work be bis. Seated at the Speakers' table were Professors Harvey Cushing, Henrv H. Donaldson, lohn A. Hartwell. Russell H. Chittenden. THE MEMBERS OF THE GOVERNING BOARD Oh THF SHEFFIELD SCIENTIFIC SCHOOL OF YALE UNIVERSITY EXTENÜ CONGRATÜI.ATIUNS TO THEIR CüLLEAG.UE THE DIRECTOR OF THE (GOVERNING BOARD PROFESSOR RUSSELL HENRY CHITTENDEN PH. D..SC.D.,LL.D. ON THE OCCASION OF THE TESTIMONIAL DINNER GIVEN IN HIS HONOR BY HIS PUPILS AND FRIENDS ON MARCH FIRST NINETEEN HUNDRED AND THIRTEEN IN NEW YORK CITY THE EVENT WHICH HAS CALl.F.D KORTH THIS PERSONAL MANIFESTATION OF ESTEEM AFFORDS AN Ol'PÜRTUNlTY TO THK MHMKERS OF THE (»VKRNING BOARD TO GIVE FORMAL EXPRES- SION TO THEIR HIGH ESTIMATK OF l'ROFESStDR CHITTHNDF.NS CONTRIBUJI'JNS TO SCIENCE AND EDUCATION AND THEIR GRATIFICATION AT THE FAVORABLE RECOGNITION WHICH HAS BEEN ACCORDEDTO HIS DISTINGUISHED SERVICES IN THE PROMOTION OF PHYSIOLOGICAL RESEARCH. THE SUCCESS VVITH WHICH PROFESSOR CHITTENDEN HAS FURNISHED INSPIRATION FOR THE LIFE WORK Ol" 0THER5 IS WORTHY OF COMMFNDATION: HIS ENERGY RECALLS THE VVORDS OF ANOTHER EMINENT STUDENT OF NUTRITION : •■THE GRFATFST J<;Y OF THOSE WHr> ARE STF.EPED IN WORK AND WHO HAVE SUCCEEDED IN FINDINi:. NEW TRUTHS AND IN UNDERST ANDING THE RELATION OF THINGS TO FACH OTHER. LIES IN WORK ITSELF.' TO THE (iRATITÜDR AND REGARD OF PROFESSOR CHITTENDEN^S PUPILS HIS COLLEACUES OF THE GOVERNING BOARD NOW DESIRE TO ADD THE CORDIAL ASSW^ANCE OF THEIR BEST WISHES AND THEIR PERSONAL GRFJETINGS. •■ «.f» Cera*ti. Greetings to Professor Chittenden by bis CoUeagues of tbe Governing Board of tbe Sbeffield Scientific Scbool I9I3] '94 S OD 3 Frank S. Meara, Graham Lusk, William T. Sedgwick. William T. Porter and Elliott P. Joslin. At the conckision of the clinner, the chairman of the committee, Dr. Hartwell, extended tu Professor Chittenden the affectionate greetings of his pupils and friends, and informed him of the estab- lishment of the Russell H. Chittenden Fund for the Adzrinceinent of Pli\siologieal Cheniistry in the Sheffield Seientific Sehool. Dr. Faces of the golcl medal presented to Professor Chittenden by the National Institute of Social Sciences Hartwell stated that the amount of the fnnd and the time of its presentation to the Yale corporation will he annonnced at an early date. Dr. Hartwell conclnded his remarks by introducing the toast- master, Dr. Meara. who officiated in the graceful and inimitable manner in which he is accustomed to preside on such occasions. Informal after-dinner addresses were then made by Drs. Cushing, Donaldson, Joslin, Levene, Lusk and Sedgwick, to which Professor Chittenden responded. The Speakers who preceded Professor Chittenden paid eloquent tribute to the personality, influence, ser\-ice, and achievements which have made Professor Chittenden the Dean of American biological chemists. Professor Chittenden replied earnestly and wilh deep feeline to the cordial tribute which had been conveved in the senti- ments of the Speakers, in the abundant evidence of warm approval with which each address was received, and in the evident heartiness of his own reception. 354 Dinner lo l'rofrssor Cliillriulrii [\\)r\\ Immedialcly aficr ilic conclusion of Professor C "liiUeiidcirs ad- dress, ilie loaslniaster announcc'd iliat ilie National InsiiUite of Social Sciences liad xoted a t^'old nicdal lo Professor Chillenden in reco^s^- nition of tlie distinciion lic lias aitained in original investigation in llic lield of plnsiological cbemistrv. Dr. H. Holljrook Curtis, sec- rclar}- oi ilie Inslitnte. made the presentation. The faces of Ihe niedal are shown on page 353-. Prof. Lafayette B. Mendel followed Dr. Cnrtis witli a presenta- tion of engrossed congratulatory resolutions A\liich had Ijeen adopted ])}• Professor Chittenden's associates in the Governing Board of the Sheffield Scientific School. (See page 351. ) The accompanying group portrait \\as made jnst after seats had been taken at the tables. The names of all in attendance, ar- ranged in taljle groiips, are appended :^ Speakers' table Harvey Cushing Russell H. Chittenden William T. Sedgwick H. H. Donaldson Frank S. Meara William T. Porter John A. Hart well Graham Lusk Elliott P. Joslin Table i. Lafayette B. Mendel, S. J. Meltzer, Jacques Loeb, Yandell Henderson, Simon Flexner, P. A. Levene, Frederic S. Lee. Table 2. Henry Hun, William Browning, H. H. Curtis, Henry Ling Taylor, W. M. Kenna, Harry Saltzstein, Frank C. Gephart, H. G. Barbour. Table 3. John Rogers, Wm. L. Culbert, R. H. Wylie, William Armstrong, Joseph A. Blake, W. L. Griswold, G. Wyckofif Cummins. Table 4. Theodore C. Janeway, J. H. M. Knox, S. P. Good- hart. Chas. H. Studio, George S. C. Badger, Joseph S. Wheelwright, N. R. Norton, Joseph H. Pratt. Table 5. Charles Norris, A. N. Richards, E. K. Dunham, John A. Mandel, H. D. Dakin, George B. Wallace, W^illiam J. Gies, Holmes C. Jackson, William C. Lusk. ^ The men at tables 1 1 and 12, when the photograph was taken, subsequently reassembled at tables 9, 10 and 2. One or two additi(3nal rearrangements account for the disagreement between the indications of the portrait and the table lists. I9I3] '94 S 357 Table 6. S. P. Beebe, G. A. Hanford, A. L. Dean, Frank P. Underhill, Benjamin White, Oswald T. Avery, Leo F. Rettger, S. R. Benedict. Table 7. Isadore Dyer, Wm. C. Wurtemberg, Robert Taylor W'heeler, Donald Guthrie, Henry H. Janeway, A. W. Elting, Charles L. Scudder. Table 8. Lewis F. Frissell, Seth M. Milliken, Robert P. Wad- hams, William P. Healy, Norman E. Ditman, W. W. Herrick, M. Heminway Merriman, Cyrus W. Field. Table 9. Frank C. Yeomans, Alfred Jerome Brown, J. L. Bendell, Isaac F. Harris, Stanley D. Beard, Frank E. Haie, E. JMonroe Bailey, Otto G. Hüpfel. Table 10. Israel S. Kleiner, Lewis H. \\'eed, Orville H. Schell, Simon B. Kleiner, Morris S. Eine, Victor C. ]\Iyers, Warren W. Hilditch, Henry C. Courten. Adjournment occurred at a very late hour, bnt many tarried to discuss informally the happy events of the evening and to talk over " old times" in Chittenden's laboratory. '94 s. -Vc^z^' York City. SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE Tenth anniversary meeting and dinner The tenth anniversary of the estal)Hshment of the Society for Experimental Biology and Aledicine was celebrated in Xew York on the iQth of February. The fifty sccond regulär scientific meet- ing was held, at 4 p. m.. at the College of Physicians and Surgeons, in the lecture room adjoining the main biochemical laboratory. After the conclnsion of the scientific meeting. at about 7 ]). m., the members adjonrned to the Hofbräu Honse (39th Street and Broad- way), where, in a body, they feasted on a beefsteak dinner prepared nnder the auspices of a committee of which Prof. Graham Lusk was chairman. The scientific session was the most interesting and important in the history of the society. The nature of the proceedings is shown by the appended copy of the official program : G. X . Calkiiis: Further light on the conjugation of paramecium. — JJ\ H. Maincaring and J. Bronfcnhrciuier: On lysis of tubercle bacilli (II) ; '^'On chemotherapeutics of tuberculosis. — U\S. Halstead: Hyper- trophy of the thyroid ; Partial occlusion of the aorta by bands of living tissue. — G. H. A. Cloivcs: Hay fever, with demonstration. — *J. /. Ringer: Further studies on the fate of fatty acids in the diabetic organ- isni. — R. M. Pcarcc and P. F. Williams: Experience with Abderhalden's test for pregnancy. — L. L. JJ'oodruff: The kernplasma relation during the life of a pedigreed race of O.vyfliricha f alias. — Richard JJ'cil: A new factor in anaphylaxis. — E. E. Biiffcrficld: The reaction between oxygen and hemoglobin. — *F. S. Lee and S. Everinglmin: The myo- neural junction in fatigue. — F. H. Pike: A demonstration of the effects of electrical Stimulation of the labyrinth of the ear. — E. L. Scott: The relation of pancreatic extract to the sugar of the blood. — A. F. Hess: The pancreatic lipase of infants in acute intestinal disturbances. — JV. H. Park, L. ]]'. Fainiilcucr and E. J. Banzhaf: Influence of protein concentration on absorption of antibodies in sul)cutaneous injections. — * On the ofificial program, but not abstracted in the Proc. Soc. Ex[>. Bio!, aiid Med.. 1913. x. pp. 65-122. ;8 J3' c^ Portrait of the Foundcr ot the Society for Experimental Biology and Medicine. Reproduced from \'olume II ( 1904-05 ) of the Society's Proceedings 1913] / Ninetccn O. Tlircc 361 G. C. Robinson: The inflnence of the vagits nerves 011 tlie faradized auricles in the dog heart. — B. S. Oppenheimer and H. B. Williams: Pro- longed complete heart block with freqiient changes in the idio-ventricu- lar complexes. — C. J. Wiggers and E. F. DnBois: Methods for the pro- duction of temporary valvulär lesions.— .^. /. Gold färb: The influence of the central nervons System on regeneration ; The effect of salinity lipon regeneration. — B. T. Terry: Variations in the amount of trans- formed atoxyl (trypanotoxyl) produced by varying the strength of atoxyl inciibated with blood. — IV. J. MacNeal and A. F. Chace: Some observations on bacteria of the dnodenum. — A. F. Colin: The effects of morphin on the mechanism of the dog heart after removal of one vagus nerve. — T. S. Githens: The influence of temperature on the mini- mal dose of strychnin and the onset of tetanus in the frog. — '■'/. E. McWhorter and F. Prime (by invitation') : Cinematographic demon- stration of the growth of tissues. — '''F. S. Lee: Cinematographic dem- onstration of the beating heart. — "7?. Bnrton-Opitz: Demonstration of the vasomotor nerves of the liver. — *//. B. IVilliams: Demonstration of the electrovagogram. — Wm. de B. MacNider: The difl^erence in the efifect of Grehant's anesthetic and of morphin-ether on the total output and composition of the urine in normal dogs. — Sntherland Simpson: The rate of growth in the dog. — Andrew Hnnter: The influence of ex- perimental cretinism upon nitrogenous metabolism in the sheep. — De- Witt Stettcn and Jacob Rosenblooni: ]\Ietabolism studies in a case of hypopituitarism, with infantilism of the Lorain t}pe. — /. P. Atkinson and C. B. Fitspatrick: On the presence of pressor substances in experi- mental immunity. — G. H. A. Clowcs, Francis C. Goldsborongh and F. West: On a complement-deviation reaction exhibited in pregnancy. — G. H. A. Clozces and Francis C. Goldsborongh: On the antitryptic reac- tion exhibited in pregnancy. Prior to adjournment an election of officers for 191 3-' 14 oc- curred, with the following results : President, Janics Ezcing (re- elected ) ; vice president, Cyrns U\ Ficld; secretary, Hohnes C. Jackson; treasurer, Charles Morris (reelected). The new members elected were Russell L. Cecil, Gary Eggleston, K. George Falk, Davenport Hooker, Paul E. Howe and Charles J. West. The dinner was a very enjoyable event. The accompanying group Portrait shows the condition of the party at midnight. All * On the official program, but not abstracted in the Proc. Soc. E.vp. Bio!, and Med., J')i3, X. pp. 65-122. ?62 Society fov Exf^criiucuhil Hioloj/y aiuf Mcdiclnc April iliose at tlic rcadcr's cxnxnic Icfl aiid ri^lii wlio did not gel inlo the piclurc wcrc under ihe table wlieii ihe jjluHograpli was taken.'' The ])resi(lent, Prof. James r^wing-, ably and entenainingh- eon- cliKied llie afler-dinner proceedings. Infonnal speeclies were made. at the call of the President, l)y the distinguished fonnder, Dr. S. J. iMeltzer. also \)\ 1 )rs. (iraham T.usk. I'"rederic S. Lee. G. H. A. Clowes. Jac(|ues Loeb. William II. Park and William J. Gies. The Speakers felicitated Dr. Meltzer on the happiness of the idea that led Ihm to found the societ}' ; they also complimented him on the societ}''s past ser\ice. and on its rigor and effectiveness at the tenth anniversary of its birth. There was a strong note of con- gratulation of the society itself on the prospect of steady growth in efficiency and nsefnlness. The names of the members present at the meeting or at the dinner, or both, are appended : J. P. Atkinson John Auer T. H. Austin F. W. Bancroft S. P. r.eebe Jacob üronfenbrenner E. E. Butterfield G. X. Calkins G. H. A. Clowes A. E. Cohn Riifus I. Cole J. W. Draper E. F. Du Rois E. K. Dunham A. B. Eisenbrey C. A. Eisberg Haven Emerson James Ewing L. W. Famulener Cyrus W. Field C. B. Fitzpatrick Simon Flexner N. B. Foster W^illiam J. Gies T. S. Githens A. J. Goldfarb W. S. Halstead Isaac F. Harris Alfred F. Hess Paul E. Howe H. C. Jackson Walter A. Jacobs H. H. Janeway Don R. Joseph Ludwig Käst L S. Kleiner R. A. Lambert Frederic S. Lee P. A. Levene Isaac Levin Charles C. Lieb Jacques Loeb \\\ F. Longcope (iraham I,usk W. G. MacCallum W. T. AFacXeal F. H. ^IcCrudden A. R. Mandel John A. ^Mandel W. H. Manwaring S. T. ^leltzer G. '^r. Meyer H. O. :\Iosenthal John R. Ahndin j. B. ALn-phy A". C. Apvers Hideyo Xoguchi Charles Xorris B. S. Oppenheimer A. M. Pappenheimer William H. Park R. ^L Pearce F. H. Pike A. L Ringer G. C. Robinson Pevton Rons E.'L. Scott ( r. G. Scott H. D. Senior M. Sittenfeld Edna Steinhardt H. A. Stewart ^ Drs. Auer, Bancroft, Dunham, Eisenbrey, Field, Hess, Jackson, Mandel brothers, Norris, Oppenheimer, Park, Pearce. Senior, Swift, Wadsworth, Wal- lace, Wood. Anticipating the fate of the Michigan editor in the Roosevelt water- wagon case, we wish to add that \ve do not l3elie^"e this situatiiMi implics any- thing more than the facts themselves indicate. c\ . — I 3 o CO I9I3] Nineteen 0. Three 365 H. F. Swift B. T. Terry D. D. Van Slyke A. B. Wadsworth George B. Wallace Richard Weil C. J. West C. J. Wiggers Anna W. Williams H. B. Williams Francis C. Wood The Society for Experimental Biology and Medicine has been an important influence in the development o£ biological and medical science, particularly in New York. It has stimulated aspiration, quickened activity, increased productivity, afforded a congenial and ready means of expression, and opened a suitable Channel for com- munication, during a period of awakening in the biological and medical sciences in New York. It continues in this röle as an in- fluential factor in the advancement of science in this country. The growth of the society is indicated by the appended tabula- tion of its total membership at the end of each successive academic year since its foundation in 1903 : Year Total Increase Year Total Increase Year ; Total Increase 1903 1904 1905 1906 19 55 87 119 36 32 32 1907 1908 1909 1910 140 162 185 205 21 22 23 20 1911 1912 1913: (Feb. 19) 222 239 255 17 17 16 Biological chemists may be interested in the following Statistical summary relating to the Society for Experimental Biology and Medicine : Of the seven men at the Conference in Prof. Graham Lusk's home preliminary to Organization, on January 19, 1903, four were biological chemists. The society was formally organized at a meeting in the bio- chemical laboratory of Columbia University, at the College of Physicians and Surgeons, N. Y., on Feb. 25, 1903. Two of the three authors of the Constitution, and two of the first five officers, were biological chemists. The folloAving members of the American Society of Biological Chemists are members of the Society for Experimental Biology and Medicine : J. J. Abel, J. G. Adami, H. M. Adler, C. L. Aisberg, J. P. Atkinson, E. J. Banzhaf, S. P. Beebe, F. G. Benedict, S. R. Benedict, W. N. Berg, 366 Society for Experimental Biology and Mediane [April F. J. Birchard, Russell Burton-Opitz, R. H, Chittenden, A. C. Craw- ford, H. D. Dakin, E. K. Dunham, C. W. Field, Otto Folin, N. B. Foster, C. Stuart Gager, R. B. Gibson, William J. Gies, Shinkishi Hatai, R. A. Hatcher, P. B. Hawk, Paul E. Howe, W. H. Howell, Reid Hunt, Andrew Hunter, H. C. Jackson, W. A. Jacobs, Walter Jones, J. H. Kastle, I. S. Kleiner, Oskar Klotz, J. B. Leathes, P. A. Levene, Jacques Loeb, A. S. Loevenhart, Graham Lusk, A. B. Macallum, J. J. R. Mac- leod, W. deB. MacNider, J. A. Mandel, F. H. McCrudden, L. B. Mendel, G. M. Meyer, J. R. Murlin, V. C. Myers, F. G. Novy, T. B. Osborne, Franz Pfaff, A. N. Richards, A. I. Ringer, T. B. Robertson, Jacob Rosenbloom, William Salant, P. A. Shaffer, H. C. Sherman, Torald Sollmann, L. B. Stookey, A. E. Taylor, F. P. Underhill, D. D. Van Slyke, G. B. Wallace, H. G. Wells, C. G. L. Wolf. Of the 775 Communications to the Society for Experimental Biology and Medicine at its first fifty-two meetings, 430 — more than half — were largely er entirely biochemical in character. NiNETEEN O. ThREE New York City METHODS FOR THE ELECTROMETRIC DETERMI- NATION OF THE CONCENTRATION OF HYDRO- GEN IONS IN BIOLOGICAL FLUIDS K. A. HASSELBALCH (Finsen Institute j Copenhagen, Denmark) (WITH PL ATE 3) The great importance of the reaction of the medium in many biological processes has long been appreciated and has led to a series of more or less successful endeavors to measure its degree. It is only within the most recent years, hovvever, that the methods of measurement have been so far perfected as to enable us to say that the "true reaction" of biological fluids can now be measured with sufficient accuracy for most purposes. This is due, in the first instance, to the insight into the nature of the question which has been derived from the electrolytic dis- sociation theory : the " true reaction " of a liquid is not determined by its concentration of free acid or alkali, but by its concentration of hydrogen and hydroxyl ions — or, practically speaking, by the concentration of hydrogen ions alone, for the product of the two is a constant. Since the dissociation of acid or base in a liquid, and thereby also its hydrogen-ion concentration, is in many respects dependent on the nature of the dissolved substances, the true reac- tion of the liquid cannot be determined by merely measuring the quantity of alkali or acid that must be added to a certain quantity of the liquid in order to effect a particular change of color in the indicator used. As is now known, such a titration shows only that, at the moment, a certain hydrogen-ion concentration, to which the indicator reacts, has been reached, the original hydrogen-ion con- centration of the liquid remaining unknown. Thus, the determination of the true reaction of a liquid requires some procedure by which the concentration of the hydrogen ion is 367 3(38 Hydro gen Ions in Biological Fluids [April not altered. Only two methods of the latter kind are in practical employment, namely, the colorimetric and the electrometric. The colorimetric method is based on the above-mentioned fact, that a series of indicators shows certain color nuances with known hydrogen-ion concentrations, which miist be determined electro- metrically, so that the electrometric determination of the hydro- gen-ion concentration miist at any rate be considered as the funda- mental method. The colorimetric method has been indicated by Friedenthal and Salm ;^ its field has been considerably widened and its trustworthiness assured by the thorough investigations and im- provements of S. P. L. Sörensen and his collaborators. I shall not discuss the technical details of the method but merely refer to Sören- sen's latest smnmary of his work.^ We owe the electrometric method orginally to Nernst.^ It was first applied to biological fluids by Bugarsky and Liebermann, ^ and by Höber.^ It is based on the fact that a hydrogen-saturated metal electrode in a hydrogen-saturated liquid gives rise to a difference of Potential between the electrode and the liquid, which is dependent on the hydrogen-ion concentration according to known laws. The determination of this difference of potential thus makes it possible to determine the hydrogen-ion concentration of the liquid. The experimental method generally employed to measure the difference of potential between the hydrogen-saturated electrode and the hydrogen-saturated liquid has been so often described in its main features, most recently by Sörensen*^ in the above-cited work, that it needs no attention here. The present paper deals with the difficulty of obtaining the condition presupposed by the method, vis., Saturation of the electrode and liquid zvith hydrogen, without any alteration in the hydrogen-ion concentration of the liquid. Biological fluids, as is well known, usually contain volatile acids (or bases) which determine, in great part, their hydrogen-ion con- centration, so that the normal electrometric method, by which liquid and electrode are saturated with a current of hydrogen bubbled ^ Friedenthal and Salm: Zeitschr. f. Elektroch., lo, 1904; 12, 1906; 13, 1907. ^Sörensen: Ergebnisse der Physiologie, 12, 1912. ^ Nernst : Zeitschr. f. physikal. Chemie, 4, 1889. * Bugarsky and Liebermann : Pflüger's Arch., 72, 1898. ° Höber: Pflüger's Arch., 81, 1900. ' Sörensen : Loc. cit. I9I3] K. A. Hasselbaich 369 through the fluid, cannot be used. Let us take an extreme case and see what even a slight carbonic-acid tension — according to ordinary ideas — may mean for the hydrogen-ion concentration o£ a liquid. Sea-water and the surrounding atmosphere have the same carbonic acid tension — ca. 0.04/100X760 = 0.3 mm. On driving all the carbonic acid f rom the sea-water we should cause the hydrogen-ion concentration to sink from ca. lo"^ to ca. lO"^ or, using Sörensen's terminology/ the hydrogen-ion exponent, pn-, would rise from 8 to 9. If, therefore, in this case we saturated the liquid and elec- trode with a current of hydrogen, quite an erroneous result would be obtained. A similar error, though less in amount, would also arise if we had recourse to the auxiliary method applied in such cases at the beginning of this Century, namely, if the hydrogen-saturated elec- trode remained in contact with the liquid and we waited until the Potential became constant, i. e., until equilibrium had been attained in the diffusion between the liquid and the hydrogen atmosphere. For instance, a sample of sea-water (kept in a bottle for nearly a year), whose p-a: was in reality 7.55, showed /'h- = 7.74 on using this method. The error may be reduced if, with Michaelis,^ we take a small quantity of hydrogen and let the electrode only just touch the surface of the liquid. But the most satisfactory method of proceeding seems to me the f ollowing :^ A current of pure hydrogen, saturated with moisture, is led through the vessel containing the electrode until the latter has be- come saturated with hydrogen. The experimental liquid, which is stored in such a way that it retains its natural tension of volatile acid (or base), is now led into the vessel in such a quantity that the electrode reaches more or less deeply into the fluid (see below) and the vessel is then closed. By shaking the vessel, the establish- ment of diffusion equilibrium between liquid and hydrogen, and attainment of constancy in the measured potential, are accelerated. This constancy, however, has been obtained by the loss of part of the volatile acid (or base) from the liquid to the hydrogen and ^Sörensen: Bloch. Zeitschr., 21, 1909. ^Michaelis: Ibid., 18, 1909; 46, 1912. 'Hasselbaich: Ibid., 30, p. Z17, 1910; 38, p. 77, iQiS; 49, P- 45o, 1913- 370 Hydrogen Ions in Biological Fluids [April would, therefore, indicate a too alkaline (or too acid) reaction of the fluid. The liquid is now renewed without changing the gas- mixture around the electrode, and shaking is repeated. It is easily Seen that electromotive constancy may now be obtained without any, or at least without any appreciable, alteration of the tension of the volatile acid (or base), i. e., without alteration of the original hydrogen-ion concentration of the liquid. This procedure may be repeated, if necessary, until the renewal of liquid no longer causes any alteration in the measured potential. For blood, urine, and probably the majority of biological fluids, a Single renewal is sufficient. Sea-water and similar Solutions, which are poor in "reaction regulators "^^ (Henderson^'^), are eo ipso far more susceptible to the change in carbonic-acid tension resulting from the method of measurement and would require three to four or, according to circumstances, even a larger number of renewals of the liquid before constancy is reached. In such cases it is easier, and more correct, to extrapolate graphically from the first three measurements (i. e., after two renewals of the liquid) in order to get the final value. The procedure described here permits one inconsiderable error, for which a correction may be made, if necessary. When diffusion equilibrium between the hydrogen and the liquid has been obtained, none of the components are, strictly speaking, any longer saturated with moist hydrogen at the existing barometric pressure but at a somewhat lower pressure. The potential changes, however, in ac- cord with the logarithm of the hydrogen pressure, so that, e. g., a fall in the hydrogen pressure from 760 to 700 (and a greater fall is practically inconceivable) would drop the measured potential to a value ca. i milli-volt too low. An error like this lies very near the limit of error of the whole method but may be eliminated, as already mentioned, by analysis of the hydrogen mixture and by calculation. Solutions which are poor in "reaction regulators," but whose hydrogen-ion concentration (owing to the volatile acid or base they contain) must necessarily be measured in the above-mentioned way (if it cannot be measured colorimetrically), have been found to '" Compounds which, by their presence, diminish the effect on the hydrogen- ion concentration of changes in the Proportion of acid or base. " Henderson : Ergebnisse der Physiologie, 8, 1909. 1913] K. A. Hasselbaich 371 present the difficulty that the potential changes in an unaccountable nianner when the liquid in the electrode vessel is at rest. For this reason I have proposed, as a normal method in measuring biological fluids, that the shaking should be mechanical and permanent, even during the reading of the electrometer. The electrode vessel is seen in Plate 3, Fig. i. The arrows indicate the direction and extent o£ the movement. While the shaking takes place, the electrode is constantly immersed in the liquid. It is easily seen that this electrode vessel, by another arrange- ment of the T-tube, may also be used in measurements which permit hydrogen to be led through the liquid. This property of the vessel may be of use, e. g., in efforts to control the correctness of the electrode by measurement of " Standard Solutions " of a known hydrogen-ion concentration. Fig. 2 (Plate 3) shows an electrode vessel used by me for small quantities of fluid, especially in determinations of the hydro- gen-ion concentration in 2-3 c.c. of human blood to which some hirudin is added. The blood may be taken f rom the lobe of the ear ; it is saturated in a glass syringe by rotation with about 20 c.c. of the alveolar air of the individual. The electrode, F, is saturated with a current of hydrogen flowing in the direction A— > B -» D -^ E. D is a groove in the inner part of a ground glass stopper lubricated with Vaseline and, during the flow of the hydrogen, it is turned so as to be opposite the hole E in the outer wall of the apparatus. The Saturation with hydrogen being completed, D is turned, as shown in Fig. 2 (Plate 3), and the cock B, which must be quite free from Vaseline, is turned around so that the first portion of the liquid from the syringe passes through A and down into the rubber tube C. The electrical connection between the liquid in the elec- trode vessel and the Solution of potassium chlorid (Fig. i, Plate 3) takes place along this route. When cock B is then turned as shown in the figure (Fig. 2, Plate 3), and cock H (which is carefully lubricated) is opened, the liquid rises in the electrode vessel as high as the side-tube ; H is now closed, B turned around, and the syringe disconnected. The shaking of the apparatus and the reading of the electrometer may now be started, When we are dealing with blood or other fluids containing dis- 372 Hydrogen Ions in Biological Fluids [April sociable oxygen Compounds, electromotive constancy is not attained until the moment when the layer of liquid into which the electrode projects is completely reduced. In such cases it may be useful, by saving time, to apply the Suggestion of Michaelis,^^ namely, to let the electrode just touch the surface of the liquid. I am of the opinion that by following the lines indicated above, we shall be able to measure the hydrogen-ion concentration of bio- logical fluids in many cases where it has hitherto been considered impossible, or where we have had to be satisfied with rough approxi- mations. There are undoubtedly numerous questions in biology and pathology which these improvements in method may help to solve. " Michaelis : Loc. cit. A METHOD FOR THE DETERMINATION OF TRYPTOPHAN DERIVED FROM PROTEIN JESSE A. SANDERS and CLARENCE E. MAY (Chetnical Laboratories of Indiana University, Bloomington, Ind.) Introduction. Tryptophan is a protein cleavage prodiict that is never obtained abundantly. So far as we know the tryptophan yield has been determined quantitatively in the case of but two proteins, namely casein^ and wheat ghadin.^ Only traces of tryp- tophan can be obtained from other proteins; and some proteins, especially gelatin, fail to yield it, if the indications of the usual test with glyoxyhc acid and sulfuric acid are rehable. Although tryptophan cannot be abundantly obtained from pro- teins, considerable importance is attached to it because it is produced in the tryptic digestion of protein and, in putrefaction, yields indol. The quantity of indican in urine indicates, in a general way, the extent of intestinal putrefaction. One usually accepts that con- clusion without considering the details of the tryptophan trans- formation, which involves the necessary presence of tryptophan precursors in the original protein molecules ; the degree of digestion of the particular proteins that yield tryptophan; the conversion of tryptophan into indol rather than skatol ; followed by the absorption of indol, its oxidation to indoxyl, its esterification and its excre- tion in the urine in the form of the potassium ethereal sulfate. Owing to the evanescent nature of tryptophan, its Isolation from tryptic digestion mixtures has been the subject of many investiga- tions. Although Hopkins and Cole, Abderhalden, and others, have ^Abderhalden: Zeit. f. physiol Chem., 190S, xHv, p. 23; Abderhalden and Samuely: Ibid., p. 276. (100 gm. of gliadin yield about i.o gm. of tryptophan; 100 gm. of casein yield 1.5 gm. of tryptophan.) ^ Osborne and Clapp : Amer. Jour. Physiol., 1906, xvii, p. 231; Osborne and Guest : Jour. of Biol. Chem., igii, ix, p. 426. (Hydrolysis of gliadin; revised gliadin-tryptophan figures.) 373 374 Determination of Tryptophan [April used the mercury stilfate-sulfiiric acid method^ in work on casein, this treatment has always been reported as giving figures somewhat lower than actual valnes. Tryptophan diminishes in quantity after a time, and may disappear, during the progress of tryptic digestion. In this laboratory we have found that the mercury sulfate-sulfuric acid method of Hopkins and Cole does not completely precipitate the tryptophan present in the digestion mixture. After precipitating the tryptophan-mercury-sulfate product from a casein digestion mixture, fikering, neutrahzing with calcium hydroxide and remov- ing the calcium sulfate and insoluble calcium salts, we obtained a filtrate that gave a characteristic tryptophan test with glyoxylic and sulfuric acids. Obviously the Hopkins-Cole method did not com- pletely precipitate tryptophan. Because of the smallness of the amounts of tryptophan usually derived from proteins, this method is necessarily dependent on the use of relatively large quantities of protein. The tedious nature of the methods for the purification of large amounts of proteins led us to endeavor to devise an accurate process involving the use of small amounts of protein. * The Solution of the problem seemed to depend on perfecting a method for the quantitative determination of small amounts of indol. We desired to use i^-napthoquinone mono-sodium sulfonate, such as Herter* employed in his work on indol, but could not find it on the market. We prepared a substance that reacted with indol, giving a deep violet colored Solution such as Herter obtained, but the substance formed by the combination of indol with our supposed ;8-napthoquinone mono-sodium sulfonate was not soluble in Chloro- form. Herter used Chloroform to extract the indol-containing Com- pound. Further use of our reagent was abandoned. It is probable that we had an isomer of Herter's reagent differing mainly from his in its reaction with Chloroform. We prepared our reagent by cautiously oxidizing " Eikonogen," the Photographie developer, by means of concentrated nitric acid. The oxidation was quite satis- factory but the_ substance obtained was evidently an isomer, bearing the sulfonic acid radical on a benzene nucleus other than the one holding the quinone linkages. * Hopkins and Cole: Jour. of PhysioL, 1901-2, xxvii, p. 418; Ibid.j 1903, xxix, p. 451. (Mercury sulfate-sulfuric acid method; Isolation of tryptophan.) * Herter and Foster: Jour. of Bio!. Chent., 1905-6, i, p. 257; Ibid., 1906-7, ii, p. 267. (ß-napthoquinone reaction with indol.) 1913] Jesse A, Sanders and Clarence E. May 375 General method. We studied the production of indol in the tryptic digestion of casein, the tryptophan yield from which is known approximately. We used small amounts (i. 0-1.75 gm.) of casein, digesting them with strong pancreatin Solutions free from trypto- phan, as determined by negative response to the glyoxylic-sulfuric acid test. The digestive periods differed in length. At the end of each, the mixture was neutralized, reinforced with neutral salts, and then made alkaline to one of several degrees of alkalinity. After sterilization in an autoclave, the mixtures were inoculated with mixed fecal bacteria from the stools of an individual on a mixed diet. No bacteria were isolated for purposes of Identifica- tion. The organisms were allowed to develop in the digestion mixtures at 37° C. for periods of different length. The reaction mixtures, after neutralizing and making them alkaline with a known amount of alkali, were distilled with steam until about 700 c.c. of distillate had been obtained. The distillate was diluted to 1,000 c.c. and an aliquot portion was treated with 0.2 per cent. sodium nitrite and conc. sulfuric acid solutions. A control Solution containing 0.25 per cent. of indol (Kahlbaum) was treated in the same manner. Each nitroso-indol Solution was then allowed to stand until the maximum color developed.^ We used the Wolf colorimeter for the tinctorial comparisons, and found that even with the small amounts of indol obtained (see figures later) an error of 3 per cent. was very easily detected by difference in the intensity of the resulting colorations. We experienced some difficulty, at first, in mixing definite amounts of the indol Solution with the nitrite and sulfuric acid Solu- tions, and water, which would give uniform shade and intensity of color. Later we obtained very constant results by taking an aliquot portion of the indol Solution, adding the nitrite Solution and enough water to fill the cylinder of the apparatus almost to the 100 mark, then adding the conc. sulfuric acid Solution and sufficient water to fill to the mark. On mixing uniformly, a very faint though distinct ' Moraczewski : Zeit. f. physiol. Chem., 1908, Iv, pp. 42-47; Chem. Ahstr., 1908, ii, p. 2578. (Colorimetric determination of indol in feces. The abstract of the original article contains an error that should be corrected: the sodium nitrite Solution has a concentration of 0.2 per cent. instead of 2.0 per cent. See also, Levene and Rouiller : Jour. of Biol. Chem., 1906-7, ii, p. 481. A bromine-tryptophan colorimetric method for the determination of tryptophan.) 37^ Determination of Tryptophan [April color developed which reached its maximum intensity in about half an hour. Details of the experiments. About 500 c.c. of skimmed milk were diluted in a precipitation jar with five volumes of water. The casein was precipitated by the addition of 12.5 c.c. of 10 per cent. acetic acid Solution. The casein was repeatedly washed with water by decantation and then dissolved in Standard sodium hydroxid Solu- tion (enough to dissolve the casein without leaving a large excess of alkali). The liquid required about 150 c.c. of n/2 sodium hydroxid Solution to produce a permanent alkalinity, using azolitmin paper as indicator. After dilution to a definite volume and filtration, two nitrogen determinations were made by the Kjeldahl method. It was found that each 100 c.c. of the Solution contained 0.8755 ö"^- o^ casein. Of the remaining Solution 500 c.c, were neutralized with phenolthalein as the indicator and treated with 0.4 gm. of sodium carbonate for each 100 c.c. volume of the neutral liquid. Then 25 c.c. of a saturated pancreatin (commercial) Solution and xylene, as a preservative, were added. Incubation was continued at 2)7° C. for 24 hours, when an equal portion of the pancreatin Solution was added; a third portion was added at the end of 48 hours. The incubation was then continued for forty-four days. Steam was passed through the alkaline Solution to remove the xylene. The digestion was apparently complete ; common tests for tryptophan indicated its presence. The mixture was neutralized and reinforced with neutral salts, such as Hopkins and Cole used in their work — 5 gm. Rochelle salt, 0.2 gm. ammonium phosphate and o.i gm. magnesium sulfate, per liter. No gelatin was added. The total volume was now made up to one liter and divided into four equal portions. Each portion contained cleavage products corresponding to 1.0944 gm. of casein. Portion A was sterilized in an autoclave and inoculated with a 24 hour slant agar growth of intestinal bacteria.^ Portion B was • The method of inoculation was as follows : The bacteria were grown first in ordinary broth inoculated from feces. After 24 hours, agar slants were made in the usual way. When these were 24 hours old, the organisms were detached by means of sterile water and a sterile wire, and the liquid containing them was poured directly into the flask to be inoculated. In order to establish a definite degree of alkalinity, the digestion mixtures, after steam distillation, were I9I3] Jcsse A. Sanders and Clarence E. May 377 treated with sodium carbonate (0.4 gm. per 100 c.c), sterilized and inoculated with some o£ the same 24 hour growth of bacteria. Portion C was made alkaline with sodium carbonate to 0.8 per cent. ; portion jD to i.o per cent. Both were inoculated as in A and B. After four days of incubation, the four flasks were reinoculated with fresh 24 hour cuhures of the bacteria and again incubated. After eight days' incubation, flask A was removed from the oven. Active indol-producing bacteria were present in the mixture. The original putrefaction mixture, neutral to litmus and giving a faint odor of indol, was made alkaline with sodium carbonate (to 0.4 per cent.). With steam distillation, all the indol passed into the first 300 c.c. of distillate. Determination of indol. A Standard indol Solution was made by dissolving 0.25 gm. of the pure substance in a liter of water. Twenty-five c.c. of the original distillate from flask A, diluted to about 90 c.c, were treated with 10 drops of a 0.2 per cent. sodium nitrite Solution, and six drops of conc. sulfuric acid Solution, diluted to 100 c.c, and mixed uniformly. The liquid was allowed to stand until the maximum rose-red color of the nitroso-indol developed, when it showed the same intensity of color as that produced by 1.3 c.c. of the Standard indol Solution diluted in the same manner to 100 c.c The total distillate contained 3.9 mg. of indol. In flask B, incubation was continued for nine days after the second inocula- tion. At the end of that time, indol-producing bacteria were still active. The reaction-mixture being distinctly alkaline, no alkali was added prior to steam distillation. All the indol appeared in the first 650 c.c of distillate, which was diluted to 700 c.c. and thor- oughly mixed. Of this Solution 50 c.c. contained as much indol as 1.2 c.c. of the Standard Solution. The total indol content of flask B was 4.2 mg. Flask C was incubated twenty-six days. The mixture smelled strongly of indol and was alkaline in reaction. No additional alkali was added. A distillate of 1000 c.c. was obtained, each 50 c.c. of which contained as much indol as 2.05 c.c. of the Standard indol Solution. The total content of indol in this putre- faction mixture was 10.25 mg. Flask D was incubated twenty-five titrated with n/io hydrochloric acid Solution and then neutralized quantitatively. The required weight of sodium carbonate was then added to the neutral Solu- tion to give the desired alkalinity. 378 Determination of Tryptophan [April days. It contained indol and was alkaline in reaction. Of i liter of distillate obtained by steam distillation, each 50 c.c. contained the quantity of indol present in 2.0 c.c. of the Standard Solution, in- dicating that the indol in the putrefactive mixture amounted to lo.o mg. A summary of the analytic data is appended. Flask Casein, gm. Digestion, days Bacterial action, days Indol, mg. A B C D 1.0944 1.0944 1.0944 1.0944 44 44 44 44 8 13 26 25 3-9 4.2 10.25 10.00 The yield of indol could be derived from 1.7872 gm. of trypto- phan in the case of sample C and from 1.7436 gm. of tryptophan in the case of sample D. The weight of casein corresponding to the amount of indol found must have yielded the calculated weight of tryptophan. This being the case, 100 gm. of casein yield either 1-593 g"^- of tryptophan (C) or 1.633 g"^- of tryptophan {D), as the minimum amounts. Hopkins and Cole claim that intestinal bacteria form small amounts of indol-acetic acid and other indol-containing substances, but we have not found these in our putrefactive mixtures, although they may have been formed in very small amounts, f or which reason we give the two results as indicating the minimum amounts of indol- yielding radicals in casein. Our results are as high as those ob- tained by other investigators. It is likely, of course, that the method will be improved by the further study we hope to give it, especially in its application to other common proteins. The present paper presents only preliminary results. The method, as outlined, is slow but it promises to be a satis- factory process for the determination of one of the cleavage products of protein material that hitherto has been difficult to de- termine quantitatively. PHYSICAL CHEMISTRY OF MUSCLE PLASMA^ FILIPPO BOTTAZZI (Physiological Institute, University of Naples, Italy) My experiments have been made on striated muscles of oxen, dogs, Scyüium stellare and Dentex vulgaris, and on piain muscles {M. retractor penis) of oxen. In the case of the dogs, the muscles were removed after flushing the blood vessels with 0.9 per cent. Solution of sodium chlorid (sometimes cooled to 4-5° C). In nearly all cases the muscles were preserved in dry vessels at low temperatures. They were freed from fatty and connective tissues, then minced, thoroughly pounded with quartz sand and infusorial earth, and plasma obtained in a Buchner press, generally at a maximum pressure of about 350 atmospheres. In some experi- ments the irritability of the animal (Scyllium) was abolished by gradually cooling it to about — 2° C, so that on cutting off the body musculature no contraction ensued. The muscle plasma (about 600-800 c.c.) was collected in dry vessels, centrifuged for an hour and preserved in a refrigerator. The plasma of striated mammalian muscle was always deep red in color and rather turbid; that of fish muscle was less colored. The plasma of piain muscle was always opalescent and almost color- less. The microscopic examination, made with powerful apochro- matic objectives, revealed no trace of morphologic Clements or granules in the centrifuged plasma, which always appeared to be perfectly homogeneous. But ultramicroscopic examination revealed the presence of innumerable very small and highly brilliant granules, mixed with a relatively small number of coarse particles, which have nothing to do with the granules, being composed of fat, glycogen and nuclear or sarcoplasmic f ragments. The existence of the ultra- ^ Presented at the eighty-first meeting of the British Association for the Advancement of Science, in Dundee, September, 1912. In these researches I was aided by my assistant Dr. G. Quagliariello. 379 380 PJiysical Chemistry of Muscle Plasma [April microscopic granules was never observed before; this is the most important result of my investigations. The number or concentration of the granules in the original plasma is so great that the iiltramicroscopic field appears almost uniformly luminous — the individual granules cannot be distinctly Seen. But when the plasma is diluted with Ringer Solution, the granules are separated, and then appear as distinct brilliant cor- puscles endowed with lively Brownian movements on a darkish homogeneous background. They are not precipitation-particles of a dissolved muscle protein, because they do not disappear under the action of dilute alkali. Precipitation of such protein might be caused by lactic acid produced in the muscles, but in that case the particles would be dissolved by alkali — we do not know of any acid- precipitated proteins that are not resoluble in alkalies. No ordinary reagent causes the granules to disappear at a low temperature. Moreover, acid increascs the number of particles, by precipitating a special dissolved muscle protein. The granules are present in almost equal number in plasmas ex- pressed from muscles which have been cooled to a low degree and which, therefore, are non-irritant, t. e., from muscles in which acid production is greatly diminished. The concentration of the granules is greater in the plasma of striated muscle than in that of piain muscle. Normal muscle plasma is, then, a Suspension of ultramicroscopic granules in a liquid zvhich, besides containing mineral salts and ex- tractives, certainly holds protein in a state of true Solution. Ac- cordingly, the plasma, f reed from the granules, is an optically homo- geneous fluid, but on adding to it a weak acid Solution, or on heating it at 55° C, additional particles appear — true precipitation- particles of a dissolved muscle protein, which may be termed myo- protein, while we may give the name myosin to the protein of which the plasma granules are made. The existence of other muscle pro- teins in the muscle plasma has not been proved. The observed plasma granules are apparently a degradation or cleavage product of material in the myofibrils, piain or striated, and preexist in all muscle plasma. Their preexistence is not only easily conceivable, but we are also obliged to assume it, when we reflect 1913] Filippo Bottassi 381 that both materlals — fibrillar and sarcoplasmic — exist in muscle fibres in two distinct phases, which of course must also remain distinct in the plasma. The fibrillar phase being represented in the plasma in the form of granules, these are probably constituent ele- ments of the fibrils, in harmony with the views of Heidenhain. The granulär material tends to flocculate spontaneously ; but spontaneous agglutination and Sedimentation of the granules occurs very slowly, because of their smallness and the high viscosity of the Suspension fluid. Dilution with water, or with neutral, faintly acid or alkaline Solutions, dialysis, or heating to about 30° C, accelerates the process; but it also occurs, in from about 12 to 24 hours, as I have Said, when all accelerating action of physical or chemical agents is excluded. This aggregation, followed by precipitation, of the granulär material is essentially the so-called "spontaneous coagidation" of muscle plasma or extract; it is therefore neither an enzymic-coagulation nor a heat-coagulation of dissolved protein. I have never observed phenomena like those described by Kühne — of nearly instantaneous clotting of cold muscle plasma when raised to room temperature. Precipitation of the granules is greatly accelerated by heating muscle plasma to between 38° and 54° C, when, after a few minutes, a heavy precipitate is produced, from which there separates a clear yellowish-red fluid, muscle serum. This phenomenon, which many authors Interpret as one of heat coagulation of a dissolved protein, is, on the contrary, the effect of rapid aggregation and pre- cipitation of the suspended granules. When their concentration is very great, massive clotting of the plasma occurs. The precipitate which appears during the first 24-48 hours of dialysis of plasma is composed of the granulär material, and is not formed from a dissolved protein. Sometimes the plasma trans- forms itself into something like a blood coagulum. Heat-coagulation of dissolved myoprotein is a continuous process, which does not appear to be complete even at 80° C. As we cannot deny that it begins at a temperature as low as 50° C, we are bound to admit that the precipitate of granules, formed at 54°-55° C, probably also contains a little myoprotein. In Opposi- tion to von Fürth and others, I have observed that the (dissolved) 382 Physical Chemistry of Muscle Plasma [April myoprotein is totally precipitated by strong and prolonged dialysis. As this precipitation process is also a continuous one, I do not deny that it begins during the first 24-48 hours; therefore the pre- cipitate of granules which forms early during dialysis may also contain a little myoprotein. A trace of dissolved protein can always be found in plasma wiiich has been dialyzed continuously for several months. It is probably serumalbumin, which cannot be wholly eliminated. Muscle pigments (hemoglobin, MacMunn's myohematin) are partly removed from the plasma by the agglutinated granules — ad- sorbed by them, and are also precipitated in some degree with myo- protein by prolonged dialysis. The granules and myoprotein, freed from electrolytes by suffi- ciently long dialysis, move toward the anode, when put under the influence of a strong electric current; they carry electronegative charges. For muscle plasma I have determined the quantity of plasma as per Cent, of fresh muscle, the total solid and total protein contents and ash yield, the specific gravi ty, lowering of the freezing point, electric conductivity, viscosity, surface tension and chemical reac- tion. Tables 1-3 contain the results of these determinations. TABLE I General data pertaining to muscle and muscle plasma Plasma No. Muscle Plasma Muscle Weight, kg. Pressure, atm. Weight, gm. Per Cent, of muscle Scyllium: Striated Ox: Striated Dentex: Striated Ox: Piain Dog: Striated Ox: Piain 7 8 9 5 II 12 1.067 0.938 0.894 I.184 0.829 I.190 350 350 50 350 350 50-350 666 592 217 580 339 562 62 63 24 50 40 47 The plasma varied in quantity from 40 to 63 per cent. (volume), for pressures which never exceeded 350 atmospheres. The dry residue from piain muscle plasma was less than that from striated muscle plasma. The total protein content was relatively low — less than that of blood serum. Muscle plasmas are very rieh in their I9I3] Filippo Bottazzi 383 TABLE 2 Percentage data for water, total solid and total protein contents, and ash yield, of muscle plasma A. Piain muscle Muscle plasma Water Total solids Total protein Ash Organic solids mious protein 3:Ox 4: Ox 5:Ox 6:Ox 12: Mixed — Fraction a Fraction b Fraction c 93-14 97-57 93.808 94.126 93-27 92.76 93-60 93-99 6.86 6-43 6.192 5-874 6.73 7-24 6.40 6.01 3-63 3-37 3-iS 2.75 3-37 4-13 2.90 2.67 1.32 1-30 I.148 1-37 1.30 I.91 1.76 1.894 1-754 2.06 B. Striated muscle 2: Ox 91.086 8.914 4-53 1-739 2.64s 8 Ox 92.57 7-43 3.65 0.85 2.93 9 Dentex 83-90 16.10 4.10 1.82 3-i8 7 Scyllium Scyllium 90.36 9-64 3-04 3-36 1.80 4.80 loa: Scyllium 2.38 1 1 : Dog 87-37 12.63 3.85 1.50 7.28 TABLE 3 Data pertaining to physico-chemical properties of muscle plasma A. Piain muscle 6 Lower- Surface Muscle Specific ing of the Electric conduc- Viscos- tension at25° Chem- ical Remarks regarding ¥n gravity freezing tivity ity (P 25°) reaction the plasma CS point (A) (K180) (Ch-io7) 3 Ox 1.026 Normal, fresh 4 Ox 1.024 0.806° — — — — Normal, fresh 5 Ox 1-023 0.761° — 4-13 73-15 7.44 Normal, fresh 6 Ox I.02I 0.730° 0.0144 3-04 76.76 6.18 Normal, fresh 12 Mixed 1.024 0.804° — 4-48 73-34 7.30 Fraction a 1.026 0.812° — 5-98 69-34 — Expressed at 50 atm. Fraction b 1.025 0.812° — 3-68 74.67 — Expressed at 200 atm. Fraction c 1.023 0.810° — 2.96 76.27 — Expressed at 350 atm. Striated muscle 7 Scyllium 1.027 2.455° 0.0147 1-59 II. I Muscles were not cooled 8 Ox 1.027 0.868° 0.0107 1.75 78.19 31.4 Normal conditions 9 Dentex 1.049 1.196° 0.0120 2.82 76.22 12.S From muscles of cooled animal 10 Scyllium Scyllium: Frac. loa 1.024 1.024 2.337° 2.494° 0.0149 0.0150 3.71 1.71 72.9 73.1 S.65I 6.50 j From three Scyllia (cooled to from -2° to -3° C.) II Dog 1.039 1.088° — 2.24 68.90 10.6 Three hours after expres- Dog (i) 1.037 1.016° — — 70.00 35.0 Data obtained the fol- lowing day 384 Physical Chemistry of Miiscle Plasma [April yield of ash and content of organic non-protein substances. Gen- erally, the dry residue and the protein contents are inversely pro- portional to the pressure at which the plasma is expressed. Since the specific gravity (1,021-1,027, as a rule) is quite near that of blood serum, I believe the salts and extractives, lipoids, glycogen, etc., help to accoimt for it. The osmotic pressure is always very high (A =0.730-1.088° C. for mammals; A =2.337-2.494° C. ior Scyllium; A = 1.196° C. for Dcntex), higher than that of the blood. The reaction is always acid (Ch- 10'^ = 5.65-12.5 ; but there are also higher values : ChIO^=3I-4 f- e.). Acidity is lower in plasma of piain muscle and of cooled striated muscle, higher in the plasma of striated mammalian muscle. As a rule, the hydrogen- ion concentration increases with time; but as this augmentation is rather feeble, I believe, with Fletcher, that the maximum production of acid substances occurs in muscles soon after their Separation from the body. The high osmotic pressure of muscle plasma is probably due mainly to the substances that determine the acid reaction. The low electric conductivity (Kjso = 0.0107-0.0144 for mam- mals; 0.0147-0.0150 for Scyllium; 0.0120 for Dentex) and high viscosity (p25o= 1.59-2.82 ; 3.04-5.98 for piain mammalian muscle) of the plasmas are explained by their corpuscular composition. But the very high viscosity of the plasma of piain muscle is probably caused by some particular protein derived from the connective tissue. Zw The surface tension (100-^=68.90-78.19) is generally higher than that of the blood serum. My results are, for the most part, in Opposition to those obtained by previous authors. But the new interpretation of phenomena like those of spontaneous coagulation and heat coagulation, etc., was suggested to me mainly by the granulär Constitution of muscle plasma. As I stated above, this is the most important result of my investigations, a result which hereafter must be recognized by all who study problems pertaining to the chemistry and physical chem- istry of the Contents of the muscle fiber. 1913] Filippo Bottazzi 3^5 I have endeavored to corroborate my hypothesis, that the granules are disintegrated fibrillar material, by trying to stain the granules with some pigment which would selectively color the muscle fibrils ; and also by attempting to show that the granules possessed double refractive power. My attempts have been unsuccessful, however, although this was not unexpected, because of the ultra- microscopic dimensions of the granules. FASTING STUDIES II. A note on the composition of muscle from fasting dogs^ H. C. BIDDLE AND PAUL E. HOWE (Laboratory of Physiological Cheniistry, University of Illinois, Urhana, Illinois) The variations in the composition of the different forms of muscle in the normal individual have received considerable atten- tion. A comparison of the nitrogen and moisture contents of heart and striated muscle reveals a lower percentage of nitrogen and a higher percentage of moisture in the heart muscle than in the skeletal muscle. The proportions of these and other constituents in the fasting muscle have not been studied extensively. The changes which occur in the composition of muscle during fasting are significant for the Solution of general problems relating to the effects of fasting. In a study of the influence of fasting upon the creatine content of dog muscle, determinations were made of the proportions of nitrogen, moisture, fat and creatine in normal and fasting muscle ; and also of the nitrogen and creatine in heart muscle. In these preliminary experiments particular attention was paid to the per- centages of nitrogen and creatine : an attempt was made to show the relation between the nitrogen and creatine contents of muscle, so that the ratio of creatine-nitrogen to total nitrogen might be used as an index of the changes due to pathological conditions. This factor should be more significant than the percentage of creatine in fresh muscle, and as accurate as that for the creatine content in muscle on a fat- and moisture-free basis. These results, as well as the variations in the creatine content of muscle, will be discussed in a later paper.^ ^ Presented before the Columbia University Biochemical Association, December 6, 1912; Biochemical Bulletin, 1913, ii, p. 288. ^Howe and Hawk: Presented before the recent annual meeting of the American Physiological Society, but not abstracted in the proceedings. 386 1913] H. C. Biddle and Paul E. Howe 387 The procedure employed in determining some variations due to fasting was as follows : muscles (M. sentit endinosus and M. hiceps femoris) of normal animals were analyzed for moisture, fat, phos- phorus and creatine ; and, in one case, the heart was analyzed for nitrogen and creatine. Muscles of the same kind were removed aseptically from normal dogs under ether anesthesia^ and analyzed for nitrogen and creatine. After a prolonged fast the corresponding muscles of the opposite legs of the "operated" dogs, as well as the remaining muscles on the same side, were analyzed for total nitrogen and creatine, and in two cases for moisture and fat. With such a procedure the changes which resulted from fasting were studied on the same individual, also in fasted animals as compared with different normal controls. The "operated" dogs recovered readily and the wounds healed rapidly even when the fast was begun immediately after the Operation. The analytical methods employed were as follows: Total nitrogen was determined by the Kjeldahl process ; moisture by dry- ing in a vacuum over sulfuric acid at room temperature ; fat by the ether-extraction process, and creatine by the Folin procedure as modified for meat by Emmett and Grindley.* The creatine was extracted according to the methods of Grindley and Woods^ and of Mellanby.^ The table on page 388 contains the more significant data. A consideration of this data shows an increase in the percentage of moisture, and a decrease in the percentages of nitrogen and crea- tine, in the striated muscle as a result of fasting. From the data on a Single normal heart and a single fasting heart there appears to have been a decrease in the nitrogen and an increase in the creatine content of the fasting heart. We also note that the changes which take place in the same fasting individual, as con- trasted with the fasting changes when compared with different con- trol animals, are approximately the same. 'We wish to thank Dr. O. O. Stanley, of the University of Illinois, and Dr. C. T. Moss, of the Michael Reese Hospital, of Chicago, Illinois, for their aid in the removal of the muscles. * Emmett and Grindley: Jour. of Biol. Chem., igoy, iii, p. 491. ^ Grindley and Woods : Ibid., igoö-'o;, ü, p. 309. ' Mellanby : Jour. of PhysioL, 1908, xxxvi, p. 453. 388 Fasting Studies [April Data pertainlng to the composition of muscle front fasting dogs Dog Kind of muscle Moisture, per Cent. Nitrogen, per Cent. Fat, per cent. Creatine, per Cent. E. D. Normal leg Normal leg Normal heart Normal leg Fasting leg Fasting heart' Fasting legi» Normal leg Fasting leg" 73-4 73-4 3-42 3-51 2-95 3-34 2.82 2.6s 2.95 3-99 3.6l 2.4^ 2.28 0.56 0.31 0.33 B. 0.23 81.2 0-34 0.31 C. A. 81.8 0.49 0.19 0-39 The increase in the moisture content of fasting muscle may be associated, in part, with the decrease in the fat content; in normal animals there is a decrease in the percentage of moisture associated with an increase in the fat content of muscle.^^ The increase in the moisture content of fasting muscle may also be due to changes in the colloidal State or the molecular condition of the cellular con- stituents. This increase in moisture is more significant when we consider that there is apparently a greater decrease in the cytoplasm than in the nuclei of the cells as a result of a fast;^^ the nucleus and the connective tissue, the substances which would then pre- dominate, normally contain less water than the cytoplasm. The increase in the moisture content of fasting muscle has been noted by other investigators. The lower absolute nitrogen content of fasting muscle, when considered on the basis of fresh muscle, becomes an increased rela- tive nitrogen content, when the values for nitrogen are calculated ^Content of phosphorus = o.i6 per cent. 'Content of phosphorus = 0.21 per cent. 'A sixty-four day fast which resulted in original weight. The animal received 320 c.c. ^'A twenty-one day fast. " A fifteen day fast, which resulted in a loss of 38 per cent. of the original weight. The animal did not receive water. " This f act, together with certain other deductions, has been corroborated by data in a personal communication from Professor P. F. Trowbridge of the University of Missouri. "Morgulis: Archiv für Entwicklungsmechanik der Organismen, 1911, xxxii, p. 169. a loss of 62 per cent. of water daily. of the 1913] H. C. Biddle and Paul E. Howe 389 on a moistiire- and fat-free basis. An alteratlon in the direction of increased nitrogen content is more in harmony with the changes that actually take place in the muscle. The relatively greater de- crease in the volume of the cytoplasm than of the nucleus, and the apparent relative increase in the connective tissue, are modifications in the direction of a higher nitrogen content of the muscle. The proteins which predominate in the connective tissue and in the nuclei of muscle contain higher percentages of nitrogen than do the pro- teins which make up the major portion of the cytoplasm of the cells. In addition to changes in the chemical nature of fasting muscle, certain physical modifications arise: normal muscle is firm to the touch and, when hashed, may be readily handled without sticking to the fingers ; fasting muscle, on the other hand, is soft to the touch and, when hashed, adheres tenaciously to the fingers. While the lowered percentage of nitrogen in the fresh skeletal muscle, as a result of fasting, may be due to diminutions in the Contents of moisture and fat, the difference in the composition of heart and skeletal muscle cannot be explained in this way. If the nitrogen content of normal striated and heart muscle be calculated to the moisture- and fat-free basis, there still remains a greater per- centage of nitrogen in the striated muscle. That there are differ- ences in the relative proportions of the soluble protein and the stroma in heart, and in striated and smooth muscle, has been shown by Saxl,^'* who finds that seven-eighthsof the skeletal muscle consists of soluble proteins while but one-third of the heart muscle is of this nature. A careful differential study of the proteins of fasting muscle should throw some light upon the nature of the disintegrative processes which take place in the tissues as a result of fasting. Such a study is contemplated. " Saxl : Beiträge s. ehem. Physiol. u. PathoL, 1907, ix, p. i. SOME NOTES ON THE FORM OF THE CURVE OF CARBON-DIOXIDE EXCRETION RESULTING FROM MUSCULAR WORK FOLLOWING FORCED BREATHING^ G. O. HIGLEY (Ohio Wesleyan University, Delaware, Ohio) (WITH PLATE 4) In an earlier research^ an attempt was made to determine how soon after the beginning of work an increase in the production of carbon dioxide begins to show itself in the expired air. This time (the latent period) was at first found to vary from three to fourteen seconds. Now, clearly, such periods are not long enough to cor- respond with the time required for the carbon dioxide formed in the muscles at the first muscular contraction to reach the outside air. It must first diffuse into the blood from the tissues where it is formed, then traverse the venous half of the systemic circulation, the right side of the heart, and the arterial half of the pulmonary circulation, and finally diffuse into the air of the alveoli before any of it can appear in the breath. From the conclusions of Stewart and others it appeared that from fifteen to twenty seconds is the least possible time required for the blood to traverse this distance, to say nothing of the diffusion time. It was finally found that the sudden increase in the rate of excretion of carbon dioxide, after the beginning of work, was due primarily to a better Ventilation of the lungs, while the continuation of the increase was due to the Ventilation of the blood and tissues as well. A recognition of this fact led to the following modification of the method for the de- termination of the latent period of carbon dioxide excretion : ^ This paper was accepted for publication by the officers of Section VIII, d. Eighth International Congress of Applied Chemistry, and was read before the Section at a stated meeting on September 11, 1912; Biochemical Bulletin, 1912, ii, p. 153. ^Higley and Bowen: American Journal of Physiology, 1904, xii, p. 311. 390 UJ I- < O > CO _l < ü UJ X o o CO ^ o H W CÜ u d X w c i H X <; o w Q 1 ^ n u w m ( ) Pi H W J o 1913] G- O. Higley 391 After the " normal " rate o£ excretion at rest had been de- termined, the subject began forced breathing at a predetermined rate, continuing this for a minute or so until the curve of carbon dioxide excretion had apparently assumed its permanent direction. At this point, at a signal from the experimenter, the subject began to drive the bicycle as in the preceding experiments. The effect was marked, the new rate of excretion being sharply defined from the normal rate preceding it. The further increase in the rate of excre- tion, after the beginning of work, was now not so prompt in its appearance, and came on more gradually, reaching its maximum after a minute or so, depending on the work. In these experiments the latent period of increase due to work was from seventeen to twenty-two seconds. It is evident that as the latent period will vary with the rapidity of the circulation, the rapidity of diffusion, and the rate of work, a more definite figure was not to be expected. Shortly after the publication of these results by Bowen and the writer, a communication was received from Prof. N. Zuntz, calling attention to the gradual character of the change in rate of excretion of carbon dioxide after the beginning of work (as already men- tioned) and kindly suggesting a modification of the method of carrying out the latent-period experiments. According to Prof. Zuntz, if the forced breathing were continued for five minutes instead of one minute, as already stated, the blood and tissues would become thoroughly ventilated; the direction of the curve of carbon dioxide would become parallel to that before forced breath- ing began ; and, furthermore, with the beginning of work, the carbon dioxide curve, after the latent period of twenty seconds, would change much more sharply than it did in the published record. The writer accordingly made a series of experiments in which the forced breathing was continued for from five to seven minutes before bicycle work has begun. The results of one of these experiments are seen in Plate 4 in which A is the Pneumograph record, Pqrs the carbon dioxide curve, T the Chronograph record, and M the bicycle record. The line Pq'q^'r, as in the previous paper,^ represents the rate of excretion * Higley and Bowen : Loc. cit. 392 Curve of Carbon-dioxide Excretion [April before the beginning of forced breathing; the line Pqq'q"r (broken by the arresting of the beam and the addition of foiir gram weights) represents the curve of carbon dioxide during forced breathing; r is the Position on the curve of the carbon-dioxide-writing lever at the instant when work was begun ; and s is the point where the curve changes as a residt of the zvork. This research was conducted on two subjects. It was found very difficult to maintain respiration of uniform depth for five minutes, since there is a decided tendency to make the respiration shallower. Indeed, notwithstanding the great care on the part of the subject, the Pneumograph record indicated, in some cases, a lessened depth of respiration toward the end of the forced respira- tion period. In the case of one subject the curve for rate of excretion of carbon dioxide returned, during the period of forced respiration, practically to the original value. With the other subject the return was less perfect, It would seem that as a result of the additional work of the respiratory Organs a return of the rate of excretion to the value during normal respiration could not be expected. While, therefore, the writer is able to confirm Prof. Zuntz's pre- diction regarding the sharpness of the change, as a result of work, in the curve of carbon dioxide after continued forced respiration, he can confirm only in part Prof. Zuntz's prediction on the return of the curve, during forced respiration, to the direction which it had before forced respiration was begun. This work was done in the physiological laboratory of the Uni- versity of Michigan, Ann Arbor, Michigan. THE INFLUENCE OF BAROMETRIC PRESSURE ON CARBON-DIOXIDE EXCRETION IN MAN^ G. O. HIGLEY (Ohio Wesleyan University, Delaware, Ohio) (WITH PLATE 5) Introduction. This work was suggested by that of Lombard^ on " Some of the influences which affect the power of muscular contraction." In that research, which was made with the ergo- graph, Lombard found that, in general, there was a fall of muscular power during the day, this result being noted on eighteen out of a series of twenty-three days. However, on certain days, the fall in power due to fatigue was slight and on five days the power was greater at the last experiment than at the first. These exceptions led to the suspicion that barometric changes had an influence on muscular endurance. When, later, a comparison was made between Lombard's endurance curve and the curve of barometric height, it was found that, while no constant relationship existed between the two variables, they varied in the same sense on twenty out of twenty- three days ; i. e., in general " when the barometer rose during the day, or feil less than on the preceding day, the muscular endurance either rose, or feil less than on the preceding day." It has been shown, furthermore, that while a diminution of barometric pressure increases both the respiration rate and the volume of air respired, after allowance is made for the increase of volume due to the lower pressure the volume respired is less (Speck). Now, the effect of increasing barometric pressure upon the power of the muscular System might possibly be due to some influence ^ This paper was accepted for publication by the officers of Section VIII, d, Eighth International Congress of Applied Chemistry, and was read before the Section at a stated meeting on September 11, 1912; Biochemical Bulletin, 1912, ii, p. 153. ^ Lombard : Journal of Physiology, 1892, xiii, p. i. 393 394 Carhon-dioxide Excretion in Man [April exerted through the nervous and circulatory Systems tending to increase the readiness of metabolism; if such were the case then a Variation in barometric height should be accompanied by a Variation, in the same sense, in the rate of excretion of carbon dioxide. Plan of the experiments. It seemed that a series of experi- ments carried out for a month on three healthy subjects might throw light on this question, and also give interesting results as regards the effect of other conditions on the rate of carbon dioxide excretion. A series of respiration experiments was planned, accordingly, for three subjects, A and B, students in the University of Michigan, and the writer, C. A and B were 24 and 22 years of age re- spectively, and weighed, without clothing, 158 and 159/^ pounds. C was 46 years of age and weighed, exclusive of clothing, 148 pounds. Each subject was to live his regulär daily life except that no vigorous muscular exercise was to be engaged in immediately preceding any experiment and that nothing whatever was to be eaten between meals. The plan of work is indicated in the ap- pended summary, where the data for the third part of each experi- ment are placed below those for the first and second : Subject Hour of rising Reclined First experi- ment Breakfast Time until next experiment Reclined Second experiment begun Dinner A B C 6 6 6 6:4s 7:05 7:25 7:00 7:20 7:40 7:40-8:00 8:00-8:20 8:00-8:20 4 hr. 4 hr. 4ihr. 11:4s 12:05 12:25 12:00 12:20 12:40 12:40 1:00 1:00 Subject Time until next experiment Reclined Third experiment begun Lunch Experi- menter A B C 4 hrs. 4 hrs. 43 hrs. 4:45 5:00 5:2s 5:00 5:20 5:40 5:40 6:00 6:00 c A B The routine of work was as follows : The subjects rose at 6 o'clock, reaching the laboratory at about 6:35. A reclined upon a couch at 6 :45 in preparation for the first experiment. B and C prepared all the apparatus, making the initial calibration of the balance, weighing the guard tubes, reading and recording the barometric height, the outdoor and room temperature, etc. In order to enable the experimenter to judge the better as to the physical I9I3] G. O. Higley 395 condition of each subject, mouth temperature and pulse were also taken and recorded. This routine at the laboratory was followed at 12 M. and 5 P. M. TABLE I Data showing the excretion of carbon dioxide by subjects A, B and C, in milligrams per minute. Date S ubject A Subject ß. Subject C. Dec. 7 A. M. 12 M. 5 P. M. 7 A. M. 12 M. 5 P.M. 7 A. M. 12 M. 5 P. M. 23 406 422 381 498 567 406 390 447 24 438 460 447 489 422 541 419 409 409 26 442 448 466 SI4 429 (743) 422 403 428 27 422 453 466 535 434 548 390 403 390 28 403 448 (635) 488 507 498 397 375 419 29 407 422 381 520 553 546 382 387 456 30 438 483 405 529 495 518 419 381 374 31 42 s 470 444 (647) 489 476 390 362 438 Jan. 2 433 487 480 438 (611) 570 393 422 473 3 470 436 422 416 462 508 394 377 422 4 507 435 480 537 442 553 386 448 S 469 473 442 528 466 515 410 396 6 458 466 442 525 531 449 403 476 7 46s 442 432 439 560 466 406 386 411 9 416 436 436 410 442 462 390 380 448 10 416 459 448 455 506 383 42s 473 II 456 439 426 453 422 526 363 402 337 12 T "7 446 543 388 13 14 405 462 445 476 504 398 427 16 436 496 449 469 531 440 402 396 437 17 412 453 418 460 459 18 377 407 462 459 469 364 442 435 19 472 487 422 445 474 409 403 438 429 20 469 476 436 402 455 432 402 474 419 21 462 436 493 399 442 493 406 448 434 23 428 481 429 509 442 436 396 449 429 24 422 517 495 500 459 537 409 460 429 25 422 475 402 528 486 422 468 428 26 495 561 422 422 Averages 438 462 443 472 476 501 401 414 427 Results. The results of this series of experiments are shown in Table i, in milligrams of carbon-dioxide excretion per minute. It will be noted that A's average for the midday experiments is con- siderbly higher than that for the morning and evening experiments. 39^ Carbon-dioxide Excretion in Man [April This is due, in part at least, to the fact that this subject took his heartiest meal in the morning. The excretion of carbon dioxide for B and C, on the other band, was greatest in the evening, since these subjects took their dinner at i P. M. The remarkably high excretion shown for A at the evening ex- periment of December 28 (635 mg., while the average for that hour for this subject is only 443 mg.) is explained as follows: This subject went skating in the afternoon of that day and at about 2 130 o'clock had the misfortune to break through the ice, becoming wet to the neck. On being rescued, he walked about two miles in his frozen clothing, exposed meanwhile to a strong wind at a temperature of about — 6° C. On reaching his room he took a thorough rubdown, made a change of clothing, rested for one and one half hours, and appeared at the laboratory at the usual hour for the experiments, with the result stated above. It will be noted that all of this subject's values for the following day, especially that of the evening, were much below the average, indicating a reaction from the exposure and excitement of the preceding day. The high ex- cretion of the morning of January 4 is supposed to be due to lunch eaten late on the preceding evening; that of 12 o'clock, January 26, to an exceptionally heavy morning meal ; and the low result of the evening of January 19 to an especially light midday meal. The irregularity of results obtained from B are somewhat diffi- cult to explain. Those of the morning of December 27, 30 and 31, were due to lunch eaten late the preceding evening and in the case of the two latter results, also in part to excessive haste to reach the laboratory in time for the regulär experiment. Other high results, especially those of 5 P. M., December 26, and of 12 M., January 2, were undoubtedly due to Indigestion. Passing now to a study of the relation of carbon dioxide excre- tion to barometric changes, Plate 5 will be found to embody, in the form of curves, the results already given in Table i, with time as abscissae, and milligrams of carbon dioxide per minute as ordinates; it presents curves for A, B and C, together with that for the barometer in millimeters of mercury and of the outdoor temperature in degrees centigrade. The temperature of the room was practically constant throughout the series of experiments. Three curves are Ui I- < o > ui 3 OQ _j < o s UJ I ü o m o u o — Z Z < v: \ /"-v ' \ / \ ■y^ / / / V \ / / / \ ; \ /■' ); \\ // \ \ /l ;. \ / i \: 'w/ ■N §88 o o o (p ^ c\j r^ rv. r-~. 9 o 2 u I V- w OJ 5 ■-> CVJ 1—* ^ "ü? a. O -o S >-H u ^ H -«-> v» ^ o o Ü •o CO •"2 i 3 o w •ö Q s c CD 1— t X tn !> o 3 1 — 1 Cfi p ' C/J 1 "O U( 3 z a. r^. "o (J ^^' c« CQ ^ c> E w ^^ u ';;;; o i-i ^ rt ^: ^ -^ ^ /-^ "^ u> o ^^' O C > 00 CO C/2 ■g 3 W cd c •5 <0 bi c u p o f_( *^ a "^ t/D f— ^ 't W u o s "Ö o o c Cii nj o CJ <; M ^ m iö O < -4-* U w •;^ 3 o ^ 3 0) Ol w fe> 4> !lj C/} J C fc _o N ^ ^ cu t— ( o ■o d tfl fu u- >^ W o fO hJ 'i Ä UJ u 1- 3 < U I9I3] G. 0. Higley 397 given for each subject where the necessary data were at band. In each case the morning, midday and evening ciirves are represented, respectively, by solid, long-dash and short-dash lines. Analysis of the results. Comparison of the data for haro- metric pressure and carhon dioxide excretion. Before proceeding to a rigorous mathematical investigation of the relationship between barometric change and carbon dioxide excretion, it seemed desirable to make a comparison of these two variables at a number of the dates on which especially marked barometric fluctuations took place, since in such cases the effect would be more pronounced and less likely to be masked by other varying conditions, such as amount and character of the preceding meal, character of muscular exercise, etc. To f acilitate such a comparison Table 2 was prepared ; it indicates experiment number; dates between which the comparison is made ; barometric height, rise or fall ; subject ; carbon dioxide for the two days between which comparison is made; rise or fall of excretion; and relation between barometric change and carbon dioxide excretion, whether direct or inverse. Taking first the morning values, it was found that the barometer rose between 7 A. M., December 23, and 7 A. M., December 24, from 739 to 746, or 7 mm. During the same period the excretion of carbon dioxide of the three subjects changed as follows : That of A from 406 to 438 mg. per minute, an increase of 32 mg.; that of B from 381 to 489, an increase of 108 mg., and that of C from 406 to 419, an in- crease of 13 mg. per minute. Thus with rising barometer there was an increase in the rate of excretion of carbon dioxide in the case of each subject. A similar result is obtained in four other morning experiments (two subjects). In three morning experi- ments there are two direct results each. One experiment shows two indirect results, i. e., there is a change in carbon dioxide excretion which is opposite in sign to that in the barometer. Summing up the results of the morning experiments we have the following : Eleven experiments were carried out on A, seven on B, and eleven on C. The degree of correspondence of barometric change with carbon dioxide excretion was : A, 7 cases out of 11, or 63.6 per cent. B, 6 cases out of 7, or 85.7 per cent. C, 6 cases out of 11, or 55.5 per cent. 398 Carhoiirdioxide Excretion in Man [April TABLE 2 Data obtained at 7 a.tn. Relation between barometric Barometer Excretion of carbon dioxide change and No. Date. Dec. Subject carbon dioxide excretion. Rise, Fall, Per minute, Rise, Fall, Heights, mm. mm. mm. mg. mg. mg. Direct Inverse I 23-24 739 -746 7. — A B C 406-438 381-489 406-419 32 108 13 — 32 108 13 2 26-28 745-I-72I 24.1 A B C 442-403 514-488 422-397 — 39 26 25 39 26 25 3 28-29 721 -742.1 21. 1 ' A B 403-407 488-520 4 32 Z 4 32 C 397-382 — 15 — 15 4 29-31 742.5-736 6.1 A B C 407-425 18 — — 18 382-390 8 — — 8 5 Jan. 2-4 737-I-743-5 6.4 A B C 433-507 438-537 393-394 74 99 I — 74 99 I 6 5-7 743-5-732.5 II. A B C 469-465 528-439 4x0-406 — 4 89 4 4 89 4 7 11-12 753.9-737-1 16.8 A 456-446 — 10 10 B 453-543 90 — — 90 C 363-388 25 — — 25 8 12-14 737.1-751-2 I4.I — A B C 446-405 — 41 — 41 388-398 10 — 10 9 18-19 748.2-739.3 8.9 A B C 377-472 95 — — 95 364-403 39 — — 39 10 23-24 749 -739.8 '~^ 9-2 A B 428-422 509-500 6 9 6 9 C 396-409 13 — — 13 11 24-26 739.8-758.8 19. — A B C 422-495 73 — 73 409-422 13 — 13 The direct results from the midday experiments were as foUows A, 3 cases out of 7, or 42.8 per cent. B, 3 cases out of 9, or 33.3 per cent. C, 6 cases out of 9, or 66.6 per cent. From the evening experiments the direct results were A, 3 cases out of 6, or 50 per cent. B, 4 cases out of 9, or 44.4 per cent. C, 4 cases out of 8, or 50 per cent. I9I3] G. O. Higley 399 TABLE 3 Data showing the relation of carhon dioxide excretion to barotnetric change, at noon for subject A. Barometer reading, mm. 739 746 742 722 726 742 740 736 738 742 745 743 739 732 745 753 749 753 747 748 747 740 744 743 749 738 752 758 Carbon dioxide excretion, mg. 422 460 448 453 448 422 483 470 487 436 435 473 466 442 436 459 439 462 496 453 407 487 476 436 481 517 475 561 X ■ 4,2 2.9 - 0.8 ■21.2 -16.4 - 0,8 ■ 3-2 - 7-2 ■ 4-4 - 0.4 1,8 0.2 - 3-4 -10.4 2-5 9.9 6.6 10.3 4.2 4.8 4.6 - 3-2 0.8 0,6 6.5 - 4-5 9.6 I5-I -40 - 2 -14 - 9 -14 -40 21 8 25 -26 -27 II 4 -20 -26 - 3 -23 o 34 - 9 -55 25 14 -26 19 55 13 99 ^•2 17.64 8,41 0.64 449.44 268.96 0.64 10,24 51,84 19.36 0,16 3-24 0,04 11,56 108.16 6.25 98.01 36.00 106.09 17.64 23.04 21.16 10.24 0.64 0.36 42.25 20.25 92.16 228.01 1,642,43 1-2 1,600 4 196 81 196 1,600 441 64 625 676 729 121 16 400 676 9 529 o 1,156 81 3.025 625 196 676 361 3.025 169 9,801 Products {XV) 27.078 Negative Positive 5-8 67.2 57-6 HO. 48,6 13-6 65- 29,7 151,8 43-2 253-0 80. 15.6 247-5 1,174-6 168 II. 2 190,8 229,6 32 10,4 2.2 208 142.8 II. 2 123-S I24,& 1,494,9 2,749,4 1,174,8 1.574-6 |i>642.43 ^., 127^078 ^{xy)= 1,574-6 Coefficient of correlation : Na-^a-^ 28x7.65x31.1 = +0.236 ^ . 0.2365 Regression = — ^ = 0.95 400 Carhon-dioxide Excretion in Man [April Or, out of a total of seventy-seven experiments, there was direct relationship between barometric change and carbon dioxide excre- tion in forty-two experiments, or 54.5 per cent. It will be Seen from these results that the apparent degree of correspondence, so far as it is revealed by this method of analysis, is greater in the morning experiments than in those carried out at midday or in the evening. This is probably due to the fact that in the morning not merely the digestive organs, but the whole System, is in a more uniform condition than at any other time during the twenty-four hours. Application of the method of least Squares. It now seemed desirable to subject the results obtained in this series of experiments to a more rigorous analysis than that just described, with a view of discovering what is the degree of correlation between the two vari- ables, the barometric height and the rate of excretion of carbon dioxide, during muscular rest. The data obtained in the experi- ments were, therefore, examined by the method of least Squares, which was applied separately to the three sets of data from each subject in order that the effect of different times of day might be determined separately. In Table 3 are given the barometric height and the correspond- ing carbon dioxide excretion f the problem is to find the correlation between these two quantities, and also the regression of carbon dioxide on barometric height, i. e., the amount of change in excre- tion of carbon dioxide for a millimeter change in barometric height. The means of columns i and 2 are obtained in the usual manner, by dividing the total in each column by the number of experiments {N). Having obtained these means, two additional columns are formed, giving the deviation of each Observation from the mean of its column. In columns 5 and 6 are entered the Squares of the deviations {X^ and F^). The Standard deviation {a^) is now ob- tained by dividing the sum of the Squares in the fifth column by the number of experiments, N, and extracting the square root of the quotient; the Standard deviation for 3; is, of course, found in the same manner. p;ir2 I1642.4 12/ 127078 "^ = A'^ = >i-2F- = 7.65 and ., = ^^=^-g^ = 3i.i * The data are those obtained from experiments on subject A at noon. I9I3] G. 0. Higley 401 The products XY are now collected, the negative in column 7 and the positive in column 8, and the totals determined. We have, then, 2(XF) =2,749.4—1,174.8 = 1,574.6. From this the coefficient of correlation (f ) is obtained : ^xy) 1,574.6 Na.a^- = + 0.236 28 X7.65 X 31. 1 The positive sign of this coefficient indicates, of course, that the relationship between barometric change and carbon dioxide excre- tion in this case is direct or that the two variables change in the same sense. Since a coefficient of correlation of i indicates perfect correlation, the result obtained in the series of experiments repre- sented in Table i indicates a slight degree of correlation. The probable error of a correlation coefficient of this value for a series of 25 observations is at least 0.13 so that the value of r is 0.16 ±0.13. The results of the whole series of experiments are summed up in Table 4. TABLE 4 General Summary Sub- ject Hour xi y2 N Ol 1 ppB oqaov 222 1 1 2 1 1 22 rt E ♦OS'H 222 1 1 d 1 222 ü (3 DH 222 1 1 d i 1 22 V •0 C *0 31E533B tnnipog c^ 12: 1 1 1 1 1 1 1 '(HO)«D d d d 1 1 d d 1 d d CO 0 HO^M dddd |dd2dd 2 ^O^D'^I 1 1 1 1 (^ 1 12 1 1 0 Ö ^ONSh 1 12 1 Miliz ••* *OSn3 1 1 Z 1 i Z II 1 2 5J ^ID^J 1 |Z 1 12 1 1 12 .- ^ (q) 3;e33Dv qd 222 1 i 1 1 1 2a. ■^ s rt 'Ö (u) 3jE5aov qd 222 1 1 2 1 1 22 ^ ■§ W Ö5 'ID^H 222fo i 1 0.22 1 .2 S "ID^O 2^2 1 1 2 1 1 1 1 ID'-HN dd2 1 1 22 1 1 1 C J3 5 ^OD'^'^N d d 1 1 1 d 1 d d 1 *OS''(^HM) dddd Iddldd rt ^ «3 3 *ossk d d d 1 1 d 1 1 d 1 s -4-* .IM ID^N dddd |ddi
0 0 x: w pj f» : : :^3 : NN f^ - 0 0 ö 1 .S ^ 2 » 5 'S ° M M U] Ui W rt a.^ -^-' 0 (-1 (u t; _g 0 et 428 Histon and its Preparation TABLE 5 Data pertaining to the responses of histons to protein color tests [April Nature of product tested Color test Kind Biuret Xanthoproteic MiUon Molisch Adamkiewicz Goose-blood . Thymus Salmin Arbacin Globin Scombron . . . Para-histon. . Gadus Lota NaCl ppt. NaCl and NH4OH ppt. Histon-HCl Histon-H2S04 NH4OH ppt. NaOH ppt. Ale. ppt. NaCl ppt. Histon-HCl P (red)i p p p P (violet) P P P P (violet) P P P P Weak P P P P Weak Weak P P Weak Weak Weak Weak P Ni N N N N P N Weak Weak 6. A summary of characteristic properties of histon (2). Histons are precipitated from aqueous Solutions by ammonia, and are insoluble in excess of the reagent. Bang (2) claims that the presence of salt is necessary for complete precipitation. Huiskamp (10) Claims that the presence of salt hinders the ammonia-precipita- tion of thymus histon. Histons are coagulated by boiling only in the presence of salts. This coagulum is soluble in dilute hydrochloric acid. The protein is not reprecipitated when the acid Solution is neutralized. Histons are precipitated by concentrated nitric acid Solution in the cold. The precipitate dissolves on heating and reappears on cooling. Histons are precipitated by the "alkaloidal reagents" from neutral or weakly alkaline Solutions. Neutral Solutions of histon yield precipitates with neutral Solu- tions of egg albumin and with blood serum ; also with such Solutions of caseinogen and serum albumin, if these are poor in salts. IV. THYMUS HISTON I. The nature of the histon complex. The most exhaustive studies of histon developed from a discussion of the form in which histon occurs in the tissues. Most of the investigation in this con- nection dealt with thymus histon. The researches of Malengrau (20), Lilienfeld (19), Huiskamp (10, 11), and Bang (3, 4, 5, 6), * P = positive ; N = negative. 191 3] Walter H. Eddy 429 bring out the points at issue, which may be briefly summarized as f ollows : Lilien feld first advanced the proposition that a water extract of thymus glands contains nucleohiston, precipitable by acetic acid. From his study of this substance he suggested a Constitution that is indicated by the appended tabular sequence of decomposition products : Thymus nucleohiston I ^1 . Histon Leuconuclein I I . Protein Nuclein I I Protein Nucleic Acid Malengrau (20) repeated Lilienfeld's work and claimed that the latter's precipitate was a mixture of at least two substances. Malengrau called these " nucleoalbumins A and B." He obtalned histon from each. Bang (3) entered the discussion in 1900 and, while agreeing with Malengrau that Lilienfeld's nucleohiston was a mixture, maintained that a water extract of thymus contained at least three substances : Histon, nucleic acid, and a nucleoprotein free from histon. He interpreted Lilienfeld's results as follows: Acetic acid plus water extract of thymus yields a precipitate which is a mixture of histon nucleate and nucleoprotein (Lilienfeld's nucleohiston). This precipitate, in 0.8 per cent. hydrochloric acid Solution, yields histon hydrochlorid plus a mixture of nucleic acid and nucleoprotein (Lilienfeld's leuconuclein). In 1901 Huiskamp (10) stated his belief that Lilienfeld's nucleohiston is a mixture of nucleoprotein and nucleohiston. The latter he separated by adding calcium chlorid to an extract of thymus, to a strength of 0.2 per cent., obtaining a precipitate of calcium nucleohiston and a Solution of nucleoprotein. The former yielded histon hydrochlorid with extraction in 0.8 per cent. hydrochloric acid Solution and, according to Huiskamp, a nuclein but not a nucleic acid as was maintained by Bang. In 1903-4 Bang (6) reported an extensive study of the whole problem. He summarized the results to this point as follows: Huiskamp claimed that thymus contains and yields to water, a 430 H ist 011 and its Preparation [April nudcoprotcin free from histon and a nucleohiston. Malengrau claimed that thymus contains and yields to water, A-micleoalbiimin and B-nudcoalhumin, both containing histon and the latter com- parable to Bang's complex. Bang claimed that thymus contains a nticleo protein, free from histon and extractable by water or 0.9 per Cent, sodium chlorid, and a histon micleate. Commenting generally on these views, Bang held that the thymus gland yields either a multitude of nuclein-containing sub- stances or the nucleoprotein is changed by the various processes in- volved in its Separation, or the dififerent methods of preparation produce mixtures of pure and impiire substances. Bang therefore considered it necessary^ to reinvestigate all methods. In brief his conclusions are : Malengrau's A-nucleoalbumin is an alhuminate, not a histon. It is identical with Bang and Huiskamp's nucleo- protein. Huiskamp's nucleoprotein is identical with Bang's accord- ing to the following percentage analytic data: Analysts C. per Cent. H, per Cent. N, per Cent. P, per Cent. Ash, per Cent. Huiskamp Bang Bang (2d) 50.09 7.18 49-5 6.35 16.II 16.51 15.89 0.97 1.22 0.91 3-11 2.36 2.18 The essential difference, therefore, lies between Bang's histon nucleate theory and Huiskamp's nucleohiston theory. Bang re- ported extensive studies to confirm his point of view. For com- ment on these views see p. 436. V. EXPERIMENTAL I. The cause of the water-insolubility of ammonia-pre- cipitated histon. A. Comparison of ammonia and sodium CHLORiD-PRECiPiTATED HISTON. Many preparations of thymus histon were made involving variations in the process and the use of many different sets of glands from calves. All the glands used were, with one exception, obtained immediately after slaughtering and were extracted as soon as they could be brought to the laboratory. The methods of preparation embraced the following variations : Precipitation [after LiHenfeld (19)] of water extracts with 1913] Walter H. Eddy 43 1 acetic acid and extraction of the purified precipitates with 0.8 per Cent, hydrochloric acid Solution. Precipitation (modification of Lilien feld's process) of water extracts with a few drops of conc. hydrochloric acid Solution and extraction of the purified precipitates with 0.8 per cent. hydrochloric acid Solution. Acidificationof water extracts [after Kossei and Kutscher (16)] with hydrochloric acid, to 0.8 per cent. strength, and filtering. Precipitation of water extracts with alcohol and extraction of the washed precipitates with 0.8 per cent. hydrochloric acid Solution. Precipitation of water extracts [after Huiskamp (10)] with calcium chlorid, to 0.2 per cent. strength, and extraction of the purified calcium chlorid-precipitates with 0.8 per cent. hydrochloric acid Solution. In every case the ammonia-precipitated product from the histoo hydrochlorid Solution was insoluble in water, and addition of am- monia to the acid Solution gave a precipitate which differed in no respects from that obtained by first removing free acid by dialysis and then precipitating with ammonia. The best precipitation re- sults were obtained by first carefully neutralising the acid Solution with ammonia and then adding the excess a few drops at a time, with stirring. It was demonstrated that ammonia-precipitation takes place in the absence of salts (contrary to Bang's contention), but care must be taken to add the ammonia slowlv and in small amounts. The insolubility of the ammonia-precipitated product was shown to be due neither to the action of the alcohol or ether used in wash- ing the product, nor to abnormalities in the glands used. It was found to increase with the length of the period in distilled water. Precipitates allowed to stand for several weeks in distilled water, with toluene, failed to dissolve or putrefy and ultimately became completely insoluble in 0.8 per cent. hydrochloric acid Solution. In contrast with these results, the histon obtained by saturating a histon-hydrochlorid Solution with sodium chlorid was invariably soluble in water. This solubility was not altered by washing with alcohol and ether, by drying at 45°, or even when the product was dried to constant weight at 105° C; the sodium chlorid-saturation products from both add and neutral histon-hydrochlorid Solutions 43 2 Histon and its Preparation [April were the sanie in this respect. Furthermore the precipitate obtained with sodium chlorid was always readily soluble in 0.8 per cent. hydrochloric acid Solution. The above mentioned results suggested different constitutions for histons precipitated by sodium chlorid and by ammonia. To detemiine this point pure preparations of each kind were made and analysed. B. Methods of preparation and purification. Histon hydrochlorid was obtained by the Kossel-Kutscher (16) method. A portion of the clear bluish filtrate containing it was saturated with sodium chlorid and filtered, and the resulting precipitate washed with alcohol, dissolved in water and the Solution dialyzed in a parch- ment bag for several days against running water. At the end of that time the bag was transferred to a tall jar of distilled water and the latter renewed daily until no trace of chlorin could be detected in the histon Solution with silver nitrate. The Solution was then removed from the bag, filtered and evaporated in thin layers to dryness at 45° C. Light yellow flakes were obtained. (During the dialysis and evaporation toluene was used as a preservative. ) The flakes were ground to a white powder and dried to constant weight — first in vacuo over sulfuric acid and finally at 105° C. A second portion of the histon-hydrochlorid Solution was treated with am- monia and the resulting precipitate washed free from all traces of ammonia with water. It was then washed with alcohol and ether, and dried to constant weight at 105° C. A third histon product was obtained by adding a few drops of ammonium hydroxid Solu- tion to a portion of the chlorin-free Solution of the sodium chlorid- precipitate. The precipitate thus obtained was insoluble in water. It was washed with water, alcohol and ether, and dried to constant weight at 105° C. These three preparations^ were used in the quantitative studies. In the tabulation on page 434 these preparations are designated as follows: (a) sodium chlorid-precipitated histon, free from chlo- rid admixture; (b) sodium chlorid-precipitated histon, reprecipi- tated with ammonia; (c) ammonia-precipitated histon. *Inability to find a solvent for ammonia-precipitated histon which would not hydrolyze it made it impossible to attempt the preparation of a sodium chlorid-saturation precipitate of the ammonia-histon. I9I3] Walter H. Eddy 433 C. Quantitative procedure. The methods employed in the quantitative analysis were the following: Total nitrogen was deter- mined by the Kjeldahl process. For the chlorin detemiination 0.15 gm. of the material was placed in a casserole with 100 c.c. of water, 30 c.c. of nitric acid sohition and 10 c.c. of n/20 silver nitrate Solu- tion (containing 1.52 mg. of chlorin per c.c). The liquid was boiled gently for two hours to effect complete decomposition. The Solution remained pale yellow, with silver chlorid at the bottom of the casserole. After cooling, the liquid was filtered and the silver chlorid thoroughly washed. To the filtrate and washings, i c.c. of ferric alum Solution was added, the mixture decolorized with nitric acid, and titrated with n/20 potassium sulfocyanate Solution of such strength that i c.c. equalled i c.c. of silver nitrate Solution. The ash was obtained by cautiously incinerating 0.2 gm. of the dry sample in a small porcelain crucible until all carbonaceous matter disappeared. Six hours was usually sufficient. The crucible was then cooled in a desiccator and weighed. The moisture was deter- mined by heating 0.5 gm. of the sample in a weighing bottle at 105° C. for 24 hours. All samples had been previously dried at 45° C. and powdered. The amide nitrogen, diamino nitrogen, monamino nitrogen and humin nitrogen were determined by the Osborne-Harris method.^ D. Quantitative results. The quantitative data in these experiments are summarized in Tables 6 and 7. E. DiscussiON OF the quantitative results. It will be seen that the lower nitrogen content of the sodium chlorid-precipitated histon is not accounted for by the ash difference (sodium chlorid- product averaged 1.33 per cent. and the ammonia-precipitated prod- uct, 0.76 per cent). This slight difference, combined with the Obser- vation that ammonia separates from an aqueous Solution of the sodium chlorid-precipitated product, a water-insoluble mass of higher nitrogen content with a filtrate that responds to the biuret test, suggests the presence (in the salt-precipitated product) of a protein fraction which is absent from the ammonia-precipitated product and contains relatively little nitrogen. The differences between the monamino and diamino fractions tend to confirm this * Osborne, T. B., and Harris, J. F., Joitr. of the Amer. Chem. Soc., 1903, XXV, p. 323. 434 Histon and its Preparation [April vievv. The data for the amide fractions also tend to show that the difference in nitrogen content cannot be due to combination of the ammonia nitrogen vvith the ammonia-precipitated histon. TABLE 6 Data pertaining to percentage elementary composition Preparation Histon A. Sodium chlorid-precipitated product, free from chlorid admixture Histon B. Sodium chlorid-precipitated producl, reprecipitated with ammonia Histon C. Ammonia-precipitated product N,l per Cl, per Cent. Cent. Ash, per Cent. N,i per Cent. Cl, per Cent. Ash, per Cent. N,i per Cent. Cl, per Ash, per Cent. Cent. I 16.05 0.52 — 16.85 0.00 — 17.47 0.00 — 15.87 0.56 — 16.84 — — 17.61 0.00 — 2 15.96 0.62 1.46 16.86 0.00 0.78 17.37 — 0.84 16.09 0.52 1.20 16.70 0.00 0.88 17.29 0.72 3 16.52 — 16.75 — — 17.20 — — 16.47 — 16.86 — — 17.16 — — TABLE 7 Data in duplicate pertaining to nitrogen partition Histon A Histon B Histon C Preparation I I I 2 I 2 Material taken (gm.).. Amide N 0.8035 % 1.04 1.03 13.14 0.87 16.08 0.6742 % 1.30 0.91 12.28 1.03 15.52 O.6111 % I.Ol 1.03 13.46 I.I9 16.69 0.4995 % 0.70 1.12 13.54 2.01 17.37 O.5112 % 0.99 I.18 13.61 X.42 17.20 0.4327 % 1.02 Humin N 0.99 14.17 I.19 17.37 Diamino N Monamino N Total Aside from the difference in nitrogen content the only other striking difference shown by these results is the presence of chlorin in the sodium chlorid-precipitated product and its absence from the ammonia-precipitated substance. That this is not due to failure to sufficiently purify the sodium chlorid-precipitated material was shown qualitatively in the f ollowing experiments : A portion of the water-solution of the sodium chlorid-precipi- tated product was treated with a few drops of ammonium hydroxid. ^ Nitrogen calculations on an ash-f ree basis — Histon A, preparation 2 : 16.20 per Cent, and 16.28 per cent. ; Histon B, preparation 2: 17.00 per cent. and 16.88 per cent. ; Histon C, preparation 2 : 17.52 per cent. and 17.41 per cent. I9I3] Wolter H. Eddy 435 The clear filtrate from the resultant precipitate, after acidification with nitric acid, gave a good chlorin test with silver nitrate. It therefore seems correct to assume that ammonia actually separates chlorin from the sodium chlorid-precipitated siibstance and that chlorin is present in the latter histon product in an adsorbed or com- bined condition, not as an impurity. A further investigation of the validity of these conclusions is in progress. F. Conclusions. The results of this vvork justify the follow- ing conclusions: Both sodium chlorid and ammonia precipitate, from histon-hydrochlorid Solutions, products having characteristic histon properties but differing in water-solubility, and in nitrogen and chlorin contents. The filtrates from both the ammonia- and sodium chlorid-precipitates give strong biuret reactions and are precipitable by alcohol-ether and Saturation with ammonium sulfate. The latter precipitates are not identical in properties and, while parahiston is possibly admixed with them, they also contain frac- tions of other protein matter which makes them different from para-histon and from one another. At present the nature of this protein admixture, and hence the exact nature of the difference between sodium chlorid- and ammonia-precipitated histons, is still uncc. vain. 2. An improvement in the method o£ preparing thymus histon. In following out the various methods of preparing the hydrochlorid Solution of thymus histon it was difficult to obtain a water extract of the glands which would filter clear. The glands are also particularly prone to putrefaction at the beginning of the extraction process. The latter difficulty Avas overcome by conducting the extractions in a refrigerator rather than by depending upon the use of preserv'atives such as Chloroform and toluene. Both difh- culties can however be avoided by the following modified method which is recommended as more practical than any of those out- lined in the general literature. After freeing the glands from fat with a knife, pass them through an ordinary meat grinder and pour the hash directly into a comparatively large volume of 95 per cent. alcohol. All the protein material is precipitated and all chance of putrefaction is thus 43 6 H'iston and its Prcparation [April avoided. Filter off the alcohol and extract the precipitated material directly with 0.8 per cent. hydrochloric acid Solution. A clear, bluish white, readily filterable, extract will result. Extraction with hydrochloric acid Solution should be continued for several days. In our experiments it was found necessary to repeat the extrac- tion with hydrochloric acid Solution several times to obtain all the histon, Preliminary extraction of the glands with alcohol removes none of the histon (as was detemiined by study of the extract) and, by precipitating all the proteins in the glands, the alcohol prevents the presence of protein impurities in the acid extract. The histon can then be precipitated from the acid extract by whatever method is desired, as already outlined. 3. Proteins left after removal of histon from thymus extract. Some experiments were conducted to detemiine if possible the merits of the contentions of Lilien feld, Huiskamp and Bang as to the nature of the histon complex in the cell. These views were outlined in detail on page 428, The procedure was as f ollows : Thymus glands were minced and extracted with water for 48 hours in a refrigerator. The extract was decanted, filtered, precipitated with a few drops of conc. hydrochloric acid Solution and the super- natant fluid treated with hydrochloric acid until a strength of 0.8 per cent. was present; after standing for several days, the acid Solution was filtered off and the precipitate extracted with 0.8 per cent. hydrochloric acid Solution. This process was repeated until the hydrochloric acid extract failed to give an ammonia-precipitate or a biuret reaction. The histon-f ree residue was then washed with alcohol until it was free from acid, treated with 0.3 per cent. potas- sium hydroxid Solution, in which it dissolved slowly but com- pletely, and the Solution filtered, placed in a parchment bag and dialysed free from hydroxyl ions. No precipitate resulted, the Solu- tion remaining clear. (Toluene was used as a preservative during this process.) To the neutral Solution 10 per cent. calcium chlorid Solution was now added (2 c.c. of 10 per cent. calcium chlorid Solution to each 100 c.c. of liquid). A copious precipitate resulted, the filtrate from this mass giving a good biuret test. Addition of more calcium chlorid to the filtrate failed to produce further precipitation. I9I3] Walter H. Eddy 437 The precipitate was next completely dissolved in water with the addition of a little 10 per cent. potassium hydroxid Solution and the liquid again dialyzed free from hydroxyl ions in a parch- ment bag. This neutral Solution was then reprecipitated with calcium chlorid as before, yielding again a sharply separating pre- cipitate and a clear filtrate which gave neither biuret reaction nor alcohol precipitate. The precipitate was then washed with alcohol, and ether, and dried to constant weight at 105° C. A Solution in dilute potassium hydroxid Solution, dialysed free from hydroxyl ions, gave characteristic protein tests. The dried product was then analysed for nitrogen, ash and calcium. The filtrate from the calcium chlorid-precipitated product was treated with a few drops of conc. hydrochloric acid Solution, yield- ing a copious precipitate and a water-clear filtrate, which gave no biuret test or alcohol precipitate. The precipitate was purified by Solution in 0.3 per cent. potassium hydroxid Solution and reprecipi- tation with hydrochloric acid; washing with water, alcohol, and ether, and finally drying to constant weight at 105° C. The dry product was analyzed for total nitrogen and ash. Both precipitates were rieh in phosphorus. A. B. Product precipitated Product precipitated with hydrochloric acid with calcium chlorid, in the filtrate from A Per Cent. Per Cent. Total N 7.65 15.97 Ash 7.30 5.96 Calcium None Not determined These results agree with Huiskamp's views in certain respects. Assuming that water extracts iboth a nucleoprotein and a nucleo- histon, treatment of the product with hydrochloric acid may be assumed to separate the histon from the nucleohiston, and leave a mixed residue of histon-free nucleoprotein and a nuclein. Both of these substances dissolve in 0.3 per cent. potassium hydroxid Solu- tion, but the latter is precipitable by calcium chlorid while the former is not. Furthermore, the view that calcium chlorid precipi- tates a nuclein rather than nucleic acid is borne out by the protein reactions of the product precipitated by calcium chlorid. Huis- kamp, however, claims that his nuclein fonns a calcium salt with 43 S Histon and its Preparation [April calcium chlorid and comes down as calciuni-nucleinate. In our analyses we were unable to detect even a trace of calcium in the ash. It is possible, of course, that a nucleic acid fraction was sepa- rated from the histon product by the acid treatment, which, in the presence of other protein, may have formed a protein-nucleate pre- cipitable by calcium chlorid. It might be true therefore, as Bang (6) Claims, that histon exists in the cell as a histon-nucleate. The failure to obtain any nucleic acid in these tests, and the different nitrogen figures for the product precipitated with calcium chlorid, together with the agreement between the values for nitrogen con- tent of the nucleoprotein and those obtained by both Bang and Huiskamp, suggest that histon occurs in cells as a true nucleo- histon and not as a nucleate. VI. SUMMARY OF CONCLUSIONS These studies seem to show that histon obtained from aqueous extract of thymus gland by precipitation with ammonia is essentially dififerent from histon obtained by Saturation with sodium chlorid. The histon precipitated by ammonia is insoluble in water and has a higher nitrogen content than the histon precipitated with sodium chlorid. The latter product is readily soluble in water, even after drying at 105° C, and contains an appreciable amount of chlorin in combined form. The difference relates apparently both to the con- tent of chlorin and to the presence of a protein fraction in the salt- precipitated histon which is absent from that obtained with ammonia. The preliminaiy use of alcohol to precipitate histon and other protein materials in the glands seems to offer marked advantages over the direct water-extraction method, both in öbviating putre- faction and in facilitating filtration of the aqueous extract. Bang's contention that ammonia does not precipitate histon in the absence of salts is incorrect. It is true, however, that their pres- ence facilitates the process. The data pertaining to the residues insoluble in 0.8 per cent. hydrochloric acid Solution seem to confirm Huiskamp's views rather than those of Bang, but in view of the power of nucleic acid to combine with protein and the absence of calcium from the ash of the calcium chlorid-precipitated fraction, the matter cannot be regarded as settled. iQi 3l Walter H. Eddy 439 Histon prepared by the salt-precipitation method offers excel- lent material for the study of protein-salt formation. The necessity of knovving the method o£ preparation of a given histon prodiict, in order to understand the properties ascri'bed to it, is obvious from these sttidies. The quantitative data on page 434 were obtained with the assist- ance of Mr. E. G. Griffin and those on page 437 with the aid of Mr. W. F. Hume, to whom I am greatly indebted for the effective Cooperation they have given. I wish also to acknowledge my indebt- edness to Dr. Gies for facilities and suggestions throughout the entire research. VII. SELECTED BIBLIOGRAPHY OF HISTON STUDIES 1. Ackerman, D. Zur Chemie der Vogelblutkerne. Zeitschr. f. phys- iol. Ch., 1904, xliii, p. 299. Gives Plenge's method of preparing histon from hen blood. 2. Bang, Ivar. Studien ueber Histon. Z&frf.^ 1899, xxvii,p. 463. First report of the discovery and preparation of scombron, a histon in mackerei sperm. Also presents comparisons with previously dis- covered histons, a discussion of the ammonia reaction, and first list of the " five characteristic properties " of histon. 3. . Bemerkungen ueber Nucleohiston. Ihid., 1900, xxx, p. 508. Raises the question as to the nature of Lilienfeld's nucleohiston. 4. . Reply to Kossei. Ibid., 1901, xxxi, p. 407. 5. . Reply to Kossei. Ibid., 1901, xxxii, p. 79. 6. . Chemische Untersuchungen der lymphatischen Organe. Bei- träge mir Chem. Physiol. u. PathoL, 1904, v, p. 317; iv, pp. 115, 351. Füll report of Bang's experiments, with review of the work of Malengrau and Huiskamp. 7. Eddy, W. H. A study of some protein Compounds. Biochemical Bulletin, 1912, ii, p. iii. Reported attempts to form protein salts with histon and difficulties in preparing acid-free histon. 8. Ehrström, Robert. Ueber ein neues Histon aus Fischsperma. Zeitschr. f. physiol. Ch., 1901, xxxii, p. 350. Preparation of Lota histon from the sperm of the burbot. 9. Fleroff , A. Ueber einen histonähnlichen Körper aus Thymus. Ibid., 1899, xxviii, p. 307. Discovery and preparation of para-histon from thymus ; comment on water-insolubility of ammonia-precipi- tated histon. 440 Histon and its Preparation [April 10. Huiskamp, W. Ueber die Eiweisskörper der Thymus. Ibid., igoi, xxxii, p. 145. Review of Bang's and Malengrau's claims, and introduction of calcium chlorid as a means of separating nucleo- histon from nucleoprotein. Maintains that nucleohiston is true nucleoprotein and that it yields nuclein, not nucleic acid, with re- moval of histon. 11. . Beiträge zur Kenntniss des Thymus Nucleohistons. Ibid., 1903, xxxix, p. 55. Further study of nucleohiston. 12. Kossei, A. Ueber einen peptonartigen Bestandtheil des Zellkerns. Ibid., 1884, viii, p. 511. Describes the preparation of the first his- ton : from gooseblood. 13. . (Comment on Bang's paper.) Ibid., 1900, xxx, p. 520. 14. . (Reply to Bang.) Ibid., 1901, xxxii, p. 81. 15. . Ueber ein einfachsten Eiweisskörper. Biochem. Central- hlatt, 1906-7, V, p. 35. Reports the preparation of Centrophorus and Spcorechinns histon, but gives no details. 16. Kossei, A,, and Kutscher, F. Beiträge zur Kenntniss der Eiweiss- körper. Zeitschr. f. physiol. Ch., 1900, xxxi, p. 165. A study of cleavage products of proteins and also a report of the preparation of gadus histon from cod. 17. Kossei, A., and Pringle, H. Ueber das Histon. Ibid., 1906, xlix, p. 314. Quantitative studies. 18. Lawrow, D. Ueber die Spaltungsprodukte des Histons von Leu- kocyten. Ibid., 1899, xxviii, p. 388. A modification of the method of preparing thymus histon and a study of its products. 19. Lilienfeld, Leon. Zur Chemie der Leukocyten. /&ic?., 1893, xviii, p. 473. Description of the first preparation of histon from thymus. 20. Malengrau, F. La Cellule. 1900, xvii, p. 19. A study of the nucleohiston of thymus and Separation of an A- and B-nucleo- albumin. 21. Mathews, Albert P. Zur Chemie der Spermatozoon. Zeitschr. f. physiol. Ch., 1897, xxxiii, p. 399. Preparation of arbacin histon from sea urchin sperm. 22. Miescher, F., and Schmiedeberg, O. Physiologisch-chemische Untersuchungen ueber die Lachsmilch. Archiv, f. ex per. Pathol. u. Pharmakol., 1896, xxvii, p. 100. Preparation of an albumose or histon from unripe salmon sperm. 23. Schulz, F. N. Der Eiweisskörper des Haemoglobins. Zeitschr. f. physiol. Ch., 1898, xxxiv, p. 449. Preparation of globin from horse blood. DID VON WITTICH ANTEDATE OSTWALD IN THE DEFINITION OF ENZYME ACTION? WILLIAM N. BERG It seems to be the consensus of opinion that enzyme action was first properly understood and defined by Wilhelm Ostwald about 1893. That this is a reasonable inference is evident from the following typical quotations from the literature of the subject: The word catalysis was introduced about eighty years ago by Ber- zelius. It grouped together phenomena that had up to that time re- mained unconnected. But while a great and increasing number of new catalytic phenomena were discovered in subsequent years, the concept itself remained vague until Ostwald introduced his well known defini- tion based on the conception of velocity of chemical change — a con- ception which was born with modern chemical kinetics and was un- known to Berzelius. According to Ostwald, a catalyzer is a substance whose presence hastens a given chemical reaction, although the reaction would also take place in its absence. . . .^ Wir werden also Kontaktwirkungen der Katalysatoren und der En- zyme am besten unter die von Ostwald gegebene (1893) Definition bringen können: 'Katalyse ist die Beschleunigung eines langsam ver- laufenden chemischen Vorganges durch die Gegenwart eines fremden Stoffes/^ Ein wirklicher Inhalt ist diesem Begriff erst von Ostwald gegeben worden. Er bezeichnete als Katalysator 'jeden Stoff, der, ohne im Endprodukt einer chemischen Reaktion su erscheinen, ihre Geschzvin- digkeit verändert' . . . (S. 16), Es ist das anzuerkennende Verdienst W. Ostwalds, eine der indessen ausgebildeten Thermodynamik gerecht werdende Definition der Begriff Katalyse und Katalysator gegeben zu haben.^ ^Rosanoff: "Outline of a theory of homogeneous catalysis," Jour. Amer. Chem. Soc, 35, p. 173, I9i3- *Bredig: "Die Elemente der chemischen Kinetic, mit besonderer Berück- sichtigung der Katalyse und der Fermentwirkung," Ergehnisse d. Physiologie, i, p. 139, 1902. * Oppenheimer : Die Fermente und ihre Wirkungen, 3 Aufl., p. 159; Leipzig, 1910. 441 442 The Definition of Enzyme Action [April Partly on accoimt of their obscure places of publication, and partly on account of fanlty references in the literature, Ostwald's original definitions were f ound with some difficulty : Katalyse ist die Beschleunigung eines laugsam verlaufenden chem- ischen Vorganges durch die Gegenivart eines fremden Stoffes. . . .* Man nennt die Stoffe, welche solche Änderungen der Geschwindig- keit bewirken, Katalysatoren, und zwar positive und negative, je nach- dem sie Beschleunigungen oder Verzögerungen hervorbringen. Der Begriff der Katalysatoren hat erst in neuerer Zeit diese bestimmte Defi- nition erfahren (Ostwald 1894). . . .^ Since Ostwald himself dates the definition of catalysis at 1894, it would naturally be assumed that that date is the correct one. While engaged, about five years ago, in a study of enzyme ac- tion, the attention of the writer was drawn to a paper publisht by von Wittich,*^ not alone because of the ingenuity of the experi- mental work described therein, but especially because of von Wittich's striking conclusion (p. 469) regarding the nature of peptolytic action: Es bleibt daher nichts anderes übrig, als anzunehmen, dass die Säure allein hinreicht, um jene bekannte Umwandlung des Fibrins ein- zuleiten, dass aber die Gegenwart des Pepsins letztere wesentlich beschleunigt. The "Umwandlung" referred to is the transformation of fibrin into peptone. von Wittich had observed that fibrin was slowly transformed into peptones by hydrochloric acid and that this transformation took place much more rapidly in the presence of pepsin. Other investi- gators had made the same Observation, but without correctly under- standing the role of the enzyme, as von Wittich apparently did. These facts were mentioned about four years ago by the writer, who believed that von Wittich had antedated Ostzvald in the defini- tion of catalysis. Until recently, lack of opportunity prevented a sußiciently careful search of the literature on this particular point. * Ostwald : Ztschr. f. physikal. Chemie, 15, p. 706, 1894. 'Ostwald: Grundriss der allgemeinen Chemie, 3 Aufl., p. 515; Leipzig, 1899. ' von Wittich : " Weitere Mittheilungen über Verdauungsfermente," Archiv f. d. gesammte Physiologie, 5, p. 435-469, 1872. 191 3] William N, Berg 443 It was for this reason that the name of Ostwald was held in reserve when the following Statement was made : A rather early insight into the nature of peptolysis was given by von Wittich, who concluded that the pepsin simply accelerates a reac- tion which the acid alone will bring about more slowly/ In his conclusions, von Wittich plainly states that the transforma- tion of fibrin into peptone by pepsin-hydrochloric acid is a reaction similar to the transformation of alcohol into ether by sulfuric acid. This is evidence of the fact that when he defined peptolytic action, he did so not unknowingly, but with a füll understanding of the facts. It is reasonable to suppose that von Wittich had a broad, general understanding of enzyme action as the basis for his defini- tion, because he had previously publisht work on diastatic, proteo- lytic, glucosid-splitting and various other enzymes.^ In one of these papers (1870, p. 352) he mentions the " katalytische Wirk- samkeit" of a diastase in such a way as to leave no doubt that he understood enzyme action to be catalytic. Two years later (1872, p. 465) he stated his belief in the similarity between the action of pepsin in peptic digestion and the action of sulfuric acid in the transformation of alcohol into ether, in the following terms : Der Vorgang erscheint mir dem ganz analog bei der Ueberführung des Alcohol durch Schwefelsäure in Aether, Wie sich hier zunächst Aethylschwefelsäure bildet, wie diese bei 140° C. auf ein zweites Mole- cul Alcohol wirkt und dasselbe in Aether und Wasser zerlegt, wie die sich bildenden Wasser und Aether überdestilliren und die Schwefel- säure in ungeschwächter Wirksamkeit zurücklassen, so dass sie immer neuzufliessenden Mengen Alcohol in denselben Verhältnissen in Aether und Wasser zu spalten vermag, so bildet sich bei der Pepsinwirkung zunächst dessen Verbindung mit der freien Säure, die ihrerseits durch Contact das Fibrin in jene leicht lösliche und diffusible Form die Pep- tone umwandelt. Wie aber bei allen Contactwirkungen die Mengen der durch sie gebildeten Stoffe von der Grösse der Contactfläche abhängt, so bedingt auch hier die letztere, d. h., die Menge des ver- wendeten Pepsins die Menge der entstehenden Peptone. Wie aber die 'Berg: "A comparative study of the digestibility of different proteins in pepsin-acid Solutions," Atner. Jour. of Physiology, 23, p. 423, 1909. * von Wittich : " Ueber eine neue Methode zur Darstellung künstlicher Verdauungsflüssigkeiten," Archiv f. d. gesammte Physiologie, 2, p. 193, 1869. Also, Weitere Mittheilungen über Verdauungsfermente, Ibid., 3, p. 339, 1870. 444 The Definition of Enzyme Action [April Schwefelsäure keine dauernde Verbindung mit dem Aether eingeht, kein integrirender Theil des sich neubildenden Stoffes wird, wie sie unter günstigen Bedingungen sich von jenem wieder trennt, um immer neue Mengen Alcohols in gleicher Art zu verändern, so geht auch bei dem Verdauungsvorgang weder die Salzsäure noch das Pepsin in eine bleibende Verbindung mit der Peptone ein und entfernt man letztere, so vermag dieselbe Menge der Säure dieselbe Menge Pepsin, immer neues Fibrin in ähnlicher Weise zu zerlegen. Man kann die Peptone von dem Pepsin und den noch vorhandenen Parapeptonen scheiden, , . . On the third page after this paragraph, von Wittich concludes his paper (1872, p. 469) — füll of interesting and suggestive ex- periments — with the Statement: Es bleibt daher nichts anderes übrig, als anzunehmen, dass die Säure allein hinreicht^ um jene bekannte Umwandlung des Fibrins ein- zuleiten, dass aber die Gegenwart des Pepsins letztere zvesentlich be- schleunigt. In view of the fact that von Wittich was probably the first to correctly define enzyme action, it seems stränge that his works should have received so little attention. Several of his publications are indext and very briefly discust in Oppenheimer's^ book, but the important points in the above quotations from von Wittich are not mentioned, Cohnheim^° and Höber^^ both mention von Wittich once, in a few words, the former to the effect that von Wittich was one of the early investigators who prepared glycerol extracts of gastric mucosa, etc. ; the latter mentioned von Wittich's name on a subject other than enzyme action! In short, a reference to the fact that von Wittich defined enzyme action in 1872, twenty or more years before Ostwald, could not be found, altho it was looked for in many places besides those mentioned above. von Wittich was probably the first to show that when fibrin is immersed in pepsinogen-glycerin Solutions (1872, p. 443), or in pepsin-hydrochloric acid Solutions (1872, p. 444), the enzyme is rapidly adsorbed by the fibrin. He used the term pepsin for both pepsinogen and pepsin. He also showed, most ingeniously, that while pepsinogen would not dififuse from a glycerol extract thru a Graham dialyzer into ' Oppenheimer : Loc. cit. " Cohnheim : Enzymes, p. 2, New York, 1912. "Höber: Physikalische Chemie der Zelle und der Gewebe, 3 Aufl., p. 537, Leipzig, 191 1, 1913] William N. Berg 445 water, it would pass thru, if fibrin were placed in the water. He explains this as follows (1872, p. 443) : Der Vorgang erklärt sich, wie ich glaube, durch die Annahme, dass das Fibrin das Pepsin sehr energisch absorhirt, dass auch beim Fehlen jenes minimale Mengen diffundiren, die Diffusion aber durch die Ab- sorptionsfähigkeit des Fibrins beschleunigt und verstärkt wird. It is quite possible that the principle of this experiment might have a wide application, i. e., that the diffusibility of many sub- stances (now regarded as indiffusible) might be greatly increased by placing material in the diffusion medium which was not soluble in the medium but which could combine with the substance whose diffusibility was under investigation. The above mentioned observations by von Wittich, on the diffusibility and absorption of pepsin by fibrin, have recently been used by several investigators. Beginning with the work of Abder- halden and Steinbeck,^^ Abderhalden and his co-workers have publisht a long series of researches on the adsorption of enzymes by proteins. In none of these publications that the writer has seen is there any allusion to the fact that the principle involved was not new. The name of von Wittich was not mentioned. From their papers one might justly infer that Abderhalden and Steinbeck be- lieved that they had discovered the adsorption of pepsin by pro- teins, when they were, in fact, using a principle discovered in 1872 by von Wittich. To a lesser extent this criticism applies to Hedin, ^^ in whose several publications von Wittich is not mentioned, altho many of the experiments and conclusions of Abderhalden and his co-workers, and of Hedin, can be found in von Wittich's paper. In a very interesting manner Chodschajew^^ discusses the work of von Wittich on the diffusibility of pepsin, some of which Chodschajew repeated. Dauwe^^ mentions von Wittich as the first to observe the adsorption of pepsin by fibrin. Washington, D. C. "■^ Abderhalden and Steinbeck: "Beitrag zur Kenntnis des Pepsins und der Salzsäure," Ztschr. f. physiolog. Chemie, 68, p. 293, 1910. ^ Hedin : " Observations on the action of trypsin," Joiir. of Physiol., 32, p. 468, 1905 ; Biochemical Journal, 2, p. 81, 1907. "Chodschajew: " Les enzymes sont-elles dialysables ? " Archives de Physiol- ogie normale et pathologique, 1898, p. 241. " Dauwe : "Ueber die Absorption der Fermente durch Kolloide," Beiträge zur chemischen Physiologie und Pathologie, 6, p. 427, 1905. THE BIOCHEMICAL SOCIETY, ENGLAND SCIENTIFIC PROCEEDINGS Research Institute of the Cancer Hospital, Brompton RoAD, London, S. W., Fchruary 5, 191 3, at 5 p. m. — S. B. Schryver: Notes on bile acids; Investigations 011 phenomena of clot formations: (a). clotting of calcium cholate, (b) clotting of milk. — G. Barger and A. J. Ezvins: The identity of trimethylhisti- dine {histidine betaine) from various sources. Chemical Department of the London Hospital Medical College, March iß, 1913, at 5 p. m. — W. H. Hnrtley: The old and a new test for aceto-acetic acid. — /. H. Ryffel: A sensitive modifica- tion of Gmelin's test for bile pigment in iirine. — IV. M. Bayliss: The combination of amino acids with neutral salts. — R. H. A. Flimmer: The Separation of cystine and tyrosine. Physiological Laboratory, Cambridge, May 10, 1913, at 4 p. m. — C. Singer and S. B. Schryver: Some investigations on the gastric juice. — E. Graf von Schönhein and S. B. Schryver: Some properties of the bile salts. — S. Walpole: On the use of litmus paper as a quantitative indicator of reaction. — C. Funk: A com- plete chemical and physiological investigation of the vitamines from rice polishings and yeast. — F. W. Foreman: Esterification of amino acids from proteins. — A. Neville: The fatty acids of yeast. — A. Neville and E. T. Halman: Feeding experiments with Bastol. — C. G. L. Wolf: A note on the estimation of lactic acid. OFFICERS, 1913-14 Committee:^ Hon. Treasurer, J. A. Gardner; Hon. Secretary, R. H. A. Flimmer; Editors of the Biochemical Journal, W. M. Bayliss and A. Harden; Ordinary Memhers, G. Barger, A. Chaston Chapman, J. S. Ford, W. D. Halliburton, F. G. Hopkins, W. H. ^ The business of the Society is conducted by a Committee consisting of a treasurer, a secretary, the editors of the Biochemical Journal and twelve ordi- nary members. 446 1913] List of Memhers 447 Hurtley, F. Keeble, B. Moore, W. Ramsden, E. J. Russell, J. Lor- rain Smith and T. B. Wood. PROVISIONAL SCHEDULE OF MEETINGS, 1913-14 May 10 — Physiological Laboratory, Cambridge. June II — Institute of Physiology, University College, London. July 12 — Rothamsted Experimental Station, Harpenden. Oct. 10 — Pathological Department, St Thomas's Hospital, London. Nov. 13 — Physiological Laboratory, King's College, London. Dec. 9 — Lister Listitute, London. Feb. II. — Guy's Hospital, London. March 12 — Botany Department, Lnperial College of Science, London, S. W. LIST OF MEMBERS' AcKROYD, Harold, Esq., Great Shelford, Cambridgeshire. Armstrong, Dr. E. F., 27 Lastern Avenue, Reading. Annett, H. E., Esq., B.Sc, The Heath, Walton-on-Thames, Surrey. Bainbridge, Prof. F. A., Durham College of Medicine, Newcastle-on- Tyne. Barger, Dr. G., 107 Tyrwhitt Read, St. John's, London, S.E. Bateson, W., Esq., F.R.S., The Manor House, Merton, Surrey. Bayliss, Prof. W. M., F.R.S.,.St. Cuthbert's, Hampstead Heath, Lon- don, N.W. Blackman, Dr. F. F., F.R.S., St. John's College, Cambridge. Blackman, Prof. V. H., F.R.S., Imperial College of Science and Technology, South Kensington, London, S.W. Brown, Prof. Adrian J., F.R.S., West Heath House, Northfield, near Birmingham. Brown, Dr. David, 13 Clarence Drive, Harrogate. Brown, Dr. Horace T., F.R.S., 52 Nevern Square, Kensington, Lond. Buckmaster, Dr. G. A., 113 Haverstock Hill, Hampstead, London. Bywaters, Dr. H. W., The University, Bristol. Cameron, A. T., Esq., M.A., University of Manitoba, Winnipeg, Can. Candy, Hugh C. H., Esq., 6 Gordon Square, London, W.C. Cathcart, Dr. E. P., i Bruce Street, Hillhead, Glasgow. Chapman, A. Chaston, Esq., F.I.C., 8 Duke Street, Aldgate, E.C. ^A copy of the ofificial register published after the " annual general meeting" on March 13, 1913. 448 The Biochemical Society, England [April Chapman, Dr. H. G., The University, Sydney, New South Wales. Chick, Miss Harriette, D.Sc, Lister Institute, Chelsea Gardens, London, S.W. Close, Col. J. K., I.M.S., Allahabad U.P., India. CooPER, E. A., Esq., B.Sc, Arborfield, Woodcote Valley Road, Purley, Surrey. Gramer, Dr. W., The University, Edinburgh. Growther, Dr. C, The University, Leeds. CusHNY, Prof. A. R., F.R.S., 8 Upper Park Road, Hampstead, Lond. Dakin, Dr. H. D., 819 Madison Avenue, New York Gity. Dale, Dr. H. H., 140 Thurlow Park Road, Dulwich, London, S.E. Davis, W. A., Esq., 7 Garlton Bank, Harpenden, Herts. Dean, Prof. H. R., The University, Sheffield. Dewitz, Dr. J., yy Lorryerstrasse, Devant-les-Ponts, Metz, Germany. DixoN, Prof. W. E., F.R.S., Museums, Gambridge. Doree, Dr. G., 58 Gore Road, S. Hackney, London, N.E. Douglas, Dr. G. Gordon, St. John's Gollege, Oxford. Dyer, Dr. Bernard, 17 Great Tower Street, London, E.G. Ellis, G. W., Esq., University of London, South Kensington, S.W. Evans, Dr. G. Lovatt, University Gollege, London, W.G. EwiNS, A. J., Esq., B.Sc, 144 Ribblesdale Road, Streatham, London. Eyre, Dr. J. V., South Lastern Agricultural Gollege, Wye, Kent. Fletcher, W. M., Esq., Trinity Gollege, Gambridge. Ford, J. S., Esq., Abbey Brewery, Edinburgh. Foreman, f. W., Esq., School of Agriculture, Gambridge. Fowler, Dr. Gilbert J., Rufford, 143 Dickinson Road, Rusholme, Manchester. Frankland, Prof. Percy F., F.R.S., University, Edgbaston, Bir- mingham. Funk, Dr. Gasimir, Lister Institute, Ghelsea Gardens, London, S.W. Gardner, J. A., Esq., M.A., 26 Horbury Grescent, Notting Hill Gate, London, W. Garrod, Dr. A. E., F.R.S., 9 Ghandos Street, Gavendish Square, Lond. GoADBY, Dr. K. W., 46 Harley Street, London, W. GoLDiNG, John, Esq., University Gollege, Reading. GoLLA, Dr. f. L., 46 Welbeck Street, London, W. Green, H. H., Esq., 36 Prince's Square, Glasgow, S.D.O. Grey, E. G., Esq., B.Sc., Lister Institute, Ghelsea Gardens, London. Gunn, Dr. J. A., 9 Ghalfont Road, Oxford. Haas, Dr. Paul, ii Westbourne Park Road, London, W. 1913] List of Members 449 Hall, A. D., Esq., F.R.S., The Development Commission, Queen Anne's Chambers, London, S.W. Halliburton, Prof. W. D., F.R.S., Church Cottage, 17 Marylebone Road, London, N.W. Harden, Prof. A., F.R.S., 5 Cambridge Gardens, Marlborough Road, Richmond, Surrey. Hardy, W. B., Esq., F.R.S., Newnham Lea, Orange Road, Cambridge. Hartley, Dr. Percival, Lister Institute, Chelsea Gardens, London. Haslam, Dr. H. C, Lyndhurst, Orange Road, Cambridge. Hele, Dr. T. Shirley, Emmanuel College, Cambridge. Henry, Dr. T. A., 70 Doneraile Street, Fulham, S.W. Hill, T. O., Esq., 9 Bloomfield Terrace, London, S.W. Hill, Dr. A. Croft, 169 Cromwell Road, London, S.W. Hopkins, Dr. F. O., F.R.S., Saxmeadham, Orange Road, Cambridge. Hudleston, L. J., Esq., 68 Parliament Hill, Hampstead, London, N.W. Hurtley, Dr. W. H., St. Bartholomew's Hospital, London, E.C. Hutchinson, Dr. H. B., Rothamsted Experimental Station, Harpen- den, Herts. Irvine, Prof. James C, The University, St. Andrew's, N.B. Jones, W. Neilson, Esq., University College, Reading. Keeble, Prof. F., F.R.S., University College, Reading. Kennaway, Dr. E. L., Ouy's Hospital, London, S.E. Laidlaw, P. P., Esq., 38 Gubyon Avenue, Herne Hill, London, S.E. Lander, Dr. O. D., Royal Veterinary College, London, N.W. Lauder, Dr. Alex., 13 George Square, Edinburgh. Ledingham, Dr. J. C. O., Lister Institute, Chelsea Gardens, London. Ling, A. R., Esq., F.I.C., 74 Oreat Tower Street, London, E.C. Lowry, Dr. T. M., 130 Horseferry Road, London, S.W. Macara, Thos., Esq., F.I.C., 13 Ridge Road, Stroud Green, London, N. Maclean, Dr. Hugh, Pathological Department, St. Thomas' Hos- pital, London, S.E. Maclean, Mrs. H., Lister Institute, Chelsea Gardens, London, S.W. Mann, Sydney A., Esq., Qaybury Asylum, Woodford Bridge, Essex. Marsh, J. E., Esq., F.R.S., Merton College, Oxford. Martin, Prof. C. J., F.R.S., Lister Institute, Chelsea Gardens, London. Martin, C. H., Esq., The Hill, Abergavenny. McCay, Major D., I.M.S., The Medical College, Calcutta, India. McKenzie, Dr. Alex., Birkberck College, Bream's Buildings, London. Mellanby, E., Esq., St. Thomas' Hospital, London, S.E. Milroy, Dr. J. A., Queen's University, Belfast. Milroy, Prof. T. H., Queen's University, Belfast. 450 The Biochemical Society, England [April Moore, Prof. B., F.R.S., The University, Liverpool. Mond, Robert, Esq., Combe Bank, near Sevenoaks. Monier- Williams, Dr. G. W., Oiessington Place, Chessington, Surrey. Mottram, V. H., Esq., The University, Liverpool. Nierenstein, Dr. M., The University, Bristol. Neville, A. D., Esq., ii de Freville Avenue, Cambridge. Norris, R. V., Esq., M.Sc., i6 Roseneath Road, Clapham Common, London, S.W. O'Mara, James, Esq., Dunlica, College Road, Dulwich, London, S.E. OsBORNE, Prof. W. A., The University, Melbourne, Australia. Osler, Prof. Sir William, Bart., F.R.S., Norham Gardens, Oxford. Page, H. J., Esq., B.Sc, bei Frau Wwe. A. Pleesow, Mommsenstrasse i6 III, Gr. Lichter felde West, BerHn, Germany. Paine, Sydney G., Esq., Imperial College of Science and Technology, South Kensington, London, S.W. Paton, Prof. D. Noel, The University, Glasgow. Perkin, Prof. A. G., F.R.S., The University, Leeds. Pickering, Spencer U., Esq., F.R.S., Harpenden, Herts. Plimmer, Dr. R. H. A., 59 Queen's Road, St. John's Wood, London. Pope, Thomas H., Esq., The University, Edmund Street, Birmingham. Priestley, J. G., Esq., Little Missenden Abbey, Great Missenden, Bucks. Pyman, Dr. f. L., Carlee, Seiborne Road, Sidcup, Kent. Ramsden, Dr. W., Pembroke College, Oxford. Ransom, Dr. f., Grange Road, Cambridge. Renall, M. H., Esq., B.Sc, 76 Connaught Road, Roath, Cardiff. RoAF, Dr. H. E., 44 Rotherwick Road, Hendon, London, N.W. Rogerson, Harold, Esq., Tavistock, Longslands Park Road, Sidcup, Kent. Rosenheim, Dr. O., King's College, Strand, London. RoTHERA, A. C. H., Esq., The University, Melbourne, Australia. Rowett, J. Q., Esq., Perry Mount, Mayow Road, Forest Hill, London. Russell, Dr. E. J., Rothamsted Experimental Station, Harpenden. Ryffel, J. H., Esq., Medical School, Guy's Hospital, London, S.E. Sadler, W., Esq., Macdonald College, Quebec, Canada. Schryver, Dr. S. B., 48 Overstrand Mansions, Battersea Park, Lond. Scott, Dr. S. G., 20 Charlbury Road, Oxford. S'enter, Dr. G., Medical School, St. Mary's Hospital, Paddington, London, W. Simpson, A. G., Esq., M.A., Coxes Lock Mills, Weybridge. Slator, Dr. Arthur, 174 Ashley Road, Burton-on-Trent. 1913] List of Members 45 1 Smith, A. R., Esq., F.I.C., Whitehall Soap Works, Leeds. Smith, H. L., Esq., F.I.C., King's College for Women, Kensington Square, London, W. Smith, J. Henderson, Esq., M.A., Lister Institute, Chelsea Gardens, London, S.W. Smith, Prof. J. Lorr.\in, F.R.S., The University, Edinburgh. Spriggs, Dr. E. L, 48 Bryanston Street, London, W. Starling, Prof. E. H., F.R.S., 40 West End Lane, London, N.W. Underwood, J. E., Esq., B.Sc, The Homestead, Ulundi Road, Black- heath, London, S.E. Vernon, Dr. H. M., 5 Park Town, Oxford. Vincent, Dr. Ralph, i Harley Street, London, W. ViNES, Prof. Sydney H., F.R.S., Headington Hill, Oxford. Walker, Dr. E. W. Ainley, University College, Oxford. Wallis, Dr. R. L. Mackenzie, St. Bartholomew's Hospital, London. Walpole, Dr. G. S., Wellcome Physiological Research Laboratories, Herne Hill, London, S.E. Ward, Percy G., Esq., University of London, South Kensington, Lon- don, S.W. Wedgewood, A., Esq., Caius College, Cambridge. Weizmann, Dr. Ch., 57 Birchfields Road, Rusholme, Manchester. Wheldale, Miss Muriel, Newnham College, Cambridge. Whitley, E., Esq., 13 Linton Road, Oxford. Whymper, R., Esq., 9 Jerningham Road, New Gross, London, S.E. Wolf, Dr. C. G. L., Southacre, Cambridge. Wood, Prof. T. B., School of Agriculture, Cambridge. WoRLEY, F. P., Esq., M.A., Mansfield House, Clifton Gardens, Maida Vale, London, W. Young, Dr. W. J., Institute of Tropical Medicine, Townsville, N. Queensland, Australia. SCIENTIFIC MEETINGS OF THE COLUMBIA UNI- VERSITY BIOCHEMICAL ASSOCIATION, AT THE COLLEGE OF PHYSICIANS AND SURGEONS, NEW YORK* Proceedings reported by THE Secretary, ALFRED P. LOTHROP I. NINTH MEETING The ninth scientific session of the Columbia University Bio- chemical Association was held at the Columbia Medical School, at 4:15 p. m., 011 February 7, 1913.-^ Abstracts of the papers are here presented (pages 453-461) in two groups: (A) Abstracts of the papers on research by non-resident members^ and (B) abstracts of papers from the Columbia Biochemical Department and affili- ated laboratories. The appended summary facilitates reference to the abstracts (63-72).^ A SUMMARY OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE SUCCEEDING ABSTRACTS (63-72) A K. George Falk and Marston L. Jacob J. Bronfenbrenner and W. H. Hamlin. The action of manganous Manwaring. Resistance in tuber- sulfate on castor-bean lipase. (66) culosis. (63) E. Newton Harvey. The temperature BuRRiLL B. Crohn. The enzymic limits of phosphorescence of lumi- power of duodenal Contents as a nous bacteria. (67) means of diagnosis of the func- A. Hymanson (by invitation). Me- tional activity of the pancreas. (64) tabolism studies of amaurotic family K. George Falk (by invitation). The idiocy. (68) occurrence of a urease in castor Max Morse. A micro-Kjeldahl appa- bean. (65) ratus. (69) * Scientific meetings are held regularly on the first Fridays of December, February and April, and on the first Monday in June. * Proceedings of the eighth meeting were published in the last number of the Biochemical Bulletin, 1913, ii, p. 284. * Members of the Association who were not officially connected with the Columbia Biochemical Department when the research was conducted. ®For abstracts 1-44 see Biochemical Bulletin, 1912, ii, p. 156; for abstracts 45-62, Ibid., 1913, ii, p. 285. See also page 462. 452 I9I3] Alfred P. Lot Itrop 453 B of the convulsiva action of potas- Max Kahn. The calcium content of ^ ^^J!^"^ sulfocyanate. (71) tuberculous areas in lung tissue. R- Ottenberg, D. J. Kaliski and S. S. /„ -v Friedman. Expenmental agglutina- Charles C. Lieb. On the localization ^ive and hemolytic transfusions. (72) A. ABSTRACTS OF PAPERS ON RESEARCH BY NON-RESIDENT MEMBERS* 63. Resistance in tuberculosis. Jacob J. Bronfenbrenner and W. H. Manwaring. (Rockefellcr Institute for Medical Re- search, Nczv York City.) Tubercle bacilli, injected into the peritoneal cavities of tuberculous guinea-pigs, occasionally de- generate and develop into the non-acid resistant forms described by Deycke and Much,^ and others. Under certain conditions, there may be a complete disappearance of the bacilli from the peritoneal fluids within as short a period of time as three hours. Whether this disappearance is due to an actual lysis of the tubercle bacilli, or to other causes, we have not yet determined. As evidence in favor of lysis we have observed that all of the normal control guinea pigs, injected intraperitoneally with the test suspen- sions of tubercle bacilli, died from a fulminating type of visceral tuberculosis, within a period of from three to four weeks, while most of the tuberculous guinea pigs, receiving the same test doses, have survived for longer periods of time A few of these tubercu- lous guinea pigs, however, have died within twenty-four hours after the intraperitoneal tests, suggesting an anaphylactic reaction. We have obtained a similar rapid disappearance of tubercle bacilli from the peritoneal cavities of tuberculous rabbits, from tuberculous rats, and from tuberculous dogs. The question now arose as to the mechanism of this heightened peritoneal resistance. From the similarity between this phenome- non and the Pfeiffer reaction attempts were made to determine whether or not the specific antibodies, upon which the intraperitoneal *Members of the Association who were not officially connected with the Columbia Biochemical Department when the research was conducted. * Deycke and Much : Beitrag s. Klinik f. Tuberk., 1910, xv, p. 277; Much and Leschre: Ibid., 1911, xx, p. 405; Kraus and Hofer: Deutsch, med. Wochen- schr., 1912, xxxviii, p. 1227; Wiener klin. Wochenschr., 1912, xxv, p. 1112. 454 Procecdings Columbia Biocheniical Association [April lysis may be stipposed to depend, are present in the circulating fluids. To test this, giiinea-pigs, rabbits and dogs were made tubercu- lous by inoculating them subcutaneously with tubercle bacilli. After intervals of from five to eight weeks, the animals were bled and their blood tested in vitro and in znvo. In a number of these experiments direct transfusion of the blood was made from the tuberculous animals into normal animals, an amoimt of blood often as great as three quarters of the total blood-volume being thus passed into the circulating System of the normal animals, the normal animals having been previously bled to free them as much as pos- sible from normal blood. The transfused animals were subsequently tested by intraperitoneal injections of tubercle baciUi. Neither in the test-tube experiments, nor in normal animals injected subcutaneously, intravenously or intraperitoneally with tuberculous serum, nor even in normal animals directly transfused with large quantities of the unaltered blood of tuberculous animals, has the reaction thus far been obtained. Therefore, the substances responsible for the heightened peritoneal resistance do not, appar- ently, exist in appreciable quantities as circulating antibodies, at least at the stage of the disease studied. The heightened tubercu- lous resistance is apparently due to substances held in fixed tissue cells. Evidences of tuberculolytic substances have, however, been obtained in the peritoneal fluids of tuberculous guinea-pigs, soon after the introduction of tubercle bacilli. If these fluids are with- drawn, centrifuged free from form Clements and then introduced into the peritoneal cavaties of normal guinea-pigs, tliey confer upon the normal peritoneal cavities a slight power of destroying tubercle bacilH. It is suggested, therefore, that fixed tuberculolysins are set free by the peritoneal cells in response to the presence of tubercle bacilli, and that these lysins account for the heightened resistance to intraperitoneal reinoculation with tubercle bacilli. 64. The enzymic power of duodenal contents as a means of diagnosis of the functional activity of the pancreas. Burrill B. Crohn. {Pathological Lahoratory, Mt. Sinai Hospital, New York City.) The Einhorn duodenal pump was used, in these studies, to obtain duodenal material. This Instrument consists of a small acorn-shaped metallic capsule (perforated), to which is I9I3] Alfred P. Lothrop 455 attached a thin rubber tube, 80 cm. long. The capsule and attached tube are swallowed at night to the point marked 80 cm. In the morning 200 c.c. of milk are drunk by the patient and two hours later the contents of the duodenum are aspirated. During the night the capsule is propelled through the pylorus by peristalsis. The milk acts as a test meal in stimulating pancreatic secretion. The material obtained is tested quantitatively for the strength of its pancreatic enzymes. The methods employed are : For amylasc, a modification of the Wolgemuth starch test; lipase, the ethyl buty- rate test; trypsin, the Fuld-Gross casein test and tests with Permi gelatin tubes, Mett tubes and coagulated tgg albumen cubes. Normal values for the strength of the pancreatic enzymes in the duodenum were first noted in repeated tests of a normal adult. Figures were then obtained similarly in pathological cases of in- terest. The following results were recorded : Acute pancreatitis, marked deficiency of the enzymes; chronic pancreatitis, partial inter- ference with the strength of the enzymes; diabetes mellitus, in- creased strength of enzymes; hypertrophic cirrhosis of the liver, hypersecretion of the pancreas, enzymes very active; gastric dis- eases, enzymes normal ; achylia gastrica, enzymes normal. In this latter group of cases it was impossible to demonstrate the occurrence of a milk-coagulating enzyme, the conclusion being that also the normal pancreas does not contain rennin or any other milk-coagulat- ing enzyme. Enzymes were absent in cases of tumor of the head of the pancreas hut present in stone impacted in the diverticulnm of Vater. 65. The occurrence of a urease in castor bean.*^ K. George Falk {hy invitation) . {Harriman Research Laboratory, Roosevelt Hospital, New York.) A castor bean preparation, husk and oil- free, when allowed to stand in an aqueous Solution of urea, caused the formation of ammonia, as shown by the distillation of the ammonia in a current of air at the ordinary temperature. A castor bean preparation, heated with water and then similarly treated, formed no ammonia. 66. The action of manganous sulfate on castor-bean lipaseJ K. George Falk and Marston L. Hamlin. {Harriman Re- 'Falk: Journal of the American Chemical Society, 1913, xxxv, p. 292. ' Falk and Hamlin : Ibid., 1913, xxxv, p. 210. 456 Procecdings Columbia Biochemical Association [April scarch Lahoratory, Roosevelt Hospital, New York.) The hydro- lytic action of castor bean lipase on ethyl butyrate was decreased in aqueous Suspension and entirely inhibited by heating the aqueous Suspension one to two hours in a water bath. Suspensions that had been partially deactivated by Standing showed an increase in lipolytic power when allowed to act in the presence of small amounts of manganous sulfate ; those that had been completely deactivated by heating showed a sHght but consistent activity in the presence of manganous sulfate, and the activity was still greater if, to the water Suspension of lipase, manganous sulfate was added and the Solution allowed to stand fifteen to twenty hours before testing. These results suggest the tentative hypothesis that, in view of the common function of manganese as an oxygen carrier, the lipase in castor bean is formed from a zymogen, by oxidation aided by an oxygen carrier, with or without simultaneous hydrolysis. 67. The temperature limits of phosphorescence of luminous bacteria. E. Newton Harvey. (Physiological Laboratory, Princeton University, Princeton, N. /,) Light production by many organisms has been observed at relatively high and low tempera- tures, above or below which we should not expect biochemical processes to continue. According to Panceri, Phyllirrhöe (a naked snail of the Mediterranean) is luminous at 75° C, while Pyrosoma (a pelagic ascidian) still glows at 60°. On the other band tissues of the South American fire-fly, Pyrophorus, phosphoresce at — 100° C. (Dubois) and Pseudomonas javanica (a protozoan) at — 20° C. (Eijkmann). The following temperatures have been recorded for luminous bacteria : Organism Observer Minimum Maximum Bacterium phosphorescens Bacterium phosphoreum Light bacteria Lehmann Molisch Tarchanoff -I2OC. - 5" - 7» 28» 37» 50« The temperatures recorded are not excessively high or low, yet the variations in the results suggest that further observations are desirable. Luminous bacteria isolated from fish were grown on absorbent I9I3] Alfred P. Lothrop 457 cotton saturated with beef-broth-peptone-glycerol culture medium. Free access of air between the cotton fibers supplies the conditions for a brilliant light and at the same time an excellent means of handling the bacteria. A wisp of cotton strongly phosphorescent with bacteria was placed in a very thin walled glass tube about 2 mm. in diameter and attached to a thermometer bulb as for melting- point determinations. On slowly raising the temperature, the Hght (to a dark-accustomed eye) becomes dim at 30°, very dim at 34°, and disappears at 38°. On lowering the temperature the light weakens at 0°, is very dim at — 7° and disappears at — ii-5°. These values agree best with those given by Lehmann for Bacterium phosphorescens and do not greatly exceed the usual temperature hmits of activity of organisms. Bacteria raised to 38°, and then cooled, phosphoresce only very dimly, but, as first obsen'ed by Macfayden (an experiment which I have repeated), glow brilhantly at room temperature even after an exposure to liquid air. 68. Metabolism studies of amaurotic family idiocy. A. Hymanson (by invitation) . (Chemical Lahoratory, Beth Israel Hospital, New York City.) Two cases of amaurotic family idiocy were kept under Observation until death. The metabolism of nitrogen, sulfur and phosphorus was carefuUy studied. It was found that both absorption and retention were normal or above normal. The digestive System does not seem to be at all deranged in this fatal disease. 69. A micro-Kjeldahl apparatus. Max Morse. (Boardnian Laboratories, Trinity College, Hartford, Conn.) This is a com- bination of the apparatus designed by Fritz Pregl^ [Plate 6] for total nitrogen determination in small quantities of material, and the fume absorber devised by Folin and Denis.'' The apparatus may be equally serviceable with that devised by Sy.^^ With such a means of eliminating fumes, the determination of total nitrogen may be carried on wherever the water-pressure is sufficient to maintain an ordinary filter-pump in action; in conjunction with the 'Pregl: Abderhalden's Handbuch der biochemischen Arbeitsmethoden, 1912, V, p. 1344- ' Folin and Denis : Journal of Biological Chemistry, 1912, xi, p. 503. '"Sy: Journal of Industrial and Engineering Chemistry, 1912, iv, p. 680. 45^ Proceedings Columbia Biochemical Association [April micro-amino-nitrogen apparatus of Van Slyke, it makes an admir- able outfit for work with small amounts of protein. During digestion, the tiibe with the bulbs is inserted into the absorber, B, and the fumes from the 200 mm. Jena test-tube, /, are carried to the sink through the pump, PM, via tube S, which may have a half dozen openings, so that a number of determinations may be conducted at the same time. Since much water vapor is lost during digestion, Pregl passes steam from a flask, /, to the bottom of the Jena test-tube, J, through the long small-bore tube, A, which is fused into the bulb through which it passes. It is quite necessary to pass steam in this way. Neutralization is effected with sodium hydroxid Solution poured down tube A, after the apparatus has cooled; if the hydroxid is added immediately after digestion, tube A will frequently crack. After neutralization, the upper end of tube A is closed by a clamp or glass rod introduced into the rubber connection and distillation is effected by connecting tube R, which dips into the decinormal acid in the beaker P. This should be done, as a matter of fact, before neutralization, so that no am- monia is lost. In place of distilling by heat, Folin's method for Urea may be introduced, by driving an air current through A, and the only precaution necessary is that the current be sufficiently strong to drive all of the gas into the distillation tube, R. The bulb at the lower end of the distillation tube, R, prevents back- suction, as sometimes occurs. The Jena test-tube fits loosely the collar of the apparatus above it, and the water-vapor condenses sufficiently around it to insure an air-tight Joint. There is Variation in Jena test-tubes of 200 mm. length, but it is small, so that the apparatus fits practically any such tube. The apparatus is made by the Emil Greiner Company, New York City. B. ABSTRACTS OF PAPERS FROM THE COLUMBIA BIOCHEMICAL DEPARTMENT AND AFFILIATED LABORATORIES 70. The calcium content of tuberculous areas in lung tissue. Max Kahn. Wherever the tubercle bacillus lodges it induces a deposition of calcium salts, which hinders the ingress of other bacilli. The body in general becomes poorer in lime salts. It was c tu Q •a c n! p C H o < Uh ^ T) < c CL, 03 CM ^ < u kJ Pl, ffi >> < ^ Q •ö J 4 ; 7s. 6d. net. W. Keffer & Sons, Ltd., Cambridge, Eng., 1913. Glycosuria and allied conditions. By P. J. Cammidge. Pp. 467 — 4X6^; $4.50 net. Longmans, Green & Co., New York; Edward Arnold, London, 1913. The Chemical Constitution of the proteins: Part II, Synthesis, etc. 2d ed. (One of the Monographs on Biochemistry.) By R. H. A. Plimmer, Univ. reader and ass't prof. of physiological ehem., Univer§ity Coli., London. Pp. 107 — 4^ X 71/2; $1.20 net. Longmans, Green & Co., 1913. Studies from the Rockefeiler Institute for Medical Research. Volume XVI; 1913. (29 reprints.) CoUected papers: Institute of Physiology, University College, London. Edited by Ernest H. Starling, Jodrell professor of physiology. Volume XVII; 1912-13. (32 reprints.) CoUected papers: Physiological Laboratory, Kings' College, University of London. Edited by W. D. Halliburton, professor of physiology. Volume XII;i9i3. (12 reprints.) Sigma Xi Quarterly. No. i. Vol. i (March, 1913)- Pp- 30. Editorial committee: J. McK. Cattell, D. C. Miller, H. B. Ward, S. W. Williston. Pub- lished by the Society of the Sigma Xi, H. B. Ward, corresponding secretary, Champaign, 111. Bulletin of the American Home Economics Association. No. i, Series i (Nov., 1912). PubHshed quarterly by the American Home Economics Associa- tion, Benjamin R. Andrews, secretary, 525 W. I20th St., New York City. CoUected papers: Laboratory of physiological chemistry, Sheffield Sci- entific School, Yale University. 1911-1912. (35 reprints.) Practical physiological chemistry. A book designed for use in courses in practical physiological chemistry in schools of medicine and of science. By Philip B. Hawk, professor of physiological chemistry and toxicology in the Jefferson Medical College of Philadelphia. Fourth edition, revised and en- larged. Pp. 475— 4J^ X 8 ; $2.50 net, P. Blakiston's Sons & Co., Philadelphia, 1912. The protein dement in nutrition. (One of the International Medical Mono- graphs.) By Major D. McCay, professor of physiology, Medical College, Cal- cutta. Pp. 216 — 4X7, with 8 füll page portraits of human subjects; $2.00 net. Longmans, Green and Co., New York; Edward Arnold, London, 1912. Oxidations and reductions in the animal body. (One of the Monographs on Biochemistry.) By H. D. Dakin, The Herter Laboratory, New York. Pp. 135 — ^41/^X8; $1.40 net. Longmans, Green and Co., 1912. Researches on cellulose. III (1905-1910). By C. F. Gross and E. J. Bevan. Pp. 173 — 3y2X6; $2.50 net. Longmans, Green and Co., 1912. OFFICERS OF THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-1913* OFFICIAL REGISTER, MAR. 31, 1913 William J. Gies: Professor and Chairman of the Staff; Consulting chemist, New York Botanical Garden ; Pathological chemist, First Division, Bellevue Hospital; Member of the Faculties of N. Y. Teachers College and N. Y. College of Pharmacy. [B.S., Gettysburg College, 1893 and M.S., 1896; Ph.B., Yale University, 1894 and Ph.D., 1897. Instructor, i898-'o2; adjunct Pro- fessor, 1902-05; Professor, 1905-.] Paul E. Howe: Assistant Professor. [B.S., University of Illinois, 1906; A.M., 1907 and Ph.D., 1910. Assistant Professor, 1912-.] Alfred P. Lothrop: Associate and De partmental Registrar. [A.B., Oberlin, 1906 and A.M., 1907; Ph.D., Columbia, 1909. Assistant, i9o8-'o9; instructor, i909-'i2; associate, 1912-.] Nellis B. Foster: Associate; Associate Physician, New York Hospital; Chemist, St. Luke's Hospital. [B.S., Amherst College, 1898; M.D., Johns Hopkins University, 1902. Instructor, i9o6-'o8; associate, 1908-.] Walter H. Eddy: Associate and Secretary of the Staff. [B.S., Amherst Col- lege, 1898; A.M., Columbia, 1908 and Ph.D., 1909. Assistant, 1908-'! 0; associate, 1910-.] Herman O. Mosenthal: Associate; Assistant Attending Physician, Presbyterian Hospital; Assistant Physician, Vanderbilt Clinic; Instructor in medicine. [A.B., Columbia, 1899 and M.D., 1903. Assistant, I9o8-'o9; instructor, 1909- '12; associate, 1912-.] Emily C. Seaman: histructor. [B.S., Adelphi College, 1899; A.M., Columbia, 1905 and Ph.D., 1912. Tutor, i909-'io; instructor, 1910-.] Max Kahn : Instructor ; Director of the chemical and physiological laboratories of Beth Israel Hospital. [M.D., Cornell University Medical College, 1910; A.M., Columbia, 1911 and Ph.D., 1912. Instructor, 1912-.] Louis E. Wise: Instructor. [A.B., Columbia, 1907 and Ph.D., 1911. Instructor, 1912-.] Edgar G. Miller, Jr. : Assistant, 1911-. [B.S., Gettysburg College, 1911.] Frederic G. Goodridge: Assistant, 1912-. [A.B., Harvard University, 1897; M.D., Columbia, 1901.] Arthur Knudson : Assistant, 1912-. [A.B., University of Missouri, 1912.] Ethel Wickwire: Assistant, 1912.-, [A.B., Tri-State College, 1909.] TuLA L. Harkey: Assistant, 1912-. [A.B., Colorado College, 1909.] Benjamin Horowitz: Assistant, 1913-. [B.S., Columbia, 1911 and A.M., 1912.] Christian Seifert: Laboratory assistant, 1898-. Stella Waldeck : Recorder, 1908-. Blanche E. Shaffer: Laboratory assistant, summer Session, 1912. Joseph S. Hepburn: University fellow, 1912-13. [A.B., Central High School, Philadelphia, 1903 and A.M., 1908; B.S., University of Pennsylvania, 1907 and M.S., 1907.] * The work of the department was inaugurated in October, 1898, by Prof. R. H. Chittenden (lecturer and director), Dr. WilHam J. Gies (instructor), Messrs. Alfred N. Richards and Allan C. Eustis (assistants), and Christian Seifert (laboratory assistant). COURSES OFFERED BY THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-13 (Abbrevtations: C, Conference; D, demonstration ; L, lecture; Lw, labora- tory work; R, recitation.) ORGANIC CHEMISTRY 51. Elementary ORGANIC CHEMISTRY. (First half year. Medical School.) Introductory to course 102 (52). (Required of first year students of medkine.) L, I hr. D, I hr. R, 2 hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Drs. Wise and Goodridge, and Messrs. Miller and Knudson. NUTRITION (PHYSIOLOGICAL AND PATHOLOGICAL CHEMISTRY) 101(2) — Grad. General biological chemistry. A course in the elements of normal nutrition. (All year. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies, Dr. Lothrop and Messrs. Miller and Knudson. 101(2) — B. T. General biological chemistry. Specially adopted to the needs of secondary school teachers of biology. {All year. Medical School.) L, I hr. Lw, 4 hr. Dr. Eddy. 101:102 — ^T. C. General physiological chemistry. A course in the ele- ments of normal nutrition. {Each half year. Teachers College, School of Practical Arts.) L, 2hr. R, i hr., each section (2). Lw, 5 hr., each section (2). Prof. Gies, Dr. Seaman and Misses Wickwire and Harkey. (This course is designated " Chemistry 51 " and " Household Arts Education 125 " in the Teachers College Announcement.) This course is designated " Chemistry s 51 " in the Teachers College Division of the Summer School Announcement. The course was given last summer by Prof. Gies, Dr. Seaman and Miss Shaffer. 102 (52) — Med. General physiological chemistry. (Second half year. Medical School.) A course in the elements of normal nutrition. (Required of first year students of mediane.) L, 2 hr. R, i hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Dr. Wise, and Messrs. Miller and Knudson. This course is designated " 5" — 104 " in the Medical Division of the Summer School Announcement. It was given last summer by Prof. Gies and Dr. Smith. 104. General pathological chemistry. Lectures on nutrition in disease. (Second half year. Teachers College, School of Practical Arts.) L, i hr. Prof. Gies. (This course is designated "Chemistry 52" in the Teachers College An- nouncement.) 209-210. Chemistry of nutrition. (All year. School of Pharmacy. Re- quired of candidates for the Degree of Doctor of Pharmacy.) L, i hr. Prof. Gies. 213-214. Advanced physiological chemistry, including methods of re- search in nutrition. (All year. Teachers College, School of Practical Arts.) L, I hr. Lw, 5 hr. Prof. Howe, Dr. Seaman and Mr. Horowitz. (This course is designated " Household Arts Education 127 " in the Teachers College An- nouncement.) 217-218. BlOCHEMICAL methods OF RESEARCH, INCLUDING CLINICAL METHODS and urinary analysis in GENERAL. (All year. Medical School.) L, i hr. Lw, 7 hr. Profs. Gies and Howe, Dr. Lothrop, and Messrs. Miller and Hepburn. 219-220. Nutrition in health. A laboratory course in advanced physio- logical chemistry. (All year. Medical School.) L, 2 hr. Lw, 14 hr. Profs. Gies and Howe, and Dr. Lothrop. Courses in Nutrition (continued) 221-222. Nutrition in Disease. A laboralory course in advanced patholog- ical chemistry. {All year. Medical School.) L, 2 hr. Lw, 14 hr. Prof. Gies. 223-224. Nutrition in Disease. {All year. Medical School.) L, 1 hr. Profs. Gies and Howe, and Drs. Foster, Mosenthal, Kahn and Goodridge. 225-226. Advanced physiological and pathological chemistry, including ALL PHASES OF NUTRiTioN. {All year. Medical School.) Research. C, i hr. (individual students). Lw, 16 hr. Profs. Gies and Howe, and Dr. Lothrop. 1 TOXICOLOGY 231-232. Effects and detection of poisons, including food preservatives AND adulterants. {All year. Medical School.) Lw, 6 hr. Prof. Gies and Mr. Miller. BOTANY 235-236. Chemical physiology of plants. {All year. New York Botan- ical Garden.) L, i hr. Lw, 5 hr. Prof. Gies. BACTERIOLOGY 241-242. Chemistry of microorganisms: Fermentation s, putrefactions and the behavior of enzymes. An introduction to sanitary chemistry. {All year. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies. SANITATION 105. Sanitary chemistry. {Second half year. Teachers College, School of Practical Arts). L, i hr. Lw, 3 hr. Dr. Seaman and Miss Harkey. (This course is designated "Chemistry 57" and "Household Arts Education 129" in the Teachers College Announcement.) BIOCHEMICAL SEMINAR 301-302. Biochemical Seminar. {All year. Medical School.) i hr. Prof. Gies. RESEARCH IN BIOLOGICAL CHEMISTRY Biochemical research may be conducted, by advanced workers, independently or under guidance, in any of the departmental laboratories. LABORATORIES FOR ADVANCED WORK IN BIOCHEMISTRY The laboratories in which the advanced work of the biochemical department is conducted are situated at the College of Physicians and Surgeons, Teachers College, New York Botanical Garden and Bellevue Hospital. Each laboratory is well equipped for research in nutrition and all other phases of biological chemistry. BIOCHEMICAL LIBRARY Prof. Gies' library occupies a room adjoining the main biochemical labora- tory at the College of Physicians and Surgeons and is accessible, by appoint- ment, to all past and present workers in the Department. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION The Biochemical Association holds scientific meetings regularly on the first Fridays in December, February and April, and on the first Monday in June. These meetings are open to all who may be interested in them. SUMMER SCHOOL COURSES Summer session courses are mentioned in the foregoing references to Courses 101-102 and 102 (52). Prof. Gies will have charge of both courses next summer. He will also conduct a special lecture course in nutrition. The labora- tories will be open for research throughout the summer. ANNOUNCEMENTS Professional Assistance Offered to Biological Chemists The Columbia University Biochemical Association will be glad to coöperate confidentially with all who desire the Services of biological chemists and with all who seek positions in biological chemistry. Address inquiries to William J. des, 43 j West sgth St., New York. Joumalistic New JOURNAL. Physiological Researches. To appear at irregulär intervals. Edited by Burton E. Livingston, Manager, Johns Hopkins University ; Daniel T. MacDougal, Carnegie Institution of Washington ; Herbert M. Richards, Columbia University. " The recent rapid advance of physiological science has been accompanied by a realization of the Community of interest and uniformity of method which characterize the physiology of plants and of animals, and it has seemed highly desirable that the general physiological field thus indicated should possess an organ of publication in which its more comprehensive and technical papers might appear. This need is emphasized by the fact that pres- ent facilities for publication in physiology are generally taxed beyond their capacity and papers are consequently subject to long delays in appearance. It has therefore been decided to inaugurate a new series of scientific papers which will embrace contributions towards the advance of fundamental physiological knowledge." " The plan of publication of the new series, for which the title of Physiological Researches has been adopted, is one in which practical ownership is vested in the contributors. It is hoped that the project will receive the interest and support of biologists of all classes. ^ (Each volume will contain about 450 pages ; each number will contain but a Single contribution ; and the numbers will be issued irregularly) . Pub- lication of the first contribution may be expected in a short time. Sub- scriptions will be received by the volume, the price being $5.00 per volume, payable in advance. Subscriptions to volume I, which are received prior to the date of publication of the first research, may be made at the reduced price of $4.00. At the date of the appearance of the first research the price will automatically become the regulär one. Remittances should be made payable to Physiological Researches, and all correspondence should be addressed to Physiological Researches, Station N, Baltimore, Maryland, U. S. A." (Editors' announcement.) Reduced subscription price of the Journal of Biological Chemistry. The directors of the Journal of Biological Chemistry have announced that "beginning with the February issue of 1913 (Vol. 14, No. i) the subscription price of the Journal to domestic subscribers will be reduced f rom $4.00 to $3.00 per volume ; to f oreign subscribers, $3.25. Any one engaged in biochemical work who subscribes for the Journal at this rate (beginning with Vol. 14) may secure Volumes 1-13 for $20.00, plus cost of transportation. The price at which a complete set has hitherto been sold is $50.00. Subscribers for the Journal who wish to complete their files may secure early volumes for $1.50 each, plus cost of transportation. Address : Alfred N. Richards, Secretary, University of Pennsylvania. Meetings of Societies and Congresses Tenth International Congress of Agriculture: Ghent, Bel- gium, June 8-13. Secretary-general, Dr. P. de Vtiyst, 22 Avenue des Germaines, Brüssels. American committee: Dr. L. O. Howard, mem- ber of the International Commission on Agriculture and chief of the Bureau of Entomology; and Dr. A. C. True, director, Mr. John Ham- ilton, specialist in farmers' institutes, Dr. C. F. Langworthy, chief of nutrition investigations and Dr. J. I. Schulte, assistant agriculturist, of the Office of Experiment Stations. Second International Congress for the Teaching of House- hold Economy: Ghent, Belgium, June. General Secretary, Miss Deleu, 19 Rue Willems, Brüssels. Third International Congress of the Associations of Agri- CULTURAL Women: Ghent, Belgium, June. General Secretary, Miss Van Aarschot, 38 Rue du Pepin, Brüssels. American Medical Association, Annual meeting: Minneapolis, Minn., June 17-20. General secretary, Geo. H. Simmons, 535 Dear- born Ave., Chicago. General Meeting of the International Association of Botan- ists : Copenhagen, June 27. Secretary-general, J. P. Lotsy, Haarlem, Holland. CONGRES INTERNATIONAL POUR LA PROTECTION DE l'eNFANCE: Brüssels, July 23-26. General Secretary, Henry Jaspar, 93 Avenue de la Toison d'Or, Brüssels. Seventeenth International Congress of Medicine: London, Aug. 6-12. General secretary, Dr. W. P. Herringham, 13 Hinde St., London, W. FouRTH International Congress on School Hygiene: BuflFalo, N. Y., Aug. 25-30. Secretary-general, Prof. Thomas A. Storey, Col- lege of the City of New York. Ninth International Physiological Congress : Groningen, Hol- land, Sept. 2-6. American Secretary, Prof. W. T. Porter, Harvard Medical School. Third International Congress of Refrigeration : Washington, D. C, Sept. 15 (opening meeting) ; Chicago, Sept. 17-23 (business and scientific meetings). Secretary-general, Mr. J. F. Nickerson, 431 So. Dearborn St., Chicago. The Biochemical Bulletin The Biochemical Bulletin is a quarterly biochemical review. It publishes results of original investigations in biological chemistry and presents miscellaneous items of personal and professional in- terest to chemical biologists. Original contributions to research, preliminary reports of investigations, abstracts of papers, addresses. reviews, descriptions of new methods and apparatus, practical sug- gestions to teachers, biographical notes, historical sümmaries, bibliographies, quotations, news items, proceedings of societies, personalia, views on current events in chemical biology, etc., are solicited. Siihscription prices. Vol. I : $6.00 (No. i, $1.50 ; No. 2, $2.50 ; No. 3, $2.00; No. 4, $1.50). Vol. II: $2.75 (domestic) ; $3.00 (foreign) ; $6.00 after July i, 1913 (No. 5, $1.25 ; No. 6, $1.00). Remittances, manuscripts and correspondence should be addressed to the Biochemical Bulletin, 437 West 59th St., New York City. Vol. II July, 1913 No. 8 Biochemical Bulletin Edited, for the Columbia University Biochemical Association, by tho EDITORIAL COMMITTEE: HERMAN M. ADLER. JOHN S. ADRIANCE. CARL L. ALSBERG, D. B. ARMSTRONG, CHARLES W. BALLARD. LOUIS BAUMANN, GEORGE D. BEAL, STANLEY R. BENEDICT, WM. N. BERG, JOSEPHINE T. BERRY, ISABEL BEVIER, A. RICHARD BLISS, CHARLES F. BOLDUAN, SAMUEL BOOKMAN, SIDNEY BORN, J. BRONFEN BRENNER, GERTRUDE S. BURLINGHAM, RUSSELL BURTON-OPITZ, A. T. CAMERON, HERBERT S. CARTER, ELLA H. CLARK, ERNEST D. CLARK, BURRILL B. CROHN, LOUIS J. CURTMAN, WILLIAM D. CUTTER, C. A. DARLING, NORMAN E. DITMAN, EUGENE F. DuBOIS, JAMES G. DWYER, WALTER H. EDDY, A. D. EMMETT, ALLAN C. EUSTIS, BENJ. G. FEINBERG, RUTH S. FINCH, HARRY L. FISHER, KATHRYN FISHER. NELLIS B. FOSTER, C. STUART GAGER, MARY C. de GARMO, MARY E. GEARING, SAMUEL GITLOW, WILLIAM J. GIES, Treasurer, A. J. GOLDFARB, H. G. GOODALE, F. G. GOODRIDGE. ROSS A. GORTNER, ISIDOR GREENWALD, JAMES C. GREENWAY, LOUISE H. GREGORY, MARSTON L. HAMLIN, R. F. HARE, TULA L. HARKEY, E. NEWTON HARVEY, P. B. HAWK, MICHAEL HEIDELBERGER, JOSEPH S. HEPBURN, ALFRED F. HESS, BENJAMIN HOROWITZ, PAUL E. HOWE, Secretary. LOUIS HUSSAKOF, HENRY H. JANEWAY. MAX KAHN, JOHN L. KANTOR, EDWARD C. KENDALL, J. E. KIRKWOOD, ARTHUR KNUDSON, MATHILDE KOCH, MARGUERITE T. LEE, CHARLES C. LIEB, BURTON E. LIVINGSTON, ALFRED P. LOTHROP, Chairman, DANIEL R. LUCAS, MARY G. McCORMICK, LOUISE McDANELL, GRACE MacLEOD, C. A. MATHEWSON, H. A. MATTILL, CLARENCE E. MAY, GUSTAVE M, MEYER, E. G. MILLER, JR., MAX MORSE, H. O. MOSENTHAL, B. S. OPPENHEIMER. RAYMOND C. OSBURN, REUBEN OTTENBERG, OLIVE G. PATTERSON, W. A. PERLZWEIG, W. H. PETERSON, E. R. POSNER, P. W. PUNNETT, ALFRED N. RICHARDS, ANNA E. RICHARDSON, WINIFRED J. ROBINSON, ANTON R. ROSE, JACOB ROSENBLOOM, WILLIAM SALANT, OSCAR M. SCHLOSS, H. von W. SCHULTE, FRED W. SCHWARTZ. C. A. SCHWARZE, EMILY C, SEAMAN, FRED J. SEAVER, A. D. SELBY, A. FRANKLIN SHULL, CLAYTON S. SMITH, MATTHEW STEEL, CHARLES R. STOCKARD, MARY E. SWEENY, A. W. THOMAS, PHILIP VAN INGEN, EDWIN D. WATKINS, WILLIAM WEINBERGER, JULIUS W. WEINSTEIN, CHARLES WEISMAN, WILLIAM H. WELKER, H. L. WHITE, DAVID D. WHITNEY, ETHEL WICKWIRE, LOUIS E. WISE, WILLIAM H, WOGLOM, L. L. WOODRUFF, HANS ZINSSER. NEW YORK Entered as second-cIaiB matter in the Post Office at Ivancaster, Pa. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Members PROF. R. H. CHITTENDEN, First Director of the Columbia University De- partment of Biological (Physiological) Chemistry; Director of the Shef- field Scientific School of Yale University PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Switzerland PROF. SAMUEL W. LAMBERT, Dean of the Columbia University School of Medicine DR. JACQUES LOEB, Member of the Rockefeiler Institute for Medical Re- search; Head of the Department of Experimental Biology PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. LEON ASHER, University of Bern, Switzerland PROF. FILIPPO BOTTAZZI, University of Naples. Italy PROF. ROBERT B. GIBSON, University of the Philippines, P. I. PROF. VLADIMIR S. GULEVIC, University of Moscow, Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. D. McCAY, Medical College, Calcutta, India PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden. — C. Stuart Gager. College of the City of New York. — ^Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Goldfarb. Columbia University: Departments. — Anatomy: Alfred J. Brown, H, von W. Schulte; Bacteriology: James G. Dwyer; Biological Chemistry: Walter H. Eddy, Nellis B. Fester, William J. Gies, F. G. Goodridge, Tula L. Harkey. Joseph S. Hepburn, Benjamin Horöwitz, Paul E. Howe, Max Kahn, Arthur Knudson, Alfred P. Lothrop, Edgar G. Miller, Jr., H. O. Mosenthal, Emily C. Seaman, Chris. Seifert, Ethel Wickwire, Louis E. Wise; Botany: E. R. Alten- burg, C. A. Darling, Fred D. Fromme; Cancer Research: W. H. Woglom; Chem- istry: A. M. Buswell, R. P. Calvert, Gustave Egioff, H. L. Fisher, P. W. Punnett, A. W. Thomas; Clinical Pathology: Edward Cussler, Peter Irving; Diseases of Children: Herbert B. Wilcox; Gynecology: Wilbur Ward; Medicine: T. Stuart Hart, I. Ogden Woodruff; Pathology: B. S. Oppenheimer, Alwin M. Pappen- heimer; Pharmacology: Charles C. 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Professional Assistance Offered to Biological Chemists The members of tlie Columbia University Biochemical Association will coöperate confidentially with any one who desires tbe Services of biological chemists or who seeks a position in biological chemistry. Address inquiries to William J. des, 4^y West 59th St., Nezv York. EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the collaboration of the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockef eller Institute for Medical Research PROF. WILDER D. BANCROFT, Cornell University, Ithaca DR. WALTER L. GROLL, Elisabeth Steel Magee Hospital, Pittshurgh, Pa. DR. CHARLES A. DOREMUS, 55 W. 53d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station, Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR. LEWIS W. FETZER, U. S. Dep't of Agricultnre, Washington, D. C. PROF. MARTIN H. FISGHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. PROF. J. E. GREAVES, Utah Agricultural College, Logan DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. M. HARDISTY, McGill University, Montreal, Canada DR. J. A. HARRIS, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. K. A. HASSELBALCH, Einsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, A^. Y. Univ. and Bellevue Hospital Med. College PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan PROF. LAFAYETTE B. MENDEL, Yale University PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven DR. AMOS W. PETERS, The Training School, Vineland, N. J. PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SP AAR, City Hospital, Trincomalce, Ceylon PROF. UMETARÖ SUZUKI, University of Tokyo, Japan MISS ANNA W. WILLIAMS, University of Illinois, Urbana, III. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switserland DR. JULES WOLFF, Pastair Institute, Paris BiocHEMiCAL Bulletin Volume II JULY, 1913 No. 8 lfbrar' New Vr;;.: ßOTA ■ AN INVESTIGATION TO DETERMINE THE ACCU- t. RACY OF A MODIFIED MEIGS METHOD FOR THE QUANTITATIVE DETERMINATION OF FAT IN MILK, WITH A DESCRIPTION OF AN IMPROVED FORM OF APPARATUS WALTER LEWIS CROLL (wiTH PLATE 7) (Robert Hare Chemical Lahoratory, University of Pennsylvania) Owing to the importance of determining accurately and quickly the amount of fat in a given quantity of milk, special efforts have been made to devise a method that would embody both these requi- sites. These efforts have resulted in methods based upon many principles, and some are brilliant examples of chemical and mechan- ical ingenuity. The Separation of the fat has been accomplished by the use of differences in specific gravity, by saponification, extrac- tion with ether, colorimetry, and absorption; but, to the present time, no rapid method, which is sufficiently accurate for legal, pediatric and biological work, has been devised. Today the most accurate method and, indeed, the only one suitable for most legal ' work, is the Adams paper-coil method, with the Soxhlet extraction » apparatus. This method, while extremely accurate, requires at least twenty-four hours for its execution and considerable care in its manipulation. A COMPARISON OF WELL KNOWN METHODS The Adams method. In this investigation the Adams method was carried out as follows: First a homogeneous mixture was 509 510 Quantitative Determination of Fat in Milk [july obtained, then some of it was placed in a small Erlenmeyer flask having a 5 c.c. bulb-pipette fitting snugly in its mouth. About 4 or 5 gm. were removed with the pipette and dropped into the center of a coil of specially prepared, fat-free, absorbent paper. The sample was weighed by difference. The paper coil containing the milk was transferred to a hot-air oven, the temperature of which was constantly below 100° C, and permitted to dry for three or four hours, after which it was removed to a glass desiccator, kept there at least twelve hours, over conc. sulfuric acid, and then trans- ferred to a Soxhlet extraction apparatus, where it was extracted for twelve to sixteen hours with absolute ethyl ether which had been redistilled after Standing over metallic sodium for from five to six days. After completing the extraction, the excess of ether was distilled into the upper part of the apparatus and the ethereal Solu- tion of fat quantitatively transferred to a weighed evaporation dish, using redistilled ether as a rinsing fluid. The ether was evaporated over a safety water-bath (the dish being protected from dust by an inverted funnel), at a temperature below 33° C, to prevent loss from ebullitlon of the ether. After the ether had been evaporated, the fat was placed in a hot-air oven and dried for three hours at a temperature ranging between 95°-ioo° C, then dried over sulfuric acid to constant weight, which usually required about twenty-four hours. The weight was then recorded and the percentage of fat calculated. Of the above method there are many modifications, but none of them are rapid methods. Miscellaneous methods. In the method devised by Soxhlet,^ the drying of the milk is accomplished by the use of gypsum. Froidevaux^ precipitates the protein and fat by means of acetic acid and calcium phosphate. The precipitate is collected quantitatively on a filter paper, dried and extracted in a Soxhlet apparatus with absolute ether. Le Comte^ suggests the use of sodium sulphate as desiccating material. In the modification suggested by Rieter,* gypsum and later some Fehling Solution are used to precipitate the 1 Soxhlet : Polytech. Jour., 1879, ccxxxii, p. 461. 2 Froidevaux : Jour. de Pharm, et Chem., 1897 (6), vi, p. 485. 3Le Comte: Ibid., 1901 (6), xiii, p. 58. * Rieter : Schweiz. Wochschr. Pharm., 1903, xli, pp. 39 and 53. 1913] Walter Lewis Croll S^i protein and fat. The precipitate is collected quantitatively on filter paper, dried, and extracted with absolute ether. "Rapid" methods. Nearly all the so-called rapid methods depend upon centrifugation. In these methods accuracy is sacri- ficed for speed and the results obtained are, at best, only approxi- mate. For commercial and pediatric uses they are of sufficient precision to answer most requirements. An example of these meth- ods is the widely used Babcock process, with the special scale for reading off the contained fat, and various other modifications dif- fering only in the shape of the tube or the scale. Another rapid but cruder method is the well known Feser lactoscope process. Woosnam,^ following the idea of Schmid, suggests the follow- ing method : 25 c.c. of milk and 28 c.c. of conc. hydrochloric acid sol. are placed in a special apparatus and heated on a water-bath until a slight browning occurs, when the flask and contents are cooled, an ether extraction made, and the volume of fat-ether mixture is read. A definite portion of this mixture is then taken, evaporated to dryness in a weighed glass dish and the dry fat weighed. Following the method of Krug and Hampe^ as a basis, Arndt'^ describes a process involving special extraction apparatus. The milk is desiccated with kaolin and dry sodium sulfate, and then extracted with ether in the apparatus. As a modification of the usual butyrometer method, Gerber and Craandijk^ recommend the following: Into a beaker of 5.5 c.c. capacity place 4-5 gm. of previously weighed, well mixed milk, introduce the beaker and contents into the butyrometer, add 10 c.c. of warm water, place the butyrometer in a water-bath at 6o°-'jo° C. until complete Solution has occurred. To the liquid add i c.c. of amyl alcohol, 10 c.c. of sulfuric acid sol, (sp. gr., 1520-1525), close the butyrometer, shake until the contents are well mixed, and then place it in a water-bath until the greater part of the fat separates. Finally, centrifuge twice and read the scale. ^Woosnam: Analyst, 1897, xxii, p. 91. 6 Krug and Hampe : Ztschr. f. angew Chemie, 1894, p. 683. ''Arndt: Chem. Centr., 1897, "> P- 636 (Forsch.-Ber. üb. Lebensm. u. ihre Gez. 2. Hyg., etc., iv, p. 231). * Gerber and Craandijk: Milch-Ztg., 1898, xxvii, pp. 35 and 273. 512 Quantitative Determination of Fat in Milk [july Richmond and Rosier^ advise treatment of milk with definite proportions of 90-91 per cent. sulfuric acid sol. and amyl alcohol, and, after cooling to about 25° C, extracting with 20 c.c. of petroleum ether. This method is not very accurate because of the fact that petroleum ether itself frequently leaves an evaporation- residue, probably an isomeric substance, hence the percentage by this method is likely to be too great. Utilizing the tragacanth process devised by Rusting,^° Bon- nema^^ describes an ether-extraction method, — following prelimi- nary treatment with potassium hydroxid sol. Tragacanth is added to facilitate the Separation of the water and the ethereal Solutions, The results are fairly good but not as accurate as those obtained with the paper-coil method. In the Ram-Fouard^2 method, milk is treated with a special reagent containing potassium hydroxid, ethyl alcohol, amyl alcohol and ammonium hydroxid. The milk is placed in a special 50-60 c.c. flask with 10 c.c. of this reagent, the flask is immersed in boiling water for ten minutes, then removed, and enough distilled water added to carry the liquid into the neck of the flask, when the flask is placed in water at 40° C, and the fat allowed to separate. The average milk fat has a specific gravity of about 0.90 at 40° C. ; hence, one-quarter the amount of fat recorded volumetrically gives the number of grams of fat per liter of milk. Hanget and Marion^ ^ speak favorably of a method in which they use a special reagent consisting of 100 c.c. of n/6 ammonium hydroxid sol. neutralized to litmus by the addition of lactic acid and made up to 150 c.c. with distilled water, and 435 c.c. of absolute ether, 420 c.c. of absolute alcohol and 40 c.c. of a Solution of i gm. of methyl violet (5B) in 1,000 c.c. of absolute ether. After con- traction has occurred, the volume is measured and 38.9 c.c. of abso- lute ether are added per liter. A special apparatus is used, by the aid of which the percentage of fat is read off on the scale at 2)7° C. ö Richmond and Rosier : Analyst, 1899, xxiv, p. 172. lORusting: Chem. Centr., 1898 (ii), p. 393 {Nederl. Tijdschr. v. Pharm., Chem. en Tox., 1899, x, p. 163). 11 Bonnema : Chem-Ztg., 1899, xxiii, p. 541. i2Leze (Ram-Fouard method) : Ann. de Chim. anal. appL, 1899, iv, p. 371. 13 Hanget and Marion : Ibid., 1902, vii, p. 297. 1913] Walter Lewis Groll 513 The refractometer is utilized by Wollny^^ on an ethereal Solution of the fat obtained by extraction after the application of a special reagent. The refractive power of the ethereal Solution of fat is determined at 17.5° C. Maccagno and Mizzi^^ have devised a special apparatus in which they treat the milk with a definite quantity of a Solution containing ethyl alcohol, amyl alcohol and ammonium hydroxid. The per- centage of fat is read off on the scale of the apparatus at a tem- perature near the boiling point of water. By the Sichler " sinacidbutyrometer method,"^® the casein and lactalbumin are dissolved by means of a Solution of sodium phos- phate containing a small amount of sodium tri-citrate. The fat is dissolved in isobutyl alcohol, which is called " sinol " in this method. In the Pilsner method/'^ the butyrometer is filled with 11 c.c. of alkaline Solution (5 gm. of sodium phosphate, 15 gm. of neutral sodium citrate, 30 gm. of sodium chlorid, 65 gm. of sodium hydroxid dissolved in 600 c.c. of distilled water and filtered), 0.5 c.c. of isobutyl alcohol and enough sudan iii to color, and 10 c.c. of milk and, after thoroughly mixing, the butyrometer and contents are placed in a water-bath at 58°-62° C. and the temperature kept above 55° C. In a half hour the butyrometer is removed, centri- fuged and read. All the foregoing procedures, which include the important methods, are open to criticism either because of the time consumed in making the determinations or because of inaccuracy in the final results. THE MEIGS METHOD The original Meigs method. The Meigs method, described in this article, is neither extremely rapid nor simple but, everything considered, it possesses certain advantages over each of the above 1^ Hals and Gregg (Wollny method) : Milch-Ztg., 1902, xxxi, p. 433. 15 Maccagno and Mizzi : Cheni. Centr., 1906 (3 qt), p. 633 (Rev. intern, falsific, 1906, xix, p. 55). 16 Cornalba (Sichler method) : Chem. Centr., 1905 (ii), p. yy (Stas. sperim. agrar. ital., 1905, xxxviii, p. 227). 1'^ Kundrat and Rosam (Pilsner method) : Chem. Centr., 1907 (i qt.), p. 513 {Milchwirtschaft. Zentr., 1907, iii, p. 20). 514 Quantitative Determination of Fat in Milk [July methods that make it preferable In biochemical and pediatric work. The time consumed is as short as in any of the above methods, except the Babcock or other simple centrifugation methods, the simpHcity of the apparatus compares favorably with that of any other method, and the degree of accuracy with ordinary care and skill is so great that it entirely suffices for all except the most precise work. This method was described by Dr. Arthur V. Meigs^^ before the Philadelphia County Medical Society, on Feb. 22, 1882, It is, in brief, as foUows : Approximately 10 c.c. of milk, after being care- fully weighed, are transferred to an ordinary 100 c.c. graduated cylinder, 20 c.c. of distilled water added and, after this, an equal amount of ether (0.720 sp. gr,). The ground-glass Stopper is inserted and the bottle shaken for five minutes. Then, after care- fully removing the stopper, 20 c.c. of ethyl alcohol are added, the Stopper re-inserted and the cylinder shaken again for five minutes. This, as soon as settling occurs, gives two distinct strata, the upper of which contains little but ether and fat, the lower contains the other constituents of the milk. The upper Stratum is now drawn off with a small pipette and transferred to a weighed glass dish. Then 5 c.c. of ether are added and pipetted off, five successive times, and these 25 c.c. are added to the ethereal Solution first removed. This is done to wash off the thin layer of fat and ether which was left behind at the first pipetting. The dish and contents are now transferred to a safety water-bath, protected from dust, the ether evaporated, and the residue placed in a hot-air oven, heated at a temperature below 100° C, and finally dried in a desiccator over sulfuric acid until the weight is constant. Dr. Meigs stated in his report that the idea for this method was gained from a paper by Hallock.i9 The results we have obtained by this method were fairly accu- rate but, in order to be absolutely sure that no lactose or protein material was weighed as fat and to be certain that in both this and the Soxhlet method the end-products were identical, we decided to dissolve the first fat product in absolute ether, filter through a small hardened filter paper, as in the Soxhlet method, into a weighed dish, evaporate the ether, dry the fat and weigh. 18 Meigs : Philadelphia Medical Times, 1882, xii, p. 660. 1^ Hallock : Amer. Jour. of Pharmacy, 1874, xlvi, p. 477. 1913] Walter Lewis Croll 5^5 Results obtained with the original Meigs method. While the results first obtained by this process seemed to compare very favorably with those obtained by the Soxhlet method, a series of investigations was begun to determine whether this was a mere coincidence or due to the efficiency of the method. For this pur- pose there were secured twelve samples of human milk f rom women in various stages of lactation and six of cow milk, from as many dairies so widely separated that in no two cases were the milks from the same herd. In all determinations the milk for both the Meigs and the Soxhlet methods was taken from one specimen, and at the same time, so that the samples were as nearly uniform as it was possible to get them. In every weighing the figures were recorded to O.Ol mg, The results are given in the accompanying table. The average difference in the results of the eighteen determinations recorded in the table is 0.0234 per cent. The data in the table show that the method is useful for the determination of fat in both cow and human milk, and that it is applicable to all stages of lactation and to all kinds of milk. The degree of accuracy is such that it can be used in practically all work ; the differences between the results obtained by this method and those by the Soxhlet, as may be seen from the individual determina- tions given in the table and from the average difference for the series, are so small that they fall within the limits of experimental error. A modification of the Meigs process. In the manipulation, it was found that the use of the pipette was exceedingly tedious, required a great deal of skill and practice, and that, even under the most favorable circumstances, the possibility of error was great. To obviate this danger, we devised a simple inexpensive piece of apparatus (Plate 7). If only ordinary care is exercised in its use, the possibility of error is practically eliminated, This apparatus renders use of the pipette unnecessary, there is no need for careful attention to the protein layer or to the end of the pipette, and the speed of the whole process of removing the ethereal Solution and washing away the last traces of fat is markedly increased. 5i6 Quantitative Determination of Fat in Milk [July Comparative data pertaining to the content of fat in milk A. Human milk Days of lactation Fat by the Soxhlet method Mean by the Soxhlet method Fat by the Meigs method Mean by the Meigs method Difference in favor of the Soxhlet method lO 3-254 3.266 3.260 3.228 3.241 3.234 +0.026 10 S-413 5-422 5-417 5-400 S-398 5-399 +0.018 4 s-278 S-2I5 S-246 S-22I S-I83 5.202 +0.044 33 6-I5S 6.IS3 6.154 6.149 6.151 6.150 +0.004 9 3.713 3-701 3-707 3.686 3.690 3.688 +0.019 3 2.391 2.40s 2.398 2.401 2.384 2.392 +0.006 2 3-958 3-943 3-950 3-923 3-935 3-929 +0.021 4 3-042 3.041 3.041 3-032 3.020 3.026 +O.OIS II 3-910 3-900 3-905 3-892 3-897 3-894 +O.OII 5 2.795 2.797 2.796 2.767 2.781 2.774 +0.022 6 3-642 3.627 3-634 3.622 3-618 3.620 +0.014 12 4-363 4-359 4.361 4-333 4-362 4-347 +0.014 B. Cow milk 1* 3-372 3-376 3-374 3-359 3-3SI 3-355 +0.019 2^ 2.478 2.4S3 2.480 2.422 2.399 2.410 +0.070 3* 3.148 3.146 3.147 3.122 3-130 3-136 +0.011 4* 3-759 3.736 3.747 3-722 3-714 3-718 +0.029 S* ?-954 2.925 2.939 2.900 2.846 2.873 +0.066 6* 3.235 3.249 3.242 3-238 3.220 3-229 +0.013 * Sample number. BiocHEMicAL Bulletin (Vol. II) Plate 7 ^---F ^ CROLL: NEW APPARATUS FOR USE WITH THE MEIGS METHOD FOR THE DETERMINATION OF FAT IN MILK. I9I3] Walter Lewis Croll 5^7 In the drawing of the apparatus (Plate 7), A indicates a 100 c.c. graduated measuring cylinder with a ground-glass stopper; B is a rubber stopper; C, a glass tube of small bore, which is welded to another tube, D, at the points where it passes through the latter. The lower portion of tube C is beut sharply on its length and the open end is ground. Tube D is enlarged at the top to facihtate the introduction of ether; at its lower end there is a bulb, SB, with small holes in its walls, which allow the ether poured into tube D to spray against the sides of the cylinder, thus washing them. F is a side arm of small-bore tubing welded to D near the Upper end. In forcing out the ethereal Solution of fat, a finger is held over the Upper end, D, and the Operator blows air through F, the Solution being ejected through E into a weighed glass dish. In case it is desired to increase the force of the washing spray, a finger may be placed over the Upper end of D and, by blowing into F (just as when driving out the ethereal Solution of fat), the pressure can be increased as much as may be desired. Before starting to force out the ethereal Solution of fat, the opening of the lower end of tube C is brought on a level with the surface of the protein layer by rais- ing or lowering the entire glass piece through the rubber stopper, B. The shaking is done, of course, with the ground-glass stopper inserted tightly into the cylinder just as when the pipette is used, and, after a few minutes, the stopper is carefully washed with absolute ether and the special apparatus inserted. Advantages o£ the modified Meigs method. The improved Meigs method possesses the following advantages over the Soxhlet : (i) The time in weighing out is shortened and the danger of acci- dental loss is diminished, for no absorption of the milk by fat-free material is necessary. (2) Two or three hours are saved, as no drying is required. (3) The extraction requires ten minutes for its execution against three hours as the minimum for the Soxhlet process. (4) The apparatus is simple and inexpensive, while the Soxhlet is neither. The advantages over the other processes have already been men- tioned and are even greater than those over the Soxhlet method. The Meigs method has been tested against the Soxhlet process on six samples of cow milk, with the aid of the special apparatus 5i8 Quantitative Determmation of Fat in Milk [July dcscrihed ahove, and the differences in favor of the Soxhlet method have been f ound to ränge between + 0.007 P^'' cent. and + 0.059 per Cent., or about the same as when the pipette was used, but with miich more rapid and less difficult manipulation. I wish to express my thanks to Prof. John Marshall and Dr. Wm. H. Welker for suggesting this investigation, and for the kindly interest shown by them throughout its progress. THE OCCURRENCE OF ARSENIC IN SOILS J. E. GREAVES (Utah Experiment Station, Logan) Introduction. KunkeP showed the presence of arsenic in many rocks and waters, while Czapek^ states that traces are nearly always present in soils. Herzfeld and Lange^ found arsenic in certain German raw sugars, and traced it to the lime which had been used in the manufacture of the sugar. Headden^ found some virgin prairie soils relatively rieh in arsenic, an Observation in accord with my own experience. I have found arsenic to the extent of 4 parts per million in virgin soil ; and, as in the cases ref erred to by Headden, it did not result from smelter fumes or any such source, but was derived from the decay of native rocks. On the other band, Headden found arsenic in some cultivated orchard soils to the extent of 138 parts per million. He claims that in many places arsenic is accumulating in sufficient quantities to become injurious to Vegetation. Francois,*^ however, considers there is little danger of the earth becoming unfit for Vegetation from the proper use of insecticides. Grunner,^ who found arsenic to the extent of from 0.026 per cent. to 1.426 per cent. in the Reichenstein soil, is not so optimistic. It appears, however, that it is not so much the total quantity of arsenic present as the form in which the arsenic occurs, that determines toxicity. Little work has been done on this phase of the question. I have therefore determined the quantity of arsenic, both total and water-soluble, in many of the orchard soils of West- ern America, and it is the purpose of this paper to consider briefly a few of these results. Experiments. The water-soluble arsenic was determined by 1 Kunkel: Zeitschr. f. physiol. Chem., 1905, xHv, p. 511. 2 Czapek : Biochemie der Pflanzen, 1905, ii, p. 862. 3 Herzfeld and Lange: Chem. Abs., 1911, v, p. 2342. * Headden : Proc. Col. Sei. Soc, 1910, ix, p. 345. ^ Francois : Rev. de chim. ind., 1912, xxiii, p. 124. ^ Grunner: Landw. Jahrb., 1910, xl, p. 517. 519 520 Occiirrence of Arsenic in Solls [July extracting, for eight days, 500 grams of soil with 2000 c.c. of carbon-dioxide-free distilled water, and then using an aliquot part, while the total arsenic was determined by extracting the soil with nitric and sulfuric acids, and applying the Marsh method so modi- fied that the iron did not interfere.'' The results are given in the accompanying table ( i ) as parts per million of dry soil. TABLE I Data pertaining to the quantities of arsenic in soils: parts per million Total Water sol. Per Cent. Total Water sol. Per Cent. Total Water sol. Per Cent. arsenic arsenic soluble arsenic arsenic soluble arsenic arsenic soluble 102 0.92 0.9 36 3-48 9.67 16 0.72 4-50 79 6.20 7.85 33 5.16 15.63 15 1.74 11.60 64 4.1 6.41 32 3-92 12.25 13 0.84 6.46 63 6.88 10.92 32 1.07 3-34 12 3.85 32.08 63 1.02 1.62 24 3.56 14.83 12 0.70 5-83 62 3-38 5-45 24 1.32 6.33 II 3-40 30.91 60 I.OO 1.67 20 1.08 5-04 II 1.36 12.36 SS 8.87 16.13 19 6.08 30.20 10 0.88 8.80 50 4.68 5.36 19 Traces 9 3-13 34.78 45 4-3 9-55 18 1.36 7-56 5 3-08 61.60 45 1.8 4.00 18 0.48 2.67 5 1.08 20.80 40 5-2 13.06 17 0.36 2.12 5 1.08 21.60 39 0.68 1.74 16 2.50 15.60 From the data in the table it may be seen that some orchard soils contain large quantities of arsenic and some carry compara- tively large proportions of it in the water-soluble form; but there is no uniform relationship between the total arsenic in soils and the water-soluble arsenic. If, as is most likely the case, the injury to plants is due to the water-soluble arsenic, greater injury would result in a soil containing only 5 parts per million of total arsenic with 61.60 per cent. of the arsenic water-soluble, than in a soil con- taining 102 parts per million, in which only 0.9 per cent. of the arsenic is water-soluble. It may be seen, further, that one of the above soils, which contains 63 parts per million of total arsenic, has only 1.62 per cent. in soluble form, while another soil, having exactly the same amount of total arsenic, contains 10.92 per cent. in soluble form. Arsenic in soil is not confined to the surface. One soil, obtained "^ Greaves : Jour. Amer. Chem. Soc, 1913, xxxv, p. 150. 1913] J- E. Greaves 521 at a depth of three feet, yielded 11 parts per million of total arsenic, 30.91 per Cent, of which was water-soluble. It may be safely concluded, from the above data, that some virgin soils contain arsenic in large quantities, but that the propor- tion in a soil is no index of the amount which is soluble in water. The latter is probably governed by many factors, e. g., kind of soil, water-soluble salts in it, and form in which the arsenic was applied to the soil. The latter factor has been tested by applying the same quantity of different forms of arsenic to different portions of the same soil and then determining the quantity of water-soluble arsenic present in the soil. The soil used was a typical bench soil — a sandy loam — fairly high in content of calcium and iron, and supplied with an abundance of all the essential Clements of plant food with the exception of nitrogen, which was low as is characteristic of the arid soil. The proportion of water-soluble salts in the soil was low. The influence of the soil on the solubility of the arsenical insecti- cide was determined as follows: Quantities of lead arsenate (21.96 per Cent, of arsenic), Paris green (47 per cent. of arsenic), zinc arsenite (31.25 per cent. of arsenic) and arsenic trisulfide (60 per Cent, of arsenic), were added to 100 gm. portions of soil in quanti- ties sufficient to give 112 mgm. of arsenic per 100 gm. of soil. The soil and arsenic, together with 2 gm. of dry blood, were placed in sterile tumblers, covered with Petri dishes, the water content made and kept at 18 per cent.; and then each mixture was incubated at 28° C. for three weeks. At the end of this time the soil was trans- ferred with 1000 c.c. of carbon-dioxide-free distilled water, to large acid bottles. The mixture was left in these bottles with occa- sional shaking for eight days, then filtered and the arsenic deter- mined in an aliquot part. In another set each quantity of insecticide was mixed with a 100 gm. portion of soil and 2 gm. of dry blood, and the water-soluble arsenic determined as above, without incuba- tion. All determinations were made in duplicate. The data are given in Table 2, as mgm. of water-soluble arsenic in 100 gm. of soil either before or after three weeks' incubation (112 mgm. of arsenic had been added to each.) 522 Occurrence of Arsenic in Soils [July TABLE 2 Data pertaining to ihe water-soluble arsenic in soils mixed with arsenical insecticides Treatment Lead arsenate Paris green Zinc arsenite Arsenic tri- sulfide Incubated three weeks. Water- soluble arsenic determined . . . Water-soluble arsenic deter- mined direct mgm. 14-3 20.2 mgm. 80.80 82.00 mgm. 36.9 31-7 mgm. 50.0 5-6 Average 17-3 81.40 34-3 27-3 These results show that there may be a great difference in the quantity of water-soluble arsenic existing in the same soil to which various forms of arsenic have been added in equivalent quantities ; and that even a soil comparatively rieh in iron and calcium, to which arsenic has been added in large quantities, may have a high water-soluble content of arsenic. It is much higher when arsenic is added in the form of Paris green, than when added in the other forms mentioned. The solubility of the Compounds, with the excep- tion of arsenic trisulfide, is not greatly changed on standing in a soil containing a large quantity of decomposing organic matter. The results also show the superiority of lead arsenate over any of the other arsenical insecticides. Any injurious effect of such Compounds on plants must be proportional to the available amount of water-soluble arsenic, not to the total arsenic. In the use of an arsenical insecticide it should be the rule to select a Compound which would remain insoluble for the greatest possible length of time. Lead arsenate possesses this advantage in high degree. Summary. Some virgin soils contain arsenic in appreciable quantities which comes from the decay of the native rocks. Many cultivated orchard soils contain it in large proportions, but there is no uniform relationship between the total quantity of arsenic in different soils, and the water-soluble arsenic of these soils. A soil containing over 100 parts per million of total arsenic contained much less water-soluble arsenic than did a soil carrying only 5 parts per million of total arsenic. The solubility of the arsenic found in a soil is governed largely by the salts in the soil and the form in which the arsenic is applied. Different portions of the same soil, 1913] J- E. Greaves 523 to which equivalent quantities of various so-called insoluble arsen- ical Compounds had been added, showed great dissimilarities in water-soluble arsenic content. The portion to which Paris green was added contained four times as much water-soluble arsenic as did a portion of the same soil to which an equivalent quantity of lead arsenate had been applied. Arsenic trisulfide, when first applied to soil, is less soluble than lead arsenate, but as time progresses, at least in some soils, the arsenic trisulfide becomes more soluble. For this reason lead arsenate is probably safer than any of the other arsen- ical insecticides. FURTHER NOTES ON THE RELATIONSHIP BETWEEN THE WEICHT OF THE SUGAR BEET AND THE COMPOSITION OF ITS JUICE J. ARTHUR HARRIS and ROSS AIKEN GORTNER (Biochemical Laboratory of the Station for Experimental Evolution, Cold Spring Harhor, L. I.) (WITH PLATE 8) 1. Introductory remarks. In an earlier paper^ we discussed in terms of correlation and regression the relationship between the weight of the sugar beet and the composition of its juice (total soHds, sucrose and purity). At that time (October, 1912) we were unaware of any previous attempt to deal with the problem by corre- lation methods. It was with considerable pleasure, therefore, that we found that only a few months before (January, 191 2) Andrlik, Bartos and Urban^ had considered some of the same problems on the basis of data derived from pedigreed strains, and had even gone so f ar as to form correlation tables. They have not, however, calcu- lated the constants which are essential to a füll understanding of the tabled data. For this reason, and because their conclusions differ from our own, we have thought it worth while to take up the problem again. 2. Analysis of data. Although they discuss six series, Andrlik, Bartos and Urban give only one table from which a correlation 1 Harris, J. Arthur and Ross Aiken Gortner ; On the relationship between the weight of the sugar beet and the composition of its juice: Proc. Columbia Biochem. Assoc, Biochemical Bulletin, 1913, ii, p. 287, and Journ. Industrial and Engineering Chem., 1913, v, p. 192. 2 Andrlik, K., V. Bartos and J. Urban ; Über die Variabilität des Gewichtes und des Zuckergehaltes der Zuckerrübenwurzeln, und über die gegenseitigen Beziehungen dieser beiden Merkmale: Zeitschr. f. Zuckerindustrie in Böhmen, 1912, xxxvi, p. 193. 524 I9I3] /. Arthur Harris and Ross Aiken G ortner TABLE I Weight of root in grams 525 Sugar content in percents 300 to 350 3SO to 400 400 to 450 450 to SOG SOG to 550 550 to 600 600 to 650 650 to 700 700 to 750 75° to 800 800 to 850 850 to 900 Totals 16.I-16.5 16.6-17.O 17.I-17.S 17.6-18.O 18.I-18.S I8.6-19.O 19.1-19-5 19.6-20.0 20.1-20. 5 20.6-21.0 I 4 7 4 I 2 3 5 12 20 15 8 2 4 I I 4 15 38 35 22 2 I 2 7 13 33 30 28 8 I 2 8 48 32 30 17 I I I 5 10 25 23 14 6 2 5 7 17 19 14 7 I I 6 17 15 16 2 I 2 3 4 I 4 2 I I I I 2 7 30 76 211 179 138 47 10 Totals . . . 19 69 1x8 123 138 87 70 57 II 7 I I 701 coefficient can be determined.^ From their summary, reproduced in slightly modified form in the above table, we deduce, using the con- venient formula ''«.= '2{w's')/N—ws LÜ D OQ _l < Ü I Ü o CO L -m _^ „"^ < ■ — • ^ - ' ' - - ^ < " 1» / l 4 / Vi »* \ \ c »: «_ \ •* \ »: »* k? r V J 1 \ w. H ' \ '^ i w 1 r " « »t »» * 1 1 •i 1, 1 w 1 a ■ L H W w CQ oi < o D m W ffi H IX C H X . O o w rt wH' Pi r^. r- c W CA U K H O H ■- o O tn > c c/i f- - et w t -^ ö c/> t; ^£ .^ ^ Sa ^ CO O ;z; L> O 2w O) H ffi t- < H ■*-» J O W Q —^ c^ 2; .. < C£^ w; :z; H- C^ O C7 o ^ < CA) 1 — k P4 fC < X "(uaiuoj^ jESng uB3p\[ 1913] J- Arthur Harris and Ross Aiken G ortner 527 value of the data been completely destroyed by the reduction of all frequencies to percentages. We must confess that these evidences of gross carelessness in the treatment of their numerical data lessen our confidence in the conclusions they and their reviewers have drawn from it. Turn now to three other series of analyses by Andrlik and Urban made for a different purpose.*^ They give the weight, sugar content and nitrogen'^ content of 36 beets grown from normal- nitrogen-content mothers and the same data for 36 roots grown from high-nitrogen-content parents. The first of these we designate as Series A, the second as Series B. They also give 100 analyses of beets from the same parent individual. We refer to these as C. For weight and sugar the correlations are : For Series A, r„, = — 0.278 ± 0.104 For Series B, r^, = — 0.456 ± 0.089 For Series C, r^, = — 0.031 ± 0.067 All the relationships are negative. The probable errors are high because of the small number of beets given, but the two first cases may perhaps be regarded as statistically significant in comparison with their probable errors. They are in excellent agreement with the correlations published in our first paper. We have also deduced the coefficients for weight and nitrogen, n. They are : For Series A, r„„ = — 0.234 ±0.106 For Series B, r„„ = - 0.393 =*= oo95 For Series C, r„„ = + 0.054 =^ 0.067 One of the constants is positive in sign but less than its probable error. The two negative values are fairly large and possibly sig- nificant in comparison with their probable error. Still another series of data^ of quite a different sort affords, it 8 Andrlik, K. and J. Urban ; "Über die Variabilität des Stickstoffgehaltes in Zuckerrübenwurzeln : Zeitschr. f. Zuckerindustrie in Böhmen, 1912, xxxvi, p. 513- ''■ Milligrams of nitrogen in 100 grams of root. 8 Novotny, K. ; Ein Beitrag zur Betrachtungen über die Beziehungen zwischen dem percentuellen Zuckergehalte und dem Gewichte der Rüben : Zeitschr. f. Zuckerindustrie in Böhmen, 1912, xxxvi, p. 269. 528 Notes 011 the Siigar Beet and its Juice [July seems to us, the strongest evidence for a negative relationship between the weight of the beet and its sugar content in commercial cultures. Novotny selected from his laboratory record books analyses of samples of beets which had been divided into two sub- samples, one of large and one of small beets. Altogether there are 2y of these pairs of samples, taken from 1892 to 1910. With only two exceptions (in which the sugar percentage was identical in the two cases) the sugar content of the lighter was higher than that of the heavier fraction.^ 3. Summary and discussion. There is a more or less wide- spread idea that the percentage sugar-content of large beets is lower than that of small roots. This belief, which has often been opposed, was placed on a scientific basis for American commercial cultures by an earlier study, in which we showed that there is a significantly negative (and sometimes numerically very substantial) correlation between the weight of the root and its total solids, its percentage sugar-content and its coefficient of purity. The strongest evidence in Support of our conclusions (so far as sugar content is concerned) is furnished by the data of Novotny. Recently, however, wide cur- rency has been given to the conclusions that there is no necessary negative relationship between weight of root and composition of Juice, and that when analyses are made of a series of beets derived from the same mother plant no such correlation is demonstrated. The data upon which this Statement has been made seems to be six series by Andrlik, Bartos and Urban. Of these the figures for one series only are given in a form really suitable for Statistical analysis. This gives a low positive correlation. The tables for the other five series contain so many obvious inconsistencies that they cannot be used. But three other series by Andrlik and Urban, also from beets derived from an individual mother but analyzed for a different purpose, all give negative correlations between weight and sugar content. One of these is numerically insignificant ; the other ö Notwithstanding the fact that his data bear evidence without a single ex- ception against the Statement that there is no negative relationship between weight of root and composition of juice, Novotny attempts, by the use of a formula which seems to us to have no theoretical justification, to show that the difference in sugar content between large and small beets is becoming smaller and that his results are consequently in accord with those of Andrlik, Bartos and Urban. igi3] /. Arthur Harris and Ross Alken Gortner 529 two are of the order r = — 0.300. Thus, of the four series of data which can be scientifically analyzed, three give correlations which are negative in sign while one is positive. One negative and one positive coefficient are very low; two of the negative coefficients are more substantial, agreeing fairly well with the values previously pub- lished by us. Surely such facts as these form a very slender basis for the con- clusion (widely circulated by uncritical reviewers) that in beets of the same strain there is no negative correlation between weight and sugar content ! Nevertheless one must recognize the possibility of the correctness of the conclusion. Should it prove to be valid, the Suggestion follows that the negative correlation demonstrated in commercial cultures has a genetic origin, i. e., that strains character- ized by large root size are also characterized by low sugar content, and that when these strains are intermingled and intercrossed in field cultures there results a negative correlation between the weight of the individual beet and the sugar content of its juice. Such a result would be of the greatest interest to breeders. NOTE ON THE RELATIONSHIP BETWEEN BARO- METRIC PRESSURE AND CARBON-DIOXIDE EXCRETION IN MAN Higley's application of the product-moment correlation method to the question o£ the influence of barometric pressure on carbon- dioxide excretion in man^ seems to deserve some extension. For- tunately, this is possible on his published data. Instead of inquiring merely whether there is a correlation be- tween barometric pressure and carbon-dioxide excretion, one may profitably consider (a) whether the volumes excreted by the same individual at different periods in the day are correlated, and (b) whether the amounts excreted by different individuals on the same day are correlated. The correlation between the volumes respired by the same indi- vidual at different times of Observation on the same day might be due either to internal physiological conditions,^ or to Variation f rom day to day in external conditions (barometric pressure, or some other environmental factor). A correlation between the volumes excreted by different individuals on the same day would necessarily be due to some common external condition. Designating Higley's subjects by a^ b and c, and the three Ob- servation periods by m^^morning, w = noon and ^=evening, I find the following results.^ A. For correlations between different Observation periods for the same individual: For a, rmn = + 0.30 ±0.12, rme= — O.I5=bO.I3, rne = -|- 0.00 ± O.I3 1 Higley : Biochemical Bulletin, 1913, ii, p. 393. 2 These might conceivably tend to bring about either a positive or a nega- tive correlation. 3 In determining the correlations, the means and Standard deviations were necessarily recalculated for each product-moment, since several observations are wanting. Thus N varies from coefficient to coefficient. 530 I9I3] J' Arthur Harris 53 1 For b, rmn = — 0.53 ±0.10, rme = + 0.19 ±0.14, ^ne = O.O/rtO.Iß For c, rmn = + 0.04 ±0.13, rme = + 0.21 ±0.13, ^ne = + O.O/ ± O. I4 B, For correlations between the excretions of different indi- viduals on the same day : For 7 A. M., ro&== — 0.00 ±0.14, roc = + 0.46 ±0.10, nc= + 0.00 ±0.14 For 12 M., ro&= + 0.20 ±0.12, roc = + 0.31 ±0.12, r6c== + 0.08 ±0.13 For 5 P. M., foö = 4- 0.15 ±0.13, roc = + 0.07 ±0.14, nc = + 0.03 ±0.14 In each case three of the values are negative and six positive in sign, They thus tend to confirm Higley's Suggestion concerning the relationship between common external conditions and volume of carbon-dioxide excreted. But the probable errors are high (because of both the lowness of most of the coefficients and the smallness of the number of observations) and the constants are very irregulär. For trustworthy conclusions larger series are necessary. When these are available, the correlations here suggested may, I believe, be of supplementary value. J. Arthur Harris The Carnegie Institution of Washington, Station for Experimental Evolution, Cold Spring Harbor, Long Island, N. Y. THE BLEACHED FLOUR DECISION May I be permitted to answer the editorial signed " M. C," on "The Bleached Flour Decision," which appeared in the April issue of the BiocHEMicAL Bulletin (p. 487) ? While I was associated with Professor Alway, at the Nebraska State Experiment Station, we had occasion to spend the greater part of a year investigating the subject of the bleaching of flours by the Alsop process, i. e., the use of nitrogen peroxide.^ At the Start I may State that the work was not done under any grant from the miliers, or others interested in the process, but was under a grant from the Adams Fund ; and that all conclusions were based solely upon the experimental evidence. I may also add that at this time I have no interest in the matter except to see that facts are stated. I. The "poison" to which " M. C." refers is the nitrite-reacting substance (NaNOg ?) which is present in bleached flours and gives the reaction with the Griess-Ilosvay reagent. This "poison" is present in bleached flours not to exceed 6-10 parts per million, and usually in much smaller quantities. A greater proportion may be introduced by "over bleaching," but in over bleached flours a dirty yellow color is produced and the flour is ruined commercially. When a flour which has been bleached is baked, all, or nearly all, of the nitrite-reacting substance is destroyed, and it very rarely happens that loaves of hread made from bleached flours and un- bleached flours can be distingiiished from each other by the nitrite test. .This is especially true if the baking has been done in a gas- fired Oven, for under such conditions bread made from unbleached flour will give the nitrite test. Aside from the nitrite test, there is only one other distinguishing feature, since the baking quality of the flour, the expansion of the gluten, and the odor, taste, weight, lightness, and texture of the 1 See Bull. No. 102, Nebr. Agric. Exper. Sta. (The Effect of Bleaching upon the Quality of Wheat Flour). 532 1913] Ross Alken Gortner 533 bread are the same, whether made from bleached or unbleached flours; but, in all cases, the bread made from high grade bleached floiir is whiter and more inviting in appearance. Eating bread made from bleached flour has, as I personally know, produced no ill effects, even when the test lasted for months at a time. The saliva of normal human beings almost always contains nitrites, and the quantity of nitrites secreted by the average indi- vidual in 24 hours in the saliva is far in excess of the quantity which we found in the entire loaf of bread highest in nitrites. In order to obtain the medicinal dose of the '' poison" (NaNOz), it would he necessary to eat at least a poiind loaf of bread each day for a year. 2. "M. C." also adds : "If it is not to conceal inferiority so that a higher price can be had for the flour, why do miliers use the process?" In the first place our experiments show that low grades of flour cannot be successfully bleached, and, when bleached, can in no case be confused with a high grade flour, for the color of the small particles of bran in a low grade flour is not affected by the nitric fumes and the bread produced from such a flour has an uninviting color. It is merely a question as to where the wheat comes from, that causes bleaching. At the last trial in St. Louis one of the witnesses for the Government testified and protested against the use of the bleachers, although he was himself a milier. On cross examination he admitted that he had formerly used the process but had aban- doned it. On further questioning it developed that, while he was in the "yellow wheat belt," he had used the bleacher but now, in Kentucky, the wheats gave a white flour, and he did not need to use the bleacher to get the same resitlt. The wheats of Nebraska, Kansas, Iowa, etc., have a dark yellow oil which, when milled, gives a very yellow tint to the flour. This answers the " WHY " : it is because the farmers of Nebraska, Iowa, Kansas, etc., wish to produceas saleable a flour as do their neighbors. The influence of the minute quantities of NO2 changes this yel- low oil into a colorless Compound : very probably it is the old reaction of oleic acid changing into elaidic acid. 3. I am heartily in accord with the Pure Food and Drugs Act, even if I do seem to fall in the class mentioned in the final para- 534 The Bleached Flour Decision [July graph by "M, C." But why do we need to go to such extremes? To quote "M. C." — ^the fifth section reads: "An article shall be deemed adulterated — if it contains any added poisonous or other added deleterious ingredient which may r ender such article injurious to health." Why not prohibit the addition of salt to the bread? Stefansson teils us that, in the Arctic when he was unable to pro- cure salt, he experienced dizziness and all of the cravings due to the lack of a customary narcotic, but his health was far better after the cravings were over, and that, on returning to civilization, salted foods were decidedly distasteful to him and he had to acquire a taste for them again. Certainly, if the salt taken in bread were increased in amount several hundred fold, physiological disturb- ances might occur, and poisonous Symptoms be observed. The only way we can use a law is to give it a sane interpreta- tion. I do not protest against the branding of the flour " bleached," That is all right ; we have a right to know what we are buying, but I do protest against forbidding its inter-state transportation when no harmful effects have ever been demonstratedj even by the widest Stretch of Imagination. RoSS AlKEN GORTNER Carnegie Institution of Washington, Station for Experimental Evolution, Cold Spring Harbor, L. /., N. Y. EMIL CHR. HANSEN FUND Pursuant to the last will and testament of the late Professor Emil Chr. Hansen and bis wife, a Fund bearing bis name bas been establisbed, tbe Statutes of wbicb were ratified by tbe Danisb Gov- ernment on June 17, 191 1. At proper intervals, as a rule every two or tbree years, beginning in the year 191 4, a Gold Medal bear- ing Hansen's effigy and accompanied by a sum of (at least) Five hundred dollars (2000 Kroner) is to be awarded on the donor's birthday, the 8th of May, to the author of a distinguished publica- tion on some microbiological subject that has appeared of late years in Denmark or elsewhere. The Fund is committed to the administration of the Chiefs of the two departments of Carlsberg Laboratory, together with a Danish biologist elected by the governing body of Carlsberg Laboratory. ^ The person to whom the Medal is to be assigned shall be desig- nated by a Committee composed of the above mentioned Trustees of the Fund, together with at least two f oreign microbiologists, who, at the request of the said Trustees, will have accepted appointment to membership in the Committee. Professeur et Dr. Calmette, Lille ; Geh. Ober-Med.-Rath Prof. Dr. Gaffky, Berlin; and Professor Theobald Smith, M.D., Boston, have become members of the Committee. It is proposed to award the medal, in 19 14, to a scientist in the field of Medical Microbiology {comprehending the morphology, biology and mode of action of the microbes generative of disease in the human or animal body). All Communications regarding the Hansen Fund should be sent to, and all further particulars will be given by, the President of the Board of Trustees. Board of Trustees: Professor C. O. Jensen, Dr. med., Serum Institute of the Royal Veterinary and Agricultural College; JoHS. Schmidt, Dr. phil., Physiological Department of Carlsberg Laboratory; Professor S. P. L. Sörensen, Dr. phil., President of the Board of Trustees, Chemical Department of Carlsberg Laboratory. Copenhagen, Valby, Denmark, June, 1913. 535 BIOLOGICAL CHEMISTRY IN THE PHILIPPINES There are, at present, no laboratories of biological chemistry in the Philippines, biit there are well equipped chemical and biological laboratories at the University of the Philippines in Manila, at the Agricultural College in Los Baiios and at the Bureau of Science. The course in biological chemistry in the College of Medicine and Surgery is under the control of the Department of Chemistry in the University, and the small biochemical laboratory is temporarily used for pharmacy. It is planned in the near future to put the biochemistry in the Department of Physiology. Research in the Department of Pharmacology has been in the biochemical field, and this laboratory is well equipped for the work. Research in the Islands is largely concerned with local and tropical problems. Aron's work on Philippine rice and on beriberi, and Freer's investigations of tropical sunlight are recent examples. Gibbs has studied the chemistry of the mongo bean, an important article of diet in the Orient. Coöperative work, by the Board for the Study of Tropical Diseases of the U. S. Army and the chemists in the Bureau of Science, on the protective principle in rice polish- ings, may be mentioned. Andrews has reported the analyses of samples of milk from native women, whose infants have died of beriberi — puppies, suckled by these women, developed the disease. Shaklee is at present carry ing out an extensive series of experi- ments on low nutrition under tropical conditions. The satisfactory conditions for work, particularly the large amount of available material out here, should appeal to the biolog- ical chemist. The Bilibid prisoners can be used as exactly controlled subjects for metabolism work; and normal fresh human organs are obtainable at the not infrequent prison executions. There is an abundance of autopsy material. Cases in the Philippine General Hospital can be studied experimentally. Animals, such as dogs and monkeys, are easily obtained. The native laboratory assistants are quickly trained, and make good technicians. There is an excellent 536 1913] R- B. Gibson 537 library at the Bureau o£ Science. Finally, living conditions are good and salaries are liberal. It is to be hoped that the near future will find more biological chemists in the Islands. R. B. Gibson Department of Physiology, College of Mediane and Surgery, University of the Philippines. DOCTORATES IN BIOLOGICAL CHEMISTRY Conferred by American Universities, ig 1 2-' 13 ^ The names of recent reciplents of the Ph.D. degree in biochem- ical science, with the subjects of the dissertations, are arranged below in university groups : Columbia University.^ — Joseph Samuel Hepburn: Biochemical studies of cholesterol. — Benjamin Horowitz: A study of the action of ammonia on thymol. — Edgar Grim Miller, Jr.: Studies in patho- logical chemistry; (I) Enzymes as possible factors in the develop- ment of edema, (II) The lecithin content of the blood in syphiHs, (III) Studies on dental caries. — Anton Richard Rose: Biochemical studies of phyto-phosphates. — George Gilmore Scott: A physiolog- ical study of the changes in Mustelus canis produced by modifica- tions in the molecular concentration of the external medium. — Clayton Sidney Smith: A study of the influence of cold-storage tem- perature upon the composition and nutritive value of fish. — Charles Weisman: Biochemical studies of expired air in relation to Ventila- tion. Comell University. — George Ellsworth Thompson: An experi- mental study of photoactive cells with fluorescent electrolytes. — Eleanor VanNess VanAlstyne: The absorption of protein without digestion. Harvard University. — Chaimcey J. Vallette Pettibone: The quantitative estimation of urea in urine. Johns Hopkins University. — Lon A. Hawkins: The influence of calcium, magnesium and potassium nitrates upon the toxicity of certain heavy metals toward fungous spores. University of Chicago. — Aaron Arkin: The influence of chem- ical substances upon immune reactions, with special reference to ^Additional information regarding the Columbia doctors (and masters) in biological chemistry is given on page 579, where may also be found the names of successful Ph.D. candidates in botany, chemistry and zoology, whose minor work was done, in part, in the Columbia departraent of biological chemistry. 538 I9I3] P. H. D. 539 oxidations. — George Lester Kite: The relative permeability o£ the surface protoplasm of animal and plant cells. — Shiro Tashiro: Chemical change in nerve fiber during passage of a nerve impulse. — 'Arthur Lawrie Tafum: Studies in experimental cretinism, University of Missouri. — Leroy Sheldon Palmer: Study of the natural pigment in the fat of cow milk. University of Wisconsin. — William Harold Peterson: Forms of sulfur in plants. — Roy Lee Primm: Some phenomena associated with cellulose fermentation. — Nellie 'Antoinette Wakeman: Plant pigments other than chlorophyl. Washington University. — Jacob Richard Schramm: A contri- bution to our knowledge of the problem of free-nitrogen fixation in certain species of grass-green algse, with special reference to pure- culture methods. — Charles Oscar Chambers: The relation of algae to dissolved oxygen and carbon dioxide, with special reference to carbonate. Yale University. — Robert Bengis: The synthesis of amino acids related to adrenalin. — Howard Bishop Lewis: The behavior of some hydantoin and thiohydantoin derivatives in the organism, together with a study of certain related sulfur Compounds. — Ben Harry Nicolet: Some derivatives of amino-malonic acid and their biochem- ical interest. — Ruth Wheeler: Nutrition experiments with mice. Universities which conferred Ph.D. degrees in the natural and exact Sciences, but at which there were no biochemical candidates, are named below : Boston University University of California Brown University University of Cincinnati Clark University University of Illinois George Washington University University of Iowa Indiana University University of Michigan Massachusetts Institute of Technology University of Minnesota New York University University of Nebraska Princeton University University of Pennsylvania Stanford University University of Pittsburgh Tulane University University of Virginia Vanderbilt University 540 Doctorates in Biological Chemistry [july Comparing the foregoing list with the data published a year ago,2 we note the main points of interest shown by the appended summary : Number of Ph.D. degrees awarded by American universities to biochemical candidates, 1913 and 1913. 1912 1913 Total Women 1912,, 1913 Brown University i o i 0 o Columbia University il 7 18 l 0 Cornell University 5 2 7 o 1 Harvard University i i 2 o o Johns Hopkins University l l 2 o O University of California 5 o 5 i 0 University of Chicago 8 4 I2 l o University of Illinois 5 0 S o 0 University of Michigan 2 o 2 o O University of Missouri o i i 0 O University of Wisconsin 4 3 7 o l Washington University o 2 2 o o Yale University 6 4 10 l i Total 49 253 74 "4 y P. H. D. 2 Biochemical Bulletin : 1912, i, p. 546. 3 The decrease from 49 in 1912 to 25 in 1913 accords with a general tend- ency: the number of doctorates in the "natural and exact sciences" decreased from 273 in 1912 to 231 in 1913, whereas the doctorates in the " humanities " increased from 209 in 1912 to 230 in 1913 — totals of 482 in 1912 and 461 in 1913. TWELFTH SCIENTIFIC MEETING OF THE COLUM- BIA UNIVERSITY BIOCHEMICAL ASSOCIATION, AT THE COLLEGE OF PHYSICIANS AND SURGEONS, NEW YORK, JUNE 2, 19131 Proceedings reported by THE Secretary, ALFRED P. LOTHROP The twelfth scientific session (fourth " anniial" meeting) o£ the Columbia University Biochemical Association was held at the Columbia Medical School, at 8.15 p. m., on June 2, 1913.^ Ab- stracts of the papers are presented here (pages 542-558) in two groups : (A) Abstracts of papers on research by non-resident members^ and (B) abstracts of papers from the Columbia Bio- chemical Department and affiliated laboratories. The appended summary facilitates reference to the abstracts (86-107).* A SUMMARY OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE SUCCEEDING ABSTRACTS (86-107) A Max Kahn. MetaboHsm studies of five cases of endarteritis obliterans ("Hebräische Krankheit"). (88) Daniel R. Lucas. On the content in expired air of protein detectable by the anaphylactic reaction. (89) Max Morse. On the comparative physiology of creatin and Creatinin. (90) Matthew Steel. Influence of elec- tricity upon metabolism. (91) A. F. Blakeslee and Ross Aiken GoRTNER. On the occurrence of a toxin in juice expressed from the bread mould, Rhisopus nigricans (Mucor stolonifer). (86) Ross Aiken Gortner. Studies on melanin : V. A comparison of cer- tain nitrogen ratios in black and in white wool from the same animal. (87) 1 Scientific meetings are held regularly on the first Fridays of December, February and April, and on the first Monday in June. ~ Proceedings of the ninth, tenth and eleventh scientific meetings were published in the last (April) number of the Biochemical Bulletin at pages 452, 461 and 486. 3 Members of the Association who were not officially connected with the Columbia Biochemical Department when the researches were conducted. * Previous abstracts were published in the Biochemical Bulletin : 1-44, 1912, ii, p. 156; 45-62, 1913, ii, p. 28s; 63-72, 1913, ii, p. 452; 73-85, IQIS, ii, p. 462. 541 542 Proceedings Columbia Biochemical 'Association [July B Louis Berman and William J. Gies. Studies of intracellular chemistry: A differential stain for mucins and mucoids. (92) Walter H. Eddy. Two new histons. (93) Walter H. Eddy. Histon nucleo- protein : A protein salt. (94) Frank R. Elder. Further experi- ments on the preparation of modi- fied collodion membranes for use in dialysis experiments. (95) Samuel Gitlow. Comparative studies of the permeabihty of collodion and collodion-fat membranes. (96) TuLA L. Harkey. Further studies of edema : On the absorption of water by white lupin seeds. (97) TuLA L. Harkey. Further studies of edema: On the postmortem absorp- tion of water by tissues from well nourished and fasting animals. (98) Paul E. Howe and William J. Gies. A preliminary study of the resist- ance of fasting dogs to hemorrhage. (99) V. E. Levine. Biochemical studies of selenium. (100) Helen I. Mattill and H. A. Mattill. The influenae of electrolytes on the precipitation of soluble starch. (lOl) Edgar G. Miller, Jr. Determinations of the acidity of fruit Juices. (102) Olive G. Patterson. A study of the influence of external hemorrhages on the partition of urinary nitrogen. (103) P. W. PuNNETT. The action of a high frequency current on the ac- tivity of pancreatic amylase. (104) Christian Seifert and William J. Gies. A further study of the dis- tribution of osseomucoid. (105) A. W. Thomas. A further effort to prepare a colorless biuret reagent. (106) Charles Weisman. Biochemical studies of expired air in relation to Ventilation. (107) A. ABSTRACTS OF PAPERS ON RESEARCH BY NON-RESIDENT MEMBERS5 86. On the occurrence of a toxin in juice expressed from the bread mould, Rhizopus nigricans (Mucor stolonifer). A. F. Blakeslee and Ross Aiken Gortner. {Biochemical Laboratory of the Station for Experimental Evolution, The Carnegie Institu- tion of Washington.) During a series of immunity studies, having as their aim a possible Solution of the chemical nature of sex, we observed that the "presssaft" from the aerial filaments of Rhizopus nigricans caused almost instant death when injected intravenously into rabbits. Several other species of the Mucorineae were tested and no such result has yet been obtained. The mycelium directly in contact with the substratum apparently contains as great a quan- s Members of the Association who were not officially connected with the Columbia Biochemical Department when the researches were conducted. I9I3] Alfred P. Lothrop 543 tity of toxin as do the aerial filaments, Furthermore, the toxin is present in both sexes of the fungus in large amount. We have not determined whether one sex contains a greater absolute quantity than the other. The toxicity o£ the Rhisopus extract may be better understood by saying that a Solution containing the water-soluble substances extracted from 0.045 S^- o^ ^^^ ^O^ fungus, when injected intra- venously, is sufficient to kill a 1.35 kilo rabbit in less than two minutes. Convulsions begin almost before the needle can be with- drawn, and are followed by great turgidity of the ehest and abdomen, then by relaxation of the rigid abdomen, throwing the head back- ward with cough-like movements of the diaphragm, protrusion of the eyes, and death. When a dose containing the toxin from a greater quantity than 0.045 g^i- is injected, the animal is often dead before the needle can be withdrawn, with no convulsive move- ments and only a sudden turning of the head and the sinking of the body on one side. A sub-lethal dose causes extreme lethargy, lasting for 48 hr. or more, during which time the animal moves only when forced to do so. This may be followed by complete recovery. We have not as yet elucidated the chemical nature of the toxin. Its activity is not diminished by peptic digestion for three hours, nor is it affected when its aqueous Solution is heated to boil- ing for 10 min. Apparently, therefore, it is not a toxalbumin. One of US (B) has shown that Rhizopus is nearly universally distributed and is almost certain to appear as an infection on starchy food under suitable moisture conditions, its occurrence being so common as to have earned for it the name, " Bread-mould." While connected with the Agricultural Experiment Station of the Univer- sity of Nebraska, one of us (G) coöperated in an Investigation of the origin of the "corn-stalk disease," which causes the death of thousands of cattle in the Middle West each year, and which is, in some way, connected with the use of corn stalks as fodder. At that time no known toxin could be detected in the stomachs of the diseased animals, and the direct cause of the disease has never been elucidated. The method of growth of the fungus, its wide distribution, as well as certain of the Symptoms produced by the toxin which it 544 Proceedings Columbia Biochemical Association [July contains, would seem to indicate that there may be a possible rela- tionship between Rhizopus and some of those diseases, such as Pellagra, the "corn-stalk disease," and the "horse disease" of the Middle West, the causes of which are at present unknown, but which have been supposed to be due to infected food. We are at present carrying out a series of investigations on the chemical nature of the Rhizopiis toxin, as well as its possible relation to such diseases, and we hope to be able to make a more detailed report in the near future. 87. Studies on melanin : V. A comparison of certain nitro- gen ratlos in black and in white wool from the same animal. Ross AiKEN GoRTNER. (Biochcmical Laboratory of the Station for Experimental Evolution, The Carnegie Institution of Wash- ington.) Black and white wool from a pied lamb were analyzed by Van Slyke's method, with the following average percentage results (corr. for solubilities of the bases) : White wool Black wool Ammonia nitrogen 9.32 9.46 Humin nitrogen 1.20 4.74 Arginine nitrogen 1746 16.81 Lysine nitrogen 3.90 3.97 Cystine nitrogen 2.70 3.09 Histidine nitrogen 7.00 7.04 Amino nitrogen (filtrate) 54-54 52.04 Mon-amino nitrogen (filtrate) 2.76 2.13 Total 98.88 99.27 The nitrogen content of the black wool was 15. 11 per cent. and of the white wool, 16.27 per cent. By subtracting the humin nitrogen for the white wool from the corresponding fraction for the black wool we have 3.54 per cent. of the nitrogen as due to the melanin present in the black wool, I have already shown^ that the pigment which remains when the melanin from wool is treated with boiling conc. hydrochloric acid, contains 8.48 per cent. of nitrogen. Using these data I have calcu- lated the percent of nitrogen which would be present in the black wool providing that the nitrogen due to the melanin, as given by the acid hydrolysis, were not present, at the same time correcting ö Gortner : Bull. soc. chim. de France, 1912, xi, p. 498. 1913] Alfred P. Lothrop 545 the weight of wool taken for the corresponding weight of melanin ; and I find that the percent of nitrogen in the keratin structure (white wool) would be only 15.48 per cent. It can be readily Seen that this is far too great a divergence from the white wool percentage (16.27) to be an experimental error. It is far more probable that the melanin molecule is broken down by acid hydroly- sis and that only a portion of the nitrogen is obtained as humin nitrogen, so that the correction which I used did not take into account a considerable portion of the melanin molecule which has a lower nitrogen content than the keratin structure. This suppo- sition is in agreement with the results of my previous work. 88. Metabolism studies of five cases of endarteritis oblit- erans ("Hebräische Krankheit"). Max Kahn. {Service of Dr. Charles Goodman, Beth Israel Hospital, New York.) In five male adults suffering from endarteritis obliterans of the vessels of the leg, and fed a Folin diet, the urinary nitrogen partition was normal, and the excretion of ethereal sulfate and calcium was increased. 8g. On the content in expired air of protein detectable by the anaphylactic reaction. Daniel R. Lucas. (Chemical Lab- oratory, College of the City of New York.) The lethal intraperi- toneal dose of dog serum for normal guinea pigs is about 5 c.c. for 300 grams of body weight; 4 c.c. for the same weight is usually followed by recovery, 6 c.c. by death. The lethal dose of human serum is about 10 c.c. for 300 grams of body weight. Two c.c. of dog serum or 15 c.c. of distilled water, injected intraperitoneally, in normal guinea pigs, caused shivering, twitch- ing and noisy respiration. Such Symptoms have been interpreted as evidences of anaphylactic shock but are not reliable indications of it. Proportionate amounts and the production of a lethal effect by a sublethal dose are essential for accuracy when dog serum is used. Intraperitoneal injections (2-15 c.c.) of condensations from the expired air of dogs did not sensitize guinea pigs to dog serum. Guinea pigs exposed to dog exhalations for a week, under condi- tions of very poor Ventilation, were not sensitized. Nor were any evidences of sensitization to human serum obtained when pigs were exposed for a week in the exit of a Ventilation System in a College building containing many people daily. 54^ Proceedings Cohimbia Biochemical Association [July These findings fail to support the concluslons of Rosenau and Amoss,'^ but are in accord with, and extend, the observations of Weisman.^ go. On the comparative physiology of creatin and Creatinin. Max Morse. {Woods Hole, Mass.) In a study of the absorption of the muscles in the tail of the larva of the common frog, attention was directed to the creatin-creatinin content of the muscle and also of the ehmination of these Compounds in the excretions. Numerous attempts to determine the amounts led to the general conclusion that, with the color reactions used (Folin-Benedict-Meyers method and the older Jaffe reaction), no creatin or Creatinin is demonstrable either in the fresh muscle or in the excretions. The results were negative with Weyl's test and with Kramm's and Salkowski's methods. An attempt to isolate the crystallin Compounds was futile. The f ollowing method was used : The larva was weighed and measured ; the tail was removed, ground in sand, covered with 95 per cent. alcohol and shaken for two-minute intervals on an International Instrument Company centrifuge-head shaker with four changes of water. The liquids were mixed and made up to 100 c.c. Ten c.c. portions were used for the various reactions. A Duboscq color- imeter was used, but the readings were not recognizable in the instrument. Excretions were caught by placing the larva in a large petri dish with a measured amount of distilled water, the whole being concentrated at low heat by evaporation. This Observation accords with other data showing that the ab- sorption of the tail involving the disappearance of over a gram of tissue within 12 hr. does not concern nitrogen elimination in excess of the normal for non-metamorphosing frogs. The study of the creatin-creatinin content of tails and excreta of metamorphosing frogs was inspired by the conclusions of several workers on mammalian material, which seemed to show that muscle metabolism, especially the atrophying muscle of involuting uteri, etc., involved quantitative relations of these Compounds, but Mellanby has more recently shown that this is not true for mammalian uteri. 'Rosenau and Amoss : Journal of Medical Research, 191 1, xxv, p. 35. ^Weisman: Biochemical Bulletin, 1913, ii, p. 295. (See also page 558 of this issue. Ed.) I9I3] Alfred P. Lothrop 547 Denis has failed to discover creatin and Creatinin in the urine of elasmobranchs, but aside from this work, I know of no determina- tions of the creatin-creatinin content, and the röle of these Com- pounds, in the lower chordates. 91. Influence of electricity upon metabolism. Matthew Steel. (Laboratory of Physiological Chemistry, Long Island Medical College.) Informal report. B. ABSTRACTS OF PAPERS FROM THE COLUMBIA BIOCHEMICAL DEPARTMENT AND AFFILIATED LABORATORIES 92. Studies of intracellular chemistry: A differential stain for mucins and mucoids. Louis Berman and William J. Gies. From a mixture of five parts of 0.25 per cent. sol. of safranin (Grübler) and three parts of 0.25 per cent. sol. of methyl green (Grübler), mucins and mucoids select the methyl green and are colored green. In a 0.25 per cent, sol. of safranin alone, these gluco- proteins are stained red. The following types of pure mucin and mucoid preparations, which we prepared by the methods best adapted for the highest purity of the products, stained green in the safranin-methyl green sol. : mucoids from tendon, ligament, carti- lage and bone; sodium salt of tendon mucoid; salivary mucin and a sodium salt of salivary mucin ; ovo-mucoid. Samples of the following protein products, prepared by the best methods and of the highest purity, selected safranin from such a safranin-methyl green sol. and stained red: albumin {tgg), chron- droalbumoid, collagen, deutero proteose, edestin, elastin, fibrin, gelatin (from bone, muscle, ligament), hemoglobin, metaproteins (acid albumin and alkali a^buminate), myosin, nucleoprotein (from ligament and yeast), Ossein, osseoalbumoid, peptone. Dry blood serum, tgg white and gluten also stained red in the safranin-methyl green mixture. All these protein preparations stain green in a 0.25 per cent. sol. of methyl green alone. Glucothionic acids from the various glucoproteins named above behave like mucins and mucoids in staining green in the safranin- methyl green sol. The products obtained from glucothionic acid by hydrolysis with boiling 5 per cent. hydrochloric acid sol. failed to show the selective action. 548 Proceedings Columbia Biochemical Association [July One part of tendon mucoid mixed with one hundred parts of Collagen or edestin can easily be detected by this differential stain- ing process. The technique, practical Utility, and applications of the test are now under investigation. 93. Two new histons. Walter H. Eddy. Tom-cod histon. Tom-cod milt contains a histon which, like thymus histon, is pre- cipitable in two forms: (a) an ammonia-precipitated product which is partially soluble in water, and (&) a sodium-chlorid precipitated product which is readily soluble in water. Both forms give char- acteristic histon tests. Shad histon. Shad milt contains a histon which may be precipitated in two forms: (a) an ammonia-pre- cipitated product which is insoluble in water, and {h) a complex precipitable by Saturation with sodium chlorid, which appears to be a combination of histon and a non-histon protein fraction. The products were prepared by the method already described.^ 94. Histon nucleoprotein : A protein salt. Walter H. Eddy. Continuing the work on histon-nucleoprotein that has already been described,^*^ and proceeding with the aid of the improved method recently outlined,^^ a neutral water-sol. of thymus histon (which had been precipitated by Saturation with sodium chlorid) was added to a neutral Solution of sodium nucleoprotein from yeast until pre- cipitation was complete. This precipitate was insoluble in water but soluble in 0.05 per cent. sodium carbonate sol. and could be repre- cipitated, without decomposition, by o.i per cent. hydrochloric acid sol. The 0.05 per cent. sodium carbonate sol. was precipitable by conc. nitric acid sol., by very dil. hydrochloric acid sol., by picric acid, alcohol, mercuric chlorid and neutral lead acetate but was not precipitable by ammonia, sodium phosphotungstate, potassium f erro- cyanid, potassium mercuric iodid, or albumin. It gave positive biuret, Millon, xanthoproteic and Molisch tests. In all these respects the aqueous sol. of this precipitate agreed with the water-sol. of sodium nucleoprotein. The precipitate obtained with o.i per cent. hydrochloric acid sol., after purification, was extracted for twenty four hours with 0.8 per cent. hydrochloric acid sol. Ammonia pre- *Eddy: Biochemical Bth-letin, 1913, ii, p. 435. i°Eddy: Ibid., 1912, ii, p. 121. 11 Eddy : Ibid., 1913, ü, P- 435- 1 /^».r'.5..' I9I3] Alfred P. Lothrop 549 cipitated from the extract a product which gave additional histon tests, proving the presence of histon in the Compound. An interesting confirmation of these results was obtained by repeating the preparation with histon precipitated with sodium Chlorid from a hydrochloric acid extract of tom-cod milts (see pre- ceding abstract) . This product resembled, qualitatively, the thymus- histon salt but was not precipitated from the sodium carbonate sol. by hydrochloric acid. It was precipitated readily with nitric acid and this precipitate yielded histon on extraction with 0.8 per cent. hydrochloric acid sol. 95. Further experiments on the preparation of modified col- lodion membranes for use in dialysis experiments. Frank R. Elder. In continuance of the studies, in this laboratory, of col- lodion membranes/^ we have endeavored to determine the compara- tive permeabilities of collodion-fat combinations. The tests were conducted with aqueous Solutions in small bags made by Dr. Gies' method, as recently published by Clark/ ^ from mixtures of U. S. P. collodion sol. and pure olive oil. Uniform mixtures of i part of olive oil and 3 parts of collodion sol. did not yield bags strong enough for the use intended, but satisfactory membranes could be obtained from mixtures containing i part of oil and 4 of collodion sol. Bags made from mixtures of i part of olive oil and 9 of col- lodion sol. were impermeable to chlorid, sucrose and peptone in dialysis experiments of a week's duration. Bags made from mix- tures of I part of oil and 99 of collodion sol. were occasionally im- permeable to each of these substances, though such bags frequently permitted all of them to dialyze freely when associated pigment was unable to diffuse. It is our Intention to determine, if possible, whether such diffusion differences depend upon inequalities in the distribution of oil in the bags, " age " of the membranes, etc. 96. Comparative studies of the permeability of collodion and collodion-fat membranes. Samuel Gitlow. Bags were prepared as usual (see the preceding abstract), and the tests were conducted in water. The following water-soluble dyes diffuse through piain collodion membranes used immediately after removal from the 12 Gies: Proc. Amer. Soc. Biol. Chem., 1912, ii, p. 75; Journ. Biol. Chent., 1912, xi, p. xli; Science, 1912, xxxv, p. 396. 13 Clark: Biochemical Bulletin, 1912, i, p. 198. 550 Proceedings Columbia Biochemical Association [July mold: (a) Very rapidly — auramin, bismarck brown, chrysoidin, erythrosin, martius yellow, metanil yellow, orange G, rose bengal; {h) in fifteen minutes — eosin A, phloxin, safranin; (c) in one hour — Biebrich scarlet; (d) in one day — cape aloes, fast red A, fustic extract, rhodamin ; (e) indiffusihle — benzopurpurin, malachite green, methylene blue. The following dyes (those named above) diffuse through coUodion-fat membranes used immediateiy affer removal from the mold: (a) Very rapidly — chrysoidin, eosin A, martius yel- low, orange G, phloxin, safranin; {b) in fifteen minutes — erythro- sin; (c) in thirty minutes — Biebrich scarlet, metanil yellow, rose bengal; {d) in one day — auramin, cape aloes, fast red A, rhodamin; {e) indiffusible — azolitmin, benzopurpurin, bismarck brown, mala- chite green, methylene blue. The older the bags of each kind, that is the more completely the residual ether and alcohol solvents of the collodion Solution (U. S. P. ) were removed from the membranes, the less permeable the bags became. In most cases, each particular pigment diffused through the different membranes at the same general rate, but in the follow- ing instances diffusion through collodion-fat membranes was faster or slower than through piain collodion membranes: (a) faster — Biebrich scarlet, eosin A, phloxin, safranin; {b) slower — auramin, bismarck brown, erythrosin, metanil yellow, rose bengal. Certain variations have been noted which appear to depend on irregularities in the dyes and in commercial U. S. P. collodion Solu- tions, also on the " age " of the bags. The study is in progress. 97. Further studies of edema: On the absorption of water by white lupin seeds. Tula L. Harkey. The swelling of weighed, normal, white lupin seeds in various media, as compared with the increase in weight of such seeds in distilled water, was determined at short intervals for a week or more. At the end of 24 hours, seeds gained more weight in the chemical Solutions than in the corresponding water controls, in the instances cited on the opposite page. Effects of other acids, and of electrolytes and non-electrolytes on the imbibition of water from acid Solutions, will be reported later. 98. Further studies of edema: On the postmortem absorp- tion of water by tissues from well nourished and f asting animals. I9I3] Alfred P. Lothrop 551 Data pertaining to the absorpHon of water by white lupin seeds Solution Percentage gain in 24 hours Natura Concentration, per Cent Water control Specified Solution Hydrochloric acid 0.125-0.5 0.25 -I.O 0.125-0.5 I.O -2.0 I.O 0.25 2.0 I.O -8.0 141.8 154-4* 126.I 155-9* 152. I* 135-0 136.7 144.8 148.6-173.6 156.9-156.2* 139-5-153-6 165. 6-163. 5* 150. 2* Sulphuric acid Phosphoric acid Lactic acid Oxalic acid Sodium Chlorid 1^8.^ Sucrose 137-2 153.4-164.5 Urea TuLA L. Harkey. We tested assumptions that the tissues of fast- ing dogs contain, at death, relatively less acid-yielding material than the tissues of well nourished dogs and that, on the basis of Fischer's collochemical theory of edema, the fasting tissues would therefore imbibe less water than the latter, under uniform postmortem con- ditions. Well nourished dogs and dogs which, after preparatory periods on the Standard diet, had f asted completely for 8-i i days, were bled to death, in each case f rom a femoral artery, and the more important parts treated in stoppered, wide-mouth bottles with mod- erate excesses of water. The weights of the swollen tissues were recorded at regulär intervals. Diffusion gains and losses occurred under uniform external conditions for each tissue. The results do not Support the assumptions on which the work was conducted. The percentage gains in weight were irregulär — fasting brains, for ex- ample, gaining more weight relatively than well nourished ones in some cases, or vice versa in others. There was also no apparent definite relation between the degree of swelling and the acidity of comparative extracts of the tissues. The results do not conflict with current opinions on the role of osmosis in the absorption of water by protoplasmic structures, and they harmonize with the belief that enzymes may be important fac- tors in such processes. The study is in progress. These studies of edema were suggested by Dr. Gies, and con- ducted under his supervision. 99. A preliminary study of the resistance of fasting dogs to hemorrhage. Paul E. Howe and William J. Gies. In further- * Record at 48 hours. 552 Proceedings Columbia Biochemical "Association [July ance of the preliminary study described at the last meeting/^ we have noted the effects of hemorrhages in dogs under conditions of partial fasting, after suitable preparatory periods on our Standard laboratory diet for dogs. Five animals {first group), weighing 4.7- 6.4 kilos, received water (187-315 c.c.) but no other ingredients of the preparatory diet. The partial fast was maintained for from ten to thirteen days, with losses in body weight of 18. 1-23.6 per cent. Blood amounting to 3.54-5.38 per cent. of the body weight (2.86- 4.4 per cent. of the initial weight) was removed without causing any serious Symptom. Three animals {second group), weighing 5.5- 12.6 kilos, received the Standard diet minus the meat. The only pro- tein in the food was the small quantity in the cracker meal. No dietary compensations were made for losses due to exclusion of the meat. The partial fast was maintained for thirty-three days, with losses in body weight of 16.05-24.95 per cent. Blood amounting to 4.50-5.1 1 per cent. of the body weight (3.62-3.86 per cent. of the initial weight) was removed without causing special respiratory difficulty or distress. 100. Biochemical studies of selenium. V. E. Levine. Op- posed to the current opinion that organic substances, generally, reduce alkali-selenite Solutions, we find that reduction is not induced by alcohols, phenols, saturated and unsaturated organic acids (ex- cept formic acid, lactic acid, gallic acid), amino acids, purin bases, proteins, fats and other lipins such as lecithin and cholesterol, Acety- lene, hydroxylamine and Phenylhydrazine cause very strong re- ducing effects. Acetone and formaldehyde reduce acidified Solu- tions of sodium selenite. Many car'bohydrates reduce alkaline sodium selenite to colloidal or red amorphous selenium. Inorganic substances, e. g., ferrous sulfate, stannous chlorid, zinc and hydro- chloric acid, sulfurous acid, arsenious acid, phosphorous acid, hydrobromic acid, hydriodic acid, also exhibit reducing power. Hydrogen peroxid, free halogens, nitric acid, potassium permanga- nate, and aqua regia, because of their oxidizing activity, inhibit or may entirely prevent the formation of colloidal or precipitated sel- enium (reduction). The possibility of using sodium selenite as a reagent for the detection of reducing substances was investigated. Arabinose, "Howe and Gies: Biochemical Bulletin, 1913, ii, p. 468. I9I3] Alfred P. Lothrop 553 rhamnose, xylose, galactose, glucose, fructose, maitose, and lactose promptly reduce alkaline sodium selenite. A selenite reagent con- taining 2 per cent. sodium selenite, 10 per cent. sodium citrate and 10 per cent. sodium carbonate has thus far been as efficient as the Fehling-Benedict reagent. Selenium Compounds are toxic to both plants and animals. Beginning with the most poisonous, the sequence of toxicity for the group selected was the f ollowing one : hydrogen selenide, selenium dioxide, selenic acid, sodium hydrogen selenite, sodium selenite, potassium selenocyanate, sodium selenate, free selenium. Intra- venous injection is followed by a marked fall in blood pressure; potassium selenocyanate, however, induces a considerable rise. Pul- monary edema, accompanied by exudation of large volumes of yellowish fluid, preceded death in the case of selenium dioxide, sodium acid selenite, sodium selenite and selenic acid. Respiratory paralysis set in before the heart stopped. Reduced selenium can be detected in neutral urine by the addi- tion of potassium Cyanide. Decomposition by means of hydro- chloric or sulfuric acid of the potassium selenocyanate thus formed results in the production of a colloidal Suspension of brick-red selenium. Potassium selenocyanate can be detected by the addition of ferric chlorid, followed by a drop of dilute sulfuric acid Solu- tion. The presence of potassium selenocyanate in the urine inter- feres with the Fehling-Benedict reduction test. The study is in progress. 10 1. The influenae of electrolytes on the precipitation of soluble starch.i^ Helen I. Mattill and H. A. Mattill. The origin of this investigation was an Observation by Dr. Gies that Solutions of soluble starch, when dialyzed free from electrolytes, were not precipitable by alcohol, and that the addition of a drop of dilute Salt Solution restored precipitability. It was the object of this work to determine at what concentrations of electrolyte the precipi- tation by alcohol ceased, i. e., how delicate a test for electrolytes it is, and how variations in the nature of the electrolyte affected the reaction. Two percent soluble starch Solutions were dialyzed until they gave no precipitate upon the addition of alcohol. Two c.c. of 1^ Some of the work was done in the Physiological Laboratory at the University of Utah, where it is now in progress. 554 Proceedings Columbia Biochemical Association [July the starch Solution were diluted with lo c.c. of alcohol (95 percent.) and one drop of salt Solution was added. The time required for pre- cipitation and Sedimentation served as a measure of the activity of the various electrolytes. It was shown that the cations in chloride Solutions were effective in the following order : Ba= Sr= Ca > Mg = Ce>Na = K>H> NH4 > Li > Hg" — results which are sim- ilar to ion effects upon many other emulsoids (Hofmeister's " lonen- reihen," etc.). The lower limits of precipitation were as foUows: one drop of o.i w HgClg sol. was without effect ; one drop of 0.025 n LiCl gave a very slight precipitate; o.oi n LiCl, 0.005 ^ NH4CI and 0.0025 n HCl, NaCl, and KCl were without effect; only in- complete precipitation occurred with 0.0025 n sol. of alkali-earth Chlo- rides. If alcohol of varying concentrations (from 80-10 percent) is used, the precipitability of the starch by electrolytes is rapidly decreased with decreasing concentrations of alcohol, and the differ- ence between the effects of the various cations is rendered more marked. A cursory examination of the Sulfates indicated a smaller effectiveness and one or two exceptions to the above order for Chlo- rides. In all cases the precipitation is reversible. Further work will be done on a more accurate differentiation of the cation and anion effects, and upon variations in this phenomenon, with varying de- grees of dispersion of the starch. 102. Determinations of the acidity of fruit Juices.^ ^ Edgar G. Miller, Jr. In line with Dr. Gies' proposal of the use of diluted vinegar and various " food-acid" media as dentifrices, I have deter- mined the acidity of some common fruit Juices as a preliminary to the selection of the most suitable ones for prophylactic applica- tion to the teeth. The appended data for acidity represent, in most cases, the averages of triplicate results, in c.c. of n/5 sodium hydroxid Solution (phenolthalein) per 10 c.c. of juice: Apple, 3.5; apricot, 3.8; asparagus, 0.9; banana, 2.6; beet (red), i.i; canta- loupe, 0.6; carrot, 0.8; cauliflower, 1.9; celery, 0.8; cherry, 7.6; cocoanut milk, 0.4; cranberry, 19.6; cucumber, i.o; currant, 20.4; gooseberry, 16.2; grape (white), 4.5; grape fruit, 10.3; horse radish, 9.2; lemon, 53.7; orange, 6.y; parsnip, 2.1 ; peach, 6.4; pear, 3.2; pineapple, 7.5; plum, 4.8; radish, 0.6; rhubarb, ii.i; straw- berry, 9.3; tomato, 4.2; tumip, 0.6; vinegar, 26.1 ; watermelon, 0.6. 16 Miller: Dissertation; Columbia University, 1913 (Part iii, p. 25). 1913] Alfred P. Lothrop 555 103. A study of the influence of extemal hemorrhages on the partition o£ urinary nitrogen. Olive G. Patterson. Two dogs were subjected to external hemorrhages (3.5-5.8 per cent. of body weight) under local anesthesia (cocain), and studied by some of the nutritional methods in use in this laboratory. One dog was subjected to four successive bleedings, at intervals of 11, 5 and 7 days, respectively. The general conclusions pubHshed by Hawk and Gies/'^ on the effects of external hemorrhage in dogs under general anesthesia, were confirmed. Each hemorrhage caused abso- lute increases in the renal excretion of total nitrogen and urea. The first hemorrhage in each animal was followed by an increased Out- put of Creatinin; subsequent bleedings increased the excretion of Creatinin or were without influence on it. The absolute amounts of urinary ammonia, uric acid and purins were unaffected by the hemorrhages. The results will be published in detail in the near future. 104. The action of a high frequency current on the activity of pancreatic amylase. P. W. Punnett. An attempt was made to discover what effect, if any, the high frequency current, such as is used in electro-therapeutics, has on the activity of pancreatic amylase. The current was furnished by a high frequency machine loaned to the department by the Van Houten and Ten Broeck Com- pany of this city, to whom we are greatly indebted for this courtesy. The amylase of commercial "pancreatin" (Parke, Davis & Co.) was used. The method of Sherman, Kendall and Clark^^ was employed throughout for the determination of activity. Solutions of the material were treated with the current in a beaker coated on the outside with tinfoil attached to one terminal of the machine and having a platinum electrode, connected to the other terminal, immersed in the Solution, the whole being kept in an ice bath. The length of treatment varied from 5 to 30 min., and the hot-wire ammeter gave a reading of 500 to 600 milliamperes. Controls were kept at the same temperature. Because of the fact that only ordinary distilled water was available, the enzyme did not show more than 70 per cent. of its maximum activity (its power was 418, using triple-distilled water). '^'' Hawk and Gies : Amer. Jour. of Physiol., 1904, xi, p. 171. 18 Sherman, Kendall and Clark: Jour. Amer. Chem. Soc, 1910, xxxii, p. 1073. 556 Proceedings Columbia Biochemical Association [July Although the results are not quantitatively exact, they strongly indicate that this type of electricity is in no degree harmful to pancreatic amylase; that the treatment of a fresh Solution for from 3 to 20 min. is probably beneficial, giving an increase of from 5 to 20 per Cent, in activity ; that longer treatment is without effect ; and that Solutions which stand for about 24 hr. at room temperature Mr. Punnett's report.) It may be noted that the direct current has an exactly opposite effect on enzymes, as has recently been shown by Burge.^^ Further work along these lines is desirable. (Mr. John W. Radu, Superintendent and Chief Engineer of the Van Houten and Ten Broeck Co., made some very interesting demonstrations with the electrical apparatus at the conclusion of Mr. Punnett's report.) 105. A further study of the distribution of osseomucoid. Christian Seifert and William J. Gies. Ten years ago we published data showing that osseomucoid is a constituent of the main limb bones (the only ones investigated) in thirteen species of mammals, ten of birds, two of reptiles and one of fish.^^ At that time we concluded that osseomucoid is probably a constituent of all bones. In resumption of this study, and proceeding by the original methods, we have lately separated osseomucoid from the main limb (or skull) bones of the following additional species: mammals — monkey (mangabey and spider), horse, fox, raccoon, white rat; hirds — dove, meadow hen ; fish — sculpin. The research is in progress. 106. A further effort to prepare a colorless biuret reagent. A. W. Thomas. In extension of the work described by Kantor and Gies,^^ I have endeavored to prepare a colorless biuret reagent. Cuprous thiocyanate was thought to be a good Substitute for copper Sulfate because of its white (or grayish white) color, but treatment with alkali caused decomposition of this salt, resulting in precipita- tion of a yellowish red modification of cuprous oxide ; and the Solu- tion, which contained some of the undecomposed Compounds, rapidly assumed a blue color due to oxidation. It was possible to obtain a nearly colorless reagent with cuprous iodide, but oxidation promptly brought about the inevitable result — a blue Solution. lö Bürge: Amer. Jour. of PhysioL, 1913, xxxi, p. 328; xxxii, p. 41. 20 Seifert and Gies : Amer. Jour. of PhysioL, 1903, x, p. 148. 21 Kantor and Gies: Biochemical Bulletin, 1911, i, p. 264. 1913] Alfred P. Lo f Itrop 557 The complex colorless Cyanide o£ copper, which is readily soluble in 10 per cent. sodium hydroxid soution, was tried. The Solution was colorless and it gave a strong characteristic pink to purple reac- tion when added to a dilute Solution of Witte peptone. Unfor- tunately, however, the Solution gradually turned blue, It was thought that this blue oxidation product might be reduced with an excess of potassium Cyanide to drive the cupric ion into the colorless complex ion. Experiment proved such to be the case, but this color- less Solution would not give the biuret reaction when added to pep- tone Solution. This is an extremely interesting fact from which the inference may be drawn that the biuret reaction takes place only with the positive copper ion. But if this inference is correct, how is it that one obtains a reaction with the colorless Solution made by adding K2Cu(CN)3 to 10 per cent. sodium hydroxide Solution? This fact is easily understood when we consider the dissocia- tion of this salt. The instability constant of the complex ion, (Cu"") X (CN")^ r rr\j\ = — ~' is 0.5 X lO"^'^, and the concentration of cuprous ion is approximately 3.7 X io~^ in a tenth molar Solution. But, this very small concentration of cuprous ion is sufficient to Start the reaction. In the strong alkaline Solution there is immediate oxi- dation of Cu"^ to Cu"^"^, which reacts with the protein giving some pink color; as soon as this happens more of the KoCu(CN)3 dis- sociates, forming more Cu"^->Cu'^^ which results in more of the pink product with the protein. If an excess of KCN is added, there is a driving back of the ionization of K2Cu(CN)3, positive copper ions are removed and hence no reaction occurs when protein Solution is treated with the reagent. In Order to determine whether the reagent could be prepared for use during a laboratory day without oxidation, 0.2 gm. of the white double Cyanide was added to 50 c.c. of sodium hydroxid Solu- tion, and observed for appearance of any blue color. From time to time the flask containing the Solution was opened and portions were withdrawn for the purpose of making tests upon proteins. After five hours of such treatment the reagent began to acquire a light blue color, due to the oxidizing effect of the air, the color deepening to that of the regulär reagent. If air is excluded from the bottle, the development of color proceeds more slowly. 558 Proceedings Columhia Biochemical Association [July To prepare this temporarily colorless reagent, about o.i gram of potassium cupro-cyanide may be dissolved in 50 c.c. of 10 percent (or stronger) sodium hydroxid sol. Fehling's alkaline tartrate sol. may be used. Convenient approximations of these amounts may be made in emergencies with satisfactory results. Several of the common organic salts of copper were treated with alkali. Many of them were decomposed, yielding cupric hydroxid or cuprous oxid, and a blue Solution. The citrate, salicy- late and tartrate, while soluble in alkali, yield Solutions that are respectively green, deep green and dark blue, thus rendering them useless for the intended purpose. 107. Biochemical studies of expired air in relation to venti- lation.22 Charles Weisman. Our preliminary results^^ have been fully confirmed. Eleven repetitions of Rosenau and Amoss' experiments have been made, with negative results in each case. Anaphylaxis failed to occur in control experiments, when expired- air condensations were used for the second injection as well as for the preliminary sensitization. In experiments extending the work, air was drawn through the macerated lungs from a dog and the vapors Condensed into a Drechsel bottle. The guinea pigs that were " sensitized " to the condensation-liquid failed to exhibit anaphylaxis after appropriate treatment with dog-blood serum. Air was also drawn through a Solution of Witte peptone, and the vapors Condensed, but the liquid thus obtained did not yield the biuret, Million or xanthoproteic test. Chemical examination of the condensation-liquids, obtained under aseptic conditions from the expired breath of several persons, gave the following results: Ahsent — sulfid, phosphate, bromid, iodid, amin, acetone (iodoform test), diacetic acid (Lipliawski test), and protein; present — ammonia (Nessler test), and chlorid. The ill effects of vitiated air in poorly ventilated or over- crowded places cannot be due to a volatile protein ("sensitizing substance " ) from the lungs. Biochemical Laboratory of Columbia University, College of Physicians and Surgeons, New York 22 Weisman: Dissertation; Columbia University, 1913. Pp. 97. 23 Weisman : Biochemical Bulletin, 191 3, ii, p. 295. BIOCHEMICAL BIBLIOGRAPHY AND INDEX 3. Second quarter, 1913 (April-June)^ WILLIAM J. GIES (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) Explanation of abbreviations, arrangement, notation, etc. Bibliography. Titles of papers are freely shortened, minor words ignored, common terms conveniently abbreviated or chemical symbols substituted; surnames of col- laborators are connected by hyphens; most pimctuation marks are omitted — all for the sake of condensation. Heavy faced Roman numerals indicate volumes; heavy faced Arabic numerals designate numbers and dates of issue (slanting lines separate numerals for months and days). Bibliographie items begin with em dashes. When two or more papers by the same author occur together, they are duly numbered, and separated by semicolons, but follow the same em dash. Numerals preceding italicized names of authors indicate sequence in the bibliography (index numerals) ; numerals preceded by commas, at the ends of items, indicate initial pages of the corresponding papers. Index (subjects). The numerals in the index (page 565) correspond with the numbered items in the bibliography. Pages are not indicated. Numerals held in groups by hyphens are piain abbreviations in accord with the indications of the first numeral of each such series (see footnote, p. 565). Abbreviations of words in the index are similar to those in the bibliography. Each group of index references is terminated by a semicolon; commas mark oflf subdivisions of a general index subject. Names of authors are not indexed. Journals included: Biochemische Zeitschrift (B. Z.), Zeitschrift für physiologische Chemie (Z. p. C), Journal of Biological Chemistry (J. B. C), Biochemical Journal (B. J.), Biochemical Bulletin (B. B.). Practical use of the bibliography. The bibliography is helpful from several Standpoints. Thus, if it is desired to ascertain whether the Journals included in the bibliography contain any papers (during the given quarter) on a particular subject, e. g., lipins, find the key word in its alphabetical place in the index and turn to the items in the bibliographic sequence indicated by the index numerals (i. e., in this case, 35, 50, 56, 415). The abbreviated items thus identified give the names of authors and suggest the nature of the correspond- ing papers (four papers, in the case selected for illustration), and help the reader to decide whether to examine the original publications. When the index gives a negative answer to an inquiry, a large mass of literature is removed 1 The preceding portions of this bibliography and index were published at pages 298 and 470 of this volume (Jan. and Apr. issues). 559 560 Biochemical Bihliography and Index [July from further consideration. During the intervals between publication of the indexes of Journals, Centralblätter and year books, this running bibliography directs the reader to most of the main tracks through current literature on the leading biochemical subjects. Bibliography. B. Z. — L:i-2;4/ii. — iBokorny'E'mü versch Subst a Keim Pflanz'sam, Wachstumsförd,i ; 2lbid.,4g; ^Ibid.^Sy. — ^Boro- zuikowUrs Wachstum Pflanz,ii9. — ^Doerr-Pickyitch^jns primär Toxiz Antiser u Eig'sch i Antigen,i29. — 6Glagolew'P\2.sitmh\\6.,i62. — yLand- steinerS-ptzii Immunreak u ihr kordchem Erklärbark, 176. 3-4; 4/21. — 8ßwM2:^/Oxydas i Blattrollkr Zuck'rüb,i85. — gHämäläinenSynih. )8-Glucosid d Terpenalkoh,209 ; loKonst Terpineol-35°-glucuronsäu, 220. — iiKretschmerTitr Harnsäu i Harn n vorgäng Ag-fäl,223. — 12- ArcichovskifWivk Giftst versch Konz a Sam,233. — i^LandsbergBlut- gerin,245. — i4Frank'Vork Kephahn u Trimyristin Leber,273. — 15- MayerZuck'irei Gär b Ster'iso,283. — iGDurigY erh Amphib i versch konz Lös,288. — i7Tö^^/-BrmMa-DMn(7Kohlenhydr'spar Wirk Alkoh, 296. — i8fFa/&z(wNachtrag: Verw Rotkohlausz a Indic corim Mess H*-konz,346. 5-6; 5/7. — i9A^^tfmawMFermentähn u Fermentreak d Blutser währ Gravid,347. — 2oMa3;^rBestim sog " Restreduk " d Blut, 362. — 2iObermayer-WillhelniFormo\t[t Unters Eiweisskör,369. — 22- ^rowEinfach Ext'app Extr fest u flüs Stof,386. — 23L^/iwawnKatal Lichtwirk b Samenkeim,388. — 24.B ernst ein'Elek'chem Grundl bioelek Poten,393. — 2^ScaffidiVeTh Musk'kreat b Ermüd,402. — 26Höber-Nast Vitalfärb,4i8. — 2yBang-StenströmAsph.yK u Blutzuck,437. — 28Loeb Milchsäu'bild a Traub'z, Glyc'aldeh u Dioxyacet Rind u Schw'blut, 451. — 2gGriesbachMi\chsä.u'hi\d a Kohl'hyd i lackf Blut,457. — Solshi- haraQudLTi Best Milchsäu i Harn,468. — 3i5'a/^ow^^zWirk Antisep a Toxin,483. — 32i?o^^w^/za/^rEmulsinart Enzy,486. — 33Pa/arfmoVeränd Stoffwech b Tier n Exstir Schildr u Parath,497. — ^4Lifschütz'ETk\diV Schreiber-Lenärd, Cholest,5o8. (Pp. 510.) B.Z. — LI:i-2;5/i6. — 35F^rMO«Oberfläch'sp u Lipoid leb Zel,i. — 36£ü/^rOxydat Abbau Eiw'k,26; 37Synth Tripept,45. — ^SSnapperCl- geh Blut u Vert a Ser u Blutk,53 ; 39Änd Permeab Rot Blutk d Säu'zu- sat,62; 4oNeut'rot-Pap Indic Alkal-bestim Serum,88. — ^iCostantino Biol Bedeut u MetabEiw'stof,9i. — 42£M/er-i?3;c?Zers Milchsä u Weinsä ultrav Licht,97. — 42)^incussohn¥ trm. Eigensch Blut, 107. — /\/\T,'öb Meth'ier Glykokol m Formal,ii6. — ^^Neuberg-RosenthalZnck'ir He- fegär : Carboxylas,i28. 3 ; 5/24. — 46WintersteinN2ivkos,i42,- — A-jHan- schmidtWivk Lecith b Vergift höh Tier, 171, — 4.8Bang-LarssonMikro- best Blutbest,i93. — 4gKoniko ff Best Blutreak mit elek Meth,200. — I9I3] William J. Gies 561 5oS?M&^rBlutHpoid u Phagocy,2ii. — 5iPorfAcetonitrilreak,224. — 52- PuglieseZvLS2im!&Q.iz dur Wärm u Arb erz Schweis,229. — ^■T^Schreiher Erwid,230. 4; 6/6. — ^^rWillberg'R.tsis versch Tier gegen As,23i. — 55Fwc^^Nachw u Best d Ameis'säu,253. — c^SLoeb-BeutnerBedeut Li- poid f Enst bioelek Poten'dif pflanz Org,288; 57Einfl Anaesth a Poten'dif Oberfl pflanz tier Geweb,300 — ^SSörensen-Palitssch" Salz- fehl" b colorim Mess H'-konz Meerwas,307, — S9^oracsewski- HersfeldKmü Ernähr Aussch Indol u Indican,3i4. — 6oMoracsewskiD b künst Verdau u Faul versch Eiw'k auftr Indolmeng,340. 5 ;6/i2. — 6i//^3'm«;rPräzis'ureom (Harnst'mes) z Bestim Harnst'geh Harn, Blut u Cer'sp'flüs: Über Gros Nier- u Lebertät,355. — 62Guggenheim Proteinog Amin,369. — 62)Vandevelde-V ander stricht Invert'reak gemis Hefekul,388. — 64DapperBest u Vork Milchsä i Harn,398. — 6^Salkow- skilst mög Geh d Gehir a Phos'tid steig,407. 6; 6/21. — 66MiuraBez'ieh Thy'parathy'ekt z Koh'hyd'st'wechs,423. — ö/Zaco^^^wEinfl Chloralhyd a exp Hy'glykäm,443. — 6STamuraFrüi Kumagawa-Suto Fettbest hins d Oxyd Fettsäu u unverseif Subst,463. — 6gNeubergBioch Umwand Meth'glyox i Milchsäu: Entst versch Milchsäu Natur,484. (Pp. 508.) B.Z. — LH : 1-2 ; 6/25. — 7oHa^^aKrit Zuck'best'meth Bang,i. — 71- Laquer-Snapper'E'mü CO2 a Cl-9t'wechs,44. — y2Fischerln3.'kt Äth'alk als Hämolytic dur norm Ser'alb,6o. — y^^ölts-Paechtner Alkoh'geh. Milch n Zufuhr wechs Alkoh'meng u unt Einfl Gewöhn,73. — y^Einis Wirk Pituitr u )g-Imidazoläth'am( Histamin) a Herzakt,96. — yc^Neu- bauerWirk antiglucosur Mitt: Leberglucosur,ii8. — y6GrameniskyZn- sam'hang akt u inakt Zust d Fermen u Oberfläch 'sp des, 142. 3-4 ; 6/30. — yyBournotUipsiS Pankr'cyst,i55; 78Lipas Chelidon'samen,i72. — ygLifschiitzO-produk Cholest tier Org (Pfortad-Leberv),2o6. — 80- Fwc^^Nachw Formal Pflanz,2i4. — SiBattelli-SternKmü. Anaesth a Oxydon,226; 82Einfl Aldeh a Oxydon,253. — S^SchewketFarh'rQak Gal'säu sow Tan (Gerbsäu) : Anw d Prob,27i. — 84Pescheck'N-spaT Wirk Salz,besond Na-acet a Fleischfr,275. — 85 to 200, blank. (Pp. 330.) Z.p.C. — LXXXIV : 2-3 ; 4/4. — 2oiGra/^N-ans b Füt kl Eiw'gab u gros Meng NH^-salz u Harnst,69. — 202Euler-JohanssonZusam'setz u Bild Enzym: Gl'zeit Veränd Geh a Invert u Gär'enz leb Hef,97. — 203LagT^^rMeth v St'wechs'unt'such Kaninch ; Milch a einz Nähr, 109 ; 204Wirk CO2 a St'wechs: Autol u St'wechs,ii7. — 205Fa«^^rEinw NHg-gas a Diast,i6i. — 2o6Abderhalden-HirschSyn Fähigk tier Zel: Wirk NaNOsaN-wechs, 189. — 207z^Z^yw^^Mod Hüfner Spek'phot,207. — 2o8Abderhalden-Lampe-LondonSchicks i Darm sich bild Eiw'ab- baustuf,2i3. — 2ogAbderhalden-Lampe'Emü per es verab Harnst a 562 Biochemical Bibliography and Index [July N-wechs Schw,2i8. — 2ioKashhvaharaYer\\ Harnsä z Zn-salz,223. — 2iiGra/^Erwid a Richtigst v Abderhalden-Lampe z mein Anmerk Arb V Grafe-Turban : " N-ret b Füt Harnst ",234. — 2i25'c/mmmAbsch kl Meng Hg dur Elektr ; Bemerk z Buchtala : " Quant Nachw Hg Harn u Geweb,239. 4; 4/15. — 2i3Z?«(7/fa-Co.yfawfmoMuskelch: Frei d Formol titr Am'säu'-N u Ges'extr'-N i Musk hung Tier,243. — 214- Fischer-HahnSyn 2-3-4-Trimeth'pyrrol u 2-3-4-Trimeth'-5-äth'pyr (isom Phyllopyr) ,254. — 2i^Fischer-Bartholomäus-RöseVor^]\yrm\n\'\ : Porphyrinog u Bez z Blutf'bst u Deriv,262. — 2i6Siegfriedß-Gluto- ky'sulf,288. — 2iy Abderhalden'E'in m Polaris'app komb elekt heizb Vor- ricli z Ables'g u Beob Drehv'mög konst Temp,300. — 2i8Lebedew Verest Dioxyacet m Phosphat,305. — 2igM anchot Antw z Burn: " O-ka- paz Blutf'bst ",306. — 22oL^&^c?^wAlk'gär,3o8. 5; 4/22. — 221F eulgen Verh echt Nucl'säu z Farbstof,309. — 222i?a^oc5:3;Peps-Chymos-Frag, 329. — 223/voj^f/Herst Trock'präp tier Org,354. — 2246'/^?/'/' Bemerk z Abderhalden: " Synt Fäh Organis Hund ",359. — 22^Abdcrhalden- Law/>(?Schlussbemerk z Graf Erwid (201,211) uns Krit sein N-stoff- w'vers,36i, 6; 4/29. — 226Feinberg'Blld Apomorph b Erhitz o Aufbew V Morphinlös,363. — 22yKroghlIa.rnst best Harn m Na-hy'brom,379. — 228Panser'E'mw NHg-gas a Invert,4o8. — 22gLichtzmt sBemerk z Schade- Boden: "Anom Harnsäulös (koll Harns)",4i6. 7; 5/3. — 2TpCohn- /^emWirk vollst abgebau Nähr a Verdau'kan,4i9. — 23iTÄo»u^nEinw Zuck a Verdau,425. — 232BoMwu/ar^Hervorr v Magenfunk'stör v Darm a,437. — 233CoÄw/iamPhysiol Nier'sekr,45 1 . — 234ßoj?oc^Säu'intox,468. — 235C7A/LÖS Metallverb geschwef Eiw'k: besond Berücks d Cu,478. (Pp. 428.) Z.p.C— LXXXV : 1-2 ; 5/1 5.~2s6P fei ff er-ModelskiVerh a-Am'säu u Polypep geg Neut'salz,i. — 22,7ThierfelderCerehrosid d Gehir,35. — 22ßJ ahnsonBlohmEm^ Cholest a Hämol,59. — 2T,gDox-Neidig'Enzym Spalt Hip'sä d Schim'pilz,68. — 24o/o/zanj^onTryp Verdau du Harn, 72. — 24iAbderhaldenNa.chtr z " Geh d Prot a /'Tyros u Genau Best dies Am'säu",9i. — 242Abderhalden'Notiz,g2. — 243PaM^^rEinw HCl- u NHg-gas a Diast,97. — 244Abderhalden-FodorD b Faul /-Asp'ginsäu enst Abbaust: Neu Meth Nachw y8-Alan,ii2. — 24^Abderhalden- Fronime-HirschBild y-Am'but'säu a c?-Glutam'säu unt Einfl Mik'org, 131. — 246Abderhalden-Lampe'Emü Ermüd a Geh Blutser a dialysierb, mit Triketohydrindenhydr reag Verb, 136. — 24."/ Abderhalden-Schmidt Beob u Vers m Triketohydrindenhydr, 143. — 248F^j^/^rBuchw'sam'- schal,i48. — 24gTrigtE'mü Diät a Ptyarakt,i56. 3; 5/26. — 2SoKasht- wabaraAutol Thymus,i6i. — 25iZ^w/'/a^wlnt'med metab am-ac,32i. 4; 5. ■ — 42oHolsberg'Mew meth isol tryps,335. — 42iDakin-Janney-Wakeman Cond affec form a excr form-ac; estim form-ac urin,34i. — 422Loeb- WasteneysRel infl weak a str bas on rate oxid unfert egg sea-urch, 355. — 42T,Green'waldMeta.h exp parathyr'ect dog,363; 424P cont blood 564 Biochemical Bihliography and Index [July norm a paratliyr'ect dogjßög. — 42^Johns-Baumann'FMrm: 2-meth'mer- cap-6,8-dioxypur, 2-meth'mercap-6-oxy-8-am-pur,38i. — 426Pilcher Excr N aft lig succes br ren arter,389. — 42yFengerI a P cont, siz a physiol act o fetal thyr,397. — 42STaylor-RingerUtiUz NH3 prot metab, 407. — 42gTaylor-RoseFunn metab : Uricol,4i9. — ^yyDak'm-Dudley Glyoxalas,423. 5; 6. — 43i^oo/j^3)Blood relat animals, as per comp ser prot; compar prot ser ox, sheep, bog, goat, dog, cat, guin-pig,433. — ^432pFooc?3'a^/Diabet; sar'lac-ac diab musc,44i. — 433Fo/w-Z>^wwAb- sorp N-prod; repl Abderhalden-Lampe,453 ; 434Tyrosmas cont prot; repl Abderhalden-Fuchs,457. — 4;^c^Loeb-Wasteneyslnü bas o rate oxid i fertil egg,459. — 436Ma^/t^wjImport chem dif bet egg s-urch a st-f, 465. — 427Loeb-lVaste7ieyslnü hy'ton sol o rate oxid fert a unfert egg, 469. — 42ßO sborne-LeavenworthDo gliad a zein yiel lysin hydrol,48i. — 429Myers-Voloviclnü. fev elim cr'inm,489. — 44oFoHw-AformNorm prot metab rat,509. — 44iLoeb-Wasteneysls narcos due asphyx,5i7. — 442Ringer-Frankel-JonasG\ucon'gen: Fate isobutyr, isoval a isocapro ac diabet ; int'med metab leuc a val,525 ; 443lbid. : Fate succin, mal a malon ac diabet ; int'med metab aspart a glutam ac, prol, lys, argin, ornith,539. — 444Levene-MeyerAct leucoc o hexos ; mechan lact-ac form, 551. — 445Z)a^m-F>M(i/^3;Int'med metab car'hyd a prot; mut int'conv a-am-ac, a-hydrox-ac, a-ket-ald,555. — 446-600, blank. (Pp. 405.) B.J. — VII: 3; 5. — 6oiAtkins-WallaceCrit-so\ point urin,2i9. — 602- SchmidtQuant rel i capil-anal,23i. — 6oT,MottramlIopkins-Cole mod Adamkiew test prot,249. — 6o4lValpoleLitm pap quant indic reac,26o, — 6o5Coo/>^rPrep fr anim tiss o subst eure polyneur bird induced by diet pol rice,268. — 6o6T/ijV/^Lipol act blood,275 ; 6o7Lipol act tiss,287. — 6o8Plimmer-EavesF.st tyrosin i prot b Br,297. — 6ogPlimmerSep cystin a tyrosin, 311. — öioCMV^Factor in sol a precip etiglob,3i8. — 611-700, blank. (Pp. 122.) B.B. — II : 7 ; 4. — 701 Osborne-FetzerYLe'mnch Ritthausen,334. — 702 AnonD'm'r t Prof. Chittenden ; testim b pupils,349. — 703^wowSoc Exp Biol a Med: 10 anniv a din'r,358. — yo4HasselbalchMeth. elect deter conc H' i biol fl,367. — yoc^Sanders-MayMeth deter tryptoph fr protein, 373. — yo6Bottaz:siFhys'-chem musc plasm,379. — yoyBiddle-HoweFsist ; comp musc fast dog,386, — 7o8Hi^/^3;Form curv CO2 excr result fr work fol forc breath,390. — yogHigleylnü barom pres CO2 excr,393. — 710/aw^way-Fww^Relat acap t shoc ; mechan ef artif hyp'-resp o circ, 403. — yiiDox-NeidigClea-v pyromucur-ac mold enzy,407. — yi2Hamlin Anal ash castor bean,4io. — 7i3CZar^Chem nat "tann mass" fr persim, 412. — 7i4FJc?3/Histon a prep,4i9. — yi^BergDid vWittich antedate Ost- 1913] William J. Gies 565 wald i defin enzy act,44i. — 716/^nowBioch Soc, Eng,446. — yiyLothrop Proc Col Univ Biochem Assoc,452. (Pp. 176.) Subject index. Absorp433; acapni7io; ac'aldeh274; ac'nitrilsi ; acid39, osis2ZA; Adamkiewicz-test6o3 ; alan244; alcohi7,72-j,220-S3-8o;2 aldeh82; al- kal4o; amin62; am-ac2 13-36-45 ,41 9-45 ; am-but-ac245 ; am-purin425 ; NH3205- 28-43-56,428,NH^salt,20i-62 ; amphibiö; anesth57,8i; annivers703; antigens; antisep3i; antisers; aort265; apomorph226 ; appar22,207-i7-23 ; argin443; AS54; ash403,7i2; aspart-ac244,443 ; asphyx27,44i ; autol204-50. Bang-sug-meth7o ; bar- pres709; base27i, 422-35; betonicin254-5 ; bile409; Bioch-Soc-Eng7i6 ; bioelec24, S6; bl'd20-5-9,38,43^-9,50,6i,2i5-p-38,424-3i>6o6-io,coagi3,cöc-p3&-9,seri9,246-67, sug27; brain6s,237,4i2; Br-at'n6o8; buckwh-br248. Calcif265; Ca-casein(para), 404; capiI-anal6o2 ; carbohyd 1 7,29,66,445 ; €0271,204,708-9; carbox'as45; case- mi^oZ-4-5-6; cast-bean7i2 ; cell35,2o6; cerose25i-i'; cerebron-ac4ii ; c'brosid237; cer'sp-fl6i ; chees407; Chelidon-seedyS; ChittendenRH702 ; chralhyd67; CI38-71 ; cholest34,79,238; chymos222; circ7io; coagi3; cord7,229; Col-Univ-Bioch-As7i7; compos'n52,25i, 404-5-7-9,707-12; Cu235; cork25i ; creatin25; cr'imn439; crit- S0I601; cyst77; cystinöog. Desic223; devel437; diabet432,442-j ; diast205-43-€3 ; diet249,6o5; diges6o,2o8-30-/-^-70-9; dinner702-j; dioxyacet28,2i8; dye22i-64. Egg422-3 5-6-7 ; elec49,2i7,chem24,metr2i2,704; embry4i2-27 ; emulsin32,276 ; envir437; enzymi9,32,43,76,202-39-4o-70,402-3o,7ii-5; euglob6io; excr7o8-9; ex- tr'iv257. Fast2i3,707; fat68; fatig25,246; fat-ac68; ferm't'ni5,45,202-2O-53- 80-7,401-^; fetus4i2-27; fever439; food59; f orc-br'th'g7o8 ; forma/d (01)21,44, 80,213; form-ac55,42i ; fung239. Gal-ac83; gent'bios273 ; germ't'n 1,2,3,23; gliad438; glucon'gen442-5 ; glucos28; glu'sid9; glucosur75,242 ; glu'ron-ac242 ; glutam-ac245,443 ; glutoky'sulf2i6; glyc'ald28; glycoc44; glyoxalas430 ; growi,2, 3,4,435-7- Hair26o; h'rt74,4i5; Hb2is-9; hemol72,238; hep-vein79; hexos444; hip-ac239; histam74; histon7i4; Hopkins-Cole-tes6o3 ; Hufner-spec'phot207; hydantoin-deriv4io; H"i8,58,704; HCI243-56; hydrol252,438 ; hydrox-ac445 ; hy'glycem67; hy'resp7io; hy'ton-sol437. Imidazoreth'am74; immun7; indican59; indicati8,40,6o4; indol59,6o; int'med-met4i9-42-j-5 ; intes2o8-30-^-42 ; invert63, 202-28-56-72; 1278,427; Fe-salt275; is-but-ac,is-capr-ac,is-val-ac442. Kephali4; kerat259-6o-(5 ; keton-ac258; keton-ald445 ; kidnöi, 233,426; Kumagawa-Suto- meth68. Lac-ac28-9,30,42,64-9,262,432-44; lecith47,275 ; leucin442; leucocy444; Iight23,ult'vi42 ; lipas 77-8; lipm (cid) 35,50-0,415; lipol-act6o6-7 ; Iitm-pap6o4; livi4,6i,75; lysin438-43. Mal-ac,malon-ac443 ; Manis-]apon25g; melanos242; mercap-purin425 ; Hg2i2 ; met-Hb26i ; method30,40-5-9,55-5,6i-4-5',7o,8o,203-i2- 27-4i-^-62-78,403-^-5-i3--#-6-20-J,6o4-^-9,704-5-i4 ; nieth'at'n44 ; raeth'glyox69 ; milk73,203-62 ; mold7ii; mol-w't4o6; morph226; musc25,2i 3-57,432,706-7; Mytil- edul2S7. Narco46,44i ; nerv-tis,syst4i2-j ; neut-red4o; ninhydr246-7 ; N84,20i- 6-9-11-5-25,426-33; NO263-72; nur1-ac22i; nuc'sid4i8; nutri7,33,4i,59,66,7i,84, 201-5-4-6-^-9-11-25,402-10-^23-^-40-2-5-5. Ornith443; osm-pres437; oxi- das8; oxidat36-7,68,79,275,422-35-7; oxydon8l--?; O219; ox-purin4i7. Pancr279, cyst77; paracas403-4-5-ö ; parathyr66,ect33,423-4 ; peps222-7o; permeab39; per- sim7i3; phagocyt50; P0^2i8; phos'tid65; P424-7; phyrpyrr2i4; pigm242; 2 This series of abbreviations, illustrating all others in the index, represents the following sequence of numerals; 17, 72, 75, 220, 253, 280. The numerals in bold-face type here are omitted from the abbreviations above. 566 Biochcmical Bihliography and Index [July pituitr74; plastein6; polariz2i7; polem34,53,2i 1-^-9-24-5-9-41-69,433--^; pol- riceöos; polyneuröoS; polypep236; porphyr2iS; port-vein79; potent24,56-7 ; pregnig; proceed703-i7; prolin443; prot'n2i,36,4i,6(}-i',2oS-35-4i-8i,4o8-28-3i-^- 40-5,603-^-10,705; ptyal249; purin268,4i7-25-9; put'fac6o,244; pyrid276; py- rim4i8; pyromucur-ac7il ; pyrrol2i4. Reac49,5 1,604; red-cab'gi8; reduc20,274; ren-art426; renn222; resis54; resp7o8-9-io; nce6oS; RitthausenH70i. Saliv249; salmin4o8; sar'Iac-ac432 ; scales259; sea-urch422-36; seedi,2,3,i2,23,78; seri9,38, 40,72 , 246-67,43 i,alb72; shock7io; skin266; snak266; Soc-Exp-Biol-Med703 ; Na'acet84,hy'br227,nitr2o6 ; spec'phot207 ; stain26,264; star-fi436; ster'isois; stom232; succ-ac443; sugar27,23i-8o,beet8,meth7o; surf-ten35,76; sweatS2; synth9,2o6-i4-24-55. Tannas402; tan-ac83,40i-i',mass7i3; tart-ac42; temp52; terp-alcoh9; terp'olio; test83,6o3; testim702; thiohydantoin4io; thiopurin4i7; thymus25o; thy'parathy'ect66; thyr'd277-5,427,ect33 ; tiss2i2,6o7; toxin3i; trike- tohydrindenhydr246-7 ; tnmyrisi4; tripept37; tryps240,420 ; tryptoph705; turic- in2S4-5; tyros24i,6o8-9,ase434. Ult'vi-rt42; urea6i,20i-9-i 1-27,414-6; ureom6i; ur-ac 11,210-29-68; uricol429; urinii,30,6i-.^,2i2-27-33-40,4i4-2i-39-6oi. Val- enc4o6; valin442; vit-stain26. Work52,7o8. Xanth4i7. Yeast45,63,202-74-8o-z. Zein438; Zn-salt2io-6S-8o-/. BIOCHEMICAL NEWS, NOTES AND COMMENT Contents. — I. General: Necrology, 567; in memoriam, 567; honors, 567; res- ignations, declination and appointments, 568; lectures, 570; buildings, funds and scholarships, 570; commissions, 572; miscellaneous items, 573. II. Columbia University Biocheniical Association: i. General notes, 574; 2. Proceedings of the Association, 577; Columbia Biochemical Department, 578. I GENERAL Necrology. N. H. Alcock, prof. of physiology, McGill Univ. — Francis Gotch, Waynflete prof. of physiology, and fellow of Mag- dalen Coli., Oxford Univ. — Max Kassowits, prof. of diseases of children, Vienna. — William McMurtrie, one of our leading indus- trial chemists, formerly chief chemist, U. S. Dep't of Agric, and prof. of chemistry, Univ. of Illinois. In memoriam. At a recent meeting of the N. Y. Soc. of Anes- thetists, the following memorial was drafted: " Algernon Thomas Bristow — His editorial and practical hospital support of the ad- vancement of the art of general anesthesia has time and again aroused the admiration and emulation of this body of anesthetists, and so active and influential has this work among his many follow- ers been, that this society wishes itself placed on record as consider- ing his work monumentally constructive, and its cessation as a great loss to the surgical world." Honors. Knighthood. Dr. E. A. Schäfer, prof. of physiol- ogy, Univ. of Edinburgh, has received the honor of knighthood. Order of merit. Dr. Hans Molisch, director of the Inst, for Plant Physiology, Univ. of Vienna, has been invested with the Order of the Iron Crown. HoNORARY DEGREES. Columbia Univ. : Dr. Alexis Carrel (Rockefeiler Inst.), Doctor of Science. — Princeton Univ.: Dr. Simon Flexner (Rockefeiler Inst.) and Dr. David L. Edsall (Har- vard Med. Seh.), Doctor of Science. — Univ. of Edinburgh: James Wilson (lately Sec'y of Agric), Doctor of Laws. — Univ. of Michi- 567 568 Biochemical News, Notes and Comment [july gan: Dr. Ludvig Hektoen (Univ. of Chicago) and Dr. Lafayette B. Mendel (Yale Univ.), Doctor of Science. — Yale Univ.: Dr. David F. Houston ( Sec'y of Agric. ), Doctor of Science ; Dr. Harvey Cushing (Harvard Med. Seh.), Master of Arts. Presidency of THE Amer. Medical Association. Dr. Victor C. Vaughan, pro f. of hygiene and physiol. chemistry, Univ. of Michigan, and dean of the dep't of medicine and surgery, was elected president of the Amer. Med. Assoc. at the recent annual meeting. ..." In the selection of Dr. Vaughan, the Amer. Med. Assoc. has done deserved honor to one of the eminent members of the medical profession — one who is eminent not only as a physician, but also as a chemist, as a medical teacher, and as a scientist." {Ed.: Jour. Amer. Med. Assoc., 191 3, Ix, p. 2053.) RiCKETTS prize. Dr. George L. Kite and Mr. Esmond R. Long, graduate students in the dep't of pathology and bacteriology, Univ. of Chicago, have been jointly awarded the Howard Taylor Ricketts prize ($250) for original research in that dep't. AwARDs OF medals. Tlie Hanbury medal of the Pharmaceu- tical Soc, London, has been awarded to Dr. Frederick B. Power (director, Wellcome Research Lab., London). The medal is awarded biennially for original research in the chemistry and natural history of drugs. — Dr. Harry C. Jones (Johns Hopkins Univ.) has been awarded the Edward Longstreth medal of the Franklin Inst., of Phila., for his work on the nature of Solution. Resignations, declination and appointments.^ Resignations. Dr. L. H. Bailey, director, N. Y. State Col. of Agric, Cornell Univ. (p. 569). — Dr. Geo. F. Gracey, prof. of chemistry and toxicology, Univ. of Texas. — Dr. Paul G. Woolley, dean, medical dep't, Univ. of Cincinnati. Declination. Dr. Emil Abderhalden, prof. of physiology, Univ. of Halle, has declined a call to Vienna to succeed Prof E. Ludwig as head of the Inst, for Medical Chemistry. Appointments. Berlin Univ.: Prof. Ernst Friedberger, head of the Pharmacol. Inst. Cambridge Univ.: Dr. F. H. A. Marshall, univ. lecturer on animal physiology. 1 In this summary, institutions f rom which resignations occurred are named in parenthesis. See page 574. igi3] General 569 Columbia Univ. : Drs. Wm. R. Williams, assoc. prof ., and Henry S. Patterson, assis. prof. of therapeutics (promotions). Cornell Univ. : Prof. W. A. Stocking, Jr., of the dairy dep't, N. Y. State Agric. Col., acting director of the Agric. Col. vice Dr. L. H. Bailey, resigned. General Mem'l Hosp. (N. Y. City) : Dr. Robert C. Lewis, physiol. chemist. Hamburg-Eppendorf : Prof. Emil von Dungern (Heidelberg), di- rector of the newly established institute for experimental Cancer research. Harvard Univ.: Dr. Reid Hunt (chief of the div. of pharmacology, U. S. Public Marine Service), prof. of pharmacology, vice Dr. Frans Pf äff, resigned (p. 310) ; Dr. JV. J. V. Osterhout, prof. of botany (pro- motion) ; Dr. P. G. Stiles (assis. prof. of physiology, Simmons Col.), instr. in physiology. Johns Hopkins Univ.: Dr. L. G. Rozuntree, assoc. prof. of exper.- therapeutics (promotion). Leipzig Univ.: Prof. Walter Kruse (Bonn), director of the Hy- gienic Inst., succeeding Prof. Franz Hofmann. Leland Stanford, Jr., Univ. : Dr. W. H. Manwaring (assis. in pa- thology and bacteriology, Rockefeiler Inst.), prof. of bacteriology and immunity ; Dr. F. W. Weymouth, assis. prof. of physiology (promotion) . Marine Biolog. Lab. (Woods Hole, Mass.), Session 1913: Profs. A. P. Mathews (Univ. of Chicago), R. S. Lillie (Univ. of Penn.), H. C. Bradley (Univ. of Wis.), W. E. Garrey (Wash. Univ., St. Louis), dep't of physiology. Mass. Inst, of Tech. : Dr. E. B. Phelps, assoc. prof. of biochem. research; R. G. Daggett, research assis. in sanitary chemistry; Lester F. Hoyt, assis. in water analysis, succeeding W. J. Daniels. Northwestern Univ.: Medical School — Dr. R. G. Hoskins, assoc. prof. of physiology ; School of Pharmacy — Dr. John H. Long, dean, to succeed the late Oscar Oldherg (page 476). Rockefeiler Inst. : Promotions — f rom assis. to assoc, Dr. F. B. La Forge, chemistry; Dr. /. B. Murphy, pathology and bacteriology; Dr. Martha Wollstein, pathology and bacteriology. New appointments: Dr. W. H. Brown, assoc, pathology and bacteriology ; Dr. C. G. Bull, assis., pathology and bacteriology ; Dr. F. L. Gates, fellow, physiology and pharmacology. (See page 575.) Rutgers Col.: Mr. H. Clay Lint (Kan. Agric. Col.), industrial fel- lowship, plant pathology. 570 Biochemical Nczvs, Notes and Comment [july Tulane Col. of Med, (reorganized) : Dr. /. T. Halsey, prof. of clin. therapeutics ; Dr. C. S. Williamson, Jr. (assoc. prof. of chemistry, Ala. Polytech. Inst.), assoc. prof. of industrial and sugar chemistry. (See page 575.) Univ. of Minnesota: Dr. F. J. Alzvay (prof. of agric. chemistry, Univ. of Neb. and chemist of the Neb. Agric. Exper. Sta.), prof. of soil chemistry and chief of the div. of soils; Dr. A. D. Hirschfelder (Johns Hopkins Med. Seh.), prof. of pharmacy and director of the Seh. of Pharmacy; Dr. E. P. Lyon (prof. of physiology and dean of the Med. Col., St. Louis Univ.), dean of the med. dep't and director of the dep't of physiology, in succession to Dr. F. F. Wesbrook. Univ. of Nebraska: Dr, A. A. Johnson (Western Reserve Univ.), instr. in clin. pathology; Dr. Fred Upson (Univ. of Chicago), prof. of agric. chemistry, Univ. of Neb. and chemist of the Neb. Agric. Exp. Sta., to succeed Dr, F. J. Alway. Univ. of Singapore: Dr. /. A. Campbell (assis, to Prof. Schäfer, Edinburgh Univ,), prof. of physiology. Univ. of Wisconsin: Dr. P. M. Dawson, instr. in physiology. Univ. of Wyoming: Drs. C. J. Oviatt (Mich. Agric. Col.) and A. E. Bowman (Utah Agric. Col.), extension profs. of agriculture. Washington State Exp. Sta.: Dr. Ira D. Cardiff (prof. of botany, State Col, of Wash,), director. He will remain head of the dep't of botany in the College. Western Reserve Univ. : Promotions — Dr. /. D. Pilcher, assis. prof. of pharmacology ; Dr. Paul J. Hanzlik, instr. in pharmacology ; Dr. Roy G. Pearce, instr. in physiology. Yale Univ.: Messrs. S. Goldschmidt and A. J. Hogan, lab. assis. in physiol. chemistry. Lectures. Dr. James W. Johling (Michael Reese Hosp., Chi- cago) delivered the annual address before the Minn. Pathological Soc, May 20, on The toxicity and antigenic properties of the cleavage products of bacterial proteins. — Dr. Oscar Riddle (Car- negie Inst.) lectiired, May 5, under the auspices of the Cornell Chapter of Sigma Xi, on A relation between the storage metabolism of ova and the experimental control of sex. Buildings, funds and scholarships. The committee on medical research, Amer. Med. Assoc, has awarded a grant of $250 to the dep't of bacteriology of the Hoagland Laboratory, Brooklyn, to defray the expenses of an investigation on the immunity reactions 1913] General 571 of edestin.— The British Board of Agric. and Fisheries has awarded research scholarships in agric. science of the annual value of £150, tenable for three years, to the following candidates, among others : W. Brown (Edinburgh), plant pathology; Miss E. C. V. Cornish (Bristol), dairying; E. J. Hohnyard (Cambridge), plant nutrition and soil problems; R. C. Knight (London and Bristol), plant physi- ology; H. Raistrick (Leeds), animal nutrition; Miss T. Redman (London), dairying; G. Williams (Wales), animal nutrition; 6^. P. Wiltshire (Bristol), plant pathology. — The N. Y. State Legislature has appropriated $450,000 for the Coli, of Agric, Cornell Univ., which also receives $125,000 in the supply bill. — Dr. William Duane, for six years radium-research assis. in the Curie Lab., Paris, will organize, for the Harvard Cancer Commission, a laboratory in which Cancer may be studied from the point of view of the physicist. The univ. requires $250,000 for the establishment of this laboratory. Meanwhile, experiments will be conducted in the CoUis P. Hun- tington Building. — The 111. State Legislature has appropriated $4,500,000 for the Univ. of III. for the next biennium. This in- cludes $200,000 for the Coli, of Med. — The special alumni com- mittee on the needs of the Med. Seh., Univ. of Wis., has recom- mended the construction of a medical building to house the dep'ts of physiology, physiol. chemistry, pharmacology, toxicology and bacteriology, and the State Hygienic Lab. ; also a Student infirmary. — The Vienna Society for the Investigation and Prevention of Cancer has established a lab. for experimental work on cancer, mainly in the domain of chemistry and ehem. therapeutics. It will be amalgamated with the Spiegier Inst., which has been in existence nine years. Prof. S. Fraenkel has been appointed director. TuRCK Institute, New York. Dr. Fenton B. Turck has re- moved his office and residence from Chicago to 14 E. 53 St., N. Y. City, where he devotes his morning hours to office practice and the afternoons to his Research Lab., at 428 Lafayette St. This research lab., formerly in Chicago, has been endowed by two former patients, of London, England, who have removed to New York, and reside near the laboratory. The work relates to various problems con- nected with the alimentary tract and is conducted by the Director, Dr. Turck, assisted by the following staff — Organic chemistry: A. 572 Biochcmical News, Notes and Comment [july R. Rose (Univ. of Minn,, Yale and Columbia), Arthur Knudson (Univ. of Missouri and Columbia), Katherine R. Coleman (Co- lumbia Univ.); physiol. chemistry: Vincent Greco (Univ. of Pa- lermo, Italy) ; bacteriology : Otto Maurer (K, Oberrealschule, Heil- bronn, Gemiany, and Univ. of Wis. ), W. W. Browne (Brown Univ.); general pathology: P. J. Friedman (Dept. of Health, Re- search Lab., City of N. Y.) and Earle Kister (Univ. of Toronto). Commissions. In the report for 1912 of the Council of the Amer. Med. Assoc, the appointment of a Commission on Electrical Shock was announced. This commission completed its work in the fall, preparing a chart and a book of directions for resuscitation from electrical shock, both of which were printed by the Electrical World, and distributed free in large quantities to electric-light plants, power houses, factories and other places where electrical cur- rents are in constant use. As a result of the work of this commis- sion, the Council was asked by the director of the Bureau of Mines of the U. S. Dep't of Labor to appoint a similar Commission on Resuscitation from Mine Gases. The following were appointed such a commission: Dr. W. B. Cannon (Harvard Univ.), Chair- man, Dr. 6*. /. Meltzer (Rockefeiler Inst.), Dr. Yandell Henderson (Yale Univ.), Dr. George W. Crile (Western Reserve Univ.), Dr. Reid Hunt (Harvard Univ.), Dr. Joseph Erlang er (St. Louis Univ.). — A recent ruling of the U. S. Public Health Service has been made demanding that all Interstate carriers supply certified water and ice to be used in public drinking fountains, tanks, etc. Under this ruling the various ice and water companies are com- pelled to obtain certification of their ice and water to be used on steamboats and trains. Accordingly, a commission, to be known as the Chicago Ice Commission, has been formed, composed of Drs. Ludvig Hektoen, director of the Mem, Inst, for Infec. Diseases; Edwin O. Jordan, prof. of bacteriology, Univ. of Chicago; and John H. Long, prof. of physiological chemistry, Northwestern Univ., which will undertake the examinations of ice, its source, trans- portation, delivery, etc., and certify the results when found satis- factory. — Dr. C.-E. A. Winslow (Col. of the City of N. Y.). has been appointed chairman of a Commission on the Experimental Study of Ventilation Problems, with an appropriation of $50,000 to I9I3] General 573 be expended during the next four years. The other members of the commission are : Prof. F. S. Lee and Prof. E. L. Thorndike, Co- lumbia Univ. ; Prof. E. B. Phelps, Mass. Inst, of Tech. ; Dr. /. A. Miller and Mr. D. D. Kimhall, New York. The fund is part of a gift, by Mrs. Elisabeth Milbank Anderson, to the Assoc. for Im- proving the Condition of the Poor. Miscellaneous items. Pensions in the Rockefeller Inst. Pensions for the members and assoc. members of the Rockefeller Inst, have been provided by the generosity of Mr. John D. Rocke- feller, who has, for this purpose, increased the endowment of the inst, by a special gift of about $500,000. The rules provide pensions of three-quarters of füll pay for members of the inst, who retire at the age of 65, after fifteen or more years of service, and pensions of from one-half to three-quarters of füll pay, according to the length of Service, for members and assoc. members who retire at 60 years of age. There is also a provision for total disability after ten years of service, and for widows and orphaned children, at one- half the Scale upon which members of the staff are pensioned. American Chemical Society: Biological Division. The Sect. of Biolog. Chemistry, of the Amer. Chem. Soc, will meet in Rochester on Sep. 10 and 12, when the Organization of the section into a division will be completed. Drs. W. D. Bancroft, Edw. Kremers and A. W. Dox with the officers, Carl L. Aisher g (chair- man) and /. K. Phelps (secr.), are the committee in charge of this matter (p. 480). Society of the Sigma Xi. Regent elections of officers (see page 481) — Pres, of the Leland Stanford, Jr., chapter: Prof. R. E. Swain; treas. of the Brown chapter : Prof. P. H. Mitchell. Prizes. Elie de Cyon prize. The de Cyon prize ($600) is open for the third international competition until March i, 191 5. The prize will be awarded for the best printed or manuscript work (printed since Mar. 191 3) on the functions of the internal ear, thyroid, hypophysis or pineal gland. The Acad. of Sei., Bologna, has Charge of the administration of the prize fund. — Emil Chr. Hansen Prise Fund (see p. 535). Personalia. Dr. Martin H. Fischer, who was seriously ill with appendicitis, has happily recovered. 574 Biochemkal News, Notes and Comment [July Dr. F. G. Hopkins, reader in ehem. physiology, Univ. of Cam- bridge, has been appointed a member of the Med. Research Commit. under the National Health Insurance Act, Eng. Dr. Howard B. Lewis, formerly assis. in physiol. chemistry, Sheff. Sei. Seh., Yale Univ., has been appointed one of the first ineumbents of the Seessel Fellowships for the encouragement of research in biological subjects at Yale Univ. The names of Drs. Jacques Loeh (Rockefeiler Inst.) and A. P. Mathews (Univ. of Chicago) are among those constituting the latest official list of Trustees of the Marine Biolog. Lab., Woods Hole, Mass. Prof. Graham Lusk is a member of the Board of Managers of the Biolog. Lab. of the Brooklyn Inst, of Arts and Sei., Cold Spring Harbor, L. I. Dr. W. A. Murrill, assis. director of the N. Y. Botan. Garden, is in Europe, studying types of fungi and the efifeet of tar dust on the trees planted on roads wliere the surfaee binding is of tar. Prof. Howard S. Reed, of the Virginia Polyteeh. Inst., who spent the year in Europe, was a delegate to the Tenth Intern. Congr. of Agric, in Ghent, June 8 to 12. Dr. H. M. Richards (Columbia Univ.) has been elected a vice- president of the Torrey Botan. Club and reeleeted an editor of tb.e Bidl. of the Torrey Bot. Club. He is one of the three editors of Physiological Researches (see p. 576). Dr. Victor C. Vaughan was recently reeleeted president of the Mich. State Board of Health. II. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION I. General notes Marriage: On June 19, Miss Charlotte Cecil Marie Verlage and Dr. Marston Lovell Hamlin (Harriman Research Lab., Roose- velt Hosp., N. Y.) Appointments.^ Dr. Louis E. Bisch (N. Y. Post-Grad. Med. Seh.), leeturer in edueational psychology, Teachers Col., Columbia Univ. — Dr. /. /. Bronfen Brenner (Rockefeller Inst.), director of 2 See footnote, page 568. 1913] Columbia Biochemical 'Association 575 the pathological lab'y, Western Pennsylvania Hosp., Pittsburgh. — Dr. Allan C. Eustis, assis. prof. of dietetics and nutrition, Tulane University (promotion). — Dr. R. F. Hare (prof. of chemistry), vice-director, New Mexico Col. of Agric. and Mech. Arts. — Dr. Michael Heidelberger, assistant in chemistry, Rockefeller Inst, (promotion). — Dr. Burton E. Livingston (prof. of plant physiol- ogy), prof. of plant physiology and director of the lab. of plant physiology, Johns Hopkins Univ. — Dr. Gustave M. Meyer, assoc. in chemistry, Rockefeller Inst, (promotion). — Dr. Winifred J. Robinson, assis. prof. of botany, Vassar Col. (promotion). — Dr. Oscar M. Schloss has succeeded Dr. Ira S. Wile as Con- ference physician at the Riverside Kitchen of the N. Y. Diet Kitchen Assoc. — Dr. Arthur W. Swann has succeeded Dr. Otto von Huffman as instr. in clin. pathology, Columbia Univ. — Dr. Edwin D. Watkins, assoc. prof. of gynecology, and prof. of general surgery in the dental dep't, Univ. of Tennessee (Memphis). — Dr. Wm. H. Welker, assis. prof. of physiol. chemistry, Col. of Med., Univ. of 111. (Chicago). Instructors AT SUMMER SESsioNs, 1913. Biolog. Lab. of the Brooklyn Inst, of Arts and Sei. (Cold Spring Harbor, L. I.) : Prof. David D. Whitney (Wesleyan Univ.), compar. zoology. — Biolog. Sta. of the Univ. of Montana (Yellow Bay-on-Flathead Lake) : Prof. /. E. Kirkwood (Univ. of Mont.), botany and forestry, — Marine Biolog. Lab. (Woods Hole, Mass) : Prof. L. L. Wood- ruff (Yale Univ.), embryology. (See page 579.) Miscellaneous items. Woods Hole Corporation. The latest official list of the "members of the corporation" of the Marine Biol Lab. (Woods Hole, Mass.) contains the following names of members of the Biochem. Assoc. : Carl L. Aisberg, Cora J. Beck- with, William J. Gies, A. J. Goldfarb, H. B. Goodrich, Louise H. Gregory, E. N. Harvey, Mildred A. Hoge, Jacques Loeb, Max Morse, Raymond C. Osburn, Charles Packard, A. M. Pappen- heimer, Henry J. Spencer, Charles R. Stockard, Isabel Wheeler, David D. Whitney, and L. L. Woodruff. {Biolog. Bidl., 1913, xxiv, P- 454-) Members of the Association lately admitted to the So- ciety FOR Experimental Biology and Medicine: Drs. /. /. 57^ Biochcmical Neivs, Notes and Comment [July Bronfen Brenner (West Penn. Hosp. Pittsburgh), Russell L. Cecil (Presbyter. Hosp., N. Y.), H. H. Janeway (City Hosp., N. Y.), Edivards A. Park (Johns Hopkins Univ.). Personalia. Prof. A. R. Bliss (Birmingham Med. Coli. ; Grad. Seh. o£ Med., Univ. of Alabama) has been elected second vice- president of the Alabama Sect. of the Amer. Chem. Soc. — Dr. E. D. Clark has been reelected to the editorial board of the Bull, of the Torrey Botan. Club. — The Board of Managers of the Van- derbilt Clinic (Columbia Univ.) have recently issued, in pamphlet form, the interesting "Report of the day camp of the Vanderbilt Clinic of the Coli, of Phys. and Surg.," by Dr. F. Morris Class, Attending Physician. — On July 23 an expedition for the study of marine biology, under the auspices of the Carnegie Inst., Wash., set sail from San Francisco for Thursday Island, Torres Straits, Queensland, Australia. Dr. E. Nezvton Harvey was a member of the party. — The med. col. of Phila. gave a reception, April 30, at the Bellevue-Stratford Hotel, in lionor of Prof. Philip B. Hawk, new member of the faculty of Jefferson Med. Col. — Prof. B. E. Livingston is the managing editor of Physiological Researches, the first issue of which has just appeared. He is also a member of a special committee of Sect. G (Botany) of the Amer. Assoc. Adv. Sei., to consider affiliation with the Botanical Soc. of Amer. — The Univ. of Texas gave a " One-Week's School of Home Economics," from Feb. 10-15. Among the lectures and demonstrations v^ere: Nutritive value of foods; the importance of the menu, by Miss Anna Richardson and Some problems of house building; sanitary Problems of the home; the house and how to plan it, by Prof. Mary E. Gearing. — Dr. Winifred J. Robinson has been appointed an adviser for women at the summer school of the Univ. of Wisconsin. — Mr. Carl A. Schwärze's work (N. J. Agric. Exp. Sta.) on the relation of enzymes to " peach yellows " and " little peach " is pro- gressing favorably. He is planning to study an enzymic disorder of the tomato plant: " filiform-leaf tomato." — Dr. F. J. Seaver has relinquished his editorship of the Jour. of the N. Y. Botan. Garden and will devote more of his time to Mycologia. — ^We are glad to note that Prof. E. A. Spitzka has returned from his trip abroad, with his health fully restored (p. 485). 1913] Columbia Biochemical Association S77 2. Proceedings of the Association Proceedings of the twelfth scientific meeting (page 486). Fourth annual meeting. The fourth annual meeting of the Assoc. was held at the Columbia Med. Seh., June 2, 1913, at 8.30 P. M. Dr. Emily C. Seaman occupied the chair. Abstracts of the Com- munications constituting the scientific proceedings are given in this issue at page 541. After several amendments of the Constitution had been adopted (one of them ehminating the Article on Dues), the following offi- cers were elected (1913-14) : HoNORARY OFFicERS. President, Dr. Carl L, Aisberg; Vice- presidents: Dr. Hugh Auchincloss, Dr. William B. Boyd, Prof. Mary E. Gearing; Dr. James C. Greenway, Prof. Mary E. Sweeny. Active officers. President, Prof. Stanley R. Benedict; Vice- president, Dr. Frederick T. Van Beuren; Secretary, Dr. Alfred P. Lothrop; Treasurer, Prof. William J. Gies. At the Suggestion of the Ex. Committee, Prof. Hugo Kronecker, Director of the Physiological Institute of the University of Bern, was unanimously elected an Honorary Member of the Assoc, and Prof. Lafayette B. Mendel, of Yale University, was invited to be the guest of honor at our third annual dinner, next November. The Assoc. has been highly honored by letters of acceptance from both our distinguished colleagues. The Ex. Committee's selections of Corresponding Members were enthusiastically endorsed ; the füll list is given on page iv. The secretary's official register of members, including those elected at this and all other meetings during the year, was approved as read (page iv). By unanimous vote, and in accord with the amended Constitution, the Assoc. approved the Ex. Committee's suggestions ( i ) that the Ed. Committee in charge of the Biochemical Bulletin be en- larged to include the members who are actively engaged in bio- chemical work, (2) the new Ed. Committee to take the place of the Board of Directors and the former Ed. Committee, (3) the secre- tary of the Assoc. to be the chairman of the Ed. Committee and (4) the treasurer of the Assoc. to be the managing editor of the Bio- chemical Bulletin. 5/8 Biochemkal News, Notes and Comment [July The Assoc. adjoumed at a late hour, after a very happy and stimulating meeting. Alfred P. Lothrop, Secretary 3, Columbia Biochemical Department Marriage: On June 18, Miss Harriet Beckwith Rinaker and Prof. Paul E. Howe. Appointments from the staff. Dr. Nellis B. F oster (assoc), instr. in medicine, Cornell Univ. Med. Seh., with a laboratory in the N. Y. Hosp. — Dr. Max Kahn (assoc), Consulting physician in dietetics, Beth Israel Hosp., N. Y. He will continue as assoc. here and as director of the chemical lab. of the Beth Israel Hosp. — Dr. Louis E. Wise (instr.), instr. in chemistry, Univ. of Missouri. — Dr. Edgar G. Miller, Jr. (assis.), instr. of physiol. chemistry, Univ. of Illinois (Med. Seh., Chicago).^ — Mr. Arthur Kmidson (assist), chemist, Turck Inst., N. Y. (p. 571). — Dr. Joseph S. Hephurn (univ. fellow), assis. chemist, U. S. Dep'tof Agric (Food Research Lab., Phila.). Appointments to, and promotions in, the staff. Dr. Max Kahn (instr.), associate. — Dr. Frederic G. Goodridge (assis.), instructor. — Mr. W. A. Perlzweig, assistant. — Mr. Victor E. Levine, laboratory assistant (summer Session, 1913). Laboratory at Bellevue Hospital. During the past academic year Prof. Gies, aided by Dr. Edgar G. Miller, Jr., equipped a small chemical lab. in the patholog. dep't of Bellevue Hosp., and con- ducted research and routine work there in colläboration with Drs, Charles Norris and Cyrus W. Field. Mr. Grover Tracy assisted in some of the work. It is planned to extend this Service as the needs of the hospital may determine. Members-elect of societies. Amer. Soc Biol. Chem. : Dr. Alfred P. Lothrop; Soc. Exp. Biol. Med. : Prof. Paul. E. Howe; Amer. Chem. Soc. : Dr. Walter H. Eddy; Sigma Xi : Robert P. ^ Dr. Miller had been appointed prof. of physiol. chemistry in the new med. school of the Univ. of the South, Nashville, Tenn., but Mr. Carnegie's gift of $1,000,000 to the med. school of Vanderbilt Univ., in the same city, induced the Trustees of the Univ. of the South to abandon their med. school. 1913] Columbia Biochemical Department 579 Calvert and Sidney Born; Phi Lambda Upsilon : Arthur M. Buswell and Robert P. Calvert. Awards of higher degrees at Columbia to students of bio- logical chemistry. Doctors of philosophy. Of the nineteen recipients of the degree of Ph.D. under the Faculty of Pure Sci- ence, at Columbia's last commencement, nine had taken "majors or minors," or both, in the biochemical department. The names of the candidates and their major and minor subjects are given below : Name of candidate Major Minor Minor B. G. Feinberg chemistry chemistry biological chemistry H. D. Goodale zoology zoology biological chemistry J. S. Hepburn* biological chemistry biological chemistry chemistry Benj. Horowitz biological chemistry biological chemistry education Edgar G. Miller, Jr. biological chemistry bacteriologv pharmacology Anton R. Rosef biological chemistry bacteriology chemistry Clayton S. Smith biological chemistry bacteriology physiology Edward C. Stone chemistry chemistry _ biological chemistry Charles Weisman biological chemistry biological chemistry chemistry Masters of Arts. The A.M, degree was recently conferred lipon the following advanced students in the biochemical depart- ment : Anna M. Connelly, Helen B. Davis, Mary C. de Garmo, Ula M. Dow, Gustave Egloff, Frank R. Eider, Ada M. Field, Beatrix H. Gross, Clara W. Hasslock,'^ Grace F. Hinchliff, Edward Plaut, David F. Renshaw,^ Elisabeth Rothermel, Mary E. Sweeny,^ Fred L. Thompson, Helen B. Thompson, Jennie A. Walker, Isabel Wheeler. DocTORS OF Pharm ACY. The following students of biological chemistry at the School of Pharmacy received the degree of Phar.D. : Ainslie Bück, William G. Crockett, Albert A, Muench, Herbert C. Oehlers, Elsa G. Pickhardt, Hugo H. Schaefer, and Leo Stein. Summer session. Courses. The department is conducting six courses in nutrition and biochemical methods at the summer session now in progress at Columbia. Three of these courses are given at Teachers Coli., by Prof. Gies, Dr. E. C. Seaman and Miss Tula L. Harkey; three are given at the Col. of Phys. and Surg., by Prof. Gies and Messrs. W. A. Perlzweig and Victor E. Levine. The bio- *University fellow in biological chemistry, igi2-iZ- t The degree was awarded in February, 1913. ' * The degree was awarded in October, 1912. 58o Biochemical News, Notes and Comment [July chemical lab. at the Med. Seh. is open daily for research, and will continue so throughout the summen Investigators. The workers named below have been engaged in research, in the biochemical laboratoi-y at the medical school, at various times during the vacation : Louis Berman, A. M. Buswell, Arthur D. Dryfoos, Walter H. Eddy, Mary L. Edzvard, L. L. Falke, Helen Gavin, William J. Gies, Mark J. Gottlieb, Tula L. Harkey, Max Kahn, I. J. Kligler, W. M. Kraus, Alfred P. Lothrop, Her- man O. Mosenthal, William A. Perlzweig, Nathan Rosenthal, Oscar M. Schloss, Emily C. Seaman, A. W. Thomas, M, K. Thornton, William Weinberger, Charles Weisman, G. H. Worthing. Miscellaneous items. Prof. Gies is a member of the third sec- tional committee of the Third Internat. Gong, of Refrigeration to be held in Washington and Chicago, Sept. 15-24, 19 13. He was one of the Speakers at the annual banquet of the First District Dental Soc. of the State of N. Y., at the Hotel Astor, Jan. 18. He addressed the Harlem Dental Soc, April 24, on the Prevention of dental caries. On May 9 he presented, at the 45th annual meeting of the Dental Soc. of the State of N. Y., the report of the Research Committee, embodying the results of a further investigation of the origin and physiological significance of salivary sulfocyanate (under the auspices of the Research Committee), with the Cooperation of Prof. C. C. Lieb, Drs. Max Kahn and Edgar G. Miller, Jr., and Mr. Arthur Knudson. At the conclusion of his report, and pur- suant to a recommendation in the president's annual address, Prof. Gies was elected an honorary member of the Dental Soc. of the State of N. Y. He has been invited to continue to direct the soci- ety's study of dental caries. At his Suggestion the title of The Jonr. of the Allied Societies^ has been changed to The Jour. of the Allied Dental Societies (1913, viii : Mar.). Miss Jean Broadhurst is utilizing a year's leave of absence in Special study, at Ithaca, in plant physiology and bacteriology. She has received from the Torrey Botanical Club a grant of $200 from the Esther Hermann Fund to assist her in an investigation of bacteria in milk. Miss Broadhurst was recently reelected one of the editors of the Bull, of the Torrey Bot. Club. ^Jour. All. Soc, 1912, vii, pp. 408 and 507. 1913] Columbia Biochemical Department 5^1 The department was honored during the year by the return of Prof. John S. Adriance (WilHams Col.), who reviewed the work required of first year students of medicine. Mr. A. T. Cameron (Univ. of Manitoba) was a welcome visitor in the laboratories of the department during the months of May and June, and an auditor at some of the Conferences at the close of the academic year in May. Prof. Gies recently resigned his membership in the Board of Trustees of Irving Col. for Women (Mechanicsburg, Pa.), of which he had been a member since 1900. He was recently reelected an alumni representative in the Board of Trustees of Gettysburg Col. for a term of six years. Mr. Arthur M. Buswell has been awarded a university fellow- ship in chemistry for I9i3-'i4; Mr. Frank R. Eider has been appointed alternate. Mr. Guy West Wilson, lately of the N. C. Agric. Exper. Sta., and, during the past year, research scholar at the N. Y. Botan. Garden, has recently been appointed a special agent of the U. S. Bureau of Plant Industry for the study of the relation of the chestnut blight fungus to tannin and other plant products, at Rut- gers College, New Brunswick, N. J. On May 11, Dr. Max Kahn feil headlong from a moving street car. There was no internal turmoil to account for this mishap (official). His prompt recovery was very gratifying to all his associates. Prof. Gies has been requested by the N. Y. Sabbath Committee to direct, under its auspices, a study of the physiological value, if any, of the weekly day of rest, entirely apart from religious or preconceived notions, and wholly from a scientific Standpoint. The work will be inaugurated in the fall. [Makes a noise like the beginning of a reform movement in this laboratory. Printer' s devil.] EDITORIALS We learn from a Paris correspondent that Dr. Jules Wolff, of the Pasteur Institute, is about to publish in the Annais de VInstitut Pasteur (July), a paper in which the author reviews current knowl- Peroxides and edge on the subject of the presence of peroxides nitrites in plants Jn plants. Affirmed by some, disputed by others, the question of the occurrence of these substances in plants has out- lived numerous controversies. Kastle and Loevenhart, and Chodat and Bach, have attributed to peroxides the phenomena which Aso and others attribute to nitrites. Maze has shown that the sap of all the higher plants contains nitrites. On the other band, Wolff has found that nitrites are decomposed by the more or less acid Juices of plants, and thus can induce phenomena of oxidation com- parable to those which occur with the aid of the system: peroxidase — hydrogen peroxide. Wolff doubts the importance of peroxides and peroxidases as physiologic agents because they rarely occur together in vegetahle Juices. On the other band, the number of substances that can be attacked by nitrous acid far exceeds the number that may be oxidized by the System : peroxidase — hydrogen peroxide. However, Wolff has found, especially in apples {unpub- lished data), a special peroxide which is produced only when there is a lesion of the tissue. The brown color that develops on cut sur- faces of fruit is a result of the action of this peroxide on a chromo- genic substance. Such phenomena of color ation, contrary to cur- rent opinion, are not due to oxidases. In Wolff's view, such pigmentation residts from the combined action of a peroxide and a peroxidase. X. In our April issue we presented a number of typical replies to our circular note inviting expressions of opinion on the " Mathews plan for the Organization of an American Biological Society" (p. The Mathews plan 49^)- ^^ the little space available in this con- for an American cluding number of Volume II, we present a few Biological Society additional selections from the comment for- warded to us on this subject. The October issue will continue this 582 1913] Editoriais 583 plan of facilitating open consideration and possible removal of the difficulties in the way of more effective biological Organization in this country, and will also contain a summary of the ideas expressed, in this Journal, on the Mathews plan of reorganization. James P. Atkinson, A^". Y. City Dep't of Health. I believe the plan for the Amer. Biolog. Soc'y to be a very feasible one, espe- cially as regards the combining of one or more of the Journals. It seems to me to be especially adapted to individuals who cannot afford to subscribe to many Journals, and I hope that it will be successful. R. P, BiGELOW, Mass. Inst, of Technology. The plan of Organization for the Amer. Biolog. Soc'y proposed by Prof. Mathews seems inadvisable for the f ollowing reasons : 1. Details of Organization. The society as proposed would simply be a confederation of existing biological societies, which in turn would become sections of the new society. The large size, national scope, and wide ränge of interest in the new society would involve (a) difficulties in securing suitable places for meetings, (&) long journeys and hence less regulär attendance of many members, (c) long programs and formality of proceedings; as contrasted with independent small societies meeting within a restricted area and limited in scope, which have for their main object the promotion of intimacy and good-fellowship, with opportunity for informal dis- cussion among men whose interests are alike. 2. Ohjects. (a) Cooperation in the abstract seems a rather hazy basis on which to found a new society. Moreover, when a definite need of Cooperation arises, organizations suitable for this purpose may be found in the A. A. A. S. and the Amer. Soc'y of Naturalists. (&) The starting and supporting of a Biolog. Abstract Jour. seems to involve competition rather than Cooperation. Such a publication, to be of any practical use, should segregate in one place the results of a year's work at least in each division of the subject and should be international in scope. This is the aim of L'Annee Biologique, Ergeh, d. Anat. u. Entzvickelungsges., Ergeh. d. Physiol., and the Jour. of the Royal Micros. Soc'y. Why not coöperate towards the improvement of one of these existing publi- cations, instead of starting a rival Journal, (c) The attempt to 584 Mathezvs Plan for American Biological Society [July induce all biologists of the country to subscribe to all biological Journals published in America is foredoomed to failure. For no one can pretend to keep up with the advances in all the branches of biology, and, moreover, the cost of storing the volumes would soon become embarrassing to anyone who attempts to live on a biolog- ical salary. The suggested reduction in cost would seem to depend on the formation of a society approaching in size the Amer. Chem. Soc'y, but until biology shall have as profitable applications as chemistry, this seems rather a hope than a possibility. G. W. Crile, Cleveland, Ohio. Although I appreciate the many arguments advanced in favor of Prof. Mathews' plan, I do not see my own way clear to express an unqualified commendation of the idea, as I should want to be assured that the work of this associa- tion would covfer a field which has not as yet been touched upon by any other existing society, and that could not be developed in some one of them. Arthur W. Dox, Iowa State College Agric. Exper. Station. I fail to see any special advantage to be gained by consolidating the existing biological societies, much less by organizing a new society. The chief inducement appears to be the clubbing rate which would enable the members to secure a greater number of Journals at a low cost. I am personally of the opinion, however, that the majority of the biologists, were they to increase the num- ber of their subscriptions, would derive greater benefit by subscrib- ing to a few foreign periodicals in their chosen field than by taking on a greater number of Amer. Journals. Walter H. Eddy, High School of Commerce, N. Y. City. The modern biology teacher in the secondary school is largely concerned in presenting the phases of biology that are intimately related to the results of current research. At the same time he is rarely in a local- ity where he can keep in intimate touch with such research through libraries or research institutions, and even in so rieh a Community as N. Y. City the field is so broad and the interests so varied that here too he is dependent upon Journals for most of his Information. In view of all these facts and in view of the financial impossibility of indefinite subscription to Journals, any such plan as Dr. Mathews 1913] Editoriais 5^5 suggests, which aims to concentrate and assist the efforts of the secondary school biologist to keep abreast of the times, must com- mend itself to him most forcibly. C. Stuart Gager, Brooklyn Botanic Garden. While I think that an Organization such as is proposed by Prof. Mathews has admirable features, especially if it were an initial step in the Organi- zation of the biological interests of the country, nevertheless, I do not feel that it could be successful, or that another society is now desirable. I also think that the inducement of receiving periodicals over such a wide ränge of interests as experimental psychology to bacteriology would not be very attractive to most workers, whose interests are too narrow to respond to such an inducement. It seems to me that the Mathews plan would result in an Organization which in actual fact would not prove to be much more coherent than does the present Amer. Assoc. for the Adv. of Science, with its various sections, some of which at present exist chiefly on the printed program. Paul J. Hanzlik, Western Reserve Univ. So far as the uni- fication of the various biological societies is concerned, there is already the " Federation." This should suffice. The Biolog. Ah- stract Joiir. would be a supernumerary because there are already several abstract Journals which cover adequately the field of the biological sciences. Among the most widely circulated are probably the Zent. f. Bioch. u. Biophy. and Chem. Abstracts. It would be an additional financial bürden for the Society. This is objectionable. I understand that Chem. Abstracts is a very expensive enterprise and, in spite of the very large membership (running into the thou- sands) of the Amer. Chem. Soc'y, its financial support is not adequate for its needs. It would be much more difficult for the Amer. Biolog. Soc'y with its comparatively small membership to support Journals to the extent that the Amer. Chem. Soc'y does. Therefore, I disagree with the idea of organizing an "Amer. Biolog. Soc'y" and the establishment of a Biolog. Abstract Jour., as proposed in the Mathews plan. Chas. W. Hargitt, Syracuse Univ. One feature of the propo- sition appeals to me, namely, that of the Abstract Journal. I have $86 Mathczvs Plan for 'American Biological Society [July long feit that there was a real place for such a Journal among us, and have several times urged such both orally, and in written and printed Communications. At present our biological Journals are not fully meeting the necessities for either general or prompt publica- tion, and we get next to nothing in the way of abstracts of literature or reviews of current publications. I should like to see something after the plan proposed by Mathews put into Operation so far as a Journal of this sort is concerned. Concerning the proposed " Society " I have serious doubts. Already we have more societies than we can support decently. And to add another to the list, unless it were of such a character as to meet a real need, could hardly commend general or enthusiastic support. If one or two which long ago passed into practical desuetude could be allowed to find a niche in some mortuary hall of fame it could well be possible to fill the place by a society something after the Mathews plan. I should incline to hope very sincerely that its name might be unencumbered by that overworked adjective "American" ! Pray let us have at least one society which can be trusted to stand on its own characteristic merits, without expletives, apologies, or explanations. E. M. HouGHTON, Detroit, Mich. After careful consideration of the subject, I most heartily commend the proposed plan as one that will be most desirable for those of us who are specially inter- ested in biological subjects. I hope that the movement may gather sufficient force to put the matter into practical Operation. J. S. KiNGSLEY, Tufts Col. I have been greatly interested in Dr. Mathews' plan for the Organization of a general society to Cover all sides of biology and wish that it could be consummated, but I am afraid that he has overestimated the membership and income of such an Organization and has underestimated the expenses of the Journals to such an extent that it vitiates the whole scheme. Thus I believe that a Journal of abstracts, well done, would be of great value, but I do not see in the plan any adequate provision for the payment for the abstracts, and I know from experience that there are few who can be depended upon to do the work as a labor of love. The Joiir. of the Royal Micros. Soc'y publishes such ab- stracts in a limited field, and one of the editors told me that he was paid a salary of £120 a year for about half of the zoological ab- 1913] Editoriais 5^7 stracts. Then Mathews has failed to take into account the many overhead expenses in such a series of Journals — postage, clerical work, corrections, bad accounts and a thousand other items which are inevitable. F. C. Koch, Univ. of Chicago. The Mathews plan for the Or- ganization of the Amer. Biol. Soc'y appeals to me as practicable without doubt, and as very desirable and necessary. Edwin Linton, Washington and Jefferson Col. A large num- ber of working biologists are to be found among teachers in high schools and the smaller Colleges. Many of them are obliged to conduct elementary courses in as diverse subjects as anatomy, bac- teriology, botany, physiology and zoology. It is out of the question for the majority of such teachers, or of the institutions which they represent, to subscribe for the list of Journals enumerated by Dr. Mathews on page 264 of the Biochemical Bulletin for January, 1913. To such persons, access to this entire list of Journals would add greatly to their efficiency as teachers and as workers in biology. Therefore, any plan whereby a larger number of the various bio- logical publications will be made available than is now the case to the majority of biologists should command the sympathetic con- sideration of all biologists. F. E. Lloyd, McGill Univ. I am very much impressed with the Biolog. Soc'y proposition, set forth in Mathews' paper. For a long time it has seemed to me that we have a lot to learn f rom the chem- ists, and I quite believe that some such effort as outlined by Mathews would go a long way toward unifying and stimulating effort. I shall be glad to coöperate in any way that I can. I feel that the details should be gone into pretty thoroughly, so as to get the costs down as much as possible, but an abstract Journal analogous to the Chem. Ahstracts would be of immense value. HuGH McGuiGAN, Northwestern Univ. Med. Seh. The Math- ews plan suggests improvements that seem feasible. To make it more definite, the estimated costs should be worked out by a com- mittee of publishers and scientists and an authoritative Statement of the cost presented. The Federation of Amer. Socs. for Exp. Biol. forms an excellent nucleus to commence with and is a step toward the consummation of the plan. 588 Mathezvs Plan for 'American Biological Society [July Jacob Reighard, Univ. of Mich. I take it that each American biologist is now paying for the Journals to which he subscribes as much as he can afford. Most of my friends have as many Journals as they would like. Their libraries contrast unfavorably in this respect with those of their European colleagues. I do not doubt that any plan that would give them more for the same money or for less money would be welcome. On the other band I do not know of many men who would consider the whole list of thirteen Journals of use to them. If there were opportunity for them to get the Journals they now get at less price than they now pay, they would tend to take advantage of it, — and the income of the Society would be reduced by so much. There is one argument not touched upon by Prof. Mathews which rnight have great weight in starting the movement. Every biologist pays out a good deal annually for reprints. If nearly every biologist received the Journals in his field, there would be little use in distributing reprints. Those furnished him by the Journals with- out cost might meet his needs for exchange with foreign corre- spondents. He might thus save a great deal more than he now pays out for subscriptions. I am not convinced of the wisdom of central control for all bio- logical Journals, and should prefer to see Journals left under their present control. I fear that here, as elsewhere, central control is likely to inhibit individual initiative. The interests of the chemists seem to me less diverse than those of the biologists and therefore more likely to be adequately served by a central Organization. I am, then, in favor of Prof. Mathews' plan, with the elimina- tion of the feature of central control. I believe that the advantages of the plan to the individual Journals are such as to insure their permanency without central control. Change of action and interest are the secrets of recreation, as the spirit of Service is the spirit of happiness. — Creelman. The medical facts that have been elicited and elucidated in laboratories during the past fifty years have done more to revolutionize medical practise than the bedside observations of the past two thousand years. — Janeway. INDEX In two divisions: (I) Author index (pp. 589-590) ; (II) Subject index (pp. 591-604) : The subject index {II, p. ^pi) consists of two main portions: (A) Impersonal subjecfs (p. 5pi); (B) Personal subjects (p. 59p). References, in roman numerals, to parts of the summary of Con- tents (pp. x-xiii) and the list of illustrations (p. xv), facilitate use of the volume. I. AUTHOR INDEX The names of the authors of the leading papers, and the general subjects treated by each, may be obtained promptly from the summary of Contents (pp. x-xiii). This division of the index (I) includes not only the names of the main contributors, but also the names of accred- ited authors of abstracts, editorials, quotations, etc. Abbott, JF, 317 AcTON, — , 508 Adler, HM, 216 "Al I. Phatic," 210 Alperin, D, 164 "Ana Phylactic," 210 Atkinson, JP, 583 AUER, J, 269, 279 Barn hart, JH, 491 Beal, GD, 78, 164 Benedict, FG, 134 Benedict, SR, 165 Berg, WN, ioi, 158, 441 Bergey, DH, 491 Berman, L, 547 Bevier, I, 158 Biddle, HC, 288, 386 BiGELOw, RP, 583 Bisch, LE, 165 Blakeslee, AF, 542 Bolduan, CF, 247 Born, S, 166 Bottazzi, F, 379 Bronfen Brenner, JJ, 53, 166, 453, 462 BUNZEL, H, 491 Burnett, TC, 491 Carlson, AJ, 491 Carrel, A, 493 "C. H. Esty," 508 Clark, ED, 5, 168, 412, 465 CoHOE, BA, 464 Coleman, W, 145 Conklin, EG, 493 Crane, — , 508 Creelman, J, 588 Crile, GW, 584 Crohn, BB, 454 Croll, WL, 509 Davenport, CB, 493 Davis, BJ, 158 Davis, BM, 493 Dox, AW, 330, 407, 584 589 Eddy, WH, iii, 169, 291, 419, 548, 584 Elder, fr, 170, 549 Erlanger, J, 271 Erpf-Lefkovics, TA, 233, 292 Eustis, AC, 158, IS9, 28s EwfiNG, EM, 403, 464 Falk, KG, 455 Feldhaus, FM, 319 Fetzer, LW, 282, 346 Fischer, MH, 494 Fitz, GW, 494 FosTER, NB, 206, 292 Friedman, SS, 460 Gager, CS, 585 Geiger, GA, 78, 164 GiBSON, RB, 536 GiES, WJ, 55, 170, 171, 172, 203, 293, 298, 357, 366, 468, 470, 547, 551, 556, 559 590 Index: Names of Authors [July GiTLOw, S, 549 GOLDFARB, AJ, l6l, 286 GOODEIDGE, FG, 178, 292 GORTNER, RA, 287, 463, 494. 524, 532, 542, 544 Greaves, je, 519 Greene, CW, 494 Greenwald, I, 287, 466 Gross, BH, 466 Hall, WS, 495 Halliburton, WD, 128 Hamlin, ml, 410, 455 Hanzlik, PJ, 585 Harding, VJ, 223 Hardisty, RHM, 225 Hare, RF, 173 Hargitt, GW, 585 Harkey, TL, 550, 551 Harris, JA, 287, 524, 530 HARVEy, EN, 50, 456, 464 Hasselbalch, KA, 367 Henderson, VE, 495 Henderson, Y, 146, 495 Hepburn, JS, 467 Hewlett, AW, 495 HiGLEY, GO, 390. 393 H. M. A., 211 HoROWiTz, B, 293 HosKiNS, RG, 495 HouGH, T, 148, 496 HOUGHTON, EM, 586 HowE, PE, 90, 288, 386, 468, 551 HowELL, WH, 497 HOWLAND, J, 139 HoY, WE, 464 HUXLEY, TH, 210 Hyde, IH, 498 Hymanson, A, 457 Jackson, DE, 498 Janeway, HH, 17s, 403, 464 Janeway, TC, 588 " JaUN DiCE," 2X0 Jensen, CO, 535 Jordan, EO, 498 Jordan, — , 508 Jour. Amer. Med. Assoc, 198, 208, 210, 318, 320, 568 Kahn, M, 87, 177, 178, 217, 458, 545 Kaliski, DJ, 460 Kantor, JL, 179 Kingsley, JS, 586 Koch, FC, 587 Kraus, WM, 294 Kribble, VK, zzy Landolph, F, 217 Langworthy, CF, 498 Lee, MT, 179 Leffmann, H, 198 Levine, VE, 552 Lieb, CC, 459 VON Liebig, J, 332 Linton, E, 587 Lloyd, FE, 587 Long, JH, 132 LoTiiROP, AP, 156, 180, 284, 324, 452, 486, 541, 577 Lucas, DR, 545 LusK, G, 135 McClendon, JF, 500 McCrudden, FH, 137 McGuiGAN, H, 587 Macallum, ab, 140 Macleod, JJR, 147, 499 MacNeal, WJ, 499 Mandel, JA, 150 Mann, G, 499 Manwaring, WH, 453 Martin, EG, 500 Mathews, AP, 261 Mathewson, CA, 181 Matsunaga, S, 228 Mattill, HA, 553 Mattill, HI, 553 May, CE, 373, 464 " M. C," 488 Mendel, LB, 129, 138 Miller, EG, Jr., 554 MiNOT, CS, 508 Moore, AR, 500 Morse, M, 161, 162, 288, 290, 457, 500, 546 N. B. F., 206 Neidig, RE, 407 " Nineteen O. Three," 358 NoGUCHi, H, 166, 462 VON Noorden, C, 143 " N. Y.," 490 Osborne, TB, 339 Osburn, RC, 501 Ottenberg, R, 460 Paracelsus, 332 Park, WH, 501 Parker, GH, 502 Patterson, OG, 55s Pearce, RM, 502 Pearl, R, 502 Pekelharing, CA, 297 Peters, AW, 238 " Pharmacologist," 504 " P. H. D.," 538 Potteiger, CR, 158 Punnett, PW, 555 Reighard, J, 588 Richards, AN, 275 Ripley, LB, 162 Rockwood, EW, 504 Rogers, LA, 158 Rose, AR, 21, 163 Rosenbloom, J, 64, 87, 123, 178, 182, 22g, 233, 236, 290, 291, 292, 464 RusKiN, J, 210 Ruttan, RF, 223, 225 Sanders, JA, 373, 464 Schloss, OM, 182 Schmidt, J, 535 Schwarze, CA, 183 Science, 321 Seaman, EG, 184 Seifert, C, 556 SiDGWiCK, — , 332 Smith, CS, 184, 465 Smith, EE, 243 Sollmann, T, 505 Sörensen, SPL, 535 Stanley, — , 210 Steel, M, 547 Stewart, CC, 505 Strauss, H, 141 Suzuki, U, 228 Tennyson, 508 Thomas, AW, 556 Thorndike, EL, 505 ToDD, JL, 505 Weinberger, W, 123, 182, 185 Weisman, C, 186, 29s, 558 Welch, WH, 508 Welker, WH, 70, 175, 186 WiLEY, HW, 506 Williams, AW, 158 Winterstein, E, 5 WoLFF, J, 53 Wood, FC, 506 Woodruff, LL, 465 Woodward, HE, 186 "X," 582 «Yale, 94S," 349 Yerkes, RW, 507 I9I3] Index: Impersonal Suhjects 591 IL SUBJECT INDEX. A. IMPERSONAL SUBJECTS The names of authors are given on pp. 58^^-590. Personal suhjects are indexed on pp. ^pp^6o4. General suhjects may be seen at a glance on pp. x-xiii. This sub- ject index is aimed at details that the titles of papers do not include, altho it also makes due reference to the titles. A recurrenf subject in any paper or formal section of the volume is indicated but once, as a rule, hy the numeral on the first page of its occurrence in, or on the opening or concluding page of, the section containing it. Numerous cross referenccs facilitate prompt access to details. The index ignores impersonal matters that are secondary to, or of no special interest apart from, the personal references to which they pertain {e. g., items of " biochemical news, notes, and comment"). Routine matters {e. g., common tests, ordinary reagents) are not indicated unless they appear in special settings. This comprehensive index is intended to guide the reader to the main path through any and every subject, or group of suhjects, in the volume. Absorption, 103, 550 Abstracts, 132, 285, 453, 541 Academies ; see organiz. Acapnia, 175, 403, 4^4 Acetaldehyde, 76 Acetanilid, 63 Acetic acid, 66, 76 Aceto-acetic acid, 223 Acetone, 66, 76, 80, 96, 223 Acetonitrile, 178 Acetonuria, 223, 285 Achylia gastrica, 455 Acid(ity), loi, 167, 170, 179, 181, 464, SSO, 554 Acid albumin, 547 Acidemia, 286 Acidosis, 135, 28s Acid-protease, 234, 237 Acid salts, 181 Acts of congress, 196 Adaptation, 287 Adrenalin ; see epinephrin Adsorption, 444 Adulteration, 532 Agglutinm(ation),25i,46i Air, 146, 147, 29s, 545, 558 pressure, 403 Alanin, 137, 178 Albumin, 96, 166, 547 Alcannin, 80 Alcohol, 79, 435, 462, 553 Alimentation, forced, 143 Alkali(nity), 51, 167, 170, 180 Alkali albuminate, 547 Alkali-protease, 234, 237 Alkaloids, 62 Allantoin, 2 Allergy, 182 Allspice, 133 Almonds, 182 Amaurotic idiocy, 457 Amide nitrogen, 434, 463 Amino acids, 4, 178 Amino nitrogen, 544 Aminovalerianic acid, 2 Amins, 158, 159 Ammonia (OH), 51, 53, 124, 171, 180, 285, 293, 409, 430, 455, 464, 544, 555 Amphibian larvae, 288 Amygdalin, 227 Amyl alcohol, 82 Amylase, 233, 237, 455, 555 Anaphylaxis, 158,159,256, 296, 545, 558 Anesthe^ta (tics), 176, 567 Anilin mucoid, 115 Annatto, 80 Anniversaries, 189, 309, 3S8 Anti-anit'bodies, 253 Anti-bodies, 260 Antigens, 508 Antipyrin, 63 Antitoxin, 250 Apnea, 176 Apomorphin, 63 Apparatus, 170, 290, 369, 457, 509 Appointments, 190, 200, 203, 310, 321, 324, 477, 484, 568, 574, 578 Arabans, 3 Arabinose, 174, 552 Argent-amm. mucoid, 166 Arginin, 2, 463, 544 Arsenic, 519 Arsenic trisulfid, 521 Arsenious oxid, 462 Artificial resp., 176, 464 Ascites, 142 Ash, 383, 466, 410, 434 Asparagin, 2 Aspergillus clavatus, 408 A. fumigatus, 408 A. niger, 408 Associations ; see organiz. 592 Index: Iinpcrsonal Suhjects [July Asthma, 159 Atropin, 63, 465 Auramin, 81, 550 Azolitmin, 550 Babcock test, 207 Bacteria, luminous, 456 Bacteria, milk, 580 D. phosphorescens, 456 B. phosphoreum, 456 Bacteriolysins, 250 Bardach test, 186 Barfoed test, 181 Barom. pressure, 393, 530 Barwood, 82 Bases, 181 Basic nitrogen, 463 Basic salts, 181 " Baustein," 209 Benedict method, 185 Benzaldehyde, 22J Benzene, 51, 77 Benzidin, 225 Benzoic acid, 133 Benzopurpurin, 84, 550 Beriberi, 205 Beryllium sulfate, 184 ;3-hydroxy-butyric acid, 66 j8-imidazolylethylamin, 158, 159 Betain, 2 Bibliography, 5, 46, 100, 202, 214, 298, 326, 339, 439, 470, SS9 Biebrich scarlet, 83, 550 Bile, 182 Pigments, 96 Biochemistry : abstracts ; see abstr. bibliography ; see bib- liog. courses, 203, 579 degrees ; see degrees dissertations, 538 doctorates, 538 ; see de- grees history, 243 index, 298, 470, 559 institutions •; see or- ganiz. Journals; see Journals literature, 298, 470, 559 organisations ; see or- ganiz. Biography : Aisberg, CL, 211 Howe, PE, 201 Ritthausen, H, 335 Schulze, E, I Bismarck brown, 84, 550 Biuret reagent, 179, 556 Biuret test, 181 Bladdcr, 233, 292 Bleached flour, 487, 532 Blood, 66, 146, 147, 148, 167, 175, 186, 225, 237, 292, 295, 461, 463, 466, 468, 547 pressure, 403 Body weight, 469, 552 Bone, 556 growth, 137 sarcoma, 231 Brain, 65, 242, 467 Bran, 228 Brazil wood, 80 Bread, 158 mold, 542 Breast, 87, 178, 231 Breathing, 149, 390 Brucin, 63 Buildings, 195, 313, 570 Butter, 66, 158 Butyric acid, 66 Cadaverin, 159 Caffein, 63, 68 Caisson disease, 147 Calcification, 137 Calcium, 137, 163, 178, 410, 438, 458,464,545 carbonate, 186 caseinogen, 116 hydroxid, 180 Calculi, 178 Caloric requirement, 143 Calorimetry, 139 Cancer extracts, 229 Cape aloes, 83, 550 Cappenberg method, 468 Capsules, gelatin, 318 Carbohydrates, 45, 135, 136, 143, 14s, 173 Carbon bisulfid, yy Carbon dioxid, 146, 148, 17s. 390, 393, 403, 530 Carbon tetrachlorid, 77 Carcinoma, 231, 287 Carmosin B, 81 Carrotin, 66 Carrots, 66 Caseinogen, iii Cassiopea xamanacha, 286 Castor bean, 410, 455 Catabolism, 149 Catalysis, 441 Catgut, loi Celebrations, 189, 309 Cell, 21, so, 55, 137, 138, 140, 454, 547 membranes, 3, 140 nutrition, 253 Cellulose, 3, 168, 414 Cerumen, 66 Chemotherapy, 259 Chestnut blight, 581 Chittenden's pupils, 349 Chlorid(n), 294, 434, 464, 549 Chloroform, 50, 77, 79 Chlorophyl, 80 Cholesterol, 2, 57, 67, 467 acetate, 66 benzoate, 67, 468 oleate, 67 palmitate, 67 stearate, 67 Cholin, 2 Chondroalbumoid, 547 Chondromucoid, 547 Chorion, 236, 291 Chrysoidin, 80, 550 Cinnamon, 133 Circulation, 148, 176, 403, 454, 464, 468 Cleavage ; see hydrolysis Clinic; see hospital Clitocybe illudens, 465 Clitocybe multiceps, 466 Cloves, 133 Clubs ; See organiz. Coagulation, 381 Cocain, 63 Cocoanut milk, 554 Cod, 548, 549 Codein, 63 Colchicin, 63 Cold storage, 184 Collagen, 170, 179, 547 Colleges ; see organiz. Collodion, 58, 61, 70, 290 Collodion-fat membranes, 549 Collodion membranes, 186, 549 Colloid, 290, 554 nitrogen, 87, 178 non-protein N, 178 platinuni, 161 Solutions, 161 Colon bacillus, 462 Combined sulfate, 545 " Comment, news and notes," 188, 307, 476, 567_ Commissions ; see organiz. Committees ; see organiz. Complement, 251 fixation, 167 Splitting, 166 Complimentary dinners, 309 I9I3] Index: Impersonal Suhjects 593 Composition, 29, 41, 178, 287, 288, 386, 433, 524, 558 Compressed air, 147 Congress, acts, 129, 196 Congresses ; see organiz. Conin, 63, 180 mucoid, 115 Constitution, 31 Construction unit, 209 Contraction, loi, 158, 464 Convicin, 2 Convulsiva action, 459 Copper, 166 iodid, 556 salts, 133, 556 thiocyanate, 556 Cord, 460 Corn-stalk disease, 543 Coroner's Consultants, 316 Corpuscles, 461 Correlation, 288, 399, 524, 530 Courses ; see biochem. Creatin(in), 68, 95, 124, 165, 294, 387, 464, 546, 555 Culture medium, 457 Cuorin, 67 Curcumin S, 82 Cystin, 463, 544 Cytolysins, 250 Decalcification, 137 Decidua serotina, 237 Declinations, 310, 568 Decompression, 147 Defenses, disease, 249 Degrees : honorary, 190, 567 M. A., 579 M. S., Z27 Ph. D., 538, 579 Phar. D., 579 Demography, 129 Density, 286 Dental caries, 580 Dentex vulgaris, 379 Dentifrices, 554 Deuteroproteose, 547 Development, 242, 463 Dextrinoid subst., 173 d-Mandelonitril, 227 Diabetes, 144, 217, 285, 309, 455 insipidus, 142 Diacetic acid, 96 Diagnosis, 454 Dialysis, 85, 549 Diamino nitrogen, 434 Diet(etics), 129, 131, 138, 141, 143, 145, 164, 187, 482, 551 Diffusion, 50, 55, 64, 70, 78, 164, 290, 445 Digestion, 134, i35, i44. 209 ; See enzymes Digitonin, 68 cholesteride, 468 Dinners, 309, 322, 349, 358, 477 Direct current, 556 Disease, 143, 488 Dispensary ; see hospital Dissociation, 557 Distribution, 140, 177, 556 Diver's palsy, 147 Doctorates, 538; seedegr. Dove, 556 Dropsy, 142 Drop-weight method, 186 Duodenal Contents, 454 Dwarfi'wö'Cism), 138, 139 Dyes, 78, 547, 550; see pigments Ear, internal, 573 Eck fistula, 292 Edema, 103, 141, 170, 172, 550 Edestin, 338, 547 Editoriais, 205, 329, 487, 582 Egg, 161, 182, 463 albumen, 50 white, 547 yolk, 65 Ehrlich " side chain " theory, 253 Elastin, 547 Electric, 52, 320, 367, 547, 555 charges, 382 conductivity, 383 Electrolyte, 170, 553 Electrometric determina- tion, 367 Electrons, 320, 332 Embryo, 161, 162, 463 Emulsin, 227, 408 Emulsoid, 554 Endart. obliterans, 545 Endowments, 193, 313 Endurance, 402 Energy, 104, 135, 139, 463 Enzymes, 4, 183, 227, 233, 236, 291, 292, 407, 441, 454. 463, 551, 555. 576, 582 Eosin A, 82, 550 Eosin W, yellow, 83 Epinephrin, 123, 182 Equipment, 195, 578 Ereptase, 237 Erythrosin, 81, 550 Ether, 64, 70, 79, 290 Ethereal sulfate, 545 Ethyl acetate, 67, 76, 80 Ethyl butyrate, 66 Ethylene di-amin, 180 Ethyl ether, 186 Eiidendrium, 286 Excretion, 90, 124, 134, 141, 178, 293, 294, 390, 393, 465, 530, 545. 546 Exercise, 132, 148, 396 Exhalations, 545, 558 Expired air, 146, 295, 545, 558 Extractives, 380 Fasting, 90, 139, 187, 288, 386, 468, 550, SSI Fast red A, 81, 550 Fat, 45, 57, 66, 73, 93, 135, 136, 142, 143, 164, 207, 387, 464, 509 Fat-collodion membranes, 549 Fatty acid, 66 Feces, 96, 134, 226 Feeble-mindedness, 238 Feeding, forced, 143 Fehling-Benedict test, 181, 553 Fellowships, 574 Female genitalia, 233, 236 Ferric chlorid, 181, 416 test, 177, 224 Ferric sulfocyanate, 59 Ferments ; see enzymes Fever, 144, 145, 295 Fibrin, 170, 444, 547 Fibroma, Uterus, 231 Fish, 184 Fistula, Eck, 292 Flavors, 158 Flounder, 291 Flour, 532 Food, 134, 13s, 139, 143, 182, 198, 532, 544, 551, 576 " Food colors," 78 Food-drug act, 196 Food preservatives, 130, 132, 185 Forced breathing, 390 Formaldehyde, 77, 198 Formic acid, 66, 77 Fox, 556 Freezing point (A), 383 Fructose, 174, 552 594 Index: Impersonal Sitbjects [July Fruit Juices, 554, 582 Function, 39 Functional activity, 454 Funds, 193, 195. 313, 349, 535, 570, 580 Chittenden, 349 Crocker, 313 Hansen, 535 Hennann, 580 Fungus, 542, 581 Fusarium oxysporium, 408 Fustic, 83, 550 Galactan, 3, 174 Galactose, 174, 552 Gall stones, 67, 467 Gastric acidity, 179 Gastric contents, 226 Gastric diseases, 455 Gastric juice, 141 Gelatin, 547 capsules, 3 1 8 Genitalia, 233, 236 Gentian violet, 462 Germination, 3, 42 Gies biuret reagent, 179, 181, 556 Gifts ; See funds Glacial acetic acid, 80 Globulin, 166 Glucose, 3, 68, 136, 174, 217, 292, 552 Glucosuria, 292 Glucothionic acid, 547 Glutamic acid, 137, 337 Glutamin, 2 Gluten, 547 Glycerol, 66 Glycocol, 137, 409 Glycogen, 93 Gold-beater's skin, 61 Gold orange, 81 Gout, 143 Grafted embryos, 161 Grants, 193, 483, 580 Growth, 46, 99, 130, 137, 138, 229, 242, 463 Guaiacum, 225 Guanidin, 2 Gums, 173 Heart, 66, 142, 176, 387, 404 Heat, 102, 135, i39, 148 coagulation, 381 production, 135, 145 Hematoxylin, 80 Hemi-celluloses, 3 Hemoglobin, 98, 136, i6r, 382, 547 Hemolysij(n), 250, 461 Hemorrhage, 187, 468, 551, 555 Heredity, 242 High frequency current, 555 Histidin, 2, 161, 463, 544 Histon, 169, 291, 419, 548 mitcoid, 118 nucleoprotein, 121, 548 ovo-mucoid, 121 Historetention, 142 Historical ; see biochem. Honorary degrees, 190, 567 Honors, 190, 308, 476, 567, 580 Hopkins-Cole lest, 181 Horse, 556 disease, 544 Hospitals : Assoc. out-pat. clinics, N. Y., 314 Bellevue (N. Y.), 578 Herriman disp., 195 Johns Hopkins, 193 Montefiore Home, 196 New York, 313 Phipps Psych. Clin., 4S1 Roosevelt, 317 Hot-water-bottle holder, 290 Humidity, 146, 148 Humin (nitrogen), 434, 544 Hydrion (HO, 367, 384 Hydrochloric acid, 141, 462 Hydrocyanic acid, 22y Hydrogen selenide, 553 Hydrolysis, 135, 209, 407, 414, 456, 544, 547 Hydronephrosis, 142 Hygiene, 129 Hyper-artif. resp., 464 Hyper-creatininemia, 295 Hyper-respiration, 403 Hypophysis, 573 Hypoxanthin, 2 Ice, 572 Idiocy, 457 Illuminating gas, 293 Imbibition ; see water Immune body, 251 Immunity, 247 Inanition, 90 Index, biochem., 298, 470, 559 Indican, 96, 158, 285 Indicator, 172 Indol, 375 Industrial firm : Van Houten & Ten Broeck Co., 555 Infant mortality, 485 Infection, 247, 544 Infusoria, 465 Inheritance, 529 Injury, 582 Inocybe infida, 465 Inorganic salts, 2^2 Inosite-phosph. acid, 21 Insecticide, 522 Internal ear, 573 Intestin, 405 Intestinal bacteria, 319 Intest, putrefac, 159, 286, 319 Intoxication, 258, 461 Intrabronch, press., 404 Intrapulm. air-press., 403 Involuting tail, 288, 546 lodine test, 181 lodo-eosin, 290 Ions, 167, 367, 557 effects, 554 Iron ; see f erric Iso-agglutination, 461 Iso-amylamin, 159 Iso-cholesterol, 2 Iso-hemolysins, 461 Iso-leucin, 2 Jaffe reaction, 165 Journals, 261, 297, 299, 317, 470, 559 Biochem. Bull., 299, 470, 559, 577 Bio-Chem. J., 128, 209, 299, 446, 470, 559 Biochem. Zeit., 299, 470, 559 Biolog. Abstr. J., 261, 332 Bull. Tor. Bot. CL, 574, 576, 580 Die Naturwissensch., 317 Folia microbiol., 297 /. All. Dent. Soc, 580 /. Biol. Chem,, 299, 470, 559 J. N. Y. Bot. Card., 576 /. Pharm. Exp. Therap., 198 Mycologia, 576 Nederl. Vereen. voor Microbiol., 297 Physiolog. Researches, 574, 576 Zeit, physiol. Chem., 299, 470, 559 I9I3] Index: Impersonal Subjects 595 Kephalin, 67 Keratin, 545 Kidney, 66, 141, 144, 295 Kjeldahl appar., 457 Knighthood, 567 Lactase, 237 Lactic acid, 66, 68, 96, loi bacillus, 319 Lactose, 145, 552 Larvse, 162, 288, 546 Lead arsenate, 521 Lead oleate, 66 Leaves, 183 Lecithin (an), 50, 6y platinic chlorid, 67 Lectures, 312, 327, 479, 570, 576 Carpenter, 312 Harvey, 312, 479 Herter, 312 Huxley, 313 Middleton Goldsmith, Mitchell, 479 Legal test, 223 Le Nobel test, 223 Leucin, 2, 137 Leucocytes, 289 /-Mandelonitril, 227 Liebig beef-ex., 137 Life, 19s Light bacteria, 456 Lipase, 233, 237, 455 LipiMj(oids), 45, 50. 55. 64, 170, 182, 242 solvent s, 55 Lipin-soluble substances, 55 Lipochrome, 57, 65 Liver, 66, 142, 463 hypertroph, cirrh., 455 Living matter, 140 Luminous bacteria, 456 Lungs, 146, 149, 403, 458, 558 Lupinin, 2 Lupin seeds, 550 seedlings, 229 Lymph, 148 Lysin, 2, 454, 463, 544 Magenta, 80 Magnesium, 163, 464 oxide, 410 Sulfate, 187 Maintenance, 138 Malachite green, 80, 550 Maltase, 237 Maltose, 552 Manganese, 410 Sulfate, 455 Mannans, 3 Marriages, 321, 574, 578 Martius yellow, 80, 550 Mastic, 162 " Mathews plan," 332, 490 582 Meadow hen, 556 Meals, 394 Meat, 161, 187, 552 Medaillon, 316 Medals, 309, 354,476, 535, 568 Buchanan, 309 Cresson, 476 Davy, 309 Gibbs, 476 Hanbury, 568 Hansen, 535 Helmholtz, £,77 Longstreth, 568 Retcius, 309 Medical Service, Eng., 197 Medusae, 286 Meetings ; see proc. Meigs method, 509 Melanin, 544 Members-elect ; see org'iz. Membranes, 55, 64, 70, 549 Memorial, i, 188, 307, 567 ; see funds, lec- tures, medals Mercuric chlorid, 462 Metabolism, 145, 163, 241, 457, 464, 545, 547; see nutrition Metamorphosis, 546 Metanil yellow, 80, 550 Metaprotein, 547 Method, 25, 37, 55, 64, 70, 85, 88, 112, 161, 162, 164, 165, 166, 169, 172, 177, 179, 185, 186, 198, 217, 228, 230, 233, 236, 291, 367, 373, 387, 391, 419, 454, 466, 467, 509, 525, 547, 553; see ap- paratus Methyl alcohol, 76, 80 Methylene blue, 550 Methylene violet, 81 Methyl green, 547 Methyl violet, 80 Micro-Kjeld. app., 457 Milk, 178, 207, 317, 509 580 bacteria, 580 flow, 163 Millon test, 181 Milt, 548, 549 Mold, 407, 542 Moldy silage, 330 Molecular weight, 67 Molisch test, 181 Mon-amino N, 434, 463, 544 Monardas, 293 Monkey, 556 Morbus Basedowii, 144 Morphin, 63 caseinogen, 116 mticoid, 112 nticleoprotein, 115 Mucilages, 173 Mucin, 180, 547 Mucoid, III, iiS, 547, 556 silver, 165 Multiple embryos, 161 Muscarin, 465 Muscle, 148, 231, 288,379, 386, 464, 546 biceps femoris, 387 contraction, loi, 158 plasma, 379 retractor penis, 379 seniitendinosus, 387 serum, 381 tonus, 135 Mushrooms, 465 Mutton tallow, 66 Myofibrils, 380 Myohematin, 382 Myoprotein, 380 Myosin, 380, 547 Myotonia atrophica, 464 Naphthol green, 84 Naphthol red S, 82 Naphthol yellow, 80 Narcein, 63 Narcotin, 63 Necrology, 188, 307, 476, 567 Nephritis, 142, 292, 294 Neutrality mechanism, 102 Neutral red, 51, 464 News, notes and com- ment, 188, 307, 476, 567 New York (hist), 243 Nicotin, 63 Nicotinic acid, 228 Nitrils, 178 Nitrite, 198, 532, 582 Nitrogen, 87, 93, 147, 178, 182, 292, 387, 434, 457, 464, 544, 545, 555 non-prot, 178 partition, 123, 545, 555 Peroxide, 532 ratlos, 463, 544 Nitrous acid, 532 Nomenclature, 35, 209 Non-protein N, 178, 292 596 Index: Impersonal Siihjects [July " Notes, news and ment," i88, 307, 567 Nucleic acid, 438 Nucleoprotein, iii, 547, 548 metabolism, 164 Nutrition, 21, 90, 130, 134. 137. 138, 141, 143, 145. 148, 186, 199, 203, 205, 253, 337, 457, 464, 468, 482, 545, 546, 555 com- 476, 121. 123. 139, 163, 241, 466, 550, Oatmeal, 182 Obesity, 143 Obituary ; see necrol. Officers-elect ; see organiz. Oleic acid, 67 Olive oil, 67, 137, 549 Opsonins, 252 Orange G, 81, 550 Orcein, 53 Orcin, 53 Order of merit, 308, 476, 567 _ Organizations : I. Societies — Am. As. Adv. Sei., 314, 315, 480, 576 "Amer. Biolog. Soc," 261, 332, 490, 582 Am. Chem. Soc. ; Dtv. Biol. Chem., 480, 572, 578 Am. Med. As., 568, 570, 572 Am. Physiol. Soc, 269, 271, 315 Am. Phytopath. Soc, 31S Am. Soc. An. Nutr., 282 Am. Soc. An. Produc, 282 Am. Soc. Biol. Chem- ists, 269, 275, 315, 331, 578 Am. Soc. Natur., 315 Am. Soc Pharm. Exp. Ther., 269, 279, 315 Am. Soc. Zool., 316 As. Amer. Med. Col., 480 As. Improv. Cond. Poor, 573 Austr. As. Adv. Sc, 195 Eiochem. Assoc. ; see Col. Univ. Biochem. Assoc. Organizations (con.) : Biochem. Club, Eng., 318 Biochem. Soc, Eng., 128, 209, 446, 487 Bologna Acad Sei., 573 Botan. Soc. Am., 316, 576 British As. Adv. Sei., 195 Col. Univ. Biochem. As- soc, 156, 200, 284, 321, 452, 484, 541, 574 Dental Soc, 580 Harvey Soc, 481 Hospital As., 314 111. Water Supply As., 4S1 Minn. Path. Soc, 570 Nat. Ac Sciences, 480 Nat. Inst. Social Sei., 354 N. Or. Acad. Sei., 315 N. Y. Gastro-Enter. Soc, 315, 328 N. Y. Soc. Anesthetists, 567 Pharmaceut. Soc. (Lon- don), 568 Phi Lambda Ups., 578 Royal Society, 195 Rush Society, 314 Sigma Xi, 481, 570,573, 578 Soc. Ainer. Bact., 315 Soc. Chem. Indust., 316 Soc. Exp. Biol. Med., 358, 575, 578 Torrey Bot. Cl., 574 Vienna Soc. Invest. Prev. Cancer, 571 2. Comniissions, com- mittees — Chicago Ice Commis., 572 _ Commis. Elect. Shock, 572 Commis. Exp. Study Vent., 572 Commit. on Occup. Dis., 488 Commis. on Resuscit. from mine gas, 572 Harvard Cancer Com- mis., 571 Int. Pharmaceu. Com- mis., 483 Med. Res. Commit., Eng., 574 N. Y. Sabbath Com- mit., 581 Organizations (con.) : 3. Conferences, con- gresses — Conf. Prev. Inf. Mor- tal., 485 Fed. Amer. Soc. Exp. Biol., 269, 271, 314, 331, 490 Int. Cong. (8) Appl. Chem,, 194 Int. Cong. Appl. Chem. (8): See (VIII, D), Biochem. (pharma- col.), 150 Int. Cong. (6) Gen. Med. Elect. Radiol., 314 Int. Cong. Hyg. Demog. (15) [See (II), Biet. Hyg.: Hyg. Physiol.], 129, 194 Int. Cong. (3) Refrig., 580 4. See funds, hospitals, industrial firms, re- search institutions, universities (Colleges) Organo-therapy, 208, 241 o-Tolidin, 225 Osazones, 220 Osmosis, 74, 551 pressure, 59, 140, 170 processes, 141 Ossein, 547 Osseoalbumoid, 547 Osseomucoid, 547, 556 Osteitis deformans, 137 Osteomalacia, 137 Osteoporosis, 137 Ovary, 233, 292 " Over-production " the- ory, 254 Ovo-mucoid, 111,121,182, 547 Oxidase, 183, 582 Oxidation, 54, 293, 294 Oxygen, 53, 93, 146, 147, 148 carrier, 456 consumption, 145 Palmitic acid, 66 Pancreas, 454 amylase, 555 diabetes, 217 secretion, 134 Pancreatitis, 455 p-Cresol, 319 ParaflSn oil, 61, 77 /'-Hydroxyethylamin, 159 phenyl Compound, 285 I9I3] Index: Impersonal Subjects 597 Paramecium, 465 Parathyroidectomy, 466 Parchnient, 61, 74 Parenteral introd., 257 Paris green, 521 Partial fasting, 552 " Pawlow," 317 Peach disease, 576 Pellagra, 544 Penicilliuin camemberti, 408 P. e.vpansum, 408 F. rcqueforti, 408 Pensions, 573 Pentosans, 174 Pentose, 174 Pepsin(ogen), 234, 237, 444 Peptidase, 237 Peptone, 547, 549, 558 Peristalsis, 134, 406 Peritoneal fluids, 454 Permeability, 50, 549 Pernicious anemia, 461 Peroxidase, 53, 183, 582 Peroxide, 582 Persimmon, 168, 412 Personalia ; see news, notes and comment Petroleum, 199 ether, 77, 80 Phagocytes, 288 Phagocytosis, 289 Pharmacology, 184, 459, 465. 552 Ph.D. ; see biochemistry Phenacetin, 63 Phenol, 462, 467 substances, 168, 413 Phenolphthalin, 225 Phenyl acetic acid, 286 Phenylalanin, 2 Phenylethylamin, 159 Philippines, 536 Phlorhizin, 292 Phloroglucinol, 168, 413 tannoids, 416 Phloxin, 81, 550 Phosphate, 21, 163 Phosphatids, 2 Phospho-proteins, 45 Phosphorescence, 456 Phosphoric acid, 21, 410 Phosphorus, 21, 163, 387, 457, 464, 466 Photo-synthesis, 45 Phrenosin, 165 Physiol. ehem. ; see bioch. Physostigmin, 63 Phytase, 41 Phytin, 21, 163 Phytin (con.) : lecithids, 2 Phosphate, 21 Phytosterols, 3 Picric acid, 63, 80 Picrotoxin, 63 Pigments, 60, 78, ' 164, 171, 175, 293, 382, 547, 550, 582 Pineal gland, 573 Piperazin, 180 Piperidin, 180 mucoid, 115 Plant chemistry, 5, 21 Platinum, 161 Poisoning, 330 Ponceau G A, 82 Ponceau 2 R, 82 Portraits ; see page xv Postmortem swelling, 551 Potassium, 163, 466 cupro-cyanide, 558 Cyanide, 293 hydroxid, 1 80 palmitate, 66 salts, 141 selenocyanate, 553 stearate, 66 sulfocyanate, 459 Precipitation, 553 Precipitins, 252 Pregnancy, 233, 292 Preservatives, 130, 132 Prickly pear, 173 Prizes, 190, 193, 309, 477, 568, 573 Alvarenga, 193 de Cyon, 573 Dieulafoy, 193 Gedge, 309 Hansen, 535 Lieben, 190 Nobel, 190, 308 Riheri, 193 Richards, 477 Ricketts, 568 Proceedings, 129, 150, 156, 201, 269, 271, 282, 284, 452, 486, 577; see organiz. Prolin, 2 Propionic acid, 66 Protagon, 165 Proteases, 170 Protein, 70, 93, 103, 135, 136, 138, 139, 142, 143, 160, 178, 182, 186, 257, 290, 295, 337, 373, 380, 389, 419, 464, 548, 558 Compounds, iii, 169 copper, 166 Protein (con.) : film, 52 membranes, 50 metabolism, 3 salts, III, 165, 166, 180, 439 test, 181 Proteose, 547 Protoplasm, 140 Protozoa, 161 Pseudoösazones, 220 Publications ; see journ. Pub. Health a. Mar. Hosp. Service, 196 Pub. Health Service, 196 Purgation, 285 Purins, 124, 555 Putrefaction, 159, 375 Putrefactive amins, 158 Putrescin, 159 Pyramidon, 63 Pyromucuric acid, 407 Quinin, 63 Quotations (special), 210, 332, 508, 588 Raccoon, 556 Radiant energy, 210 Raffinose, 338 Rations (Standard), 199 Rat, white, 556 Reaction, 167, 383, 551 Reagents ; see test Reception, 477, 576 Receptors, 254 Reduction, 294, 552 Refrig. ; see cold storage Regeneration, 254, 286 Regression, 288, 399, 525 Relationship, 524, 530 Renal calculi, 178 Rennin, 455 Reproduc. rate, 465 Research, 238; see fel- lowships, organiz. Research institutions : Biol. Lab., Brooklyn Inst. A. and S., 574, 575 British Board Agric. a. Fisheries, 571 Carnegie Institution (Wash.), 483, 576 Cal. Citrus Exp. Sta., 316 Franklin Inst., 568 Hoagland Lab. (Brook- lyn), 570 Institute for Dietetics (Paris), 482 598 Index: Impersonal Suhjects [July Research instits. (con.) : Intern. Bureau of Food- stufTs (Paris), 317 Rockefeller Institute, 573 S. Lond. Botan. Inst., 193 Mar. Biol. Lab. (Mass.), 574. 575 Mich. St. Board H., 574 Spiegier Inst., 571 Train. Seh., Vineland, N. J., 238 Turck Institute, 482, 571 U. S. Pub. Health Serv., 572 See funds, univ. (col.) Resignations, 190, 203, 310, 324, 568 Resistance, 453, 462, 468, SSI Respiration, 146, 176, 295, 390, 393, 403, 464. 545, 558 Center, 148 Quotient, 145 System, 148 Rest, 581 Retention (salt), 142 Retention (storage), 163 Retention (water), 141 Retirements ; see resig. Rhamnose, 552 Rhisopus nigricans, 542 Rhodamin, 81, 550 Rice bran, 228 Rickets, 137 Ripening, 418 Ritter's method, 468 Ropy bread, 158 Rose bengal, 81, 550 Rubber, 55, 64, 70, 78, 171, 195, 290 tnembranes, 186 Safranin, 82, 547, 550 Salicin, 68 Salicylic acid, 63 Salinity changes, 286 Salins, 131, 140, 143, 380, 438 Saliva, 533, 580 Salivary mucin, 180 Salomon and SaxI test, 287 Salts ; See salins Salvarsan, 462 Saponin, 462 Scarlet R, 60, 80 Scholarships, 570, 571, 581 ; see fellowships Sculpin, 556 Scyllium stellare, 379 Sea water, 286 Second wind, 149 Secretion, 134, 455 Seeds, 337, 550 Seessel fellowships, 574 Selenium, 552 acid, 553 dioxide, 553 Seliwanoff test, 181 Senility, 319 Seroretention, 142 Serum ; see blood Shad, 548 Shock, 17s, 403, 464 "Side chain " th., 253 Silage, 330 Silica, 410 Silver mucoid, 165 Six-o-six (606), 402 Skin, 66, 142, 146 Societies, 194, 200, 209, 261, 308, 314, 315, 321, 446, 477, 480, 567, 573, S7S, 578, 580; see or- ganiz. Sodium, 466 benzoate, 133 carbonate, 180 Chloride, 67, 141, 431 hydrogen selenite, 553 hydroxid, 51, 462 nucleoprotein, 548 palmitate, 66 selenate, 553 selenite, 552 stearate, 66 Sulfid, 177, 178 taurocholate, 462 Soil, 519 Solids, 383 Soluble starch, 553 Soya beans, 317 Sp. dynam. action, 130 Sp. gravity, 383 Sperm, 291, 548, 549 Spices, 143 Spirochetes, 462 " Splitting " prod., 209 Stachydrin, 2 Stachyose, 3 Stain, 38s, 547 Standard rations, 199 Starch, 172, 320, 553 grains, 52 test, 181 Starvation, 90 State med. serv., 197 Stearic acid, 67 Stock poisoning, 330 Stomach, 141, 455 Contents, 179, 226 Stone, 455 Storage (retention), 163 Strychnin, 63, 68 caseinogen, 116 mucoid, 114 ovo-mucoid, 117 Suboxidation, 292 Sucrase, 237 Svicrose, 549 Sudan G, 81 Sudan I, 81 Sudan III, 60, 67, 79 test, 181 Sugars, 173, 185, 187, 217, 319, 524 Sugar beet, 287, 524 Sulfate, ethereal, 545 Sulfites, 134 Sulfocyanate, 177,459,580 Sulfur, 178, 457, 464 Sulfuric acid, 162 Sulfurous oxide, 134 Summer Session, 203, 575, 576, 579 Surface condens., 140 Surface energy, 104 Surface tension, loi, 140, 186, 383 Swelling, 103, 170, 172, 550 Symptoms, 184, 543, 545, 553 ; see pharmacol. Synthesis, 3, 227 Tail, 288, 546 Taka-diastase, 408 Tannin, 168, 414 mass, 168, 412 Taurin, 178 Teeth, 554, 580 Temperature, 136, 146, 176, 396, 463 limits, 456 Testicle, 67 Testimonial, 349 Tests, 160, 168, 179, 181, 186, 207, 223, 225, 287, 416, 547, 552, 556 Tetra-ethyl ammonium hydroxid, 180 Therapy, 208, 241, 259 Thioacetic acid, 178 Thiophenuric acid, 409 Thiourea, 178 Thirst, 142 Thymol, 171, 293 Thymus histon, 169, 548 Thyroid, 573 Tissues, 177, 550 I9I3] Index: Impersonal Subjects 599 Toluene, 77 Tomato disease, 576 Toxicol., 158, 161, 178, 461,465; See pharmacol. Toxin, 542 Tradescantia, 183 Transfusion, 461 Treatment ; see therapy Trigonellin, 2 Trikresol, 462 Trimethyl amin, 180 Tropeolin 00, 83 Trout, 463 Trypsin, 234, 237, 455 Tryptic digestion, 375 Tryptophan, 373, 464 Tuberculolysins, 454 Tuberculosis, 453 Tuberculous areas, 458 Tubularia, 2^7 Tumor, 87, 178, 455 Turmeric, 80 Typhoid fever, 145 Tyrosin, 2, 137, 286 Ultra-filter, 290 Ultramicr. granules, 379 Universities (col.), 538 Columbia Univ., 193, 313. 327; Biochem. Dept., 201, 324, 578 Cornell Univ., 571 ; Med. Col., 313 Gettysburg Col., 581 Harvard Univ., 571 Irving Col., 581 Universities, col. (con.) : J. Hopkins Med. Seh., 199 J. Hopkins Univ., 481 London Seh. of Trop. Med., 193, 313 Syracuse Univ., 313 Univ. Cal., 314, 316 Univ. 111., 483, 571 Univ. Kan., 193 Univ. Md., 313 Univ. Mon., 575 Univ. Paris, 314 Univ. Toronto, 313 Univ. Utah, 196 Univ. Wis., 571 Yale (Sheff), 351 Uranium nephritis, 294 Urate, 467 Urea, 68, 94, 124, 136, 292, 463, S5S Urease, 455 Uremia, 142, 292, 295 Uric acid, 178, 293, 294, 463, 467, 555 diathesis, 143 Uricase, 164 Uricolysis, 164, 292 Urine, 87, 123, 141, 158, 165, 178, 182, i8s, 217, 223, 226, 285, 292, 464, 466, 545, 546, 555 Urochrome, 67, 466 Uroerythrin, 466 Uterus, 233, 292 Valerianic acid, 67 Vegetable Juices, 554 Vegetable proteins, 337 Ventilation, 131, 146, 295, 545, 558, 572 Veratrin, 63 Vernin, 2 Vicin, 2 Vinegar, 554 Vineland, N. J., Viscosity, 383 Water, 136, 141, 164, 187, 383, 387, 550, 551 absorption, 103, 550 Weigert " over produc- tion " theory, 254 Weight, 287, 524 Wheat embryo, 67 ; flour, 532 White rat, 556 Witte Peptone, 70 Wool, 544 Work, los, 13s, 147, 390 Xanthin, 2 Xanthoma (skin), 66 Xanthoproteic test, 181 X-rays, 210 Xylans, 3 Xylose, 552 Yeast, 66, 548 Zinc arsenite, 521 Zymogen, 456 II. SUBJECT INDEX (continued). B. PERSONAL SUBJECTS Impersonal subjects are indexed on pp. 591-599- The names of authors are given on pp. 589-590. This portion of the index relates primarily to directly personal items, but does not include personal references in incidental historical or similar Statements. A recurrent name in any personal item or formal section of related references is indicated by the numeral on the first of the group of pages presenting the name. AbbottAC, 314 AbderhaldenE, 568 AbelJJ, 150, 198, 280 AcreeSF, 483 AdamsonGP, 489 AdrianceJ, 581 AlbaneseM, 476 Alcock.NH, 567 AlleeCW, 312 AlsbergCL, 200, 211, 277, 311, 321, 329, 480, 485, 573, 575, 577 AlwayFJ, 570 AndersonEM, 573 ArbuthnotTS, 349 ArkinA, 538 ArmsbyHP, 277, 479 ArmstrongDB, 484 AtkinsonGF, 316 AuchinclossH, 577 AuerJ, 280 6oo Index: Personal Suhjects [July Auld.SMT, 192 AxenfeldD, 307 BachmannG, 273 BaekelandLH, 476 BaileyLH, 568, 569 BakerGF, 313 BalchAW, 478 BalfourA, 479 BancroftWD, 573 BarbourHG, 273, 280 BarclayH, 315 BargerG, 446 BarnardHE, 480 BarrettAM, 482 BartowE, 481 BashfordEF, 312 BaskervilleC, 489 BassettWH, 489 BaumannL, zyj BaxterGP, 4S3 BaylissWM, 128, 446 BealGD, 321 Beard.TH, 192 BeardRO, 479 BeattieRK, 192 BeckwithCJ, 200, 575 BeebeSP, 349 Beijerinck — , 297 BenedictSR, 321, 577 BengisR, 539 BergeimO, 478 BergeyDH, 315 BermanL, 204, 580 Bernstein — , 189 BerryJT, 485 BertrandG, 194 BigelowWD, 311, 483 BinzC, 189, 307 BirchardFJ, 192, 2jj BischLE, 484, 574 BlakeJA, 349 BlakesleeAF, igo BleulerE, 481 BlissAR, 200, 576 BockJC, 190 BogertMT, 313, 316, 322 BolduanCF, 200 BookmanS, 277 BornS, 578 BossA, 310 BourquelotE, 483 BowmanAE, 570 BoydWB, 577 BradleyHC, 569 BradleyWP, 190 BradyJB, 193 BreckenridgeJE, 480 BreedRS, 478 BrennerJJB, 574, 575 BrewerGE, 315 BrewsterJF, 191 BristowAT, 567 DroadhurstJ , 580 BrooksC, 280 BrownW, 571 BrownWH, 569 BrowneWW, 572 BruntonTL, 198 BuchbinderHE, 321 BuckA, 579 BullCG, 569 Burton-OpitzR, 200 BuswellAM, 578 CairdJK, 195 CaldwellJS, 310 Calmette — , 535 CalvertRP, 578 CalvinMV, 190 CameronAT, 581 CameronFK, 479 CampbellDH, 316 CampbellJA, 570 CannonWB, 269, 274, 572 CardifflD, 193, 570 CarlsonAJ, 274 CarnegieA, 314 CarrelA, igo, 308, 476, 477, 567 CarringtonPM, 311 CarterHS, 315, 485 Cash.TT, ig8 CecilRL, 321, 361, 575 ChaceAF, 200 ChalmersAJ, 479 ChambersCO, 539 ChapmanAC, 446 Chauveau — , 309 ChillengworthFP, 312 ChittendenRH, 129, 309, 328, 349, 477, 484 ClarkED, 203, 277, 576 ClassFM, 576 ClintonLA, 311 ClowesGHA, 362 CobbCH, 481 CoddJM, 192 ColemanKR, 572 ColemanW, 486 ConnellyAM, 57g ConnollyJM, 192 CookRJ, 204 CopelandEB, 199 CornishECV, 571 CorperHJ, 277 CrewH, 477 CrileGW, 572 CrockettWG, 579 CurtisE, 307 CurtisHH, 354 CurtissCF, 283 CushingH, 349, 482, 568 CushnyAR, 198 CusickJT, 191 Cussler.E, 204 de CyonE, 307 DaggettRG, 569 DakinHD, 277, 278, 311 DaleHH, 198 DanielsWJ, 569 Daniels WW, 188 Dastre — , 309 DavisBM, 315 DavisHB, 579 DavisHW, 191 DawsonPM, 570 Delezenne — , 309 DiekmanGC, 311 DieulafoyMme, 193 DixonWE, 198 DoerflingerWF, 489 DoIiberT, 188 DonaldsonHH, 314, 349 DoolittleRE, 311 DoivUM, 579 DoxAW, 573 DresbachM, 273 DryfoosAD, 580 DuaneW, 571 von DungernE, 569 DunhamEK, 278, 481 DunlapCB, 482 Dyerl, 34g EastEM, 315 EbsteinW,307 EcklesCH, 199 EddyWH, 322, 578 EdmundsCW, 280 EdsallDL, 130, 567 EdwardML, 580 EgglestonC, 280, 361 EgloffG, 579 EhrlichP, 308 EinhornM, 315 ElderFR, 204, 480, 579 EliotE, 315 ElyHM, 481 EmmettAD, 321 EpsteinAA, 277 ErlangerJ, 274, 572 EustisAG, 575 EwingJ, 361 FairhallLF, 478 FalkKG, 361 FalkeLL, 580 FarmerR, 321 FeinbergBG, 579 FetzerLW, 311 FieldAM, 579 I9I3] Index: Personal Siihjects 60 1 FieldCW, 361, 57S FineMS, 277 FischerE, 476 FischerMH, 479, 573 FisherHL, 200, 321 FittingJ, 192 FlexnerS, 313, 567 ForbesEB, 283 Fordjs, 446 ForresterGP, 483 FosterNB, 204, 578 FraenkelS, 571 FraserTR, 198 FreemanEM, 310 FreemanWB, 311 FreerPC, 189, 308 FriedbergerE, 568 FriedmanPJ, 572 FultonJS, 194 Gad— , 189 Gaffky— , 535 Ganz — , 310 GardnerJA, 446 de GarmoMC, 579 GarreyWE, 569 GatesFL, 569 GavinH, 580 GearingME, 321, 576, 577 GettlerAO, 311 GibsonRB, 192 GiesWJ, 150, 203, 204, 269, 278, 315, 324, 327, 349, 362, S7S, 577. 578 GilmoreJW, 312 GitlowS, 204 Gley— , 309 GodingJ, 192 GoldfarbAJ, 575 Goldschmidts, 570 GoodaleHD, 321, 579 GoodrichHB, 200, 575 GoodridgeFG, 203, 578 GorgasWC, 309 GotchF, 193, 567 Gottl'ebMJ, 580 GraceyGF, 568 GraveC, 316 GrecoV, 572 Greenwaldl, 204, 277 GreenwayJC, 577 GregoryLH, 200, 575 Grignard — , 308 GrossA, 325 GrossBH, 579 GuggenheimSR, 196 GunnJA, 198 HadleyAT, 350 HainesWS, 316 HallAD, 193 HallFA, 479 HalliburtonWD, 446 HalseyJT, 570 HamlinML, 325, 574 HanzlikPJ, 273, 281, 570 HardenA, 128, 446 HardingHA, 315. 478, 479 HardyWB, 310 HareRF, 575 HarkeyTL, 203, 579 HarperRA, 316 HarrimanMrs.EH , 317 HarrisBR, 203, 325 HarrisJA, 315 HarrisonRG, 315, 316 HartCC, 325 HartwellBL, 311 HartwellJA, 349 HarveyEN, 321, 322, 575, 576 HasslockCW, 327, 579 HawkPB, 200, 278, 349, 576 HawkinsLA, 538 HayesWvV, 315 HealdFD, 192 HeidelbergerM, 200, 575 Heilbronner — , 482 HektoenL, 316, 568, 572 HemmeterJC, 313 HendersonY, 572 Henkel — , 477 HepburnJS, 204, 538, 578 HerrimanWH, 196 HerterCA, 243 HesseBG, 194 HilgardEW, 314 HillAV, 309 HillL, 310 HinchliffGF, 579 HirschfelderAD, 570 HissPH, 476 HitchensAP, 315 HoaglandR, 478 HochA, 481 van't Hoff — , 316 HoffmanF, 478 HoffmanPM, 316 HofmannF, 569 HoganAJ, 570 HogeMA, 575 HolIickA, 316 HolmströmEA, 188 HolrayardEJ, 571 Holst A, 129 HookerD, 361 HooperEG, 191 HopkinsFG, 446, 574 HorowitzB, 538, 579 HoskinsRG, 569 HotchkissLW, 315 HoughT, 480 HoustonDF, 479, 568 HoweMA, 316 HowePE, 201, 203, 361, 578 HowellWH, 274, 277, 477 HowitzFJAC, 476 HowlandJ, 200, 479, 485 HoytLF, 569 von HuffmanO, 191, 575 HumeAO, 193 HumphreyHB, 478 HuntR, 150, 278, 280, 312, 569, 572 HuntTF, 314 HurtleyWH, 447 van ItallieL, 483 JacksonDE, 281 JacksonHC, 349, 361 JacksonHL, 478 von Jaksch — , 310 Janeway,HH, 576 JanewayTC, 349 JanneyNW, 277, 311 JoblingJW, 570 JohnsonAA, 570 JonesE, 482 JonesHC, 483, 568 JonesHO, 307 JonesW, 278 JordanEO, 478, 572 JordanHE, 316 Jores — , 478 JosephDR, 190 JoslinEP, 349 EahnM, 325, 578 KaliskiDJ, 322 KassowitzM, 567 KastL, 315 KastleJH, 311 KaufmanHM, 489 KeebleF, 447 KimballDD, 573 KirbyGH, 482 KirkwoodJE, 575 KisterE, 572 KiteGL, 312, 539, 568 Klein — , 297 KleinerlS, 277, 281 KliglerJJ, 580 KnightRC, 571 KnoopF, 479 KnoxJHM, 349 KnudsonA, 203, 572, 578 KoberPA, 277 KochFC, 277, 312 KochM, 312 KochW, 271, 278, 312 602 Index: Personal Suhjects [July KohtsO, 307 KorchoofP, 199 KosselA, 309 KrausGR, 310 Kraus WM, 204, 580 KremerE, 573 KristellerL, 27-; KroneckerH, 577 KruseW, 569 La ForgeFB, 2yy, 569 LambertSW, 349 Landouzy — , 309 LangleyJN, 198, 309 Langlois — , 309 LangmuirAC, 489 LeamingE, 315 LeathesJB, 278 LeeFS, 269, 274, 362, 481, 573 von LeubeO, 190 LevenePA, 278, 349 LevinI, 311 LevineVE, 578 LewisCH, 479 LewisHB, 539, 574 LewisRC, 569 LiebCC, 485, 580 LillieRS, 569 LintHC, 569 Lister, 188, 307 LivingstonBE, 575, 576 LloydFE, 191, 308, 313 LockwoodGR, 315 LoebJ, 309, 322, 362, 477, 574- 575 LoebM, 188 LoevenhartAS, 269, 280 LongER, 568 LongJH, 316, 481, 569, 572 LothropAP, 204, 2y7, 328, 577, 578 LowenbergH, 191 LowrySM, 311 LubarschO, 478 LudwigE, 568 LuskG, 130, 269, 274, 277, 27^, Z22, 329, 353, 358, 481, 574 LusskyHO, 192 Lüthje — , 310 LyleWG, 315 LymanGR, 191 LyonEP, 480, 570 McClungCE, 316 McDanellL, 325, 485 McDougallW, 481 McGIoneB, 310 McIntyreE, 307 McKennaCF, 489 McMurtrieW, 567 MacallumAB, 129, 275, 277 MacCallumWG, 481 MacDonaldVE, 481 MacEnauP, 483 MacKenzieMD, 191 MacksIM, 192 MacleodJJR, 277, 313 MacNealWJ, 315 MacNiderWDeB, 273,277, 280 MallettJW, 307 MandelJA, 150 MannG, 315 ManwaringWH, 569 MarineD, 484 MarshallCE, 315 MarshallCR, 198 MarshallFHA, 568 MasonGF, 480 MathewsAP, 271, 315, 481, 569, 574 MattillHA, 204 MaurerO, 482, 572 MauthnerT, 151 MearaFS, 349 MeltzerSJ, 190, 269, 274, 275, 359, 572 MendelLB, 277, 322, 349, 479, 480, 483, 568, 577 MetcalfH, 316 MeyerA, 482 MeyerGM, 575 MillerECL, 479 MillerEGjr, 203, 538, 578 MillerFR, 191 MillerJA, 573 MillerSR, 481 MitchellPH, 573 MitlacherW, 483 MolischH, 567 MooreAR, 273 MooreB, 128, 197, 447 MooreGT, 316 MooreJA, 200 MoorhouseVHK, 273 MorelyEW, 194 MorgulisS, 310 MorseEW, 282 MorseHN, 483 MorseM, 322, S75 MorseWJ, 316 MosenthalHO, 204, 325, 580 MottFW, 481 MuenchAA, 579 MüllerF, 191 MullerHJ, 485 MunkH, 188, 307 MurlinJR, 480 MurphyCV, 307 MurphyJB, 569 MurrayBL, 480 MurrillWA, 574 MyersRE, 310 NernstW, 309 von NeusserE, 188 NicholsWH, 194 NicoletBH, 539 NoguchiH, 476 von NoordenC, 312 NorrisC, 349, 361, 578 NovyFG, 48 1 NoyesAA, 483 OehlersHC, 579 OldbergO, 476, 569 OlsenH, 316 OpenhymAW, 193 Ortner — , 310 OsborneTB, 150, 278, 349, 483 OsburnRC, 200, 485, 575 OslerW, 481 OsterhoutWJV, 569 OstwaldW, 441 OtisDH, 283 OttenbergR, 203 OviattCJ, 570 PackardC, 200, 575 PagnoulA, 307 PalmerLS, 539 Pappenheim er AW, 200, 575 ParkEA, 321, 576 ParkWH, 362, 476, 481 ParsonsCL, 199, 489 PatonS, 481, 482 PattersonHS, 569 PearceRG, 484, 570 PearceRM, 314 PearceRS, 273 PearlR, 316 PeckC, 315 PepperW, 314 PerkinWH, 195, 311 Perlzweig WA, 578 PetersonWH, 539 PetrunkevitchA, 316 PettiboneCJV, 538 PfafifF, 310, 569 PhelpsEB, 569, 573 PhelpsIK, 192, 480, 573 PickhardtEG, 579 PilcherJD, 273, 570 Planten, 281 PlautE, 579 PlimmerRHA, 128, 446 I9I3] Index: Personal Suhjects 603 Po eis — , 297 Ponfink — , 477 PooleJP, 193 PorterWT, 274, 353 PowerFB, 568 PrescottSE, 315 PrimmRL, 539 ProchazkaGx\, 489 PruchaMJ, 310, 478 PunnettPW, 325 RaduJW, 556 RaistrickH, 571 RamsayW, 190, 193, 320, 476 RamsdenW, 447 RansomF, 198, 483 RaubenheimerO, 483 RawlBH, 199 RayBE, 191 RedmanT, 571 ReedHS, 574 Remsenl, 199 RenshawDF, 325, 327, 579 RettgerLF, 315 ReverdinJ, 190 RichardsAN, 278, 349 RichardsHM, 574 RichardsTW, 477, 483 RichardsonAE, 321, 576 RichetC, 309 RichterO, 190 RiddleO, 570 RiggGB, 484 RinakerHB, 578 RingerAI, 277 RitthausenH, 307, 335 RobinsonGC, 273 RobinsonJE, 193 RobinsonLA, 325 RobinsonWJ, 575, 576 RockefellerJD, 573 RockwoodEW, 349 von RombergE, 191 RoseAR, 203, 204, 321, 327, 482, 485, 538, 572, 579 RosePB, 307 RoseWC, 277 RosenauMJ, 478 RosenbloomJ, 204, 324, 325, 485 RosengartenGD, 489 RosenthalN, 580 RossiO, 481 RothermelE, 579 RowntreeLG, 569 RubnerM, 129, 312 RuddWF, 479 RudnickP, 480 RumboldC, 192 RussellEJ, 447 RyanAH, 273, 281 Sabattier — , 308 SabinAM, 489 Sachs, 196 SaylorMA, 478 SchaeferHH, 579 SchäferEA, 193, 195, 567 SchattenfrohA, 129 SchiffJH, 196 SchlossOM, 321, 575, 580 Schmidt — , 310 SchneiderEC, 273, 277 SchrammJR, 539 SchulzeE, I, 205 SchwarzeCA, 200, 576 SchwendenerS, 477 ScottEL, 191 ScottGG, 538 ScovellMA, 188 SeamanEC, 204, 577, 579 SeaverFJ, 485, 576 SedgwickWT, 349 SeemanJ, 476 ShafferBE, 204, 325 ShafferPA, 277 ShearCL, 316 ShermanHC, 278, 480, 483 ShrawderJ, 192 ShulICA, 312 SilvesterRW, 310 SIeeswijk — , 297 Smedleyl, £,77 SmithA, 484 SmithCS, 203, 204, 325, 538, 579 SmithHM, 478 SmithJL, 447 SmithT, 535 SmithWT, 478 SollmannT, 269, 280 SpauldingWJ, 481 SpenceTH, 311 SpencerHJ, 200, 575 SpitzkaEA, 485, 576 StarlingEH, 193 SteelM, 201 SteenbockH, 277 SteinL, 579 StengelA, 314 SternAR, 315 StevensNE, 192 StewartFC, 316 StilesPG, 569 StillmanRG, 322 StockardCR, 200, 575 StockingWA, 569 StockmanR, 198 StoneEC, 579 StoneRE, 192 StoverWG, 191 StraussH, 313 StrongRP, 3 1 1 SulzbergerF, 196 SweenyME, 325, 577, 579 SwainRE, 573 SwannAW, 575 TaftWH, 194, 308 TashiroS, 312, 539 TatumAL, 539 Taylor AE, 314 TerriberryWK, 322 ThatcherRW, 479 ThaxterR, 191 ThayerWS, 191 ThomasAW, 580 ThompsonFL, 579 ThompsonGE, 538 ThompsonHB, 579 ThompsonWG, 349 ThomsH, 483 ThorndikeEL, 573 ThorntonMK, 580 TorreyHB, 192 TracyG, 578 TreadwellAL, 315 TreleaseW, 316 TrueRH, 483 TschirchA, 483 TurckFB, 482, 571 TürkW, 192 TurnerBB, 311 TwichellDC, 485 TwissEM, 193 UhligEC, 489 UnderhillFP, 278, 281, 312 UpshurFW, 479 UpsonF, 570 Van AlstyneEVN , 538 Van BeurenFT, 577 Van IngenP, 200, 485 Van SlykeDD, 277 VassaleG, 476 VaughanVC, 477, 478,568, 574 VerlageCCM, 574 VivianA, 484 WadsworthAB, 481 WakemanNA, 539 WalcottHP, 194 Walker JA, 579 WallaceGB, 481 WallachO, 309 WarburgE, 308 WardHB, 316 von Wassermann A, 191 6o4 Index: Personal Suhjccts [July WatersHJ, 199, 282 WatkinsED, 575 WebberHJ, 312 WeberHA, 188 WeinbergerW, 204, 580 WeismanC, 204, 538, 579 WelchWH, 350, 480, 482 WelkerWH, 203, 204, 485, 575 WellsHG, 269, 277, z-jZ, 349 WellsLF, 481 WesbrookFF, 480, 570 WesenerJA, 316 WesslerH, 204 WestCJ, 361 WestRM, 192 WeymouthFW, 569 WheelerHJ, 190, 311 Wheelerl, 200, 575, 579 WheelerR, 539 WhitneyDD, 575 IVickwireE, 203 WiggsLB, 479 WilcoxEP, 321 WilelS, 575 WileyHW, 329 WilliamsG, 571 WilliamsHB, 273 WilliamsOT, 307 WilliamsWR, 569 WilliamsonCS, 570 WilsonEB, 315, 439 WilsonGW, 581 WilsonJ, 567 WiltshireSP, 571 WinklerH, 191 WinslowC-EA, 315, 322, 572 WinterT, 188 WiseLE, 325, 578 von Wittich — , 441 WoglomWH, 201, 321 WoUsteinM, 569 WoodTB, 447 WoodruffLL, 200, 575 WoodwardHE, 321 WoodyattRT, 277 WoolleyPG, 568 WorthingGH, 580 WrightWJ, 191 Young — , 310 ZappleFC, 480 ZinsserH, 485 OFFICERS OF THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-1913* OFFICIAL REGISTER, MAY 31, 1913 William J. Gies : Professor and Chairman of the Staff ; Consulting chemist, New York Botanical Garden; Pathological chemist, First Division, Bellevue Hospital; Member of the Faculties of N. Y. Teachers College and N. Y. College of Pharmacy. [B.S., Gettysburg College, 1893 and M.S., 1896; Ph.B., Yale University, 1894 and Ph.D., 1897. Instructor, i898-'o2; adjunct Pro- fessor, 1902-05 ; Professor, 190S-.] Paul E. Howe: Assistant Professor. [B.S., University of Illinois, 1906; A.M., 1907 and Ph.D., 1910. Assistant Professor, 1912-.] Alfred P. Lothrop : Associate and De partmental Registrar. [A.B., Oberlin, 1906 and A.M., 1907; Ph.D., Columbia, 1909. Assistant, i9o8-'o9; instructor, i909-'i2; associate, 1912-.] Emily C. Seaman: Instructor. [B.S., Adelphi College, 1899; A.M., Columbia, 190S and Ph.D., 1912. Tutor, i909-'io; instructor, 1910-.] Nellis B. Foster : Associate; Associate Physician, New York Hospital ; Chemist, St. Luke's Hospital. [B.S., Amherst College, 1898; M.D., Johns Hopkins University, 1902. Instructor, i9o6-'o8; associate, 1908-.] Walter H. Eddy : Associate and Secretary of the Staff. [B.S., Amherst Col- lege, 1898; A.M., Columbia, 1908 and Ph.D., 1909. Assistant, i9o8-'io; associate, 1910-.] Herman O. Mosenthal: Associate; Assistant Attending Physician, Presbyterian Hospital; Assistant Physician, Vanderbilt Clinic; Instructor in medicine. [A.B., Columbia, 1899 and M.D., 1903. Assistant, i9o8-'o9; instructor, 1909- '12; associate, 1912-.] Max Kahn: Instructor; Director of the chemical and physiological laboratories of Beth Israel Hospital. [M.D., Cornell University Medical College, 1910; A.M., Columbia, 1911 and Ph.D., 1912. Instructor, 1912-.] Louis E. Wise: Instructor. [A.B., Columbia, 1907 and Ph.D., 1911. Instructor, 1912-.] Edgar G. Miller, Jr. : Assistant, 1911-. [B.S., Gettysburg College, 1911.] Frederic G. Goodridge : Assistant, 1912-. [A.B., Harvard University, 1897; M.D., Columbia, 1901.] Arthur Knudson : Assistant, 1912-. [A.B., University of Missouri, 1912.] Ethel Wickwire: Assistant, 1912-. [A.B., Tri-State College, 1909.] TuLA L. Harkey : Assistant, 1912-. [A.B., Colorado College, 1909.] Benjamin Horowitz : Assistant, 1913-. [B.S., Columbia, 1911 and A.M., 1912.] Christian Seifert: Laboratory assistant, 1898-. Stella Waldeck : Recorder, 1908-. Blanche E. Shaffer: Laboratory assistant, summer session, 1912. Joseph S. Hepburn : University fellow, I9i2-'i3. [A.B., Central High School, Philadelphia, 1903 and A.M., 1908; B.S., University of Pennsylvania, 1907 and M.S., 1907.] * The work of the department was inaugurated in October, 1898, by Prof. R. H. Chittenden (lecturer and director), Dr. William J. Gies (instructor), Messrs. Alfred N. Richards and Allan C. Eustis (assistants), and Christian Seifert (laboratory assistant). COURSES OFFERED BY THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-13 {Abbrcviations : C, Conference; D, demonstration ; L, lecture; Lw, labora- tory work; R, recitation.) ORGANIC CHEMISTRY 51. Elementary ORGANIC CHEMISTRY. (First half year. Medical School.) Introductory to course 102 (52). (Required of first year students of medicine.) L, I hr. D, I hr. R, 2 hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Drs. Wise and Goodridge, and Messrs. Miller and Knudson. NUTRITION (PHYSIOLOGICAL AND PATHOLOGICAL CHEMISTRY) 101(2) — Grad. General biological chemistry. A course in the elements of normal nutrition. (All year. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies, Dr. Lothrop and Messrs. Miller and Knudson. 101(2) — B. T. General biological chemistry. Specially adapted to the needs of secondary school teachers of hiology. {All year. Medical School.) L, I hr. Lw, 4 hr. Dr. Eddy. 101:102 — T. C. General physiological chemistry. A course in the ele- ments of normal nutrition. {Each half year. Teachers College, School of Practical Arts.) L, 2hr. R, i hr., each section (2). Lw, 5 hr., each section (2). Prof. Gies, Dr. Seaman and Misses Wickwire and Harkey. (This course is designated " Chemistry 51 " and " Household Arts Education 125 " in the Teachers College Announcement.) This course is designated " Chemistry s 51 " in the Teachers College Division of the Summer School Announcement. The course was given last summer by Prof. Gies, Dr. Seaman and Miss Shaflfer. 102 (52) — Med. General physiological chemistry. {Second half year. Medical School.) A course in the elements of normal nutrition. {Required of first year students of medicine.) L, 2 hr. R, i hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Dr. Wise, and Messrs. Miller and Knudson. This course is designated " 5 — 104 " in the Medical Division of the Summer School Announcement. It was given last summer by Prof. Gies and Dr. Smith. 104. General pathological chemistry. Lectures on nutrition in disease. (Second half year. Teachers College, School of Practical Arts.) L, i hr. Prof. Gies. (This course is designated "Chemistry 52" in the Teachers College An- nouncement.) 209-210, Chemistry of nutrition. (All year. School of Pharmacy. Re- quired of candidates for the Degree of Doctor of Pharmacy.) L, i hr. Prof. Gies. 213-214. Advanced physiological chemistry, including methods of re- search in nutrition. (All year. Teachers College, School of Practical Arts.) L, i hr. Lw, 5 hr. Prof. Howe, Dr. Seaman and Mr. Horowitz. (This course is designated "Household Arts Education 127" in the Teachers College An- nouncement.) 217-218. BiOCHEMICAL methods of RESEARCH, INCLUDING CLINICAL METHODS AND URiNARY ANALYSis IN GENERAL. (All year. Medical School.) L, I hr. Lw, 7 hr. Profs. Gies and Howe, Dr. Lothrop, and Messrs. Miller and Hepburn. 219-220. Nutrition in health. A laboratory course in advanced physio- logical chemistry. (All year. Medical School.) L, 2 hr. Lw, 14 hr. Profs. Gies and Howe, and Dr. Lothrop. Courses in Nutrition (continued) 221-222. Nutrition in Disease. A laboratory course in advanced patholog- ical chemistry. (All year. Medical School.) L, 2 hr. Lw, 14 hr. Prof. Gies. 223-224. Nutrition in Disease. {All year. Medical School.) L, i hr. Profs. Gies and Howe, and Drs. Fester, Mosenthal, Kahn and Goodridge. 225-226. Advanced physiological and pathological chemistry, including ALL PHASES OF NUTRITION. {All year.- Medical School.) Research. C, i hr, (individual students). Lw, 16 hr. Profs. Gies and Howe, and Dr. Lothrop. TOXICOLOGY 231-232. Effects and detection of poisons, including food preservatives AND adulterants. (All year. Medical School.) Lw, 6 hr. Prof. Gies and Mr. Miller. BOTANY 235-236. Chemical physiology of plants. {All year. New York Botan- ical Garden.) L, i hr. Lw, 5 hr. Prof. Gies. BACTERIOLOGY 241-242. Chemistry of microorganisms : fermentations, putrefactions and THE behavior of enzymes. An introduction to sanitary chemistry. {All year. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies. SANITATION 105. Sanitary chemistry. {Second half year. Teachers College, School of Practical Arfs). L, i hr. Lw, 3 hr. Dr. Seaman and Miss Harkey. (This course is designated "Chemistry 57" and "Household Arts Education 129" in the Teachers College Announcement.) BIOCHEMICAL SEMINAR 301-302. Biochemical Seminar. {All year. Medical School.) 1 hr. Prof. Gies. RESEARCH IN BIOLOGICAL CHEMISTRY Biochemical research may be conducted, by advanced workers, independently or under guidance, in any of the departmental laboratories. LABORATORIES FOR ADVANCED WORK IN BIOCHEMISTRY The laboratories in which the advanced work of the biochemical department is conducted are situated at the College of Physicians and Surgeons, Teachers College, New York Botanical Garden and Bellevue Hospital. Each laboratory is well equipped for research in nutrition and all other phases of biological chemistry. BIOCHEMICAL LIBRARY Prof. Gies' library occupies a room adjoining the main biochemical labora- tory at the College of Physicians and Surgeons and is accessible, by appoint- ment, to all past and present workers in the Department. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION The Biochemical Association holds scientific meetings regularly on the first Fridays in December, February and April, and on the first Monday in June. These meetings are open to all who may be interested in them. SUMMER SCHOOL COURSES See page 579. BOOKS RECEIVED The BiocHEMiCAL Bulletin promptly acknowledges here the receipt of publications presented to it. Reviews are matter-of-fact Statements of the natura and Contents of the pubHcations referred to, and are intended solely to guide possible piirchasers ; the wishes or expectations of publishers or donors of volumes will be disregarded, if they are incompatible with our convictions re- garding the interests of our colleagues. The sizes of the priiited pages are indicated, in inches, in the appended notices. An introduction to the chemistry of plant products. By Paul Haas (lecturer on chemistry, Royal Gardens, Kew) and T. G. Hill (reader in vegetable physiology, Univ. of London). Pp. 401 — 4X7; $-2.25 net. Long- mans, Green and Co., 1913. Excellent discussion of the chemistry and biological significance of many of the most important plant constituents. Besides extended treatment of carbo- hydrates, lipins and proteins, chapters are devoted respectively to glucosides. tannins, pigments, nitrogenous bases (alkaloids, ptomaines, purins), colloids and enzymes. Methods of preparation, detection and quantitative determination are numerous and well described. Good snbject index. The most valuable recent contribution of its kind to phyto-chemistry. Strongly recommended to biological chemists generally — to botanists in particular. Gies. Practical physiological chemistry. By Sidney W. Cole, demonstrator of physiology, Trinity College, Cambridge. Third edition. Pp. 230 — 4 X 6>4 ; 7s. 6d. net. W. Heffer & Sons, Ltd., Cambridge, Eng., 1913. . Very useful laboratory manual. Subject treated chiefiy from static point of view. Practical throughout. Methods well selected. Quantitative pro- cedures given satisfactory attention. Special emphasis laid upon Folin's micro- chemical methods of urinary analysis. Good index. See review by Walter Jones, Jour. Anier. Chem. Soc, 1913, xxxv, p. 1064. Gies. Physiological Researches. (Appears at irregulär intervals.) Edited by Burton E. Livingston (Manager), Johns Hopkins Univ.; Daniel T. MacDougal, Carnegie Inst, of Wash. ; and Herbert M. Richards, Columbia Univ. Vol. I: No. I — The rclation of environmental conditions to the phenomenoii of perma- nent wilting in plants, by Joseph S. Caldwell. Pp. 1-56 — 4% X 75^ ; July, 1913; $0.75 ($5.00 per vol.). Physiological Researches, Station N, Baltimore, Md. Physiological Researches, unlike the conventional Journal, appears at irregulär intervals in the form of individual physiological papers, paged sequentially ; each succession of about 450 pages will be a volume unit. The papers will be numbered sequentially, in each volume and in the whole series. An editorial feature will be the publication of an author's abstract in advance of the appearance of each paper, and also as a preliminary part of each paper in its final form in the series. Although the three editors are eminent botanists, it is their intention to make Physiological Researches, as its name implics, an archive for physiology in its broadest and deepest scnsc. This new publication bcgins its career auspiciously and promises not only to rival the Ainer. Jour. Physiol. in interest and value, but also to sharc with that Journal the high credit of stimulating the advancement of physiological research. The initial paper, by Prof. Caldwell, is a masterly treatment of an interesting and perplexing subject, and establishes a Standard of merit which will doubtless characterize each issue of PJiysioIogical T^X8; $1.40 net. Longmans, Green and Co., 1912. Researches on cellulose. III (1905-1910). By C. F. Gross and E. J. Bevan. Pp- ^73 — 3^^X6; $2.50 net. Longmans, Green and Co., 1912. Sigma Xi Quarterly. Vol. i, No. i (March, 1913). Pp. 30. Editorial committee : J. McK. Cattell, D. C. Miller, H. B. Ward, S. W. Williston. Pub- lished by the Society of the Sigma Xi, H. B. Ward, corresponding secretary, Champaign, 111. Bulletin of the American Home Economics Association. Series i, No. i (Nov., 1912). Published quarterly by the American Home Economics Associa- tion, Benjamin R. Andrews, secretary, 525 W. I20th St., New York City. Abstract-buUetin of the Physical Laboratory of the National Electric Lamp Assoc. Vol. I: No. i, pp. 1-128; Jan., 1913. Cleveland, O. OFFICERS OF THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-1913* OFFICIAL REGISTER, MAY 31, 1913 William J. Gies : Professor and Chairman of the Staff; Consulting cheniist, New York Botanical Garden; Pathological chemist, First Division, Bellevue Hospital; Member of the Faculties of N. Y. Teachers College and N. Y. College of Pharmacy. [B.S., Gettysburg College, 1893 and M.S., 1896; Ph.B., Yale University, 1894 and Ph.D., 1897. Instructor, i898-'o2; adjunct Pro- fessor, 1902-05; Professor, 1905-.] Paul E. Howe: Assistant Professor. [B.S., University of Illinois, 1906; A.M., 1907 and Ph.D., 1910. Assistant Professor, 1912-.] Alfred P. Lothrop: Associate and Departmental Registrar. [A.B., Oberlin, 1906 and A.M., 1907; Ph.D., Columbia, 1909. Assistant, igoS-'og; instructor, i909-'i2; associate, 1912-.] Emily C. Seaman: Instructor. [B.S., Adelphi College, 1899; A.M., Columbia, 1905 and Ph.D., 1912. Tutor, i909-'io; instructor, 1910-.] Nellis B. Foster : Associate; Associate Physician, New York Hospital ; Chemist, St. Luke's Hospital. [B.S., Amherst College, 1898; M.D., Johns Hopkins University, 1902. Instructor, i9o6-'o8; associate, 1908-.] Walter H. Eddy: Associate and Secretary of the Staff. [B.S., Amherst Col- lege, 1898; A.M., Columbia, 1908 and Ph.D., 1909. Assistant, i9o8-'io; associate, 1910-.] Herman O. Mosenthal: Associate; Assistant Attending Physician, Presbyterian Hospital; Assistant Physician, Vanderbilt Clinic; Instructor in medicine. [A.B., Columbia, 1899 and M.D., 1903. Assistant, i9o8-'o9; instructor, 1909- '12; associate, 1912-.] Max Kahn: Instructor; Director of the chemical and physiological laboratories of Beth Israel Hospital. [M.D., Cornell University Medical College, 1910; A.M., Columbia, 191 1 and Ph.D., 1912. Instructor, 1912-.] Louis E. Wise: Instructor. [A.B., Columbia, 1907 and Ph.D., 1911, Instructor, 1912-.] Edgar G. Miller, Jr. : Assistant, 191 1- [B.S., Gettysburg College, 1911.] Frederic G. Goodridge: Assistant, 1912-. [A.B., Harvard University, 1897; M.D., Columbia, 1901.] Arthur Knudson : Assistant, 1912-. [A.B., University of Missouri, 1912.] Ethel Wickwire: Assistant, 1912-. [A.B., Tri-State College, 1909.] TuLA L. Harkey: Assistant, 1912-. [A.B.. Colorado College, 1909.] Benjamin Horowitz: Assistant, 1913-. [B.S., Columbia, 1911 and A.M., 1912.] Christian Seifert: Laboratory assistant, 1898-. Stella Waldeck : Recorder, 1908-. Blanche E. Shaffer: Laboratory assistant, summer Session, 1912. Joseph S. Hepburn: University fellow, 1912-13. [A.B., Central High School, Philadelphia, 1903 and A.M., 1908; B.S., University of Pennsylvania, 1907 and M.S., 1907.] * The work of the department was inaugurated in October, 1898, by Prof. R. H. Chittenden (lecturer and director), Dr. William J. Gies (instructor), Messrs. Alfred N. Richards and Allan C. Eustis (assistants), and Christian Seifert (laboratory assistant). COURSES OFFERED BY THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, 1912-13 (Abbreviations: C, Conference; D, demonstration ; L, lecture; Lw, labora- tory work; R, recitation.) ORGANIC CHEMISTRY 51. Elementary ORGANIC CHEMISTRY. (First half year. Mcdical School.) Introductory to course 102 (52). (Required of first year students of mediane.) L, I hr. D, I hr. R, 2 hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Drs. Wise and Goodridge, and Messrs. Miller and Knudson. NUTRITION (PHYSIOLOGICAL AND PATHOLOGICAL CHEMISTRY) 101(2) — Grad. General biological chemistry. A course in the elements of normal nutrition. {AU year. Mcdical School.) L, i hr, Lw, 7 hr. Prof. Gies, Dr. Lothrop and Messrs. Miller and Knudson. 101(2) — B. T. General biological chemistry. Specially adopted to the needs of secondary school teachers of biology. (All year. Medical School.) L, I hr. Lw, 4 hr. Dr. Eddy. 101:102 — T. C. General physiological chemistry. A course in the ele- ments of normal nutrition. (Each half year. Teachers College, School of Practical Arts.) L, 2hr. R, i hr., each section (2). Lw, Shr., each section (2). Prof. Gies, Dr. Seaman and Misses Wickwire and Harkey. (This course is designated "Chemistry 51" and "Household Arts Education 125" in the Teachers College Announcement.) This course is designated " Chemistry s 51 " in the Teachers College Division of the Summer School Announcement. The course was given last summer by Prof. Gies, Dr. Seaman and Miss Shaflfer. ^ 102 (52) — Med. General physiological chemistry. (Second half year. Medical School.) A course in the elements of normal nutrition. (Required of first year students of mediane.) L, 2 hr. R, i hr., each section (2). Lw, 6 hr., each section (2). Profs. Gies and Howe, Dr. Wise, and Messrs. Miller and Knudson. This course is designated " S — 104" in the Medical Division of the Summer School Announcement. It was given last summer by Prof. Gies and Dr. Smith. 104. General pathological chemistry. Lectures on nutrition in disease. (Second half year. Teachers College, School of Practical Arts.) L, i hr. Prof. Gies. (This course is designated "Chemistry 52" in the Teachers College An- nouncement.) 209-210. Chemistry of nutrition. (All year. School of Pharmacy. Re- quired of candidates for the Degree of Doctor of Pharmacy.) L, i hr. Prof. Gies. 213-214. Advanced physiological chemistry, including methods of re- search in nutrition. (All year. Teachers College, School of Praclical Arts.) L, I hr. Lw, 5 hr. Prof. Howe, Dr. Seaman and Mr. Horowitz. (This course is designated "Household Arts Education 127" in the Teachers College An- nouncement.) 217-218. BiOCHEMICAL methods of RESEARCH, INCLUDING CLINICAL METHODS AND URiNARY ANALYSis IN GENERAL. (All year. Medical School.) L, I hr. Lw, 7 hr. Profs. Gies and Howe, Dr. Lothrop, and Messrs. Miller and Hepburn. 219-220. Nutrition in health. A laboratory course in advanced physio- logical chemistry. (All year. Medical School.) L, 2 hr. Lw, 14 hr. Profs. Gies and Howe, and Dr. Lothrop. Courses in Nutrition (continued) 221-222. NuTRiTioN IN DISEASE. A laboratory course in advanced patholog- ical cheniistry. {All year. Medical School.) L, 2 hr. Lw, 14 hr. Prof. Gies. 223-224. Nutrition in Disease. {All year. Medical School.) L, i hr. Profs. Gies and Howe, and Drs. Fester, Mosenthal, Kahn and Goodridge. 225-226. Advanced physiological and pathological chemistry, including ALL PHASES OF NUTRITION. {All year, Medical School.) Research. C, i hr. (individual students). Lw, 16 hr. Profs. Gies and Howe, and Dr. Lothrop. TOXICOLOGY 231-232. Effects and detection of poisons, including food preservatives AND adulterants. {All year. Medical School.) Lw, 6 hr. Prof. Gies and Mr. Miller. BOTANY 235-236. Chemical physiology of plants. {All year. New York Botan- ical Garden.) L, i hr. Lw, 5 hr. Prof. Gies. BACTERIOLOGY 241-242. Chemistry of microorganisms : fermentations, putrefactions AND THE BEHAViOR OF ENZYMES. An introducHon to sanitary chemistry. {AU year. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies. SANITATION 105. Sanitary chemistry. {Second half year. Teachers College, School of Practical Arts). L, i hr. Lw, 3 hr. Dr. Seaman and Miss Harkey. (This course is designated " Chemistry 57 " and " Household Arts Education 129 " in the Teachers College Announcement.) BIOCHEMICAL SEMINAR 301-302. BiocHEMiCAL Seminar. {All year. Medical School.) i hr. Prof. Gies. RESEARCH IN BIOLOGICAL CHEMISTRY Biochemical research may be conducted, by advanced workers, independently or under guidance, in any of the departmental laboratories. LABORATORIES FOR ADVANCED WORK IN BIOCHEMISTRY The laboratories in which the advanced work of the biochemical department is conducted are situated at the College of Physicians and Surgeons, Teachers College, New York Botanical Garden and Bellevue Hospital. Each laboratory is well equipped for research in nutrition and all other phases of biological chemistry. BIOCHEMICAL LIBRARY Prof. Gies' library occupies a room adjoining the main biochemical labora- tory at the College of Physicians and Surgeons and is accessible, by appoint- ment, to all past and present workers in the Department. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION The Biochemical Association holds scientific meetings regularly on the first Fridays in December, February and April, and on the first Monday in June. These meetings are open to all who may be interested in them. SUMMER SCHOOL COURSES See page 579. CONTENTS An Investigation to Determine the Accuracy of a Modified Meigs Method for the Quantitative Determination of Fat in Milk, with A Description of an Improveo Form of Apparatus. Walter Lewis Cr oll . . 50g The Occurrence of Arsenic in Soils. /. E. Greaves 519 Further Notes on the Relationship Between the Weicht of the Sugar Beet and the Composition of its Juice. /. Arthur Harris and Ross Aiken Gärtner. 524 Note on the Relationship Between Barometric Pressure and Carbon- dioxide ExcRETioN IN Man. /. Arthur Harris 530 The Bleached Flour Decision. Ross Aiken Gärtner 532 Emil Chr. Hansen Fund. S. P. L. Sörensen 535 Biological Chemistry in the Philippines. Robert Banks Gibson 536 DOCTORATES IN BlOLOGICAL ChEMISTRY. CoNFERRED BY AmERICAN UnIVER- siTiEs, 1912- 13. P. H. D 538 Scientific Proceedings of the Columbia University Biochemical As- sociation. Alfred P. Lothrop, Secretary S4i Biochemical Bibliography and Index. William J. Gies 559 Biochemical News, Notes and Comment 567 Editorials: Including additional quotations from letters on the Mathews plan for the Organization of an American Biological Society 582 Index : Volume II, (Includes names of authors, and impersonal and per- sonal subjects) 589 Title Page for Vol. II, with Summary of Contents, List of Illustra- TioNS, etc i-xvi The Biochemical Bulletin is a quarterly biochemical review. It pub- lishes results of original investigations in biological chemistry, presents mis- cellaneous items of personal and professional interest to chemical biologists, and solicits original contributions to research, preliminary reports of investiga- tions, abstracts of papers, addresses, lectures, criticism, reviews, descriptions of new methods and apparatus, practical suggestions, biographical notes, historical summaries, bibliographies, quotations, news items, proceedings of societies. personalia, views on current events in chemical biology, etc. Subscription prices. Vol. I: $6.00 (No. i, $1.50; No. 2, $2.50; No. 3, $2.00: No. 4, $1.50). Vol. II: $5.00 (No. 5, $2.00; No. 6, $1.50; No. 7, $2.00; No. 8, $1.00). Vol. III: $2.75 (domestic); $3.00 (foreign.) ; $5.00 after Jitly i, 1914. Address remittances, manuscripts and corresoöfndence to the Managing Edi- tor, William J. Gies, 437 West 59th St., New York. 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