Bute beter ast, Ht ati ish ote He 3 i : i feet ihe it i ty siege paps be st i teehee a ease retetl te itt Halt att ; “ ofits] atstranatneh ite ti ay th ett nid val ni i rhntiaaiat ras es ‘ hii ie fi iit raises ct ca petite : Sebehestys ttt i ty it a 1 Gh Datei HMeapsapkearbtecheeto NRO sah Ore iterating er Seudpeeh hatte Pesuathsrereve poche cterneCsteTerkses eseek te Hi aid Pettit Hits ? f Saree itt it ineten i f et sets ebesszers tal! ts ifr t see ant ith esata pt Hite Wert hae Hitt nigel i ih i i i SEotee ets SSSSHE : Secees = ts2 ts SESS x se ill fy i mt sig ate ae! pests zeae eats iy ies ate ihe f e itt eh ih iat et et t 4; t 4 putea i ie fiat flit oe f rep ee ee ee ee CHEMICAL MEDICAL & SCIENTIFIC PERIODICALS AND BOOKS 29 East 2! S'St NEW YORK ain, a it, Ngee THE COLLECTING NET DIRECTORY Supplementary Directory for 1934 MARINE BIOLOGICAL LABORATORY INVESTIGATORS Addison, W. H. F. prof. horm. hist. & emb. Penn- sylvania Med. Br 329. Gosnold. Beams, H. W. asst. prof. zool. Iowa. Br 123. Dr 5. Buyse, A. instr. biol. Rochester. Br 224. Gray, Buz- zards Bay. Bissonnette, 'T. H. prof. biol. Trinity. OM 26. D 108. Cohen, Rose. teach. fel. biol. Cincinnati. Br 334. Thompson, Water. Cole, E. C. prof. biol. Williams. OM 28. D 215. Furth, J. asst. prof. path. Cornell Med. Br 222. D 101. Keller, Thelma G. grad. zool. Columbia. Br 3365. MacLeish, Millfield. Larrabee, M. G. grad. path. Pennsylvania. Br 339. Loehwing, W. F. prof. bot. Iowa State. Br 210. A 108. MacCardle, R. G. instr. zool. Temple. OM Base. K 9. Nelsen, O. E. instr. zool. Pennsylvania. OM 27. D 312. Northrop, J. H. mem. Rockefeller Inst. Br 209A. High. Robinson, M. H. Swarthmore. Br 9. Nickerson, Mill- field. Schechter, V. tutor biol. City of New York. OM 10. Dr attic. Steinbach, H. B. Nat. Res. fel. biol. Pennsylvania. Br 111. Old Edwards, School. Townsend, Grace. grad. biol. Chicago. Br 217. H 7. Tulloch, G. S. instr. biol. Brooklyn. OM 39. Elliot, North. Witschi, E. prof. zool. Iowa. Br 210. D 111. Wharton, Marguerite. grad. asst. biol. Mt. Clair State Teachers (New Jersey). Hilton, Main. Woods, F. H. asst. prof. zool. Missouri. OM 32. D 105. STUDENTS (Invertebrate Zoology) Andrews, S. R. American. Dr 5. Barton, Eleanor P. head sci. dept. Edgewood Park Junior. H 6. Beard, R. L. undergrad. asst. zool. Wesleyan. K. Bengel, W. Z. undergrad. asst. zool. De Pauw. Ka. Berliner, R. W. Yale. Densmore, School. Blood, Rozella. Kansas Med. School. Nichols, Main. Blouin, A. prof. biol. Mount St. Louis. D 208. Bolt, O. A. instr. high sch. Calvin. K 5. Bragg, J. H. Central State Teachers. Conklin, High. ae ens A. undergrad. asst. zool. De Pauw. Bridgman, Jane. Smith. W A. Browman, L. asst. instr. zool. Chicago. Baitsell, Brooks. Bumpus, D. F. Oberlin. E. Falmouth. Caldwell, Billie instr. zool. Tenn. Junior. W G. Chen, Y. grad. biol. Pennsylvania. Dr 8. Crouse, Helen V. Goucher. H 1. Danforth, Louise L. American. H 8. Dansereau, A. A. prof. bot. Montreal Col. St. Joseph Rectory. Eckels, R. P. grad. zool. Virginia. A 105. Ettensperger, Flora. Barnard. D303. Farber, S. M. Harvard Med. asst. invert. course. K 5. Gay, Elizabeth H. res. asst. zool. Johns Hopkins. Hilton, Main. Gilbert, P. W. instr. biol. Dartmouth. Ka 2. Glass, M. grad. zool. Yale. Dr 10. Gross, Charlotte C. teach. biol. Union H. S. Jersey) H 7. Hadley, Ruth G. grad. zool. Cornell. W. . Hierholzer, Carolyn A. undergrad. asst. zool. N. J. Col. for Women. W B. Jakobsen, Edith M. N. J. State Teachers. H 7. Jailer, J. W. City of New York. Dr 7. Kennedy, Kathleen M. St. Francis Xavier. K 10. Leach, Eleanor, curator labs. Wellesley. A 209. Leist, C. instr. biol. Kansas State Teachers. Hilton, Main. Sean Mary. Louise. Pennsylvania Col. for Women. ist (3; (New Merrill, J. Edwarta. Wilson. H 2. Mitchell, Helen A. Hood (Maryland). Hilton, Main. Mizelle, J. D. grad. asst. zool. Llinois. Gifford, Gov- ernment. Morland, Hazel M. Swarthmore. Lewis, Buzzards Bay. Ormsby, Louise. grad. zool. Columbia. Robinson, School. Perry, J. Acadia (Canada). Ka. Pettingill, O. S., Jr. teach. fel. biol. Bowdoin. Stoky, Gardiner. Pomerat, G. R. Clark (Mass.). Higgins, Depot. Roney, H. B. grad. asst. zool. Illinois, Gifford, Gov- ernment. Rositzke, Marion M. grad. zool. Main. Saunders, Barbara. Radcliffe. K. Schoenheit, Dorothy M. Rochester. H 9. Schultz, Helen H. asst. prof. biol. State Teachers (Virginia). H 3. Simpson, A. G. undergrad. asst. zool. Wesleyan. K. Smith, M. A. Wabash. K 5. Swartz, Daphne B. instr. biol.-bot, Bradley Poly- technic Inst. Robinson, School. Thayer, Mrs. Walter N. grad. zool. Yale. H 9. Tyson, Rebecca J. Wayne (Detroit). W D. Weishaar, Irma Ruth. instr. comp. anat. Hunter. Googins, Quissett. Welch, Dorothy I. Radcliffe. W. Wharton, G. W., Jr. Duke. Googins, Quissett. Wightman, J. C. Oberlin. Dr 2. Hunter. Hilton, THE BIOLOGIST AND HIS TOOLS The Collecting Net will be under obligation to students and teachers of the biological sciences if they will give care- ful consideration to the products sold by these firms before pur- chasing from other establishments. ‘This is a natural request and a natural thing for a reader of the magazine to do, because the number of pages and the number of illustrations that we can print is more or less in direct proportion to the support given to it by manufacturers of scientific apparatus, supplies and books. In turn, the amount of advertising space that we can sell depends upon the support given to these firms by the readers of The Col- lecting Net. We shall consider it a gracious gesture if biologists will consult our advertising pages for the tools that they need in their work; and further, mention The Collecting Net when it has been a factor in helping them reach a decision concerning these tools. The publiation of The Collecting Net is made possible by the firms listed here. ADVERTISERS IN THE COLLECTING NET : SCIENTIFIC APPARATUS AND SUPPLIES AKATos, INC. HoxeE Inc. AMERICAN INSTRUMENT COMPANY, INC. AMERICAN CHEMICAL PRopuUCTS Co. BauscH & Loms OpTicaAL COMPANY BROOKLYN BOTANIC GARDEN CAMBRIDGE INSTRUMENT COMPANY CHEMICAL PRropucTs COMPANY CLay-ADAMS COMPANY Ermer & AMEND GENERAL BIOLOGICAL SUPPLY HousE SCIENTIFIC P. Braxiston’s Son & Company, INC. Henry Hott AND COMPANY CHEMICAL CATALOG COMPANY LEA & FEBIGER INTERNATIONAL EQUIPMENT COMPANY Erin zs ING: B. M. LeEvoy, INc. New York BroLtocicAL SuPPLy Co. SPENCER LENS COMPANY WILL CORPORATION THE WistTAR INSTITUTE Car ZErss, INC. AMERICAN Book COMPANY BOOKS B. Locin & Son MACMILLAN COMPANY McGraw-HiLt Book CoMPANY Joun Wiey & Sons, INc. The Collecting Net Vol. IX. No. 1 SATURDAY, JUNE 30, 1934 Annual Subscription, $2.00 Single Copies, 25 Cents. THE BIOLOGICAL LABORATORY AT COLD SPRING HARBOR Dr. REGINALD G. Harris, Director To make available immediately, to all biologists and to all biological laboratories, knowledge and complete understanding concerning the activities and plans of any given laboratory, is so desirable RESEARCH IN BIOLOGICAL SUBJECTS AT THE SCRIPPS INSTITUTION OF OCEANOGRAPHY Dr. THomMAS WAYLAND VAUGHAN, Director The editor of THE CoLttectinG NET has in- vited me to contribute an article on the kinds of biological material available for study at the that it is always a very great pleasure to give to biologists a report of TELEPHONE: TERMINUS 5385 (2 Lmen) LONDON (ROYAL FREE HOSPITAL) SCHOOL OF MEDICINE FOR WOMEN Scripps Institution, on the work accomplished at it during the last the Biological Labora- (GRIREGEDAY OF WERE) year, and on the re- tory at Cold Spring Harbor. Lack of ex- change of such infor- mation invariably re- sults in relatively un- productive duplication of efforts. One part of the whole sphere of bio- logy develops much more rapidly than, and at the expense of other parts, until invagination may occur, and instead of enjoying life on the periphery of an enlarg- ing blastosphere, we may find ourselves in the ever closing enteron of a gastrula. Perhaps the most important happening for the past year at Cold Spring Harbor, and (Continued on page 6) Mt ce ae 4 Ler Denk “ Ya a PLE ff td HUNTER STREET, BRUNSWICK SQUARE, W.C.1. searches in progress. With reference to the biological material available for study at the Scripps Institution, about all that can be said is that representa- tives of virtually all groups of marine or- ganisms can be found either along the sea coast near the Institu- tion, in the adjacent ocean waters, or at localities which are ac- cessible from the Insti- tution by means of automobile. There are representatives of all the important groups of marine plants from bacteria to giant kelp, and of all the principal groups of marine jim ok 163% Research in Biological Subjects at the Scripps Institution of Oceanography, Dr. MafeMmase VV Vial NAN® 2). sie cies Sh, cleyellers'® over 1 The Biological Laboratory at Cold Spring Harbor, Dr. Reginald G. Harris ............ al A Method of Preparing Small Insects for Photographing, Dr. P. W. Whiting ......... 9 Notes on the Research Work at the Cornell University Biological Station .............. 10 TABLE OF CONTENTS: BOOK: TREVICW ba. fale Sesicienenitaiel a) se, a\0 upteicenpe et asta isi 10 News trom the Scripps Institution of Ocean- OLTAPIVY! as cde hacteywcet sles: ac oe) leletasulal etailaliat= wiles) »)aiiehepe 11 Courses at the Marine Biological Laboratory Eigitorials pas ee crv a cmneseyetaver sree a cteratategene 4 CHOLAE 14 TLEMISHOL IMUCHESE | yin ateiecleeteleisisie wetltetar ste ate 15, 16 Chemical Room, Dr. Oscar W. Richards ..... 17 IitHey Gt VOCE SOD ssootneosooooowocas anode 18 2 THE COLLECTING NET [ Vou. IX. No. 72 ausnonenannteetnnouasanee CeCe THE THREE PRINCIPAL BUILDINGS LOOKING SOUTHEAST FROM THE PIER At the left is the new building, Ritter Hall; in the center is the Museum-Library building; at the right behind the Museum-Library building’ is the George H. Scripps Laboratory. animals, protozoa, sponges, actinozoa, echino- derms, various kinds of worms, molluscs, crustacea, tunicates, and vertebrates, from blind gobies to sea-lions, porpoises, and whales. The whales are not easy to catch, but the sea lions and porpoises are inclined to be friendly. The sea-lions come when called. As regards any par- ticular kind of organism on which an investigator may wish to work, it is advisable to address an inquiry beforehand to the Director of the Insti- tution. Some groups of organisms are not well represented, for example, the larger echinoids, but there are several local species of star fishes and a species of sand dollar, Dendraster excen- tricus, is common. Stony corals are not available for study along the coast of southern California. A few species are found in shallow water at some places, but the polyps are small and specimens are not numerous. In moderately deep water, several species have been found, but it is doubtful if material could be maintained alive in the laboratory. There are, however, species of sea- anemones, which may be obtained in quantity. Since the Institution has a good salt-water THE SCRIPPS INSTITUTION OF OCEANOGRAPHY View of the three principal buildings looking northwest. In the center is the Library-Museum building; at the right is part of the new laboratory, Ritter Hall; at the left behind the Museum-Library building is the old George H. Scripps Laboratory. At the extreme left may be seen part of the pier. June 30, 1934 ] THE COLLECTING NET 3 supply system, the facilities are good for culture studies and other kinds of experimental investi- gations. It is desirable to have for the waters adjacent to the Scripps Institution a catalogue of the various marine organisms found in its vicinity, like that published for Plymouth, and the pre- paration of such a list is being considered. A great deal of systematic work has been done on the organisms in the waters off southern Califor- nia. Studies have been made of the bacteria by ZoBell, the diatoms by W. E. Allen, larger marine algae by Setchell and Gardiner, several groups of marine protozoa by C. A. Kofoid, for- aminifera by J. A. Cushman, M. L. Natland and others, medusae by H. B. Torrey, arrow worms by E. L. Michael, copepods by C. O. Esterly, tunicates by W. E. Ritter, appendicularia by Christine E. Essenberg, and other groups have been specially studied. A list of many of the principal publications on Pacific coast marine in- vertebrates is given in Johnson and Snook’s ‘‘Sea- shore Animals of the Pacific Coast,” pages 607- 620, a valuable book for those who wish a general idea of the marine invertebrates of the region with which it deals. Regarding the biological researches at the Scripps Institution, it is difficult to separate them from other kinds of oceanographic research. An attempt will be made to illustrate what is meant. It is known that planktonic organisms are abun- dant in places, if not continuously, along the west coast of America from southern California to Valparaiso, Chile. This does not mean that planktonic organisms are not abundant in other places. Marine animals are dependent on phy- toplankton for food. The question immediately presents itself as to why phytoplanktonic organ- isms should be abundant in the strip of water indicated. The abundance is due to the circulation of the ocean waters, a problem of dynamical oceanography. Along much of the California coast, and in places along the coast of Central America and in the Gulf of Panama, and along the coast of South America, the phenomenon known as upwelling occurs. The upwelling water contains abundant supplies of plant nutrients upon which the development of the phytoplankton de- pends. The phytoplankton is eaten by smaller animals and the smaller animals are eaten by larger ones, ultimately reaching the carnivorous marine mammals. The upwelling is caused by winds, which along the coast of California and farther south, and along the west coast of South America within the trade wind belt, shove water from near the shore seaward and cause water from below the surface to move upward to take the place of the water which had been pushed away from the shore. Therefore, the atmos- pheric circulation of the region is a cause of the abundance of life in the sea. Now to consider for a moment some of the chemical aspects of sea water. The growth of phytoplankton in the sea depends upon the presence of plant nutrients. The only means; whereby the distribution of plant nutrients in the sea may be ascertained is by chemical investiga- tions of sea water. ‘The chemical properties of sea water, necessary for the growth of plants, are influenced by many factors, some of which are organic and some inorganic. For animal life in the sea, Oxygen is necessary. In order to as- certain the amount of oxygen available for animal life chemical investigations are needed. In the eastern Pacific there is a layer, which varies in depth from 100 to 800 or 1000 meters, in which there is almost no, or actually no, oxygen. Oxygen is not uniformly distributed in the ocean. In order to understand the adapta- tions of organisms to different oxygen tensions in the sea, both studies in the field and physiologi- cal investigations in the laboratory are required. A geological problem will now be mentioned. It is known that a considerable proportion of the deposits found on the bottom of the ocean is composed of the remains of organisms. From a study of the composition of the marine bottom deposits along the Florida Coral Reef Tract and in the shoal waters of the Bahamas, E. M. Thorp has shown that the calcium carbonate bottom de- posits of the two regions are composed of 95% or more of the remains of organisms. That important deep-sea marine deposits are largely composed of organic remains is indicated by the names of some of the deposits, such as Globigerina Ooze, Diatom Ooze, and Radiolarian Ooze. It has been pretty well established that the principal difference between Globigerina Ooze aad Red Clay is that calcium carbonate remains, which may have started to fall to the bottom over areas of Red Clay, were dissolved before they reached the bottom. This presents a problem in the chemistry of sea water and the variation in the properties of sea water under different con- ditions. Additional illustrations might be given, but enough has been said to show that if we are to understand conditions in the sea, each of the dif- ferent disciplines developed for the study of the sea must be utilized. There is no biology of the sea independent of the physics, chemistry, and geological aspects of the ocean. The biological program at the Scripps Institu- tion, therefore, may be said to begin with the physics and chemistry of the ocean, after which its various biological and geological aspects fol- low. A brief account will now be given of some of the recent biological investigations at the Scripps Institution, taking up the subjects more or less topically. 4 THE COLLECTING NET [ Vou. IX. No. 72 The investigations in microbiology at the In- stitution are in charge of Dr. C. E. ZoBell, as- sisted by Mrs. C. B. Feltham. Several papers have been published during the year and others are in press. Studies of water samples along and off the coast of southern California show that a large proportion of the contained bacteria are able to ammonify numerous substances. Eight new species have been isolated which produce ammonia from low concentrations of urea in sea water, and in cultures containing 2% urea raise the pH to 9.4. Nitrosofiers and nitrifiers have been recovered with regularity from bottom de- posits in relatively shallow water, but similar methods have failed to reveal their presence at depths greater than 500 meters. Nitrate-reducers are encountered very infrequently and they are functional only under unusual conditions. Obser- vations on the influence of temperature, organic. matter, pH, redox potentials, and salinity on the activities of nitrifying and denitrifying bacteria indicate that in the sea the redox potential is per- haps the most imporant factor. Analyses of over 40 bottom deposit samples from depths to as great as 1300 meters show the existence of a large and varied microflora which is clastically active against cellulose, starch, and proteinaceous substrates. Aerobes predominate, but the ratio of anaerobes to aerobes increases with the depth of the core strata. The reduction potential of most of the cores approaches that of the hydrogen-electrode and im witro hydrogen overvoltages may be produced by microbial action. Sulphate-reducing bacteria are common in the mud samples. It has been found that, while immediately fol- lowing initial isolation marine bacteria have specific sea-water requirements, they are readily acclimatized to become physiologically indistin- guishable from fresh-water species. At least 88 new bacterial types from the sea have been characterized. Dr. ZoBell and Miss Esther C. Allen have noted that the usual sequence of events in the fouling of submerged surfaces is first the ad- herence of bacteria and kindred growths, followed by the attachment of barnacles, Bryozoa, hy- droids, and other organisms. Bacteria and, to a lesser extent, diatoms play an important role in the fouling of marine structures. One paper on the subject has been published and another is in preparation. In collaboration with Dr, Nelson A. Wells, the etiological agent of a highly fatal infectious der- matitis of Fundulus parvipinnis and other marine fishes has been isolated and identified as Achro- mobacter ichthyodernus, n. sp., and a paper pub- lished on the results. A second paper dealing with the pathology of the disease has heen ac- cepted for publication in the Journal of Infectious Diseases, Professor W. E. Allen has continued his studies of phytoplankton of the La Jolla region, devoting most of his time to the preparation of the manuscript of a report on the results of daily collections for ten years at the end of the Insti- tution’s pier. Professor Allen has also continued studies of fouling organisms along the lines in- itiated at the Institution several years ago by Professor Wesley R. Coe of Yale University. Dr. Easter If. Cupp has completed a manuscript en- titled, “A critical study of certain distinguishing characters in three closely allied plankton species of the diatom genus Nitzschia and their relations to certain environmental conditions,” and she has prepared a report on the plankton samples col- - lected by the Velerq I/I between San Diego and Panama. Dr. T. W. Vaughan during the year published a number of papers on fossil foraminifera, and he has directed the work on foraminifera and marine bottom-deposits. A publication of the Scripps Institution Bulletin, technical series, en- titled, “The temperature-and depth-distribution of some recent and fossil foraminifera in the south- ern California region,” by Mr. M. L. Natland, is one of significance both to students of the ecology of marine organisms and to geologists. During the vear Dr. [karl Myers succeeded in completely working out the life history of a local species of foraminifera known as Patellina corrugata. A short note entitled, “Multiple tests in the fora- minifera,’ was published in the Proceedings of the National Academy of Sciences (vol. 19, no. 10, pp. 893-899, 1933). The complete report, entitled, “The life history of Patellina corrugata, a foraminifer,” is now ready for publication. The results were presented by Professor C. A. Kofoid at the last meeting of the National Acad- emy, April, 1934. During the last twelve months an investigation has been made by Dr. D. L. Fox, the Institu- tion’s physiologist, and Dr. G. W. Marks, re- search associate, on the digestive enzymes of the plankton feeding Mytilus californianus, with a view to determining the food that it can utilize and must remove from the sea. Its fecal ribbons have been examined for correlative information. The findings from these investigations will be correlated with Buley’s studies, in manuscript, of the gastric contents, and Miss Austin’s studies, also in manuscript, of other features of this mol- luse. Studies on the effect of oxygen in the in- activation of the enzyme, catalase, are being con- tinued, using a number of marine animals and plants. A paper dealing with the subject en-. titled, “The inactivation of mussel catalase by oxygen,” appeared in the Journal of Biological Chemistry (Vol. 103, p. 269, 1933). Another investigation in progress is concerned with the amounts of dissolved copper in sea water, the tolerance of a number of marine mole June 30, 1934 j THE COLLECTING NET 5 luses for this toxic cation, and the possible ac- cumulation of it by the animals. The mussel has in its digestive tract an enzyme or enzymes capable of hydrolyzing the poisonous glucoside amygdalin, with the production of a re- ducing sugar (glucose) and free hydrocyanic acid. Out of 69 bacteria, 6 actinomyces, 3 molds, and 2 yeasts investigated, two-thirds of the bac- teria, and all the other forms except one of the yeasts, possess the ability while living to pro- duce HCN from amygdalin. The mussel enzyme hydrolyses amygdalin in the absence of micro- organisms. A paper dealing with the subject has been prepared. In cooperation with Dr. F. B. Sumner an in- vestigation of considerable length concerning the carotenoid pigments in certain marine fishes has been completed and published under the title, “A study of the variations in the amount of yellow pigment (xanthophyll) in certain fishes and of the possible effects upon this of colored back- grounds” (Jour. Exp. Zool., Vol. 66, pp. 263-301, 1933). - During the year seven articles have been pub- lished from the section of physiology, two are in press, another has been prepared for publication, and three are in the process of preparation. The work in fish biology, in charge of Profes- sor F. B. Sumner, who published several papers during the year, consisted in a continuation of studies of the relations of marine fishes to their environment. The chief subjects of investigation were as follows: The yellow (carotenoid) pigment of fishes, which is partly responsible for the striking ad- justments of these animals to the color of their surroundings in nature, was investigated by him, in cooperation with Dr. Fox of the physiology section. Studies were carried out upon the relations of different parts of the visual field to the color re- sponses of fishes. A new technique was devised, whereby the animals were forced to look through transparent caps which were fitted over the eyes, these caps being variously colored or partly ob- scured by opaque areas. Investigations of the oxygen consumption of fishes were continued by Dr. N. A. Wells. The relations of the metabolic rate of activity, to tem- perature, season, size, sex, and some other fac- tors were determined in part, and certain phases of the work were brought to a definite conclusion. Dr. Wells completed the manuscript of a paper entitled, “Variations in the rate of respira- tory metabolism on the Pacific killifish, Fundu- lus parvipinnis, in relation to temperature, sea- son, age, and sex.” Certain diseases of fishes were investigated by Dr. Wells in collaboration with Doctor ZoBell of the section of microbiology, as has already been stated. During the year two rather large studies of marine bottom deposits have been brought to completion. One of them is a study by Dr. E. M. Thorp entitled, ‘‘Calecareous shallow water marine deposits of Florida and the Bahamas,” and the other is an investigation by Mr. Roger Revelle of the deep-sea “bottom samples collected in the Pacific by the Carnegie. It is expected that the results of both of these investigations will be published by the Carnegie Institution of Washington. It has been stated in the introduc- tory remarks to these notes, that the organic re- mains contained in the sediments have been quantitatively evaluated. Mr. Revelle has also prepared a paper entitled, “The physico-chemical factors affecting the solubility of calcium carbon- ate in sea water,” which will be published in the Journal of Sedimentary Petrology. During the year much attention has been paid by several members of the staff of the Institution to oxida- tion and reduction in the ocean. This subject is a meeting ground of physical oceanography, geology, chemistry, and a considerable range of biological phenomena, therefore members of the staff of the Institution representing each one of those disciplines have cooperated in the inves- tigation. Dr. Robley D. Evans, assisted by Mr. Arthur Kip, is making a study of the radioactive sub- stances in sea water, in marine plants and animals, and in the bottom deposits off the coast of south- ern California. They are also making similar analyses of core samples of deep-sea sediments kindly supplied by Dr. Ph. H. Kuenen from the collections made by the Willebrord Snellius in the East Indies, and on samples supplied by the Hy- drographic Department of the Japanese Imperial Navy, collected by the ship Mansyu in the south- west part of the North Pacific. It is generally known that there is concentration of radium in marine sediments, but it is not known how the concentration is effected. It looks as if it might be accomplished by organisms. In order to test such an hypothesis the inorganic constituents of many organisms are being analyzed for their con- tent of radio-active substances. In the foregoing notes account has been taken of the work of about fourteen people, ten of them members of the Institution staff, one a tem- porary assistant, and three associated with the In- stitution in its research program. During the year about fourteen visiting investigators utilized the facilities of the Institution for biological research and about as many more came to it for confer- ence or advice regarding biological investigations. Many teachers and students from colleges and high schools also came to inspect the work of the Institution, its museum, and aquarium. During the year about thirty papers dealing with biological subjects were published by those connected with the Institution and about nineteen are awaiting publication. 6 THE COLLECTING NET [ Vor. IX. No. 72 THE BIOLOGICAL LABORATORY AT COLD SPRING HARBOR Dr. REGINALD G, Harris, Director (Continued from page one) probably one of the most far reaching actions in the history of the Laboratory, has been the in- auguration here of conference-symposia on quantitative biology. There is, of course, nothing new in conferences or in symposia, but the union of the two during a month, and the inclusion of mathematicians, physicists, and chemists in the conferring group offers a new method which, we hope, may be very useful to biological research. From the point of view of the history and philosophy of marine biological laboratories, the method introduces something which is perhaps quite as unusual as the inclusion of representa- tives of the so-called exact sciences, namely, a frankly directive force in the selection of a large number of the Laboratory’s visiting scientists in any given summer. Thus, last year it was de- cided that we should consider certain surface phenomena, and, consequently men were invited whose work dealt with such phenomena. This year we are to consider some aspects of growth. The invited participants are almost without ex- ception different men from those invited last year. Such a procedure is, as far as I know, wholly new to marine biological laboratories. The purposes behind it are not new, however. We can not but believe that, along with Professor Agassiz’s famous motto of “study nature, not books,” one of the chief reasons why marine laboratories “took,” once they were innoculated into biology, was that they then presented an easy opportunity for leaders of biological research to gather at Woods Hole or at Cold Spring Har- bor, and to adequately exchange news concerning their work and ideas. This was in the days when the total productive population of biologists in the country did not much exceed the total number of persons currently in residence during a summer at these two centers. At that time there were possibly 2,000 scientists of all kinds in the United States, and probably not over six or eight hundred who had published results of research in biology and medicine. But this has all changed very much in the past forty or forty-five years. There are now about 22,000 persons listed in “American Men of Science” (Fifth edition 1933). All of these 22,000 are supposed to be somewhat actively en- gaged in research. At least they have all pub- lished results of research work. Judging from previous editions of this same directory, over 40 per cent. are engaged in the biological and medical sciences, not including chemistry and physics. This means that in this country alone there are approximately 10,000 persons engaged in build- ing up, and changing, the knowledge upon which biology rests. lf at the same time we consider the chemists and physicists, we must add another 5,000. The speed at which these numbers are increasing is indicated by the fact that the direc- tory which had 22,000 names in 1933 had in its edition of 1927 not more than 13,500, in 1921 9,500, in 1910 5,500, and about 4,000 in 1906. From this it can easily be seen that established facts and projected hypotheses in research are continually increasing and changing. ‘This is fur- ther shown by the following facts. The authors’ index of abstracts of chemical scientific papers for one year (1932) indicates that there are about 50,000 titles. A single monthly issue of Biological Abstracts contains over 2,500 abstracts of papers on biology. I pick up at random a scientific journal, Vrotoplasma, published in Ger- many. The issue 1 am looking at was received October 30, 1933. The first paper I turn to was received for publication March 18th. I find in the November 1933 issue of Biological Abstracts an abstract of a paper published in 1927. This is in no way meant to reflect upon the important and excellent work of scientific journals, but to give an indication of how difficult it is for any- one engaged in scientific research to keep abreast of the changes, an appreciable number of which may be of significance to his plans and work. Scientific societies and individuals do much to help overcome the difficulties already indicated. But of course, neither can effectively provide it- self with a news service which approaches that in use in business, diplomacy, finance and other human activities. The result is that scientific re- search is continually working under appreciable handicaps. Not only has the number of research workers increased enormously, but other complications have entered. Outstanding among these is a great broadening of the field actually covered by bio- logical research until it includes biometry, bio- physics and biochemistry. Indeed, as research in biology becomes more mature, it becomes increas- ingly quantitative, and moves more and more into territory previously almost wholly occupied by mathematics, physics and chemistry. In these facts lie some of the reasons why, the Riological Laboratory thought that the time had come to add a directive force to other desirable functions of a marine biological station. That directive force is the conference-symposium. The procedure, as outlined in THE COLLECTING Net last year, has been found to work very well. The Laboratory invites, each summer, a small June 30, 1934 } THE COLLECTING NET 7 group of outstanding workers in one given aspect ot biological research. These men are chosen so that they will represent not only biological aspects of a given problem, but those of mathematics, chemistry and physics, the idea being to try to cover each year all of the aspects of a limited’ subject from mathematics to application to medi- cine. These men are in residence at the labora- tory for one month during the summer. Their small number is supplemented by others who come from time to time to present papers and to take part in symposia and discussions. The papers presented at these symposia, together with care- fully edited discussion which takes place after the presentation of a paper, are published as soon as possible. Last year over half of the papers were given preliminary publication in THE CoL- LECTING Net within two weeks: after they were presented, and the distribution of the volume containing all of the papers and edited discussion began six months ago. It is planned that the papers and discussion shall more and more be participated in, not only by Americans but also by scientists from other parts of the world, this being done by letter when personal attendance is not possible. I wish to stress particularly the fact that we do not consider that this is a panacea for the in- creasing difficulties of the exchange of methods and ideas in biology, but we do think that the method has important values and_ possibilities, and that its cooperative adoption by other labora- tories that may be in a position to carry on some- thing of this sort, will be of considerable advan- tage to biological research. The program for this year, which is attached at the end of this “report,” indicates to what ex- tent the theoretical aspects of the procedure are being carried into practice. There is another theoretical advantage to be gained in the conference-symposia, the actual realization of which may be judged accurately only by those taking part. It is concerned with the further education of the “‘oldster” in biologi- cal research. During the last few years, especially, as biological research has become broader in its whole scope and narrower in the research scope of almost any given investigator, there has been a marked tendency for a biological investigator to become an “‘oldster’’ in a comparatively few years. Let him begin in almost any aspect of biological research, and in a few years it 1s un- believably difficult for him to go into another. He has continually gained in experience and knowl- edge in his own work. He has kept informed in detail of the published results of others working on a problem actually the same as, or very similar to, that upon which he is engaged. These facts alone have caused him tq fail to keep abreast of other advances in biology. Indeed, the rapid in- crease in knowledge in biology almost forces an investigator to boast that he knows something about such and such, but that !he is practically a layman when it comes to other parts of biology. Hence, the research, worker is likely to continue in what he can do well. If he has had the good foresight and fortune to choose a suitable pro- blem in the beginning, he may live many years and still make timely and valuable findings. It is likely, however, that after x years, but appre- ciably before he becomes senile, (if indeed he should ever become senile), his chosen pro- blem becomes an almost exhausted mine. He may not realize this, but his colleagues are quick to see that the rock which his buckets are bring- ing to the surface contains an ever decreasing percentage of gold. In other cases the miner may know that the rock from which he has taken the gold also con- tains silver, and possibly platinum, in paying quantities, but he has been so occupied in re- covering gold, that ha does not know the modern methods of obtaining either silver or platinum. And there are others who are experts in doing just this. The analogy breaks down, of course. Bioloyi- cal research is infinitely more complicated than the extraction of gold, silver or platinum from the earth, but I am sure that readers of THE CoLLecTING Net will see the full implication of the analogy even beyond the point at which it breaks down. They will also see that conference- symposia give the “oldster” an additional oppor- tunity to become a youngster in respect to free- dom of motion in research, while he holds the very great advantage of his experience. The taking of such opportunities cannot fail to bene- fit biology enormously. Be HR (ae Ee I have indicated in several previous reports that the direction in which the all-year develop- ment of the laboratory is likely to proceed is to be found in the establishment of small labora- tories concerned with research in quantitative biology. It is our aim that these laboratories to- gether should eventually present a well-rounded attack on modern biological problems; that, while independent of each other, the investigators in charge should show the highest degree of co- operation one with another ; and that the work of these laboratories should offer a dignified and stimulating background to summer activities. It will also be recalled that since the establish- ment of our laboratory for biophysics, in 1928, under the leadership of Dr. Hugo Fricke, we have felt that the next desirable move was the establishment of a laboratory for general physi- ology. This step, however, was delayed by the coming and continuance of the depression, but even more, as present events show, by our wait- ing for a suitable man to take charge of the 8 THE COLLECTING NET f Vou. IX. No. 72 laboratory. Now that such a man is available, a laboratory for general physiology is being established. The man is Dr. Eric Ponder, pro- fessor of general physiology at New York Uni- versity. His past work and future plans, his previous and present cooperation in some of the work at the laboratory for biophysics, and the value of his work to the summer activities of the Laboratory, seem to be such that we have given up waiting for the end of the depression. He has been appointed to our all-year staff as investiga- tor in general physiology. He is already in resi- dence and conducting his work at the Laboratory. His appointment here takes effect September first. Dr. Ponder will be assisted by Mr, John MacLeod, who has been associated with him at New York University for some time. It is to be hoped that at some time in the fu- ture we may be able to establish other small lai- oratories for general physiology. No one man can possibly represent the whole of this part of biology. It now seems likely, however, that our next important need will be found in making available at Cold Spring Harbor a chemist who is deeply interested in biological applications of chemistry, or a biochemist. Here again, however, accomplishment will depend more upon the avail- ability of a suitable man than upon preconceived plans or programs. Meanwhile we are happy in the achievement of another step to which we have long looked forward. Our establishment of an all-year laboratory for general physiology is concurrent with marked ad- vance in our summer course in general physiolo- gy. Following a method successfully worked out at Woods Hole, we have this year appointed sev- eral additional instructors to the course. The in- structors are now Professors Ivon R. Taylor of Brown, Harold Abramson and Kenneth S. Cole of the College of Physicians and Surgeons, Pro- fessor Eric Ponder, Dr. Hugo Fricke, and Mr. Edward Walzl of Johns Hopkins. This adoption of the Woods Hole method in the course in general physiology resulted from conversation with the Director and with various other members of the Staff of the Marine Bio- logical Laboratory, from which it became appar- ent that not only was the method good, but more important, there seemed to be need for additional first class opportunities for students of general physiology. The enrollment in the course this year indicates that the advice was sound.t Indeed our experience over many years has clearly shown that very sound advice is obtainable from men primarily attached to other institutions. Our Board of Directors includes at least four 1The opportunities in general physiology at the Laboratory were further increased by a_ special grant of money for equipment from the Carnegie Corporation last year. members of the Board of Trustees of the Marine siological Laboratory, the Director of the Mount Dessert Laboratory, several members of the Board of the Bermuda Station, and so on; while our scientific advisory committee contains others whose connections are particularly close with marine laboratories other than that at Cold Spring Harbor. There is, probably, everything to gain, for biol- ogy, and nothing to lose, by the closest coopera- tion between the administrations of the several marine laboratories, especially those on the Atlan- tic, and it is to be hoped that advances, already achieved in this direction will be small compared to those of the near future. In this connection it is gratifying to note the gradual development of Tue CoLttectinc NET into a summer biological news organ, which even- tually, we hope, will serve regularly in each issue, all of the marine laboratories of the country. THe SUMMER PROGRAM In presenting the program for the conference- symposia at Cold Spring Harbor, I wish to stress the fact that biologists, particularly interested in a given paper, or group of papers, are invited to attend and to take part in the discussion. Living accommodations are so limited, however, that per- sons wishing to spend a night or more at the La- boratory should make arrangements in advance or plan to sleep in a hotel in Huntington, Long Is- land. Ina small number of cases, men wishing to dis- cuss a given paper, and being unable to be present when it is given, may obtain a copy of the manu- script from the Director of the Laboratory, with the understanding that the paper will be returned promptly, together with such discussion as the recipient may make. PROGRAM OF CONFERENCE-SYMPOSIA FOR 1934 (The program of each day begins at 10:20 A. M.) MONDAY, JULY 2 Huco Frreke: The Chemical-Physical Founda- tion of the Biological Activities of X-rays. W. T. Astpury!: X-ray Studies of Protein Structure. Ratpo W. G. Wycxorr: Ultra-violet Photog- raphy as a Means of Studying Crystal Growth and Cell Structure. TUESDAY, JULY 3 Orro RauN: The Physico-Chemical Basis of Biological Radiations. Oscar W. RICHARDS: Growth of Yeast. The Analysis of the 1 Paper to be read by Doctor Clark. June 30, 1934 ] DHE COLEECRING NET iy Tueo. L. Jaun: Problems of Growth in Protozoa. Population MONDAY, JULY 9 Hans MUuiier: The Structure of Liquids and Solids. GeEorGE L. CLARK: Growth. The ‘Principles of Crystal TUESDAY, JULY 10 Cuarces B. DAveNporT: Critique of Curves of Growth and of Relative Growth. Ortro RAHN: Building Materials of Cells. Fetrx BERNSTEIN: Growth and Decay. MONDAY, JULY 16 GeorGe L. CLARK: The Effects of X-radiation Upon Cell Growth and’ Structure. CuHarLes PackARD: Biological Dosimeters in Radiology. TUESDAY, JULY 17 F.S. Hammett: Natural Chemical Growth and Development. VOEGTLIN: Observations Concerning Chemistry of Cell Growth and Division. . W. CHALKLEY: (Supplementary and formal- ly prepared discussion of the above). GUDERNATSCH: Specific Chemical Factors Influencing Vertebrate Growth and Differen- tiation. Factors in the lO MONDAY, JULY 23 Nicuwotas Rasuevsky: Physico-Mathematical Aspects of Cellular Multiplication and Devel- opment. Ortro RAHN: The Growth Mechanism. TUESDAY, JULY 24 C. P. Winsor: Mathematical Growth of Mixed Population. L. G. LoncswortuH: Diffusion in Cell Models and Volume Changes Analogous to Growth. Analysis of FRIDAY, JULY 27 M. Demerec: The Gene in Relation to Growth and Development. CuarLtes R. Strockarp: Internal Constitution and Genic Factors in Growth Determination. J. W. Gowen: The Gene as a Factor in Pathol- ogy. SATURDAY, JULY 28 SewaALt Wricut: On the Genetics of Abnormal Growth in the Guinea Pig. MONDAY, JULY 30 GrorGE L. CLARK: New Facts Concerning the Growth and Structure of Plant Tissues with Special Reference to Cellulose. Victor C. Twitty: Growth Correlations in Amphibia Studied by the Method of Trans- plantation. TUESDAY, JULY 31 HaArotp C. Urey: Possible Uses of Heavy Water in the Study of Biological Phenomena. Orro RAHN: Chemistry of Death. A METHOD OF PREPARING SMALL INSECTS FOR PHOTOGRAPHING Dr. P. W. WHITING Carnegie Institution of Washington, Cold Spring Harbor, N. Y. An inexpensive and convenient device for studying, drawing, and photographing — small opaque objects by reflected light was described in Tue Cottectinc Net last summer (Whiting, Anna R., 1933, VIII (1):11. This device has been used for studies on the parasitic wasp, Hab- robracon, and is adaptable for work with other small insects. The specimens are mounted on a depression slide in water or alcohol to avoid the clearing action of such media as glycerine and balsam. Considerable difficulty has been experienced in holding the minute object in the position desired for photographing and at the same time obtaining a clear white background with nothing but the insect in view. Moreover, the wings being in part translucent are séen by transmitted light even more than by such light as is reflected from their own surface. It has been difficult to grade illu- mination and exposure in such a way as to show details of structure and pigmentation both in the wings and in other parts of the body surface that are seen by reflected light only. It is the purpose of this note to explain how these difficulties have been solved. A modification of Heidenhain’s Alum Haematoxylin method has been devised to make the wings less translucent without appreciably af- fecting the color of the remainder of the body. The specimen, killed and preserved in 95% alcohol, is transferred to aqueous iron alum (4% ) and left in this mordant for one-half hour or longer. It is then rinsed with tap water and dipped in aqueous haematoxylin (0.5%) for a few seconds. The minute hairs covering the wings take the stain very readily before it has had time to penetrate the body. Excess stain is removed by tap water. If staining has been too heavy, decolorization may be brought about by returning to iron alum for a few seconds and then restaining. It will be found that after preseryation in Iron 10 THE COLLECTING NET [ Vor. IX. No. 72 alcohol, the legs, antennae, and wings of the specimen are thrown into the most unnatural positions in which they are held by rigor mortis and back to which they will immediately spring as soon as pressure of needles or other instru- ments is released. In order to induce plasticity the specimen is placed in a drop of sugar syrup on a glass slip. Here it may be manipulated with needles into the desired position. A small piece of lens paper is then placed upon the drop of syrup and as it moistens and flattens toward the glass, the insect is compressed so that its various appendages are brought approximately into one plane. This aids subsequent focussing of the camera. The process of compression is watched under the binocular microscope in order that pressure with needles at various points may direct the appendages at will. The moistened lens paper becomes somewhat transparent but the slip may be reversed and the specimen observed more clearly through the glass. After the desired position of the insect is’ ob- tained, a small flat piece of cotton batting moistened with water is placed on the slip over the lens paper. A second glass slip is placed over the cotton with a small strip of wood (piece of a match) inserted between the two slips at each NOTES ON THE RESEARCH WORK AT THE CORNELL UNIVERSITY BIOLOGICAL STATION DR. J. G. NEEDHAM, Director Professor of Entomology and Limnology, Cornell University The work of the Cornell University Biological Station centers about fresh water animals as its location, and equipment and environment invite. Three major pieces of work nearing publication are as follows: Dr. Ting-wei Liu has completed a careful study of the head of dragonflies, in which he has traced the evolution of the enormous com- pound eyes and the corresponding reduction of the antennae. Everyone has noticed the concen- tric layers of pigment in the eyes of certain dragonfly nymphs. These mark increments of growth as do the rings in the wood of trees. Suc- cessive added increments corresponded to nym- phal instars. Miss Lillian Thomsen has completed a careful survey of the group of little biting flies known as punkies (Ceratopogoninae), worked out com- plete life histories of representatives of half a dozen genera, reared more than twenty-five local species from larva to adult, finding new genera and species hitherto unrecorded in local fauna and adding much to our knowledge of the habits and economy of the group. Apparently only the species of the genus Culicoides are human pests. Professor Needham and Drs. J. R. Traver and Yin-chi Hsu are completing a book on the biology of the mayflies, which will be the first compre- hensive treatise of this order of insects ever pub- end in order to avoid excessive pressure on the specimen. The two slips are then securely tied together and the whole immersed in water until the sugar is completely removed. The water is removed by transferring to alcohol in which, the insect hardens in the desired position. After several hours‘of hardening, the covering slip, cotton, and lens paper are removed and the specimen, which remains in approximately the position in which it was held, is floated off in alcohol into a Syracuse dish. It is then placed in alcohol on a carefully cleansed depression slide, covered with a cover glass and secured against drying or bubbles with a ring of vaseline. For photographing, the slide is placed on the stage of the microscope previously described, ex- cept that the glass part of the stage is removed in order to avoid particles of dust and to allow a small piece of white paper placed some distance below the specimen to furnish the clear white background. ; A deep focus flat field lens combined with a long bellows permits a considerable range of magnifications. Panchromatic films or plates are used in order that the yellow may be distinguished from the brown or black regions of the body. lished. The systematic part of it will be an illustrated account of the mayfly fauna of the United States and Canada, covering more than four hundred species. ; BOOK REVIEW A German Reader for Biology Students, Fiedler, H. G., and G. R. de Beer, v + 92 pp. 1933. London. $1.50. At last there is a German reader of, by and for biologists with one to five page excerpts from articles on the following subjects: the vertebrate body, the heart, plant life, action in the small intestine, sense organs, hormones, regulation of temperature and of respiration, regeneration, mitosis, heredity, microsurgical technique, the organizer, tissue culture, sex determination, Berg- mann’s law, tumor metabolism, paleontology, ex- periments with apes and other infrahuman pri- mates and the crisis in biology. The articles were written by: Hempelmann, Ubrich, Jordan, v. Bruddenbrock, vy. Buticke, Korschelt, Hertwig, Goldschmidt, Baur, Spehmann, Durken, Erd- mann, Hesse, Warburg, Seitz and Gothan, and Bertalanffy. With the exception of the article on regeneration they represent the biology of the last decade. This is certainly multwm in parvo. One might wish that the book was a little longer and the excerpts were more complete, but per- haps this brevity will whet the student’s appetite and tempt him to read all of the originals. It reminds one of the Reader’s Digest with the ad- dition of a 3500 word vocabulary. Oscar W. Richards. June 30, 1934 } THE COLLECTING NET 11 News from the Scripps Institution of Oceanography FORMER SCRIPPS INSTITUTION WORKER TO LEAD HIMALAYAN EXPEDITION A scientific expedition to the high mountain country north of Kashmir, India, planned for the spring and summer of 1935, is to be led by Dr. Ancel B. Keys, now of the Harvard University Fatigue Laboratory, formerly research assistant at the Scripps Institution of Oceanography. After taking his doctor’s degree at the Scripps Institu- tion Dr. Keys studied at the Universities of Co- penhagen and Cambridge, both of which institu- tions are cooperating in the expedition to the Himalayas. The purpose of the expedition is to study body changes of man and animals in accli- matization to high altitudes, and observations will be made at various altitudes from sea level to heights greater than 20,000 feet, the base camp to be located at about 17,500 feet. The studies to be carried out are a continuation of and in line with the studies on oxygen consumption by fishes which Dr. Keys made while working under the supervision of Prof. F. B. Sumner at the Scripps Institution. Dr. W. R. Gregg, the head of the United States Weather Bureau, accompanied by Dean Blake of San Diego, recently visited the Scripps Institution to consult with, Messrs. T. W. Vaughan and G. F. McEwen on meteorological work and statistical methods for use in the study of air conditions and long-range weather forecasting. Messrs. N. B. Scofield and W. L. Scofield, of the California Fish and Game Commission, were visitors at the Scripps Institution. Mr. N. B. Scofield is a member of the Advisory Board of the Institution. Dr. and Mrs. L. G. Carpenter of Denver, Col- orado, and Colonel and Mrs. F. R. Payne of San Diego were visitors at the Scripps Institution on Tuesday of last week. Dr. Carpenter, a dis- tinguished irrigation engineer, was especially in- terested in the work in dynamical oceanography of which Professor McEwen has charge. Dr. Gertrude Smith and Miss J. F. L. Hart, of the department of zoology at the University of British Columbia, are visitors at the Scripps In- stitution. Miss Hart has also been connected with the Pacific Biological Station at Nanaimo, B. C., which cooperates in oceanographical work with the Scripps Institution. On Tuesday evening the Engineers’ Club of San Diego was addressed by Dr. Francis P. Shepard, of the Department of Geology of the University of Chicago, on “Coastal and Subma- rine Topography in Southern California,” on the study of which Dr. Shepard thas spent many months during the past year. He has used the Scripps Institution as the base for much of his work, and has got very interesting results in his study of the submerged canyons just off the sea front in the immediate vicinity of La Jolla and the Scripps Institution. TYPE SLIDE COLLECTION FOR THE M. B. L. MUSEUM The M. B. L. has established a museum for displaying the flora and fauna of the region. In view of the large amount of worl of a cytologic nature, including that done on microscopic forms, and because of the large number of biologists who at one time or another come to Woods Hole, it would seem as if the museum might also in- clude a slide collection. Most workers make more slides of an organism or of a cytologic demonstration than they usually need. Event- ually their slides become lost or donated to some university department and it is either “requiescat in pacem” or they are accessible to but few workers. If, however, a well catalogued collec- tion of type forms and subjects accumulated in the M. B. L. museum, whence interested workers could obtain particular slides of a new micro- organism or slides demonstrating certain aspects of centriole studies, ietc., valuable comparisons could be made, and perhaps at times contro- versies avoided. This would mean some addi- tional work for the curator, Mr, Gray, but he is favorable to the idea. —L, B. J. Secretary of Agriculture Henry A. Wallace and his party were visitors at the Scripps Institu- tion early in June. Secretary Wallace consulted with Professor McEwen concerning long-range weather forecasting and the application of statis- tical methods to agricultural problems. Dr. T. Wayland Vaughan recently gave an il- lustrated lecture before Sigma Xi at the Univer- sity of California at Los Angeles on “Recent ad- vance in the study of the oceanography of the Pacific.” At the end of May Dr. W. H. Osgood, curator of zoology of the Field Museum of Natural History at Chicago, and Mr. Adriaan van Ros- sem, in charge of the ornithological collections at California Institute of Technology, were visitors at the Scripps Institution. Miss Helen Matthews, on the faculty of bac- teriology at the University of British Columbia, has arrived at Scripps Institution to spend the summer months working in the bacteriological laboratory with Dr. C, E, ZoBell, 12 THE COLLECTING NET [ Vor. IX. No. 72 Courses at the Marine Biological Laboratory THE PHYSIOLOGY COURSE Dr. W. R. AMBERSON Director of the Course; Professor of Physiology, University of Tennessee A number of changes in the staff of the physio- logy course have made possible the introduction ot new lecture and laboratory work. Dr. Gerard and Dr. Sumwalt are on leave of absence for this summer. Dr. Lawrence Chute has taken over the micro-respirometer work which Dr. Gerard has been giving for the past several years. Dr. Balduin Lucké has joined the staff on a part time basis and is offering work on the permeabhil- ity of cells to salts and water. He is also intro- ducing for the first time a section for the study of phagocytosis. Dr. Robert Chambers has joined the staff on a full time basis, and with the assistance of Dr. Sichel is offering work on micro-dissection and injection. We are fortunate in having with us, on a part- time basis, Dr. Rudolf Hober, formerly Profes- sor of Physiology at the University of Kiel who has consented to give a laboratory section on the perfusion of living tissues, particularly kidney and liver. He is also lecturing on the general field of permeability. Laboratory sections are again offered by Dr. Laurence Irving covering studies of acid-base equilibrium in sea water, and the buffering capacity of tissues. Dr. Leonor Michaelis is giving his usual laboratory introduction to oxiaa- tion-reduction potential studies. Dr. Wauiliam Amberson is again giving work on electrokinetic phenomena, and on action and injury potentials. The class has had an unusually large number of applicants and has taken its full quota of twenty-two students. As has been the custom for several years students have been allowed to elect their laboratory work from the list of ten different projects which are offered in the pres- ent summer, and individual laboratory schedules have been prepared, in which each student has been given his or her first, second and third choices. In the usual case each student will do no more than five of the ten projects offered, during the first four weeks of the course. We are attempting to instruct our students in rela- tively few problems and techniques, with the hope that they may then have a better basis for ad- vancing, in the last two weeks of the course, to a research problem suggested by the earlier work. The course opened with two days devoted to a series of lectures and demonstrations intro- ducing all laboratory projects to the whole class. Meetings of the whole class for the presentation and discussion of results are planned. THE EMBRYOLOGY COURSE Dr. H. B. GoopricH Director of the Course; Professor of Biology, Wesleyan University The chief aims of the embryology course are to provide facilities for the study of living eges and embroyos and to outline various fields of research in embryology. A wide selection of forms from the Woods Hole fauna are studied illustrating many different modes of development. Almost all observations are made on living ma- terial. It is always possible to demonstrate the maturation of the egg, fertilization and a variety of types of cleavage. Later larval stages such as the trochophore, veliger, bipinnaria, pluteus and tornaria larvae are usually available. An intro- duction to experimental methods is given on favorable material. Topics that have been pre- viously presented are artificial parthenogenesis, duration of ~fertility of sperm, fertilization reactions, separation of blastomeres, effects of various environmental factors, etc. In addition to lectures by the staff, various in- vestigators have given accounts of their own - work. Last summer such lectures were given by, Dr. Charles Stockard, Dr. C. C. Speidel, Dr. Josef Spek, Dr. E. L. Clark, Dr. Robert Cham- bers and Dr. Oscar Schotté and Dr. E. G. Conklin. This summer the section on the Echinoderms previously directed by Dr. Hoadley of Harvard will be given by Dr. Schotté of Yale. The tentative schedule is as follows: June 20-25 Embryology of fish; Forms studied ; Fundulus, Tautogolabrus, Opsanus; June 26-27 Embryology of Cephalopoda; Loligo, June 28- 30 Embryology of Coelenterata. June 30, 1934 | THE COLLECTING NET 13 July 2, 3 Fertilization and early cleavage of Annelida and Mollusca Nereis, Chaetopterus, Crepidula; July 6-11 Later stages of Annelida, Mollusca and Polyzoa, Hydroides, Nereis, Cum- mingia, Chiton, Bugula; July 13-16 Embryology of Echinodermata ; Asterias, Arbacia, Echinarach- inus; July .17-19 Experimental problems on Echinoderms ; July 20 Embryology of Coelenter- ata; July 20-23 Embryology of Tunicata Styela, Molgula, Botryllus; July 24-26 Embryology of Crustacea; Libinia; Ju/y 27 Larval stages from towing. MORPHOLOGY AND TAXONOMY OF THE ALGAE Dr. WittiAm RANDOLPH TAYLOR Director of the Course; Professor of Botany, University of Michigan This course is designed to acquaint the student with both fresh water and marine algae. The approach is essentially morphological, and the forms are studied in as close to the currently ac- cepted systematic order as possible, to elucidate taxonomic features. Advanced taxonomic pro- blems cannot be approached during the course, since too wide an acquaintance with the literature is necessary, but the elements of classification and an evaluation of the criteria involved in algal taxonomy are presented. The morphological studies offer the common types ordinarily pre- sented in general comparative morphology courses, but with more adequate material than is generally available, and continue to more numer- ous, less familiar species which lead to a more complete conteption of algal structure and evolution. Emphasis is laid upon the use of living material. While the local flora determines what' species can be studied in a living state, a few important Pacific and tropical genera are: discussed from tropical material. The ecological aspects if the local flora are observed in conjunction with the collection of material in the field. Trips are taken to locali- ties of peculiar richness, and critically selected material is brought back for detailed laboratory study. As nearby stations have been rendered unavailable by the encroachment of developed shore estates, it has become increasingly necessary to take trips farther away where the plants are still in a natural luxuriance, but it continues practicable thus to study the flora in the field. Every encouragement is given for the prosecu- tion of research studies for which students are already trained, and so far as possible the atten- tion of less matured students is directed toward suitable fields for specialization and research, so that they may continue their botanical, advance- ment using algae as material for morphological, physiological or taxonomic studies. THE COURSE IN PROTOZOOLOGY Dr. GARY Director of the Course, Professor of The course in protozoology beginning this year on June 20, is designed for graduate students wishing to begin research in protozoology or in other fields requiring a knowledge of unicellular animals, free-living or parasitic. The lectures given daily except Saturday cover the principles of general biology; the morphology of the different classes of protozoa; the parasitic protozoa and the diseases caused by them ; experi- mental protozoology and the theories of growth and cell division, of fertilization, of age and re- organization, of heredity and variation. The laboratory work gives an opportunity for the collection of material from natural sources; the study and identification of living organisms representing the main groups of the protozoa N. CALKINS Protosoology, Columbia University (seventy-five drawings) classified through genera, including ten classified through species, are re- quired) ; the cultivation of protozoa in artificial media; the determination of the pH of media and of water collected from ponds around Woods Hole. Opportunity is also given for special cytological studies of the protozoa. For this purpose, perm- anent preparations (ten) are required. In ad- dition to the ordinary techniques, osmic and silver impregnation, the Feulgen nucleal reaction, and the chondriosome methods are employed. Fresh material stained by vital dyes is used to supplement fixed material when the identification of cytoplasmic elements is desired. 14 THE COLLECTING NET [ Vor. 1X. No. 72 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel W. Parker, and An- naleida van’t Hoff Snyder Cattell. Printed by The Darwin Press, New Bedford THE COLLECTING NET SCHOLARSHIP FUND One of the primary reasons for the establish- ment of THE CoLLectiNG Net when it began publication in 1926, was to initiate a scholarship fund to assist serious students of biology in meeting their expenses at the Marine Biological Laboratory. Each summer at Woods Hole there is a group of young biologists who must during the summer earn part or all of the money that is needed to carry them through their period of work at the Laboratory. The most usual method is to wait on tables at the Laboratory mess hall. In return for their meals the students work approximately four hours. If half of this regimented time could be devoted to laboratory work and the other half to recreation, the research productivity of the student would be much greater; if the number of young investigators who must “work for their meals” is cut down, not only will it directly bene- fit the individuals concerned but also be of serv- ice to the Marine Biological Laboratory and to science. From time to time individuals and groups con- tribute to the Scholarship Fund and it would seem appropriate to have a custodian for the money other than THE CotLectinc Net. To carry out this purpose THE CoLLectinc Net Scholarship Fund Association will be initiated. The membership fee has been set at $5.00. The organ of the Association will be THE COLLECTING Net and members will receive it as part of their membership. It probably will be desirable to allow one dollar of each membership fee to cover the cost of supplying the magazine to the member and for other incidental expenses. Individuals, be they biologists, summer or year- round residents of Woods Hole are invited to support the scholarships by joining the Associa- tion. A membership of 100 would provide four $100.00 scholarships; 250 members would result in $1,000 for the Scholarship Fund. Tue CottectineG Net scholarships are beyond the experimental stage ; their worth is significantly demonstrated by the letter from Miss Lucas of the University of London which is reproduced on the front cover. The first scholarships were three in number and awarded in 1928; the re- cipients were: Miss Catherine L. T. Lucas, Mr. A. K. Parpart and Mr. K. F. M. Sichel. The Scholarship Award Committee for that year con- sisted of Professors Robert Chambers, Edwin G. Conklin and Lorande Loss Woodruff. MANUAL OF METHODS IN MARINE BIOLOGY We hope to have the privilege of printing in THE CoLLectinG Nev a series of articles telling of the marine animals available for, experimental work at Woods Hole, giving the accepted me- thods of handling the animals and their eggs. The plan is to begin with the most used forms and present the material for individuals who are unaccustomed to working with them. The type set for the articles in THe CoLLect- 1NG Net will be used later to print a small manual of methods in marine biology. This dual use of the type will make it possible to sell the volume at a relatively low price. MB. L. Calendar TUESDAY, JULY 3, 8:00 P. M. Seminar: Dr. Mary Collett: Ovarian Hormones and Basal Metabolism. Dr. Laurence Irving and Mrs. Mary Scott Welch: The Carbon Dioxide Tension in the Brain. Dr. E. S. Nasset: A Method for the Estima- tion of Changes in the Rate of Intestinal Secretion. . Dr. G. H. Parker: Transfer of Neurohumors. CURRENTS IN THE HOLE At the following hours (Vaylight Saving Time) the current in the Hole turns to run from Buzzards Bay to Vineyard Sound: Date A.M. P.M. dimes BY) ooaocd 7:07 @:45 dhl Wl oeoeoac 8:19 8:42 Atty 2 oS oc au 9:13 9:41 Ul goog cas 10:06 10:40 Trulli S40 ee serns ters 11:02 11:43 Julywesncensee LBS See Jw; Googounce 12:46 12:55 In each case the current changes approxi- mately six hours later and runs from the Sound to the Bay. It must be remembered that the schedule printed above is altered somewhat by wind conditions. Prolonged winds sometimes cause the turning of the cur- rent to occur a half an hour earlier or later than the times given above. The average maxi- rum gpeed of the current is five knots per hour. June 30, 1934 ] THE COLLECTING NET 15 PPE MS 7 Or The Genetics Society of America will hold a meeting at the Marine Biological Laboratory late in August. It is hoped that this will serve to bring to Woods Hole a number of members of the Society who have not previously visited the Laboratory. While the meeting will be largely informal, there will be one or more sessions for the reading of papers. The secretary, Dr. P. W. Whiting, will be pleased to receive the titles of papers to be presented as soon as they can be given to him. Dr. Thomas Hunt Morgan of the California Institute of Technology has recently returned from Stockholm where he received the Nobel Prize in medicine for 1933. Dr. Morgan also re- ceived the degree of Doctor of Science from Yale University in June, 1934. Dr. Oscar Schotté, research fellow in biology at Yale University, has been appointed assistant professor of biology at Amherst College for the approaching academic year. Dr. Schotté will give some of the lectures in the embryology course at the Marine Biological Laboratory dur- ing the present summer. Dr. S. O. Mast, professor of zoology at Johns Hopkins University, is giving a course in proto- zoan physiology at the Stone Laboratory, Put- In-Bay, Ohio. He and his family will not be at Weeds Hole this summer, except for Miss Louise Mast who is working in the Chemical Room of The Marine Biological Laboratory. Dr. Oscar W. Richards is leaving on July Ist for the Tortugas Laboratory. He will stop on the way to give a paper on yeast growth at the Bio- logical Laboratory at Cold Spring Harbor. Dr. Edwin P. Laug will be in charge of the Chemical Room until Dr. Richards returns to Woods Hole late in August. At its annual meeting the National Academy of Sciences awarded the Elliot Medal and Honorar- ium of $200 for 1930 to Dr. George Ellett Cog- hill, professor of comparative anatomy and member of the Wistar Institute of Anatomy and Biology, in recognition of his work “Correlated Anatomical and Physiological Studies of the Growth of the Nervous System in Amphibia.” Dr. Coghill worked in Woods Hole last summer. DNFVE Rees 7 Dr. Henry B. Bigelow, director of the Woods Hole Oceanographic Institution made the pre- sentation address for the National Academy of Sciences when the Agassiz Medal was awarded to Bjorn Helland-Hansen of the Geofysiske Insti- tut, Bergen, Norway. Mr. Helland-Hansen re- ceived the award in recognition of his work in physical oceanography, and especially for his con- tributions to knowledge of the dynamic circula- tion of the ocean. Dr. Bigelow will be at Woods Hole after July 4th. Dr. George S. deRenyi of the University of Pennsylvania Medical School, is spending the first part of the summer in the Tortugas Labor- atory. He plans to come to Woods Hole in August. Dr. George Baitsell and his family are visiting relatives in Illinois during the early part of this summer. They plan to visit Woods Hole in September. Miss Peggy Woodruff, daughter of Dr. L. L. Woodruff, is teaching at Hampden Hall, a private school in Connecticut. She is living with her family in their new home in the ‘“Gansett Woods.” The marriage of Dr. William C. Young, in- vestigator at Woods Hole for several years and assistant professor of biology at Brown Univer- sity, to Miss Ruth A. Hobby took place in Fair- haven, on June 19. Dr. and Mrs. Young are spending the summer in Honolulu. M. B. L. CLUB The M. B. L. Club held its annual meeting to elect new: officers on June 27 at 7:00 in the eve- ning. The new officers were: President, Dr. Robert Stabler; (Vice President, uncontitution- ally not elected!) Secretary-Treasurer, Dr. Fer- dinand Sichel; Assistant Secretary-Treasurer, Miss Rachel W. Parker. ; After the elections, a review of last summer’s activities of the Club and plans for the present summer weré presented. Suggestions to minimize the cost of the Saturday night dances without de- tracting from their entertainment were offered. Also, it was definitely decided that the Club no longer wanted to stand the expense of mending and caring for the raft. The offer of the Supply Department, to give the Club the use of the Whale Boat, in return for keeping the boat in good condition, was unanimously accepted and ereatly appreciated. The meeting adjourned at eight o'clock. 16 THE COLLECTING NET [ Vor. IXeiNaw72 MEMS: OF Introducing Dr. Rudolf Héber, who came to the United States from his home in Stettin, Germany, in April of this year, is now visiting professor in the Department of Physiology at the Univer- sity of Pennsylvania Medical School. A new- comer at the Woods Hole laboratory, he expresses his great pleasure. in having the opportunity to meet personally the men with whom he has been working through correspondence for many years. He deciares that he is “indebted to more hiolog- ists, particularly cellular physiologists, in the United States than in any other place in the world.” Those who meet Dr. Hober are im- mediately impressed by his gracious personality and charming friendliness. He began his scientific work in Zurich, Switzer- land in 1897 as assistant in physiology at the in- stitute of Zurich, where he remained until 1909 as ‘privatdozent.’ In 1909 he went to the Uni- versity of Kiel, Germany, and became its Director from 1915 until 1933. He is lecturing in the physiology course at the Marine Biological Labor- atory this summer, on’ the physico-chemical basis of permeability. Dr. Hober’s primary work over a long period of years has been in applying physical chemistry to physiology. He is interested in the permea- bility of cells and tissues, and is working at pres- ent with the egg membranes of the sea urchin, and the kidneys of fishes. He is assisted by his wife, Dr. Josephine Héber, with whom he has published several papers, and by his daughter who is a medical student at the University of Penn- sylvania. Mrs. Hoéber has been in active medical practice and has worked especially in public hygiene. Dr. Héber has published several books and several hundred articles, many of which have ap- peared in Pfluger’s Archiv fur die Gesamte Physiologie des Menschen und der Thiere. In 1902 appeared the first edition of Physikalische- chemie der Zelle und der Gewebe, published by Engelmann, Leipzig. The sixth edition was pub- lished in 1926. Late in 1934 the seventh edition of Dr. Hober’s Lehrkuch der Physiologie des Menschen will be published by Julius Springer, Berlin, The Penzance Players this summer are plan- ning to present a gay 1890 melodrama entitled “Gold in the Hills or The Dead Sister’s Secret” by Frank Davis. It will be ready for production some time in July. Later on in the season an- other play will be given. INTEREST THE WOODS HOLE CHORAL CLUB The Woods Hole Choral Club will begin prac- tice next week under the leadership of Prof. Ivan Gorokhoff of Smith College. During the past seven summers the Club has sung English, Russian, and German folk songs, excerpts from Gilbert and Sullivan's operas, and church music of Bach, Handel, Tschaikowsky, Kastalsky, Gretchaninoff, and others. In August a public performance is given in the Auditorium. All who like to sing good music under able leadership are invited to join the Club. This in- vitation is extended not only to students and investigators who expect to be in Woods Hole in August but also to members of the summer colony, and to residents of Woods Hole and the vicinity. It is a community chorus, not restricted to workers in the laboratory. Announcement of the time of the first rehearsal will be posted on the Bulletin Board in the Mess and the Main Building. THE MIXER AND THE M. B. L. CLUB On the evening of Saturday, June 30 at 8:00 P. M. the M. B. L. Club invites everyone con- nected with the scientific institutions of Woods Hole to the annual mixer. It is hoped that every- body will take this opportunity to become ac- quainted with their fellow workers. The M. B. L. Club has served for twenty years as a social meeting place. It was originally a yacht club, but in 1914 the present social organi- zation was created, and it has received the enthu- siastic support of the summer workers since that time. One of the main objects of the club is to fur- nish an attractive club house, with facilities for relaxation and informal entertainment. It main- tains a small lending library, and a good selection of magazines. The ping-pong table on the second floor furnishes sport for many enthusiasts, and a tournament was held last summer of which Dr. T. T. Chen was the winner. Opportunities are also offered for bridge and other games. Every Saturday night the club holds an infor- mal dance, which in past years has been one of its most pleasant and well-attended activities. The membership fee for the season is $1.50, payable to Miss Crowell in the laboratory office, and everyone not already a member is urged to join. Ropert CHAMBERS, President June 30, 1934 ] THE COLLECTING NET 17 Chemical Room of the Marine Biological Laboratory 1934 Hours: 8:30 A. M.—12; The Chemical Room supplies chemicals, glass- ware, clamps and support stands for use only at the Marine Biological Laboratory. Special ap- paratus, batteries, tools, photographic trays, bolt- ing cloth, gauges and reducing valves for gas cylinders are issued at the Apparatus Room (Brick Bldg. room 216). The Supply Depart- ment (Frame Bldg. back of Brick Bldg.) sells stationery, cheese cloth and supplies that may be used by investigators elsewhere, such as micro- scope slides, cover glasses, shell Catalogs of chemicals and apparatus may be bor- rowed from the Apparatus Room. vials, ete. Members of classes are not entitled to supplies other than those provided in their regular class work. Beginning investigators will receive sup- plies only on the authorization of the person under whom they are working for the season. Certain common tools are available at the Chemical Room for temporary loan to investiga- tors and it is necessary that they be returned within 24 hours. When needed by other investi- gators they are subject to recall and will then be collected by the janitors. Tools required by an investigator for the entire season are to be obtained from the Apparatus room. The following standardized solutions will be furnished in limited quantities during the season of 1934, Special solutions, buffers, and pH stand- ards must be ordered at least two days before they are needed. N 1.000: Acetic acid Sulphuric acid Hydrochloric acid Sodium Hydroxide N 0.100: : Hydrochloric acid Sodium hydroxide Buffer mixtures: Acetate pH3.6-5.6 Phosphate pH 5.4-8.0 Phosphate-citrate pH 2.2-8.0 (Mcllvaine) Borate pH 7.6-10.0 Indicators—Clark and Lubs series. Color tube standards—on special order. 1 :30—4:30 P. M. Sat. 8 :30—12. For other standards inquire of the person in charge at the Chemical Room. Investigators ex- pecting to use special solutions or standardized reagents after September 1 are requested to notify the Chemical Room, if possible, before August 15. The standardized reagents are not usually available before June 20 or after September 15. Carbon dioxide, hydrogen, nitrogen and oxygen must be ordered by the investigator from the person in charge at least ten days before they are needed. Unless special instructions are received orders from investigators will be filled in accordance with the Formulae and Methods published by the Chemical Room in Tue Cottectine Ner (1930, 1932) which also include stain and chemical solubilities and the composition of solutions. Copies may be purchased at THE COLLECTING NET office. Supplies no longer needed will be collected if word 1s left at the Chemical Room. Investigators are urged to co-operate with the Chemical Room by cleaning their glass-ware be- fore returning it at the completion of their work and by placing their name and date of departure on the Chemical Room Bulletin Board so that their supplies may be returned promptly by the janitors. When the investigator is continuing the same work in the same room during the next season his supplies may be retained in the room only if they are listed on a Kept Out card (furnished at the Chemical Room window) and the card left with the supplies. All supplies not so listed will be returned by the janitors. Should the in- vestigator be unable to return the following sum- mer the supplies will be returned to the Chemical Room stocks if they or the room is needed by other investigators. Small amounts of special solutions will be kept during the winter for investigators in the Chem- ical Room on request. Supplies that may be in- jured by freezing should not be left in the wooden buildings. —Oscar W. RICHARDS. [ Vor. IX. No. 72 COLLECTING NET THE i Sees kien Q : Qn a a9 Se Ses inz he 7 ABdI2ELNYWN HLUON YABL | os = BQ ANIA SWHLUWW anw !'7u7 : &S / auocigad Man QL novi #9 ath ey LVOODWYALS y Ts > ; ; 2 NoWwis avoutiva ‘I ASSWE vat / ¢ x if: NOINA NX3Lis3am oy : as ef) NIGNWT NMOL a qd / NOItLALIISNI IHAdWYDONVIDO BI9OH SQOOM AWOLVYOSBWI 1v9ID07018 0 | e Z s h 4y 72 AnI>D s109 Vi ro) ] oO y | 3790H $aoo US | } ornmssiwwod HsIL S90 ayno vLSYOD NS NIONYA NMOL ony Wusva 217E9nd 2!NIONWIT NMOL June 30, 1934 ] ____THE COLLECTING NET Ovhatl Does SP ENGs Sek le Hean Co ou? sh iat hs @ When you purchase a SPENCER microscope, microtome or other scientific apparatus—you purchase service! R E @ You expect lasting satisfaction—you expect the manufacturer to not only give you a perfect piece of apparatus but also to retain his interest in this apparatus to see that it continues to func- tion to your permanent satisfaction. @ The name of SPENCER on a scientific product is your guarantee of the best material, fine workmanship, correct scientific design together with continued service. @ t inch offices are conveniently located thruout a VISIT the United States. The home office and factory are strategically located at Buffalo within over- OUR EXHIBIT night shipping distance of 80% of the nation’s From July 2 thru scientific men and thus prompt service is assured. oy, pea es across the _ street BRANCHES: New York, Chicago, San Francisco, from the Oceanogra- Boston, Washington. phic Laboratory: iR, | PRODUCTS: Microscopes, Microtomes, Delinea- scopes, Visual Aids, Optical Meas- uring Instruments. | i @ / BUFFALO NEW YORK Micrometric Controls are sturdy yet delicate Use them when bubbling gases through solutions; maintaining special at- mospheres; in pH work; when filling small tanks from large ones; easy to adjust the flow. Different inlets for dif- ferent gases. Price $5.00. Ask for Folder MC de- scribing this and other Hoke controls. Albany St., New York City Hoke Inc. 22 BLAKISTON’S CALL ATTENTION TO RAHN—Physiology of Bacteria 42 Illustrations. 438 Pages. Cloth $6.00. By Prof. Otto Rahn, Cornell University. It co-ordinates the various simplest functions of life, studies each in itself and its effect upon others. DIBLE—Recent Advances in Bacteriology and Study of the Infections. 2d Edition. 29 Illustrations. 476 Pages. Cloth $3.50. By J. H. Dible, F.R.C.P. Prof. Univ. of Liver- pool. The author takes a broad view of the progress keeping a balance between extreme technicality and what is already common knowledge. FLEMING & PETRIE — Recent Advances in Vaccine and Serum Therapy. 5 Illustrations. 463 Pages. Cloth $4.00. By Prof. Alexander Fleming, Univ. of London; and Dr. G. F. Petrie, Lister Institute, Elstree. It records progress in technical methods, a study of specific vaccines. Plant and animal virus diseases. STITT — Practical Bacteriology, Blood-Work and Animal Parasitology. 8th Edition. 211 Illustrations. 8837 Pages. Cloth $6.00. By E. R. Stitt, Sc. D. Former Sur. Gen. U.S.N. It includes keys, tables and clinical notes. P. BLAKISTON’S SON & CO., INC. 1012 EE UN CLUE LL OUD EOESTATIS | STREET PHILADELPHIA 5 do) ov 5, RRO? we MICRO mace COVER GLASSES "i DO NOT FOG », Ask your dealer —or write “Giving dealers name) to ‘Ctay- ADAMS ll WN 25 Eos! 26th Street NEW YORK THE COLLECTING NET _ [ Vou. IX. No. 72 Summer Supplies for the Naturalist COLLECTING NETS KILLING JARS INSECT PINS BALSAWOOD SHEET CORK SPREADING BOARDS DISPLAY CASES VASCULA PLANT PRESSES MOUNTING SHEETS PLANT DRIERS INSTRUMENTS A complete line of high grade collecting and mounting equipment. Illustrated catalogs sent on request. ; GENERAL BIOLOGICAL SUPPLY HOUSE Incorporated 761-763 EAST SIXTY-NINTH PLACE CHICAGO ECOLOGY All Forms of Life in Relation to Environment Established 1920. Quarterly. Official Publication of the Ecological Society of America. Subscription, $4 a year for complete volumes (Jan. to Dec.) Parts of volumes at the single number rate. Back volumes, as available, $5 each. Single numbers, $1.25 post free. Foreign postage: 20 cents. GENETICS A Periodical Record of Investigations bearing on Heredity and Variation Established 1916.. Bimonthly. Subscription, $6 a year for complete volumes (Jan. to Dec.) Parts of volumes at the single number rate. Single numbers, $1.25 post free. Back volumes, as available,$7.00 each. Foreign postage: 50 cents. AMERICAN JOURNAL OF BOTANY Devoted to All Branches of Botanical Science Established 1914. Monthly, except August and September. Official Publication of the Botanical Seciety of America. Subscription, $7 a year for compl] volumes (Jan. to Dec.) Parts of volume at the single number rate. Volumes 1-20 complete, as available, $162. Single numbers, $1.00 each, post free. Prices of odd volumes on request. Foreign postage: 40 cents. BROOKLYN BOTANIC GARDEN MEMOIRS Volume I: 33 contributions by various authors on genetics, pathology, mycology, physiology, ecology, plant geography, and systematic botany. Price, $3.50 plus postage. Volume II: The vegetation of Long Island. Part I The vegetation of Montauk, ete. By Norman Taylor. Pub. 1923. 108 pp. Price, $1.00: Vol. Ill: The vegetation of Mt. Desert Island, Maine, and its environment. By Barrington Moore and Norman Taylor. 151 pp., 27 text-figs., vegeta- tion map in colors. June 10, 1927. Price, $1.60. Orders should be placed with The Secretary, Brooklyn Botanic Garden, 1000 Washington Ave. Brooklyn, N. Y., U.S.A. June 30, 1934 ] THE COLLECTING NET NEW McGRAW-HILL BOOKS Principles of Animal Biology. New fourth edition. By A. FRANKLIN SHULL, Professor of Zodlogy, University of Michigan. With the collaboration of George A. LaRue, Prefessor of Zodlogy, and Alexander G. Ruth- ven, President, University of Michigan. McGraw-Hill Publications in the Zodlogical Sciences. 400 pages, 6x 9, illustrated. The basic aim of this successful text is to pre- sent a body of principles which may be brought under such topics as morphology, physiology, $3.50. ecology, taxonomy, geographic distribution, paleontology, and evolution. The book has been extensively rewritten, revised and simplified. Laboratory Directions in Principles of Animal Biology. New fourth edition. By PROFESSORS SHULL, LaRUE and RUTHVEN. McGraw-Hill Publications in the Zodlogical Sciences. As before, this carefully organized series of laboratory directions stresses the important generalizations of biology. The material has 100 pages, 6x9, $1.00. been brought into accord with present-day laboratory practice and conforms to the new fourth edition of the authors’ text. Introduction to Cytology. New third edition. By LESTER W. SHARP, Professor of Botany, Cornell University. McGraw-Hill Publications in the Agricultural and Botanical Sciences. $5.00.- The revision of this standard text dealing chiefly with the structural and genetical aspects of cytology adapts the material more closely to the needs of the student with comparatively Animal Biology 567 pages, 6x9, illustrated. little experience in the subject. New illustra- tions have been added and the bibliography, re- garded as one of the most important features of the text, has been brought up to date. By ROBERT H. WOLCOTT, late Professor of Zodlogy, University of Nebraska. McGraw-Hill Publications in the Zodlogical Sciences. This text for the beginning student consistently stresses principles and broad points of view. At the same time, the book is representative of the so-called ‘‘types’” course. Throughout, the 615 pages, 6x9, ill. $3.50. aim has been to give a simple, accurate, and interesting account of the whole field of zoology. Origins of the group names are given and pronunciation of words is included. The Fresh-water Algae of the United States By GILBERT M. SMITH, Professor of Botany, Stanford University. McGraw-Hill Publications in the Agricultural and Botanical Sciences. 716 pages, 6x49, ill. $6.00. Covers all fresh-water algae of the United States, including border-like organisms. Des- criptions of the various genera include a full account of morphology and reproduction, with extensive literature references. A feature is the single comprehensive key based entirely on vegetative characters. Keys to orders and the families are also included. Send for copies on approval McGRAW -HILL BOOK COMPANY, INC. 330 West 42nd Street, New York, N. Y. Aldwych House, London, W. C. 2 bo bo THE COLLECTING NET [ Vou. 1X. No. 72 Turtox News is a monthly bulletin, established in 1923 and devoted to notes and arti- cles on Laboratory Methods, Field Work, Microtechnic, New Teaching Developments, and announcements of new Turtox Biological items. It is mailed gratis to teachers in all parts of the world. If you are not receiving Turtox News regularly ask us to place your name on the mailing list. The Sign of the Turtox Pledges Absolute Satisfaction SCIENTIFIC PERIODICALS Biological, Medical, Zoological, Botanical, etc. Complete Sets, Volumes and Odd Copies. There may be some Single Copies needed to complete your Sets, or an Important Article which you may need. Prices are reasonable. B. LOGIN & SON, Inc. 29 East 21 Street New York City Books Reduced ls toy2-3 OFF Call at our Office on Water Street THE COLLECTING NET WOODS HOLE, MASS. REMOVAL NOTICE DONNELLY’S BARBER BEAUTY SHOP SHOPPE NOW OPPOSITE PUBLIC LIBRARY Telephone 211 Falmouth, Mass. now display. us to offer the following: Preserved Specimens Demonstration Preparations Dissection mounted in museum jar 30.00 Skeletons Complete semi-cartilaginous skeletons mounted in liquid in museum jar 35.00 sues are also available. plied by Turtox. AFRICAN LUNGFISH PROTOPTERUS AETHIOPICUS The much discussed, but seldom seen, lungfish is available for laboratory study and museum A recent importation from Africa enables Size 5 to 7% inches—each $9.50 Size 12 to 15 inches—each 15.00 Size over 20 inches—each 20.00 Intermediate sizes at corresponding prices. Mounted on glass plate in museum jar 25.00 Living specimens and microscopic slides of tis- Write for complete lists. This is just one of the many unusual forms sup- GENERAL BIOLOGICAL SUPPLY HOUSE Incorporated 761-763 EAST SIXTY-NINTH PLACE CHICAGO Catering to a Discriminating Clientele The BREAKWATER HOTEL Woops Hote, MAssACcHUSETTS DINING ROOM COCKTAIL ROOM June 30, 1934 } THE COLLECTING NET 23 Leitz Ultropak Illuminator The ideal illuminator for opaque objects of low reflecting power, such as many living organisms, surfaces on leaves or plants and many other biological objects. A complete series of objectives and condensers is available for observations from lowest to highest magnifications. Special Accessories enable the observer to eliminate surface glare from moist surfaces or to observe objects immersed in liquids. UAHA This equipment and many other new Leitz | Instruments and Accessories will be ex- | hibited during August, 1934 at: R. G. THOMPSON’S, MAIN STREET, WOODS HOLE, MASS. HN LITERATURE ABOUT ULTROPAK ILLUMINATOR UPON REQUEST Peel TZ, dine: Wet. CN: 60 EAST 10th STREET NEW YORK CITY Branches: WASHINGTON, D. C. SAN FRANCISCO, CALIF. CHICAGO, ILLINOIS. LOS ANGELES, CALIF. 24 THE COLLECTING NET f Vor. IX. No. 72 IMPORTANT NEW TEXTS 2220 General Zoology By FREDERICK H. KRECKER, Ohio University The simplest and most teachable text in zoology. Stresses the aspects of the animal world of interest to the intelligent layman. Discusses the typical animal, classifies the important animal groups, presents the animals in their environment, and studies the origin of animals. Written in a lucid and popular style and very fully illustrated. An excellent text for the general course. Ready tn June 22 Elements of Modern Biology By CHARLES R. PLUNKETT, New York University A completely rewritten and considerably abridged edition of the author’s OUT- LINES OF MODERN BIOLOGY. The difficult parts have been either wholly eliminated or greatly simplified. As a result the briefer book will be elementary enough to be used in the introductory courses and yet retain the excellence of the larger text. As it will not be ready till late this summer, we urge you to post- pone the adoption of a text until you have see this one. i Ready late in August 220 HENRY HOLT & CO. ONE PARK AVENUE NEW YORK TT : \ osesu oem c REFRIGERATED CENTRIFUGES ARE IMPORTANT IN THE FIELD OF RESEARCH BECAUSE: 1. Scope of work is increased. Results are more accurate than by chemi- cal method. Bacterial contamination is prevented. Greater yield from culture material. 5. Permits use of volatile denaturants. ref lig Many widely different fields of usefulness have been opened to scientific laboratories by com- bining refrigeration with the centrifuge. The International Refrigerated Centrifuge requires no new technic and occupies but little more space in the laboratory than the ordi- nary centrifuge. With the International Refrigerated Centrifuge it is possible to control the temperature of the material between 32° and 80° fahrenheit for indefinite periods. Descriptive Bulletin upon Request INTERNATIONAL EQUIPMENT CO. BOSTON, MASS. Makers of Fine Centrifuges 352 WESTERN AVE. — June 30,1934] THE COLLECTING NET VVVVVVVTVVvVvVTVVvVvVvVvVvvVvVvIFVvvVvvvws TRINH An > E. oxyuris> E. ehren- bergu> E. geniculata (?), E. acus> E. deses. These physiological differences are at least as great and in some cases greater than the morpho- logical differences between the species. Physiologeial diffrences in nutrition between species of Huglena have also been demonstrated by Dusi (1933). As a source of nitrogen Eug- lena piscifornus will utilize peptones but not amino acids or ammonium or nitrate salts. E. deses will use peptones and amino acids, but not ammonium or nitrate salts. E. anabaena will grow in amino acids, peptones, or ammonium salts, but not in nitrates while £. gracilis, E. stellata, and F. klebsii grew with any of the four compounds as a source of N but showed more rapid growth with the organic N compounds. Loefer (1934, 1934a), however, working with two species of Chlorogonium (C. elongatum and C. euchlorum) found that these two organisms were very similar as regards pH range, and utilization of amino and non-amino organic acids and carbohydrates, However, the growth rates of the two species dif- fered under fence conditions, C. euchlorum growing much more rapidly than C. elongatwi. 7. Tue Errects oF Oxmatrion-REDUCTION POTENTIAL There has accumulated a large amount of evi- dence in bacteriology that the growth rate of bacteria is affected by the oxidation-reduction potential of the medium. Since we have no rea- son for assuming that the metabolism of bacteria is unique, we might expect that a similar relation- ship should exist among the protozoa. At pres- ent there is evidence that the growth rate of Chilomonas is affected by the oxidation-reduction potential (Jahn, 1933a). In the experiments in which this effect was obtained the reduction 38 THE COLLECTING NET { Vor. IX. No. 73 potential was the result of the balance between various amount of —SH and H2Os, and con- sequently the data are open to the explanation that the highest growth rate is due to the optimal con- centration of reduced sulphur rather than to the proper oxidation-reduction potential. This point has not yet been decided, for the numerous fac- tors which must be controlled in order to perform a crucial experiment are not easily overcome. The components of the medium which may de- termine the reduction potential and affect the organism are perhaps as varied and probably more complex than the components which deter- mine the pH and its effect upon the organism. The reduced and oxidized forms of a compound in the medium may have as diverse an effect as the dissociated and non-dissociated phases of acetic acid. In sucha case the effect of the poten- tial would be only secondary, as in the effect of pH in slightly acid cultures containing acetate. ‘This seems to be a factor which might give rise to considerable confusion. 8. OxIDATION-REDUCTION POTENTIAL Vs. OxyGEN TENSION One important factor on which the reduction potential depends is the oxygen tension of the solution. It has been suggested by Knaysi and Dutky (1934) that in the case of Bacillus mega- therium oxygen tension is much more important as a limiting factor of growth than mere reduction potential. The energy of aerobic organisms is ordinarily obtained by the reduction of molecular oxygen; ina scarcity of oxygen an organism must depend upon the reduction of other substances in the medium. Since the energy yield of oxygen reduction is very much greater than the energy yield of the reduction of other eae) sub- strates which might be present (Gerard, 1932), we cannot conclude that oxidation-reduction potential is not important unless we control the oxygen consumption in our experiments. Con- trol of Oz consumption is a difficult undertaking, but it seems to be a necessary step before we can decide the relative merits of oxygen tension and reduction potential. With anaerobic organisms, such as the tetanus bacillus, the reduction poten- tial is generally admitted to be one of the most important factors affecting the growth rate. With aerobic organisms Oy tension seems to be more important as a limiting factor, probably because of the large amount of energy available from its reduction, but the relative importance of the re- ductien potential is still unsolved. Not only is this question unsolved, but for the protozoa we have no quantitative data concerning the effect of oxygen tension on growth. It is known that for short periods of time large changes in oxygen tension have little visible effect on some protozoa (e. g., Amoeba, Hulpieu, 1930) and also that it has little effect on the rate of res- piration (Amberson, 1928) The protozoan fauna of HeS pools (Lauterborn, 1916) and of Imhof tanks (Lackey, 1925) certainly live under conditions of low oxygen tension, but we do not know how it affects their growth rates. There is an obvious scarcity of information concerning the relative degrees of aerobiosis and anaerobiosis under which growth of various protozoa will occur. 9. MECHANISM OF THE EFFECT OF O-R POTENTIAL ON GROWTH How the reduction potential may affect the rate of growth of bacteria or of protozoa if the oxy- gen consumption is constant is not yet completely understood. One theory is that the reduction potential of the medium controls the reduction potential of the protoplasm, thereby controlling metabolic processes. Many cases are known where changes in the potential of the medium are reflected in changes in the potential of proto- plasm (review, Chambers, 1933). However, this brings us to the question of what is meant by the phrase “oxidation-reduction potential of protoplasm”. Surely the phrase can not have the same meaning in a polyphasic colloidal solution as it does when applied to a homogeneous one. Col- loidal particles bear electrical charges, and the potential of the continuous phase may be quite different from those of any of the discontinuous phases. In general usage the phrase “oxidation- reduction potential of protoplasm” means the potential of protoplasm as measured with indica- tors. Supposedly it is the potential of a phase which makes up the bulk of the cell. The fact that the measurements are fairly constant and re- producible (Chambers, 1933) indicates that a definite potential is being measured. The dif- ference in the potentials of the phases is at least partly due to adsorption potentials and perhaps only a small part is caused by reversible oxida- tion-reduction systems. However, we have no direct method of evaluating these various causes of potential differences, and if we assume that oxido-reductions are controlled by electrical potentials, it might make little difference as far as these reactions are concerned how these potentials arise. Quastel (1930) and Kluyver (1931) have put forward the interesting suggestion that oxida- tion-reduction enzymes might be only the intense electrical fields at the interfaces of protoplasm. If this is true, then a change of the potential of any phase of the heterogeneous mixture may have far- reaching consequences. Also, if we accept the theory of Quastel (1930) that at least some bio- logical oxidations occur in the regions of intense electrical fields on the mosaic surface of the cell, we have a method by which the reduction poten- tial of the medium might control oxidations and Jury 7, 1934 | THE COLLECTING NET 39 subsequently other cell activities even if the inner protoplasm were to remain unchanged. An ex- ample which might have some bearing on this theory is the rate of movement of Paramecium (Andrejewa, 1931). The rate of movement in various salt solutions follows approximately the same curves as the adsorption potential of col- lodion particles. Therefore, one is lead to postu- late that the rate of movement is affected by at least one of the perhaps numerous potentials on the surface of the cell. Whether or not oxida- tions or growth might be similarly affecteu re- mains an unansewered question. 10. Ox1IDATION-REDUCTION POTENTIAL OF INFUSIONS AND NaTURAL WATERS Measurements of the Eh value of hay infusions by Efimoff, Nekrassow, and Efimoff (1928) show that shortly after inoculation the Eh value may be as low as —322 mv., that during the first few weeks it may rise 400 my. or more, and that after three months it may reach a value of +422 mv. The low initial readings were probably caused by the removal of free oxygen during rapid bacterial growth, and it is interesting to note that the sequence of forms progresses most appreciably during the first month while the potential is rapidly changing. Whether or not this is a causal relationship has not been deter- mined. The reduction potential of natural waters in the presence of appreciable oxygen is quite high. Some unpublished observations by Professor G. E. Hutchinson and myself show that the reduct- tion potential of both bottom and surface water of Lake Quassapaug (18 meters in depth) was +550 to +580 my. Low reduction potentials are probably found only in waters containing little oxygen and considerable organic matter. However, the reduction potential might be a use- ful index of what we may term the peeetve oxygen tension” of anaerobic waters, i. e., the amount of oxygen which would probably be re- duced by the chemical compounds present, such as HS. 11. GrowrtH oF INTESTINAL PROTOZOA I have pointed out previously (Jahn, 1933b) that the oxidation-reduction potential might be a factor of very great importance in culturing in- estinal protozoa free from bacteria. The elahor- ate researches of Cleveland (1928) and others who have attempted to grow ars forms free from bacteria have resulted in failure, and if we assume that such a feat is Bi all possible, then it seems as if some unknown factor in the medium has been overlooked. I have shown that the Eh value of the rat caecum is about —200 my. —a potential which is decidedly in the so-called “anaerobic” range. Sterile peptone broth and other substances which have been used as culture > fluid have a potential of about -+-200 to +400 my. Under the very strictest of anaerobic con- ditions the Eh of broth may be lowered to -60 my. (Coulter, 1928), but ordinary anaerobic methods probably never result in such a low potential. However, if bacteria are present the po- tential may be lowered to —200 or less ( Escheri- chia coli, final Eh= -270, Plotz and Geloso, 1930), and the protozoa grow quite well. If we assume that our protozoa can live saprozoically, and that they are supplied with sufficient food materials, then this information points to the pro- bability that the Eh value is the most important neglected factor. This theory is quite in harmony with our knowledge of anaerobic bacteria, and it may point the way to a technique for cultivating the intestinal protozoa free of bacteria. An- other factor which might be necessary is a high carbon dioxide tension such as probably occurs in the digestive tract, but so far we have no ex- perimental evidence that this high carbon dioxide tension is necessary, and the reduction potential seems to be more important. , 12. Tue ALLELOcATALYTIC PHENOMENON Probably the most effective stimulus to research on the factors which affect protozoan populations ” was the theory proposed by Robertson (1923). Robertson found that when two ciliates were iso- lated into the same drop of culture medium the division rate was higher than if a single cell were isolated into a drop of the same size. This dif- ference in division rate, according to Robertson, was due to the mutual contiguity of the cells, and was designated by him as an “‘allelocatalytie ef- fect’, caused by a growth-catalyzing substance liberated into the surrounding medium mostly during auclear division. This theoretical catalyst called by Robertson an was “autocatalyst of growth”. The amount of this catalyst present in isolation cultures would depend upon the number of cell divisions that had taken place; therefore, a greater number of dividing cells per volume of medium would lead to a greater concentration of catalyst, and the division rate in these cultures should be correspondingly greater. Investigations of this theory, which, if’ true,. would obviously berof extreme importance to all students of growth, was undertaken by a number of people in other laboratories. Most of the re- sults obtained: (Cutler and Crump, 1923a, b; Greenleaf, 1926; Calkins, 1926; Myers, 1927; Grimwald, 1928: Petersen, 1929; Jahn, 1929; Chejfec, 1929; Darby, 1930; Di Tomo, 1932; and Beers, 1933) could not be interpreted in favor of this theory. Woodruff, years before (1911, 1913), had also obtained data which were directly contradictory to the idea of a mutual accelerative effect in that waste products of metabolism are inhibitory to growth of the species which produced them, but Robertson apparently was not aware of 40 THE COLLECTING NET [ Vor. IX. No. 73 these experiments when he applied the theory to the protozoa. On the other hand, some of the results of other workers ( Yocom, 1928; Petersen, 1929; Dimitrowa, 1932; and Johnson, 1933) were of the type which gave a numerical result in favor of the idea that smaller volumes lead to a higher division rate. The present consensus of opinion is that the growth rate may be either higher or lower in the smaller volumes, dependent upon the conditions of the experiment. The original theory has been dropped, but the large amount of re- search which it stimulated has greatly increased our understanding of the growth factors present in protozoan cultures. 13. ALLELOCATALYSIS vs. Foop SUPPLY Several alternative theories which might ex- plain cettain of the experimental results have been proposed by various authors. Cutler and Crump (1924) offered the theory that the effect was due to differences in the food supply, together with considerable evidence that the division rate of Colpidium does vary with the number of bacteria present. In 1929, using mass cultures of the autotrophic organism Euglena, | found no evi- dence of an allelocatalytic effect. In fact, I found that the rate of growth was much more rapid in cultures in which the population density was lowest. Inasmuch as the bacterial food tactor was absent, this seemed to be conclusive evidence that the phenomenon of allelocatalysis was absent in the case of Euglena. About this time Chejfec (1929) controlled the food supply of Paramecium caudatum by giving it a specified number of bac- teria per day, and failed to obtain an accelerative effect which was independent of the food supply. This line of investigation was continued by John- son (1933) ina carefully controlled series of ex- periments with Oxytricha. When he used rela- tively small numbers of bacteria he found a negative allelocatalytic effect, probably because of insufficient food or the accumulation of protozoan waste products in the small volumes. However, when he used large numbers of bacteria, he found a positive allelocatalytic effect, probably because of the greater ability of the protozoa in the smaller volumes to reduce the supra-optimal con- centration of bacteria, thereby reducing the de- leterious effects of bacterial action. Cultures with an intermediate concentration of bacteria showed no difference in division rate in different volumes of medium. A similar investigation was performed by Smith (1932). However, the cli- max of this line of research is in the work of Beers (1933). Beers, working on the assumption that the bacterial food supply was probably the source of error, chose for his studies the carni- vore Didinium nasutwn and fed it on a known number of its favorite prey, well fed specimens of Paramecium. The use of salt solutions 1n- stead of hay infusions and the careful washing of both paramecia and didinia resulted in experi- ments that were controlled not only as regards food supply but also as regards the products of bacterial metabolism. He also performed similar experiments with Stylonyclia pustulata, which he fed on measured quantities of the green flagellate Chlamydomonas. He used three clones of each of the test organisms, and isolated 1, 2, 3, 4, and 8 individuals into the same volume of medium, varying the volume of the medium (.02 to .16 ce.) in different experiments. His data demon- strate that the rate of division of each of the test organisms was the same regardless of the initial number of organisms and regardless of the volume of medium used. In other words, there was absolutely no evidence of an allelocatalytic effect. He also found that there was no lag period during the initial stages of the culture. Thus we must conclude that when the food supply is controlled and the products of bacterial meta- bolism are eliminated and the medium is favor- able and constant, the phenomenon of alleloca- talysis is not present in cultures of Didiniwm and Stylonychia. It seems as if this is probably also true of cultures of other ciliates. 14. ALLELOCATALYSIS vs. PH The second alternative theory which was used to explain the Robertson phenomenon was that of Darby (1930) who proposed that allelocatalysis might be merely a pH effect. Since Robertson’s medium probably was not at the pH optimal for the species, and was very weakly buffered, and since the organisms tend to change the pH, it seems quite likely that the more organisms pres- ent the more rapidly the pH will be changed. If this change were toward the optimum (as Darby demonstrated it to be for P. caudatum), it seems probable that some of the results showing a posi- tive allelocatalytic effect could very easily be ex- plained in this manner. 15. ALLELOCATALYSIS VS. OXIDATION- ReEpDUCTION POTENTIAL A third alternative theory of how some of the data on allelocatalysis might be explained was offered by Jahn (1933). Since the growth rate is probably affected by the reduction potential, and since the reduction potential of the medium is changed by growth of the protozoa (Jahn, 1933a) or their bacterial food supply, then it seems as if both positive and negative alleloca- talysis could be caused by an adjustment of the medium either toward or away trom the optimum for the organism. The changes which bacteria produce in the reduction potential of the medium are well known (Hewitt, 1931). In general, aerobic forms are able to lower the potential from an Eh value of +400 to about —100 my., and anaerobic forms from about 0 to —400 mv. I have demonstrated that the flagellate Chilo- Jury 7, 1934 ] monas in bacteria-free culture is capable of chang- ing the Eh value of casein peptone as much as 300 my. (425 to 125), and of an acetate-peptone solution as much as 450 my. (425 to —25). If corrected for pH, these changes in Eh (Ith’) are still 270 and 380 my. Such changes, which are almost as great as the growth range of certain bacteria, certainly must have some effect on the growth rate of the flagellate. If this change is toward the optimum for the species, we should expect results which would give a positive alle- locatalytic effect ; if the change is away from the optimum, we should expect a negative alleloca- talytic effect. In the case of a change of 450 mv. it seems likely that the optimal potential was passed and that a sub-optimal potential was final- ly established. Inasmuch as the change was slow, we might expect a period of increasing and then a period of decreasing growth rate. Here the reduction potential is complicated by the presence of oxygen. However, if we admit that oxygen tension is more important than the reduction potential, then the above explanation of allelocatalysis may be restated in terms of oxygen tension. A change of 400 my. resulting in the decoloration of methylene blue is certainly proof of a great change in oxygen tension, and inas- much as this change occurred in flasks stoppered only by a cotton plug, it is a factor which was probably also present in the experiments of pre- vious investigators. 16. ALLELOCATALYSIS vs. COvs. TENSION A fourth theory which might explain the phenomenon of allelocatalysis in the protozoa is one which involves an adjustment of the CO, content of the medium by the organism. For many years it has been known that small amounts of CO» are necessary for the growth of various bacteria, and the lack of sufficient CO. in a freshly inoculated medium has been proposed as one of the reasons for a lag period, and for the necessity of large inoculations in order to secure growth under certain conditions (Walker, 1932, and earlier workers). So far we have no evi- dence of this kind for the protozoa, but there seems to be no reason why a similar result should not be expected. Also, the extensive experiments of Jacobs (1912) on the threshold of carbon dioxide toxic- ity might be taken to indicate that the large quan- tities of CO» produced during the rapid growth of bacteria in an infusion might cause a decrease in the division rate of the protozoa. This might be a factor in experiments such as those of John- son (1933) in which. he used large quantities of bacteria. Since Jacobs found different threshold values for toxicity in different species, this might also be a factor which controls the sequence of forms in hay infusions. THE COLLECTING NET AN 17. MATHEMATICAL DEFECTS OF AUTOCATALYSIS If a mutual accelerative effect of contiguous cells is assumed, then one should expect the divi- sion rate to increase during the early stages of a culture started from a single cell. Therefore, ac- cording to the theory, the growth equation should be autocatalytic. However, there are several mathematical objections to this conclusion. In the first place a protozoan population is not a closed system (Morgan, 1926), and therefore there could be no direct valid comparison of population and growth and the amount of trans- formed material in a chemical reaction. Secondly, the autocatalytic equation does not allow for volume changes which occur during growth of an organism or population (Snell, 1929). The third mathematical criticism which I wish to make is that if growth of a protozoan culture were to follow the autocatalytic curve, then the division rate could not possibly be increasing after the first cell division. The division rate in such a case must be defined as D=K(A—x) where K is the growth constant, A is the maximum number obtainable in a culture, and x is the number pres- ent at any given time. The value of the division rate (D) is a decreasing sigmoid function of t and a decreasing linear function of x (Jahn, 1930). Therefore, if growth of a population were to follow the autocatalytic curve, the divi- sion rate must necessarily be decreasing. 18. Murua Errect oF CONTIGUOUS SPECIES A new method of investigating the effect of contiguous cells of different species has recently been introduced by Gause (1934). By using mixed and pure cultures of Paramecium caudatum and Stylonychia mytilus under comparable conditions, Gause computed with the aid of Volterra’s equa- tion, the effect of the presence of one species on the growth of the other. He found that when both species were competing for the same food supply (8. subtilis) Stylonychia had a strong in- hibitory influence on Paramecium in that the maximum number of Paramecium was reduced almost 60% (Volterra’s a = 5.5), but that the presence of Paramecium decreased the Stylony- chia population only about 20% (Volterra’s B = .12). In other words, the presence of each Stylonychia in the mixed culture reduced the number of Paramecium which would have been present by 5.5; whereas, the presence of each Paramecium reduced the number of individuals of Stylonychia only .12. This is a method of at- tacking the problem of mixed populations which has not yet been extensively used, but it is one which offers considerable possibilities. The next step, of course, is to determine the nature of the inhibitory effects involved. In this case depele- tion of the food supply is probably very import- (Continued on Page 47) 42 THE COLLECTING NET [ Vor. TX. No. 73 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel W. Parker, and An- naleida van’t Hoff Snyder Cattell. Printed by The Darwin Press, New Bedford THE COLLECTING NET IN 1934 THE CoLLEcTING NET is extending its scope this summer to include the activities of the Mt. Desert Island Biological Laboratory and_ the Scripps Institution of Oceanography. We shall print their news items from time to time and we hope that we may occassionally have the privilege of printing their reports of seminars and lectures of especial interest to the scientific workers in Woods Hole. It seems reasonable to suppose that if a part of our twenty-eight pages could be devoted to work at biological laboratories out side of Woods Hole that THE CoLLecTtinG NET would be more interesting and of wider useful- ness. It has occurred to us that THE COLLECTING Ner might have a definite use as the unofficial and informal organ of the marine and fresh water biological laboratories of North America. We would like it to form a medium for the exchange of ideas'among biologists frequenting biological stations, and we invite contributions from work- ers out side of Woods Hole. Some one recently remarked that publication in Tur Cottectinc Net was a “trial baloon” ; « that work and theories in their formative state could be offered to other biologists for discussion ‘and criti¢ism before final publication. We would like more material of this kind. CONCERNING THE MIXER The M. B. L. Club feels that it should possibly strike a note of apology concerning the “mixer” at the clubhouse last Saturday evening. The Club was founded with the idea that it should be for the promotion of social intercourse at the laboratory. A “mixer’’ was inaugurated, at which students and investigators would have the oppor- tunity to meet one another and chat informally. ’Then ‘there followed dancing or some other en- , tertainment, and the ‘evening was always a huge success. Last Saturday’s party fulfilled only part of these requirements, as the old-fashioned mix- ing was admittedly conspicuous by its absence. A word of explanation along with our apology seems fair enough. When the orchestra was en- gaged, it was told that the music would not be needed before 10:00 P M., thus allowing a good- ly time for real “mixing” before the dancing. It also happened that, due to a slight mishap, some of the materials for the mixer were not obtain- ed at the proper time and those in charge dashed to Falmouth to make the necessary purchases. Some delay was encountered, and on returning, it was quite a shock to find that the dancing was in full swing. That is not as it should have been, but it must also be said that it is not as it was planned to have been. The new committees had not gotten themselves into proper running order before the mixer was upon them and as a result the night turned out to be more of a mix-up that a mix-er. Those concerned offer their sincerest regrets that true mixing was not in evidence, and promuse those who came to the Mixer and were disappointed, that a real Mixer will be held at the opening of the second session and if they will be on hand, we will do our best to atone for our mistakes and to make them feel rewarded. —Ropert STABLER, President The Directory of the Scientific Workers in Woods Hole will be on sale on Tuesday, or Wed- nesday. Season subscribers will receive com- plementary copies; to others the charge will be ten cents. Last week we mistakenly mentioned that Dr. Thomas Hunt Morgan “received the degree of Doctor of Science from Yale University This should have read “......from Harvard University.” We wish to offer our apologies for the above error. CURRENTS IN THE HOLE At the following hours (Vaylight Saving Time) the current in the Hole turns to run from Buzzards Bay to Vineyard Sound: Date AciM. Pais July 7 1:46 1:49 July 8 2:39 2:40 July 9 Sotey Sh 5(27/ July 10 ae les abel July 11 : 4:51 4:54 July 12 Be 5:30) S5eso July 13 6:08 6:17 July 14 6:47 6:58 July 15 7:26 7:40 July 16 8:06 8:24 In each case the current changes approxi- mately six hours later and runs from the Sound to the Bay. It must be remembered that the schedule printed above is altered somewhat by wind conditions. Prolonged winds sometimes cause the turning of the cur- rent to occur a half an hour earlier or later than the times given above. The average maxi- mum velocity of the current is five knots. Jury 7, 1934 ] THE ,COLLECTING NET 43 ITEMS OF “UNAGERSe oa Dr. Edmund Newton Harvey, professor of Autroducing physiology at Princeton University, and research associate of the Carnegie Institution was elected a member of the National Academy of Sciences at its annual meeting in April, 1934. Dr. Harvey is a trustee of the Marine Biological Laboratory at Woods Hole, and is at present in Bermuda. Dr. Ella Hoppe, of the branch laboratory of the New York State Department of Health, died in April, 1934. Dr. Hoppe spent the last four summers at the Marine Biological Laboratory, and worked on the susceptibility of eels and dog- fish to diphtheria toxin. In New York she worked on tissue cultures. Dr, A. C. Redfield, head of the oceanographic investigations of the Alfantis in the Gulf of Maine, returned on July 2nd after a week's cruise. He is at present at Harvard University on business concerning the reorganization of the zoology department there. Dr. H. P. Bell, professor of botany at Dal- housie University, is working this summer at the Federal Government Experiment Station in Kent- ville, Nova Scotia. Dr. Bell was an instructor in the botany course at the Marine Biological Laboratory last summer. Dr. A. R.. Moore, who was at the Marine Biological Laboratory in 1932, visited Woods Hole for a few days last week before taking up his work on a National Research Council grant at The Wistar Institute farm at Bristol, Penn- sylvania. Dr. Moore is head of the department of general physiology at the University of Oregon. Dr. J. H. McGregor, professor of zoology at Columbia University, who usually spends his summers at Woods Hole, will not be here this summer. He is vacationing in Northhampton, Mass., with his niece, Miss Lois Te Winkel. Dr. Ralph W. Gerard of the department of physiology at Chicago University, who last sum- mer, directed the course in physiology at the Marine Biological Laboratory, is this summer in Europe. Dr. Gerard is travelling on a fellowship supported by the Rockefeller Foundation, and is visiting other European Fellows to report on their work. Dr. James Frederic Danielli of University Col- lege, London, who has come to America, received his B. Se, degree in 1931, and his Ph. D. degree in 1933. He is a member of the Associated In- stitute of Chemists. Dr. Danielli has been work- ing under a Commonwealth Fellowship in biology at Princeton University, which he retains until 1935. His special interest is in problems connected with surfaces and colloidal systems. He is work- ing on different phases of the problem of low surface tension at the interface between oil and aqueous phases. Dr. Danielli is preparing papers concerned with the surface active substances in mackerel eggs, and the permeability of thin films with special reference to pore and_ solubility theories and specific chemical affinities in thin films. At University College, London, he com- pleted papers on surface films, structure of vita- min A, carotene derivatives, sterols, aliphatic compounds and oestrin. He recently gave a lec- ture to the students of physiology at the Marine Biological Laboratory. —R. W. P. Dr. Louise H. Gregory, associate dean and as- sociate professor of Zoology at Barnard College is expected at Woods Hole for al visit some time in July. Dr. Gregory had a leave of absence during the Spring term during which she travelled through the south and west. Dr. Albert J. Dalton, teaching fellow at Har- vard University, visited the Marine Biological Laboratory on June 29th. Dr. Dalton is work- ing on chorioallantoic graphs of young chick embryos. There is a notice on the main Bulletin of the M. B. L. of books and reprints that have been lost from the Library. Most of the reprints listed in full on the ‘Library Bulletin” as lost since the check made in 1925, are probably re- posing in the personal reprint collection of vari- ous investigators—unconsciously appropriated. A hint may not be out of place in THE COLLECTING Ner that we would like now and always the re- turn of any that turn up—the M. B. L. stamp is on each. —Librarian. 44 THE COLLECTING NET { Vor. IX. No. 73 THE BEACH QUESTION! There has been considerable discussion during the summer regarding “rights” on the so-called Breakwater Bathing Beach on the Bay Shore or Buzzards Bay side of Woods Hole. It seems worth while to draw attention to some of the points involved. The records show clearly that there is no bathing beach in Woods Hole owned by the Town and therefore accessible to all. The Breakwater Beach, including the bath houses, the so-called Laboratory Beach adjoining the Gansett property and the Nobska Beach are all privately owned. When the Breakwater Beach property, Lot X on the Town plan, was sold, certain rights were given to “any such inhabitants of Woods Hole as make it their home to use the bathing beach and passage thereto.” These rights apply to a strip of shore ten to fifteen feet wide and two hundred feet long extending in front of the bath house. The rest of the adjoining beach on either side of the bath house is privately owned. The deeds to these lots read “to the waters of Buzzards Bay.’’ However, I am told by the authorities at the Town Hall in Falmouth, that while formerly titles were given to read “to the waters of Buzzards Bay” recent decisions by the Land Court indicate that a title is good only to “Mean High Tide”. Different interpretations have been given of the term “Waters of Buzzards Bay”. One of the property owners on the Bay Shore has, he tells me, had a well known Boston lawyer look into the matter and has been told that his deed entitles him to the beach in front of his property at all states of the tide; indeed that the title to the land is such that any person in the water in front of the property would be trespassing if he had his feet on the bottom. It is generally understood, I believe, that this or some similar interpretation is claimed by the other owners of property on the beach. It is clear, therefore, at least in the opinion of the property holders, that those who bathe on any of the beaches at Woods Hole are either trespassers or guests of the owners of the adjoining property, unless they are on the small strip opposite the bath houses and are “such in- habitants of Woods Hole as make it their home”. _ Appeal has been made to certain ancient fishing rights. It may be doubted, however, if shell or any other fish are abundant enough to attract fish- ermen there. Perhaps a sufficiently liberal in- terpretation of these rights might make them ap- ply to Venus anadyomene and classify the linger- ing sunlit swains as fishermen. But the law is prosaic. It is rumoured that a wall may be built. The justification for this is again sought in an ancient custom. It would be a “cattle fence’’. And again it may be asked if cattle are sufficiently abundant on the beach and prone to trepass as to warrant the building of a stone wall. There is of course no question as to the im- portance to Woods Hole of a bathing beach. People will always want to come to Woods Hole but they will not always want to bring their families with them if they cannot be sure of enjoying the beach. The value of property here depends largely on the number of families who come to Woods Hole or want to come. The Town of Falmouth has already taken over by “Eminent Domain” Old Silver Beach and the beach at West Falmouth. Shall it take over the “bathing beach”? This of course would prove a hardship to the few who at present enjoy the private ownership of its separate portions, but is that to be weighed against the hardship to the hundreds who may be deprived of the right of even walking on the beach? If the Town of Fal- mouth had the title to the bathing beach, then im- provements in the state of the beach and the bathing facilities could be made by the Town or by private subscription. Any improvements made under the present status of the beach by public subscription would be made on property privately owned on which the public has no claim. James W. Mavor. 1Owing to the interest which has been shown concerning the Bay Shore bathing beach, it seems appropriate that we should print this survey of the bathing situation which Professor Mavor wrote for us in 1930. Limitation of time and his absence has made it necessary to include this material without consulting the author. THE M. B. L. CLUB On the evening of June 30, the M. B. L. Club held the ““Mixer.’’ As has been done in the past to simplify the process of meeting strangers everyone had his or her name pinned in sight. The evening was mostly confined to dancing and consuming refreshments during the intermissions granted by the Orchestra. The Club feels that because of the expense in- volved it can provide for an orchestra only on every other Saturday. This leaves the interven- ing Saturday evenings for entertainments of a varied nature. In order to continue the Wednesday night vic- trola concerts which were much enjoyed last summer at the Club, a music committee has been formed. People who have records which they wish to loan may see Mrs. Schweitzer who is chairman of the committee. Jury 7, 1934 ] THE COLLECTING NET 45 A BIOLOGIST’S IMPRESSIONS OF THE GALAPAGOS ISLANDS Dr, WILLIAM RANDOLPH TAYLOR Professor of Botany, University of Michigan Galapagos Islands.” Galapagos Islands. by Dr. Taylor. On Friday, June 29, Dr. W. R. Taylor of the University of | Michigan gave an illustrated lecture in the auditorium of the Ma- | rine Biological Laboratory on ‘‘A Biologist’s Impressions of the Dr. Taylor was invited to accompany Mr. G. A. Hancock of California on a combined vacation and scientific trip to the west coast of Central and South America, and to the The following account is based upon short hand notes taken by Miss Rachel W. Parker and has been checked The cruise was made on a well-planned and strictly utilitarian cruises belonging to Mr. Han- cock. There was a small laboratory on board with room for four scientists to carry on their work among the packing boxes and in storage spaces. A launch fitted with motor-operated hoist dredge, and a drum with steel sounding line was a valuable part of the equipment. The Galapagos Islands lie on the equator and just south of it, and are almost exactly south of New Orleans. At its greatest length the archi- pelago is nearly equal to the distance between New York and Provincetown. There are more than twenty-five islands in the group and about six of them are distinctly larger than the rest. They were discovered by the early Spanish ex- plorers, and the white beaches became a conven- ient stopping-place where buccaneers could repair their ships. These temporary settlers left behind them goats, cattle, and dogs which gradually populated the island with livestock. Whalers came there for giant tortoises, and later the islands were used as penal colonies. At present, they are not isolated but well-visited by scientific parties, yachtsmen, and fishermen in search of tuna. Scientific interest in the Galapagos Islands has been continuous since the time of Darwin, because of the curious diversity of fauna and flora found there. Dr. Taylor collected algae of the region, and noted that some appeared to have been brought by ocean currents from the south, off the Peruvian and Chilean coasts. One of the first islands visited had a rocky shore with almost vertical cliffs, but in spite of this forbidding front and a strong surf, Mr. Han- cock managed a successful landing. High up on the rocky ledges were many nesting Frigate birds. The males were a glossy black, colored with iri- descent green and purple, and had huge red blad- ders beneath their bills. The white sand beaches were crowded with a noisy group of nesting blue- footed boobies the size of small geese. On the northern shore the vegetation consisted of dense shrubbery with foliage unfortunately affected by a long dry season. Dr. Taylor secured excellent photographs both of the scenery on the islands and of the wild life there. These were difficult to obtain because of the wariness of the animals and the inaccessibility of their habitat. On the shores of Albemarle little collecting was carried on, though this was one of the most pic- turesque islands of the group. The cove which the boat entered was deep, narrow and long, and the surrounding rocks were rough and jagged, showing stratification. Albermarle is a favorite resort for fishermen. Here cormorants and gulls were found as well as a few pelicans. At Narborough island one of the most interest- ing shores was explored. There were many small volcanic craters on this island. On the rough steep rocks lizards were discovered, bright- ly colored and quick in action. Marine iguanas were an especially interesting form to observe. They were black in color and often attained a length of three and one half feet, weighing from ten to fifteen pounds. They feed on seaweed deep in the water. After shedding their skins, they become more bright and attractive with a beautiful arrangement of sharp scales. The mar- ine iguana is not excitable, and may be caught rather easily by picking it up by the tail; it does not bite. The land iguana on South Seymour has a short tail and a long heavy body, weighing as much as twenty-five pounds. They are pugnac- ious, and though they will eat bananas or celery out of the hand, they may be incited to use their small teeth which have an amazing grip. They also differ from the marine iguanas in being more wary. On this island a sea turtle was found in an inland salt lagoon and when dissected a quant- ity of red seaweed was found within. The flatter portions of the islands are covered with different forms of cacti which exhibit a tangle of brilliant flowers when in bloom. Man- groves, which ordinarily grow in mud in Florida, root here between the rocks. The shrubbery is dense and curious in shape. Sea lions occur far from shore as well as on the rocks; the females are inoffensive but the males are more pugnacious. On Charles Island there have been several settlements. In the hills curious caves were exca- 46 THE COLLECTING NET [ Vor. IX. No. 73 vated from the natural soil by heat and water, and have been used to live in, possibly by mem- bers of a penal colony. The only permanent settlement is on Chatham Island where the gover- nor resides with an army of three men who are almost never called on to fire their guns. There is one lighthouse which is illumined only when boats are expected. Chatham Island used to be a penal colony but is now a proprietary farm. At various times Norwegians were induced by real estate men to settle on the islands. Tractors and electricity were brought in, but there was no oc- cupation for settlers but fishing and they gradual- ly disappeared. Indefatigable Island is characterized by large intercostal flats covered with sand dunes, grass, and lava from small evenly-placed volcanic cones. Gnarled and twisted trees and two species of cacti The huge tortoises to be found here are dark brown or black, and remark- able in size, being large enough to carry a man. The Manta ray is found in the Galapagos Islands, grow on the island. and one specimen measuring fifteen feet from tip to tip was harpooned on the expedition and towed in by ropes to the cruiser. THE CAPACITY OF THE BRAIN Dr. LauRENCE IRVING AND Mary Scott WELCH Department of Physiology, University of Toronto | Changes in Oy tension do not affect Oz uptake of tissues as long as the Oz tension is above the characteristically low essential minimum. If the CQOy» tension is increased, however, the CO» out- put of the tissue is diminished, and the ratio: CO, output Os utilization pression of the R.Q. is great or small according to the COz capacity of the tissues. Consequently, if two different tissues are subjected to the same increase in CO. tension, that “which suffers the greater depression of its R.Q. is the one with greater COz capacity. On this basis the R.O. of the brain and of the hind leg of dogs was examined by analysis of arterial and venous blood. It was found that 10 minutes after an increase in the CO» tension of arterial blood the R. Q. of the leg was still de- pressed, while that of the brain was approximate- ly normal. Evidently the brain became saturated with CO. more rapidly than the leg. This result conforms with our knowledge that the buffering power of muscle is considerably re- (R.O.) is depressed. The de- inforced by the presence of phosphocreatine, which is insignificant in brain. But it does not prove the existence of smaller cerebral COz ca- pacity because of two possibilities. The blood flow and the metabolic CO, production of brain’ at rest are both probably greater than in the leg and both factors would accelerate equilibration and restoration of the normal R. Q. of the brain. The facts indicate that the COs tension,in the brain rises more rapidly in response to arterial changes than in the leg. The stimulating effect of COs upon respiration is said to act directly upon the respiratory centre; and blood flow through the brain is rapidly accelerated by increasing the carbon dioxide tension, while flow is not increased in muscle. The rapid change in cerebral COz ten- sion is consistent with the idea that it is the stimu- lus for respiration and for changes in cerebral circulation. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 3.) OVARIAN HORMONES AND BASAL METABOLISM Dr. Mary E. CoLLettT Associate Professor of Biology, Western Reserve University The basal metabolism of normal women show a rythmical variation corresponding to the phases of the menstrual cycle. Since the maxima correspond to the time at which follicular fluid or corpus luteum secretion are probably formed, it seems possible that these hormones directly or in- directly increase the basal metabolism. Preliminary observations have been made on two women with defective ovaries who were taking theelin and extract of corpus luteum. In general we have found that such doses are fol- lowed by a transient rise in metabolism of 4 to 7 cc. of oxygen per minute. The effects fade grad- ually after the third day. A further series of ob- servations is needed, preferably upon a woman whose ovaries have been completely removed. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 3.) Jury 7, 1534 } THE COLLECTING NET 47 PROBLEMS OF POPULATION GROWTH IN THE PROTOZOA (Continued from Page 41) ant. However, the experiments of Woodruff (1913) suggest that waste products of one species might play an appreciable role as an accelerant of growth of the other species. Also, the work of Darby (1929) suggests that pH may be im- portant, and we might find plausible arguments for other specific factors, most of which will eventually have to be investigated. LITERATURE Alexander, Gordon. 1931. The significance of hy- drogen ion concentration in the biology of Euglena gracilis Klebs. Biol. Bull. 61: 165-184. Amberson, W. R. 1928. The influence of oxygen tension upon the respiration of unicellular or- ganisms. Biol. Bull. 55: 79-91. Andrejewa, E. W. 1931. Zur Frage iiber die physi- kalisch-chemische Bestimmung der Korrelationen einiger physiologischer Prozesse bei Paramecium caudatum. Arch. f. Protist. 73: 346-360. Beers, C. D. 1933. The relation of density of popu- lation to rate of reproduction in the ciliates Didinium nasutum and Stylonychia pustulata. Arch. f. Protist. 80: 36-64. Bodine, J. H. 1921. Hydrogen ion concentration of protozoan cultures. Biol. Bull. 41: 73-77. Bodine, J. H. 1924. Some physiological actions of cyanides. J. Gen. Physiol. 7: 19-24. Breslau, E. 1926. Die Bedeutung der Wasserstof- fionenkonzentration fiir die Hydrobiologie. Verh. d. Internat. Verein fiir Theoretische und Ange- wan. Limnologie 3: 56-108. Calkins, G. N. 1926. Biology of the Protozoa. Chambers, R. 1933. An analysis of determina- tions of intracellular reduction potentials. Cold Spring Harbor Symposia on Quantitative Bio- logy 1: Chejfec, M. 1929. Die Lebensdauer von Paramec- ium caudatum in Abhangigkeit von der Na- hrungsmenge. Acta. Biol. Experimentalis 4: 73-118. Cleveland, L. R. 1928. The separation of a Tri- trichomonas of man from bacteria; its failure to grow in media free of living bacteria; mea- surements of its division rate in pure cultures of bacteria. Am. J. Hyg. 8: 256-278. Collett, M. E. 1919. The toxicity of acids to ciliate infusoria. J. Exp. Zool. 29: 443-472. Coulter, Calvin B. 1928. Oxidation-reduction equi- libria in biological systems. I. Reduction poten- tials of sterile culture bouillon. J. Gen. Physiol. 12: 139-146. Crane, M. M. 1921. The effect of hydrogen ion concentration on the toxicity of alkaloids for Paramecium. J. Pharmacol. and Exp. Therap. 18: 319. Cruess, W. V. and P. H. Richert. 1929. Effect of hydrogen ion concentration on the toxicity of sodium benzoate to microorganisms. J. Bact. 17: 363-371. Cutler, D. W. and L. M. Crump. 1923a The rate of reproduction in artificial culture of Colpidium colpoda. Biochem. J. 17: 174-186. Cutler, D. W. and L. M. Crump. 1923b. The rate of reproduction in artificial culture of Colpidi.m colpoda. Part II. Biochem. J. 17: 878-886. Cutler, D. W. and L. M. Crump. 1924. The rate of reproduction in artificial culture of Colpidium coipoda, Part III. Biochem. J. 18; 905-912. Darby, Hugh H. 1929. The effect of the hydrogen iou in cuncentration on the sequence of protozoan forms. Arch. f. Protist. 65: 1-37. Darby, Hugh H. 1930. Studies on growth accelera- tion in Protozoa and yeast. J. Exp. Biol. 7: 308-316. Dimitrowa, A. 1932. Die fordernde Wirkung der Exkrete von Paramoecium caudatum Ehrbg. auf dessen Teilungsgeschwindigkeit. Zool. Anz. 100: 127-132. DiTomo, M. 1932. Ricerche sul comportamento di Paramecium caudatum in un dato volume di cultura liquida. Boll. di Zool. 3: 137-140. Dusi, H. 1933. Recherches sur la nutrition de quel- ques Euglénes. II. Ann. de l’Inst. Pasteur 50: 840. Eddy, Samuel. 1928. Succession of protozoa in cul- tures under controlled conditions. Tr, Am. Micr. Soc. 47: 283-319. Efimoff, W. W., N. J. Nekrassow, and Alexandra W. Efimoff. 1928. Die Einwirkung des Oxydations- potentials und der H-Ionenkonzentration auf die Vermehrung der Protozoen und Abwechselung ihrer Arten. Biochem. Z. 197: 105-118. Elliott, A. M. 1933. Isolation of Colpidium stria- tum Stokes in bacteria-free cultures and the re- lation of growth to pH of the medium. Biol. Bull. 65: 45-56. Elliott, A. M. 1934. Effects of certain organic acids and protein derivatives on the growth of Colpi- dium. Arch. f. Protist. (in press). Gause, G. F. 1934. Uber die Konkurrenz zwischen Arten. Zool. Anz. 105: 219-222. Gerard, R. W. dations. 192-194. Greenleaf, W. E. 1926. The influence of volume of culture medium and cell proximity on the rate of reproduction in infusoria. J. Exp. Zool. 46: 143- 167. Grimwald, E. 1928. Recherches sur les facteurs du développement des cultures de micro-organismes. L’action de la substance allélocatalitique appar- aitelle dans les cultures du Colpidium cotpoda Ehrb., Acta. Biol. Experimentalis 3: 81. Hall, R. P. 1933. On the relation of hydrogen-ion concentration to the growth of Euglena ana- baena var. minor and E. deses. Arch. f. Protist. 79: 239-248. Hargitt, G. T. and W. W. Fray, 1917. Paramecium in pure cultures of bacteria. J. Exp. Zool. 22: 421-454. Hewitt, L. F. 1931. in bacteriology County Council. Hulpieu, H. R. 1930. The effect of oxygen on Amoeba proteus. J. Exp. Zool. 56: 321-361. 1932. Energy relations in cell oxi- Canad. J. Chem. and Metallurgy 16: Oxidation-reduction potentials and biochemistry. London 48 THE COLLECTING NET [ Vo. IX. No. 73 Jacobs, M. H. 1912. Studies on the physiological characters of species. I. The effects of carbon dioxide on various protozoa. J. Exp. Zool. 12: 519. Jahn, Theo. L. 1929. Studies on the physiology of the euglenoid flagellates. I. The relation ot the density of population to the growth rate of Eug- lena. Biol. Bull. 57: 81-106. Jahn, Theo. L. 1930. Studies on the physiology of the euglenoid flagellates. II. The autocatalytic equation and the question of an autocatalyst in growth of Euglena. Biol. Bull. 58: 281-287. Jahn, Theo. L. 1931. Studies on the physiology of the euglenoid flagellates. III. The effect of hy- drogen ion concentration on the growth of Eug- lena gracilis Klebs. Biol. Bull. 61: 387-399. Jahn, Theo. L. 1932. The effect of temperature and the acetate radical on growth of Euglena graci- lis. Anat. Rec. 54: (Suppl.) 22. Jahn, Theo. L. 1933. Studies on the oxidation-re- duction potential of protozoan cultures. I. The effect of —SH on Chilomonas paramecium. Pro- toplasma 20: 90-104. Jahn, Theo. L. 1933a. Changes in the oxidation- reduction potential and pH of bacteria-free cul- tures of Chilomonas. Anat. Rec. 57: (Suppl.) 40. Jahn, Theo. L. 1933b. Oxidation-reduction poten- tial as a possible factor in the growth of intes- tinal parasties in vitro. J. Parasitol. 20: 129. Johnson, W. H. 1933. Effects of population den- sity on the rate of reproduction in Oxytricha. Physiol. Zool. 6: 22-54. Jones, E. P. 1929. Paramecium infusion histories. I. Hydrogen ion changes in hay and hay-fiour infusions. Biol. Bull. 59: 275-284. Jordan, E. O. and I. S. Falk. 1928. The newer knowledge of bacteriology and immunity. Univ. of Chicago Press. Kluyver, A. J. 1931. Chemical activities of micro- organisms. London. Knaysi, G. and S. R. Dutky. 1934. The growth of Bacillus megatherium in relation to the oxida- tion-reduction potential and the oxygen content of the medium. J. Bact. 27: 109-120. Kostir, W. J. 1921. The comparative resistance of different species of Euglenidae to citric acid. Ohio J. Sci. 21: 267-271. Lackey, J. B. 1925. The fauna of Imhof tanks. Bull. New Jersey Agric. Exp. Sta. No. 417. Lauterborn, R. 1916. Die sapropelische Lebewelt. Verh. d. Naturh.-Med. Ver. Heidelberg N. F. 13. Loefer, J. B. 1934. Relation of hydrogen-ion con- centration to growth of Chilomonas and Chloro- gonium. Arch. f. Protist. (in press). Loefer, J. B. 1934a. Effect of certain carbohydrates and organic acids on growth of Chlorogonium and Chilomonas. Arch. f. Protist. (in press). Luck, J. M., G. Sheets, and J. O. Thomas. 1931. The role of bacteria in the nutrition of protozoa. Quart. Rev. Biol. 6: 46-58. Lwoff, A. and Nadia Roukhelman. 1926. Variations de quelque formes d’azote dans une culture pure d'infusoires. C. R. Acad. Sci. 183: 156-158. Morgan, T. H. 1926. Genetics and the physiology or development. Am. Nat. 60: 489-510. Myers, E. C. 1927. Relation of density of popula- tion and certain other factors to survival and reproduction in different biotypes of Paramec- ium caudatum. J. Exp. Zool. 49: 1-438. Oehler, R. 1916. Amoebenzucht auf reinem Bo- den, Arch. f. Protist. 37: 175. Oehler, R. 1920. Flagellaten- und Ciliatenzucht aut reinem Boden. Arch. f. Protist. 40: 16-26. Petersen, W. A. 1929. The relation of density of population to rate of reproduction in Paramecium caudatum. Physiol. Zool. 2: 221-254. Phelps, Austin. 1931. Effect of H-ion concentra- tion on the division rate of Paramecium aurena, Science 74: 395-396. Plotz, H. and J. Geloso. 1930. Relations entre la croissance des micro-organismes anaerobies et le potentie! du milieu de culture. Ann, Inst. Pas- teur 45: 613-640. Quastel. J. H. 1930. The mechanism of bacterial action. ‘tr. Far. Soc. 26: 853-864. Rahn, O. 1932. Physiology of bacteria. Richards, O. 1929. The correlation of the amount or sunlight with the division rate of ciliates. Biol. Bull. 56: 298-305. Robertson, T. B. 1923. The chemical basis of growth and senescence. Roskin, Gr. and Ed. Dune. 1929. Zur Frage uber die Wirkung des Chinins auf die Zelle. Arch. f. vrotist. 66: 346-354. Sherman, J. M. and G. E. Holm. 1922. Salt effects in bacterial growth. Il. The growth of Bact. coli in relation to H-ion concentration. J. Bact. 7: 465-470. Smith, G. A. 1932. Strength of culture medium as a tactor in fission rate of Paramecium caudatum, Anat. Rec. 54 (Suppl.): 100. Snell, G. D. 1929. An inherent defect in the theory tnat growth rate is controiled by an autocata- lytic procces. Proc. Nat. Acad. Sci. 15: 274. Walker, Harold H. 1932. Carbon dioxide as a fac- tor affecting lag in bacterial growth. Science 76: 602-604. Wang, C. C. 1928. Ecological studies of the sea- sonal distribution of protozoa in a fresh-water pond. J. Morph. 46: 431-478. Wann, F. B. and E. F. Hopkins. 1927. Further studies on growth of Chlorelia as effected by hydrogen- ion concentration. Bot. Gaz. 83: 194-201. Woodruff, L, L. 1911. The effect of excretion pro- ducts of Paramecium on its rate of reproduction. J. Exp. Zool. 10: 557-581. Woodruff, L. L. 1912. Observations on the origin and sequence of the protozoan fauna of hay in- fusions. J. Exp. Zool. 12: 205-264. Woodruff, L. L. 1913. The effect of excretion pro- ducts of Infusoria on the same and on different species, with special reference to the protozoan sequence in infusions. J. Exp. Zool. 14: 575-582. Yocom, H. B. 1928. The effect of the quantity of culture medium on the division rate of Oxytricha. Biol. Bull. 54: 410-416. Jury 7, 1934 ] THE COLLECTING NET 49 Fa OO OOOOOOOOOOOOOOO OOOO OOOSOSOOSOSOSGIOOOOGOOGOGOOOOSOOGO000000008 Leitz Ultropak Illuminator The ideal illuminator for opaque objects of low reflecting power, such as many living organisms, surfaces on leaves or plants and many other biological objects. A complete series of objectives and condensers is available for observations from lowest to highest magnifications. Special Accessories enable the observer to eliminate surface glare from moist surfaces or to observe objects immersed in liquids. TIVOINUOINUULUUTAUUCUUULUAUUU IOUT Instruments and Accessories will be ex- Iubited during August, 1934 at: This equipment and many other new Leitz R. G. THOMPSON'S, O O O @) O O O O O O O O O O O O O O O O O O O O O O O © © O) © © © ©) O O O O O O O O ©) O O O O O O © © © © O © O O © © O ; MAIN STREET, ' ‘ WOODS HOLE, MASS. ' YWIUAIVNVUIVLLNLIULVLVLUIUUUIOLVLAUOLULUUUULVLUUIUUULOUUVLISUUUOUULUUULVUU UU LITERATURE ABOUT ULTROPAK ILLUMINATOR UPON REQUEST Beeb Z; InewMepn CN: 60 EAST 10th STREET © NEW YORK CITY Branches: WASHINGTON, D. C. SAN FRANCISCO, CALIF. CHICAGO, ILLINOIS. LOS ANGELES, CALIF. DOCONDNDODNDONNNNDNDONDNNNNDNONOOOOONOOOOOOO0O00000000000000000000 OO99000000000 50 THE COLLECTING NET [ Vor. IX. No. 73 NEW YORK CAST GIVES PREMIER PERFORMANCE ON MONDAY Tickets 50c, $1.00, $1.50, $2.00; Free Transportation to Silver Beach and Back Again On Monday, July 9th a new play is being given at the Beach Theatre for the benent of THE CoL- LectiING Net. “Tight Britches” by John Taintor Fotte and Hubert Hayes is presented by Laurence Rivers, famous producer of “The Green Pas- tures’ and other plays. It is the story of a boy of the Smoky Mountains, who had the deter- mination to be a preacher, but was surrounded by people totally unable to comprehend him. Not until the end of the play did Aunt Vistie begin dimly to understand him as she remarked: “You wuz jes’ too big fer yer britches.” In order to enable as many people as possible to attend the special benefit performance of this play at the Silver Beach Theatre on Monday eve- ning, July 9th, free transportation will be pro- vided for all scientific workers and residents of Woods Hole. This Broadway play is of excep- tional interest in that it is being produced by a New York cast including Jean Dixon and other Vheatre Guild players. The players are Jean Dixon, “Once in a Lifetime” and “Dangerous Corner”; Joanna Roos, Shepherd Strudwick, John Miltern, William Ingersoll, Arthur Hughes, Kathleen Conegys, Mary Orr, Pierre de Ramey, Doan Borup and John Forbes. Staged by Mir- iam Doyle. The Silver Beach Theatre of- fers seventy-two seats at fifty cents each, and one hundred and eight seats at one dollar each. The total expense for seeing this play would thus come to less than one would pay for transporta- tion and a moving picture outside of Woods Hole. Of other seats there are one hundred and four- teen at $1.50 and seventy at $2.00. Patrons and patronesses are being obtained, among whom is Mrs. Annie Nathan Meyer, a playwright of some note. Those having cars which they will volun- teer for the evening should communicate with Tue Cottectine NET office. As the play opens Marthy Palmer, mother of Ulys is found to have consumption, and the news brings great distress to the people in her household. Aunt Vistie Edney, who has a sharp tongue but a soft heart, meets the emer- gency with annoyance at the extra work it will mean for her. ‘Hard headed and practical; she is best described by another character in the play: “Always pilin’ bresh to kivver up the still.” ’ Ulys faces the situation by asking Sallie Tabor to care for his mother. Sallie is seventeen, ‘pretty, smouldering, with an almost sullen manner, the outcome of her unfortunate parentage and reputa- tion. Her vices are largely on the surface. The inner woman is proud, loyal, and admirable.’ Ulys’ family questions his motive in having Sallie live at the house, though he professes that he only hopes to make a better girl of her. Aunt Vistie’s comment on his desire to preach to the lowest of people is characteristic: ‘If ye lay down with dogs, y’ll git up with fleas.’ His motives are even more closely questioned by Lou Cabe who openly wants to marry him. Ulys becomes the victim of a tense conflict between his own determination to preach, his Aunt Vistie’s equal- ly strong determination to make of him a farmer and his forgivable human love for a girl. “Tight Britches” is one of the leading plays of this season in New York, and it is an unusual opportunity for the people of Woods Hole to have it made available to them at such reasonable prices and with free transportation provided. Be- side the contribution which can be made to THE CottectinG Net, your attendance at this play will provide an evening of unusual worth and entertaining value. The Tickets Are for Sale at the Office of The Collecting Net in Woods Hole Jury 7, 1934 } THE COLLECTING NET | WILEY BOOKS IN ZOOLOGY Published June, 1933 TEXTBOOK OF GENERAL ZOOLOGY By Winterton C. Curtis, Professor of Zoology, and Mary J. Guthrie, Associate Professor of Zoology; with the collaboration of Katharine R. Jeffers. In- structor in Zoology; all at the University of Mis- souri. A widely-used college textbook now available in a thoroughly revised and rewritten Second Edition. 588 pages 6 by 9 $3.75 LABORATORY DIRECTIONS IN GENERAL ZOOLOGY By Winterton C. Curtis, Mary J. Guthrie, and Far- ris H. Woods, Assistant Professor of Zoology, Uni- versity of Missouri. Revised in accordance with the new edition of the Textbook, 206 pages 6 by 9 $1.50 Published May, 1933 AN INTRODUCTION TO THE VERTEBRATES By Leverett A. Adams University of Illinois. This text has three purposes. 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It is the result of complete product control of lenses and mount- ings from the initial mathematical calculations to the final inspection. Knowledge of the optical instrument needs of science, in- dustry and education, years of experience, skilled workmen who have made precision optics their life work—are all factors in the production of optics of B & L quality. Even the preduction of the optical glass is completely controlled in the B&L Glass Plant.* Optical excellence has been combined with mechanical pre- cision and purposeful design to produce a new series of micro- scopes — the G Series, and the result is a research instrument of exceptional rigidity, stability, balance and performance. Every microscopist should haye complete information on these new microscopes. It will be sent to you if you will write to Bausch & Lomb Optical Co., 670 St. Paul Street, Rochester, N.Y. *Bausch & Lomb is the only manufacturer of scientific instruments in the United States that produces its own optical glass. B & L Microscope G G 1 E with mechanical stage, research letage and binocular eyepicce, Accessories are available which will equip this model for practically any type of research microscopy. FOR THE:SCIENCE OPTICAL IN Vol. IX. No. 3 SATURDAY, JULY 14, 1934 Annual Subseription, $2.00 Single Copies, 25 Cents. PRESENT NEEDS IN BIOLOGICAL OCEANOGRAPHY Dr. W. E. ALLEN Assistant Professor of Biology, Scripps Institution of Oceanography At the Symposium on Oceanographic Problems of the Ecological Society of America at its December meetings in Philadelphia in 1926, I delivered a paper which gave approximate expres- sion of my ideas on general biological problems at that time. Its title was “Pressing Needs in the Field of Biologi- cal Oceanography.” Publish- ed as Bulletin 13 ( Non-Tech- nical) of the Scripps Institu- tion of Oceanography. Now, IT have been honored by an invitation to bring this discus- sion up to date. Inasmuch as this publication is out of print, and not widely distri- buted, it seems desirable to quote certain parts of it for reconsideration or renewal. The views given here, as well as in the original paper, are purely individual. DEFINITIONS I have not changed my views as to a practical definition of biological oceanography, although (Continued on Page 63) ON THE SIGNIFICANCE OF THE VERTEBRATE ERYTHROCYTE Dr. WiLttAmM R. AMBERSON significance of MW. HB, H, Calendar TUESDAY, July 17, 8:00 P. M. Seminar: Mr. Henry I. Kohn: ‘‘The chlorophyl unit in photosynthe- sis;’” Dr. Eric G. Gall: “A potentiomet- ric study of phthiocol, pigment of human tubercle bacillus.” Dr. S. E. Pond: “Alteration in cal- cified tissues during drying (and their significance).” Dr. Arthur K. Parpart: “Solvent water in the erythrocyte.” Dr. William R. Duryee: ‘‘The re- lationship between the oxygen consumption and water content of the organism.” FRIDAY, July 20, 8:00 P. M. Lecture: Prof. John Runnstr6m: “Initiation of development and the metabolism of the sea-urchin Professor of Phystology, University of Tennessee I propose to speak to you this evening on the the would like to point out at the beginning that in my title there appears the qualifying word “on”, and that it is impossible for anyone to elucidate vertebrate erythrocyte. [| the full significance of this highly specialized cell within the confines of a one-hour lecture. I shall therefore ap- proach the subject from one particular point of view, simi- lar to that adopted by Profes- sor Joseph Bareroft when, in 1922 he delivered a lecture before the Harvey Society in New York City on “The Rai- son d'etre of the Red Cor- puscle.” It has become a classic in this field of thought. It represents one of the few attempts to arrive at some sort of philosophical position with regard to the significance of the red cell. The literature is crowded with detailed techni- cal studies of this or that special function, but there has been relatively little attempt to deal with the broader aspects of the problem. TABLE OF CONTENTS | Present Needs in Biological Oceanography, INeveals) Ohi IDOE So mgennnanncnagensoenmae 69, 70 Bie Vila a Allen ......-+++-++s++s++++++-+-5T at Desert Island Biological Laboratory...... 71 On the Significance of the Vertebrate Eryth- % : : | rocyte, Dr. Wm. R. Amberson ............. 57 The Biological Laboratory, Cold Spring Har- | The Chemical Nature of the Amphibian Or- WIS GIES SOT asp eO Dari Aue too UGH O DOLE 2 Peirabicgeye, TB a) Be eR 72h yl oly perea ieneipiees rec cee 66 The Cold Spring Harbor Biological Labora- | Water Changes in Trout Eggs at the Time tory Directory for 1934 ................... 73 | of Laying, Dr. L. Irving and Jeanne F. A Method for the Estimation of Changes in Manery .....- 1... eee e eee eee eee 67 the Rate of Intestinal Secretion, Dr. E. S. RCE OM Ae CMW eral slits iletny anes) siscatel tere. ofsie) eYoucy sabe 68 UN FISSO Dart cysic, che, syoccialotalas at rnereL ee etn eee te ce Cae 74 a < a * * THE MARINE BIOLOGICAL LABORATORY Jury 14, 1934] THE COLLECTING NET 59 As an introduction to this discussion I shall read a statement, partly direct quotation, partly paraphrase, which will put betore you tne gist of uarcroit’s argument ot twelve years ago. As | go on through the lecture I shall attempt to point vut to you those elements of his argument which still appear to be valid, and indicate other parts in which our present point of view must depart from his position. Barcroft argued as follows: “In the interior of the red blood corpuscle the hemoglobin exists in a world of its own. By this device nature has at a stroke increased the ef- ficiency both of the blood and of the hemoglobin. “It has saved the blood from possessing physical properties which the hemoglovin would otherwise confer upon it. If we assume that the concentration of the hemoglobin in the whole blood is 14% we arrive at the calculation, on the basis of data of Adair, that, at the hydrogen-ion concentration of blood, this amount of hemoglo- bin in solution would exert a colloidal osmotic pressure of nearly 140 mm. Hg.—a pressure higher than the arterial blood pressure, and per- haps 6-10 times as high as the hydrostatic pres- sure of the bleod on the capillary wall. Under such conditions the derangement of function would be so overwhelming as to demand a com- pletely different type of organism. Were the osmotic pressure ot the blood proteins 140 mm. Hg., the capillary pressure would have to exceed that figure in order that urine free from protein should be filtered through the glomeruli. But for the capillary pressure to reach five times its pres- ent value the power of all those organs which form the vascular system would require to be magnified to the same degree. All of these troubles are avoided by packing the hemoglobin in a corpuscle. “Another difficulty which is avoided is that of the increased viscosity of the blood which would follow upon the presence of the hemoglobin in solution. “It is a matter of common knowledge that if blood be laked hemoglobin appears in the urine; it is less commonly known that hemoglobin ap- pears in the lymph as well. The scheme of pack- ing the hemoglobin in the corpuscle then permits the meshes of the capillary lining-cells to be large relatively to the size of the hemoglobin molecule, and these correspondingly permeable to such sub- stances as must pass through them. “The device of the corpuscle has, on the chemi- cal side, saved the hemoglobin from being dis- solved in a solvent (NaC1) of less than maximal efficiency, and from functioning at a hyrodgen- ion concentration in which the remarkable proper- ties of hemoglobin as a respiratory pigment could not be properly displayed. A phosphate solution such as exists in the inside of the corpuscle is the most efficient medium known at present for the solution of the hemoglobin. Its functioning is facilitated by the slightly more acid reaction which prevails within the cell as compared with that of the plasma.” In criticism of this argument we must first of all face the fact that Barcroft made a_ serious error in his calculations of the colloidal osmotic pressure of a 14% solution of hemoglobin. In the lurst slide ] have taken a curve recently secured by Prot. H. S. Wells, of Vanderbilt University, who a year ago very kindly made some readings for us on the colloidal osmotic pressure of hemoglo- bin solutions. We see here a plot of his recent data, and of earlier data of Adair. Barcroft, in his calculations, used even earlier measurements by Adair, and appears to have made the error of using values which are valid only in the presence of a minimal amount of salt. They are not valid in the presence of such concentrations of salt as we know occur in blood. If we trace out on this curve the colloidal osmotic pressure corresponding to a 14% solution of hemoglobin we notice that the. true value is about 800 mm. of water, or 60 mm. Hg. only. This value represents the maximal possible col- loidal osmotic pressure of such a hemoglobin con- centration when dissolved in a salt solution is- osmotic with mammalian blood. The actual effec- tive colloidal osmotic pressure cannot possibly be so high, for reasons which will become more clear later. Briefly stated we may here say that hemo- globin penetrates with fair ease through the en- dothelial walls of many of the capillaries, as do the normal blood proteins, hence the effective colloidal osmotic pressure is less than that deter- mined in a solution outside of the body. We have recently made viscosity determinations on hemoglobin solutions and on whole blood, and have found that the liberation of the hemoglobin from the red cell produces little change in this physical property. Actually a 14% hemoglobin solution comes out to be slightly more viscous than normal cat or dog blood, but the evidence shortly to be considered will show that this dif- ference is of no practical significance. Chemical difficulties undoubtedly arise when the red cell is eliminated. Some of these are not so crucial as Barcroft believed. On the other hand we have now detected other chemical factors which he did not appreciate. Our evidence bearing upon this problem has been largely secured from a series. of animals, chiefly mammals to date, from which the normal blood has been removed and replaced by a solu- tion which we call “Hemoglobin-Ringer.” This fluid is made by laking a certain quantity of red cells in distilled water, then adding sufficient salts to bring the final solution to isotonic strength. It is possible, by the introduction of this solution through a jugular cannula simultaneously with a bleeding from the carotid artery, to remove all or 60 THE COLLECTING NET [ Vor. IX. No. 74 nearly all of the normal blood from cats, dogs, and rabbits. We have also carried out similar experiments on a few fish. After such removal of blood life continues for many hours. When | made our preliminary report here last summer our maximum longevity was about 15 hours after the bleeding. Since then we have increased the length of life to 36 hours in our best experiments to date. We have reason to hope that, with fur- ther improvements in our solution, we may be able to get still longer continuance of life. (Motion picture film showing recovery of con- sciousness in cats and dogs after replacement of normal blood by hemoglobin-Ringer. ) In this film you will note both the strength and the weakness of our evidence. The animals shown are not our best; we had to photograph those which were in the laboratory when the photographer arrived. You will note that the dyspnea observed during the bleeding (and part- ly due to the ether) persists for some time, even after the recovery of consciousness. More re- cently, with the use of higher hemoglobin concen- trations, we have eliminated most of this abnor- mality, and at the present time most of our ani- mals show no significant changes in respiration after the operation. The animals here seen show some difficulties in walking but on the whole possess remarkably good muscular coordination. They are able to move about the laboratory, to jump from considerable heights, to land on all fours when dropped up- side down and to execute swift movements, as when the cat attacked the dog. They frequently show a marked inclination toward sleep. Cats recover sufficiently to purr when petted, and dogs will wag their tails. On the whole dogs take the operation rather badly, but in some of our best experiments it has been impossible to distinguish between our experimental cases and normal animals. For the benefit of those who did not hear my preliminary report on this subject last summer (cf. CortectinG Net July 8, 1933) I wish to show one set of records having to do with the oxygen consumption of these animals before and after the removal of the blood. In the next slide we see the original records from which the oxy- gen consumption was determined. A dyspnea following the removal of the blood is evident. In the following slide are shown the values calcu- lated for the oxygen consumption in a number of determinations before and after the removal of the blood. You will note that there is a slight de- cline in the values with time, but there is no sud- ‘den change in the oxygen consumption when blood has been replaced by hemoglobin-Ringer. In many experiments we have found nearly iden- tical values before and after the operation. It is difficult to raise the hemoglobin concen- tration within the animal to equal that in the perfusing solution. This difficulty has become more and more apparent as we have increased the concentration of hemoglobin in solution. The concentration of hemoglobin circulating. in the animal rises for some time as the bleeding pro- ceeds, but never reaches full 100% of that pres- ent in the original hemoglobin-Ringer, even when only 7% of hemoglobin is present. When the hemoglobin is raised to about 14% the last bleed- ing sample shows somewhere around 12% only. This loss of hemoglobin concentration may be in part due to an increase in blood volume; in part it must be referred to other factors, shortly to be discussed, which bring about the reduction of the hemoglobin concentration with time. We have used, in succession, 5%, 8%, 12%, and 14% hemoglobin in solution. We have found that the animals live longer and longer the higher the hemoglobin concentration is made. In our best case the original solution contained nearly 14% hemoglobin, and life continued for 36 hours, contrary to Barcroft’s prediction that the physical properties of such a solution would make life im- possible. The spleen is an organ of importance.in these experiments. In normal animals the red cells at first diminish rapidly with each successive bleed- ing sample. There comes a time; however, when, over a number of samples, no further fall occurs. Inspection of the exteriorized spleen ‘shows that it exhibits first a gradual and then-a much more rapid diminution in size. While it is shrinking rapidly, toward the end of the bleeding, this pla- teau is observed in the curve of red blood cell number, showing that the effect is. due to the emptying of the splenic reservoir. The completeness of the removal of .the ned blood corpuscles may be checked in several ways. Dr. Pankratz, of our University, has made a number of histological studies, and has found that most tissues, including the marrow of the long bones, have been swept practically clean of red cells when as much as one liter of hemo- globin-Ringer has been passed through the body of a six-pound cat. The spleen can not be com- pletely emptied in this way; it holds on to some cells for a very long time. We have recently secured a new type of data bearing upon the completeness of the removal of the red cells. In splenectomized dogs we have often observed that in each successive bleeding sample, the red cells are reduced from those in the preceding sample by a definite percentage, nearly constant over the whole experiment. Re- cognizing the exponential character of the re- moval curve we may plot logarithms of red cell content in the various samples against total amount of fluid passed through the body before each sample (= a time scale). The resulting points arrange themselves in a straight line, whose slope is a measure of blood volume. : Jury 14, 1934 ] THE COLLECTING NET 61 By this method we have arrived at values for blood volume in the dog which range between 6 and 7% of the total body weight (spleen exclud- ed). These values are considerably lower than those recently reported in the literature for the same animal, and secured by indirect dye methods. We have made simultaneous direct and indirect measurements of blood volume, using brilliant vital red as the dye for the latter, and have con- sistently secured higher values by the indirect than by the direct method. We are led to believe that the literature figures obtained by the dye method are in error. Recently we thought it might be possible to simulate our conditions of bleeding in an in vitro control. We therefore took the figure of 720 cc. of blood, calculated as the blood volume of a cer- tain large dog, and measured out 720 cc. of ox blood into a reservoir, into which we then intro- duced, and from which we simultaneously with- drew, fluid at a rate and in a manner entirely similar to the animal bleeding. In the graph one observes that the red cells diminish in this con- trol by the same relationship which I have des- cribed for the whole animal, and that the slopes are very nearly identical. This identity establisnes the essential correctness of our blood volume measurements. We have made a preliminary reconnaisance of the manner in which both red and white blood corpuscles. return after the bleeding. Contrary to our hopes of getting indefinite survivals by re- placement of red cells, we have found only a slight rise in their number, after bleeding is com- plete, and these mostly come from the spleen. The white cells reappear much more quickly. Dur- ing the bleeding the percentage of red cells de- clines much more rapidly than does that of the whites. Thus in the table you will note that in a typical experiment the red cells were reduced, in the last bleeding sample, to 0.9% of those origi- nally present in the blood, whereas the whites were 7.2%. Ten hours later the reds had in- creased to 2.5%, but the whites had risen to 66.3% of their original count. The white cells are evidently much more easily mobilized. Such regions as the lymph nodes, and extravascular sites, must hold considerable quantities of white cells, which can later make their way back into the circulating hemoglobin-Ringer. As our duration of life increased we became aware that the circulating fluid drawn just before or at death was no longer of the same color as that of the original hemoglobin-Ringer, even after complete oxygenation. This observation led us to search for the presence of colored derivatives of hemoglobin. Spectroscopic examination of fluid taken at death showed that there are considerable quantities of methemoglobin present. In the next slide we can follow the rate of disappearance of oxyhemoglobin, and formation of methemoglobin, in an animal which gave us our best experiment to date. Endeavoring to carry out Barcroft’s “impossible” experiment, we used a solution in which the hemoglobin was slightly more than 13%. You will note the discrepancy, previously discussed, between the hemoglobin content of the original solution, and that of the last bleeding sample, which had about 12% only. The concen- trations of oxyhemoglobin at various times over the 36 hour experiment were determined by the Van Slyke, both in vitro and in vivo. Similarly total pigment was read by the spectro-photometer. In vitro at 38° C. total pigment remains constant, but oxyhemoglobin disappears at a fairly rapid rate, being transformed. into colored derivatives, mainly methemoglobin, which, at the end of the 36 hours, constitute about 40% of the total pig- ment. In vivo both total pigment and oxyhemo- globin decline, with methemoglobin constituting an ever larger fraction of the remaining total, until death occurs when the oxyhemoglobin has fallen to about 3%. Methemoglobin, you will recall, is the oxidant form of hemoglobin, electrochemically speaking, but contains no oxygen which can be readily made available for tissue respiration. It is, therefore, physiologically useless. When oxyhemoglobin, the physiologically use- ful fraction, is sufficiently diminished, the animals show all the signs of asphyxia. They lose con- sciousness and exhibit marked dyspnea and_ in- crease in heart rate, and shortly die. At autopsy the various organs of the body appear to be es- sentially normal in color, form and size, with the exception of the kidney. We have come more and more to appreciate the baleful effects of our solution upon the kidneys, as our longevity has increased. The kidney swells, and becomes al- most black in color. rom it may be expressed a fluid rich in methemoglobin, which appears to collect in this organ. The next two slides are photomicrographs of cat kidney sections taken some 30 hours after re- placement of blood by hemoglobin-Ringer. Cy- tological detail cannot be made out. The sections are fixed and stained by a method which shows the presence of iron in the tissues. The glo- merular capillaries stain very black, and are part- ly clogged with masses of hemoglobin granules. Much hemoglobin has passed out into Bowman’s capsule, and even higher concentrations appear to be present in the lumina of the nearby proximal convoluted tubules, as if some water had been re- absorbed. The tubule cells show little evidence of containing hemoglobin. Many of them, how- ever, show pathological changes undoubtedly pro- duced by it. The second slide shows a cross- section of about half a hundred collecting tubules from the kidney medulla. The lumina are filled with a darkly staining material, whereas the rest 62 THE COLLECTING NET of the section is clear. Hemoglobin is obviously readily eliminated through the glomerulus. Last summer at this laboratory Miss Dorothy Webster, Mr, Frank Engel, Dr. Edwin Laug, and I made an attempt to get information about the physico-chemical conditions which determine this loss of hemoglobin into the urine. Using per- fusion preparations of the frog kidney we allow- ed hemoglobin-Ringer containing about 1% hemo- globin to enter through an aortic cannula, We collected the urine formed through cannulae in the ureters. We studied the influence of the pH of the perfusing fluid upon the passage of hemo- globin into the urine, adjusting the reaction with phosphate buffers. We found that there is a very striking influence of pH upon hemoglobin elimination. At more acid values hemoglobin comes through in relatively high concentration (about 30% of the perfusion fluid value at pH 6.5) whereas in the more alkaline range relative- ly little appears (about 10% at pH 7.5). The effect is reversible in the same kidney when the perfusion pH is changed from acid to alkaline, or vice versa. We are still uncertain as to how far a differential reabsorption of water by the tubules may be responsible for this effect, and are con- tinuing the study, but believe that the phenomenon is glomerular in origin, and possibly connected with the dissociation state of the hemoglobin. The observation of such ready loss of hemo- globin into the urine led us to attempt the closure of this particular exit by removal of the kidneys. But even in nephrectomized animals both total pigment and oxyhemoglobin decline with time, and death ensues. We now recognize that while the kidney is an important exit point, it is not the only one. Histological studies show that hemoglobin is being taken up by cells of the reti- culo-endothelial system. In particular, the Kupffer cells of the liver become loaded with hemoglobin. Very recently we have begun studies of the acid-base equilibrium in our animals; to date we have relatively few experiments. In the last slide you will see values secured in one experiment. The normal blood of the cat gave pH 7.18. This acid condition is produced by the ether, as many workers have observed, possibly due to the pro- duction of lactic acid. Fresh hemoglobin-Ringer gave a value of 7.32. Circulating hemoglobin- Ringer, taken from the artery 15 minutes after the end of the bleeding, gave 7.23; a venous sample at the same time gave 7.16. Fifteen hours later the arterial sample gave 7.01, the venous 6.99; an hour later the animal died. A terminal acidosis is obviously present, in part due to the loss of buffering power as the hemoglobin con- centration declines. Very early in our work we found that hemo- globin-Ringer solutions containing the same amount of bicarbonate that is found in blood [ Vor. IX. No. 74 plasma are rapidly fatal. We were forced to re- duce the amount to 50 mgm. per cent. Hemo- globin in solution is itself quite an effective buff- er. We have the distinct impression that hemo- globin functions better in a medium slightly more acid than normal blood plasma, just as Barcroft argued. In summary of our experimental observations as they bear upon the significance of the erythro- cyte, we may now take the following position, Hemoglobin in solution is able to perform its respiratory role with sufficient efficiency to permit the continuation of all vital functions, including consciousness. The maximum possible colloidal osmotic pressure for a 14% hemoglobin solution is only 60 mm. Hg., and the effective colloidal osmotic pressure, because of the ready passage of some hemoglobin through the endothelial walls of the capillaries, is probably not more than half of this ; fluid exchanges in our experimental animals save only in the kidney, are normal. Such a solu- tion has a viscosity about equal to that of blood. It is not possible, therefore, to argue that the vertebrate erythrocyte exists in order to prevent a high colloidal osmotic pressure or a high vis- cosity. Its significance is best approached through an- other line of evidence. We observe that it is im- possible to maintain the concentration of hemo- globin present in solution within the circulating blood. It leaves the blood stream by various exits. These are: (1) An escape of hemoglobin into the urine, less at more alkaline reactions, greater at more acid values, but always appreciable at any pH compatible with life. (2) An escape into the tissue fluids and lymph. (3) A fixation of some of the hemoglobin by cells of the reticulo-endothelial system. (4) The conversion of hemoglobin into phy- siologically useless derivatives, the chief of which appears to be methemoglobin, although bilirubin and other colored compounds are also probably present. With the reduction in concentration of the hemoglobin circulating in solution, the buffering capacity of the fluid is diminished. An acidosis ensues which facilitates the loss of the remaining hemoglobin, particularly into the urine. Fluid balance in the kidney is destroyed, this organ be- comes edematous, exhibits high concentrations of hemoglobin in glomeruli and tubules, and shows definite pathology of tubule cells. When the oxy- hemoglobin concentration has fallen to 2 or 3% the animal enters a terminal asphyxia, character- ized by dyspnea, increased pulse, and loss of con- sciousness, shortly resulting in death. The vertebrate erythrocyte by holding hemo- globin within membranes impermeable to it, pre- vents its loss through these various exits. On the Jury 14, 1934 } THE COLLECTING NET 63 chemical side it keeps the hemoglobin in its re- duced state, electrochemically speaking, so that it may combine with Os in a readily reversible man- ner. Our animals appear to survive best when the circulating fluid is somewhat more acid than that normally present in the blood. These observa- tions appear to support the view that the slightly acid reaction of the red cell interior is of physio- logical importance in increasing the efficiency of Oz and COs transported by the blood. PRESENT NEEDS IN BIOLOGICAL OCEANOGRAPHY (Continued from page 57) I think that the following slight modification may be better than the original statement, i. e. The ocean is a mass of salt water covering most of the earth’s surface. Any information which concerns this mass of water and its contents may be said properly to belong to the science of oceanography. Any of this information which deals with living components of the ocean mass is biological oceanography. Several times in the seventeen years since | transferred my attention from freshwater plank- ton to marine phytoplankton (ocean pasturage), | have examined single drops of sea water under the microscope in order to get a better under- standing of the space relationships of minute organisms appearing in abundance great enough to attract notice. Ina few’ cases, | have found, in a single drop, the numbers of organisms visible under low magnifications to be some tens or scores. Of course, it is easy to get some hun- dreds into a single drop by using different methods of concentrating, e. g. centrifuging. In a single drop thus taken from the sea one can observe that moving organisms create or dis- turb movements of particles of water within the drop, that they affect the position and activity of other organisms, and that they must cause nu- merous chemical changes in the characteristics of the drop. Even the non-motile organisms may be seen to impede or deflect movements of their motile neighbors or of water particles, and it 1s certain that they also exert an influence on chemical changes in the drop. Thus, an examina- tion of a freshly-taken drop of sea water may show vividly that the living units involved in the composition of its mass are just as important as the unit particles of water, or the unit particles of other substances to be found within its boundaries. Of course, the above mentioned relationships were just as intricately entangled before the drop was taken from the sea, as well as being still further complicated by relationships to contiguous drops and neighboring masses. In reality, the conditions in the single drop of sea water could not be properly understood by studying water particles only, or chemical characteristics only, or living things only. But most biologists lack the ability to make delicate physical and chemical studies accurately, even when they have sufficient interest. So a biologist would naturally, give his attention to the organisms in the drop, even though knowing that their characteristics are in- fluenced by non-biological features. When he turns his attention to similar relation- ships in the ocean his interest in the strictly biolo- gical aspects must be concentrated (because of the vast extent of space and mass) and he may entirely forget that the characteristics of water particles and associated particles of other sub- stances are still more intricately concerned with the occurrence and characteristics of organisms than is the case in a drop of water. In that ex- treme he ignores the bounds of biological ocean- ography and becomes a mere biologist. In other words, biological oceanography must always in- clude with its biology a distinct recognition of the oceanic mass, and the problems of its char- acteristics and modifications in relation to or in conjunction with the living components as they occur in, Nature. 3ut, recognition of the realities of interrela- tionships between the living and non-living parts of the oceanic mass does not make the field easy to approach and explore. In no other realm of investigation is man held so nearly helpless by the barriers of space and time. The greatest and most fundamental oceanic problems are all pro- blems requiring careful investigation in four dimensions. This is most painfylly apparent in studies of biological oceanography, in which kaleidoscopic flashes of vital activities are seen amid the comparatively steady glow of physical phenomena. For relatively simple illustration take the fol- lowing: At a depth of sixty meters below a point five hundred miles directly east of Cape Hatteras, an observation of physical phenomena taken at 3 p. m. on December 30, 1926, would yield records which one might reasonably expect would approximate records taken at the same point at other hours, or days, or years. It would not even be surprising to find them the same at some other specified time. Not so in case of biological observations. There are likely to be differences in the total numbers of individuals of animals and plants, differences in the number of species found, differences in identity of species, 64 THE COLLECTING NET differences in the relative numbers of individuals of each species, differences in sizes of individuals, differences in their stages of development, and differences in their vigor. Furthermore, in immediate aspects, life in the sea is much more profoundly influenced by chemical and physical conditions than those con- ditions are influenced by living things. Where- fore, although one may get a good working knowledge of physical phenomena without giving thought to biological phenomena, he cannot go far in study of marine life without realizing the necessity for knowing a great deal about the physics and chemistry of the sea water in which it is observed. Consequently, a program of biological oceanography, however limited, must contain provision for physical and chemical studies of considerable moment. IMPORTANCE OF THE FIELD While most people may agree that biological oceanography is a fascinating field for investiga- tion, some may not understand that it has much claim to attention as long as so much remains to be done! on land, or in easily accessible fresh water. On this account{ it may be permissible to note briefly some of the claims recognized by students of marine life. First, there is the claim of the unsolved pro- blem upon the person who is able to recognize it. There would be no science, and there would be little progress in science, if all human beings ignored this claim. The most wonderful and use- ful discoveries in the history of the world have had their origin in attempts to solve problems which seemed petty, or useless, or hopeless to all except men with the urge and the will to know. Solution of some of the apparently insignificant problems of the sea may open the way to magni- ficent achievement. Second, there is the claim of the new field to service in supplementing the old. Many of the ideas derived from investigation of life on land, or in fresh water, may be clarified, or fortified, or rectified by evidence derived from study of life in the sea. Third, there is the claim of commercial in- terests. From food, clothing, and jewels to agricultural fertilizers, there is a long list of biological products of the sea, any and all of which may be better controlled and handled for human uses if more is known about the lives of their producers, and the conditions affecting their production. Fourth, there is the claim to aid in interpreting the history of the earth, the history of life on the earth, and the history of mankind, through com- parison of present forms and conditions of life ‘with those of the past. [ Vor. IX. No. 74 CONDITION OF THE FIELD In view of the establishment of the Woods Hole Oceanographic Institution, and of the be- ginning or strengthening of marine biological work in many localities throughout the world in the past eight years, my remarks on the condition of the field of biological oceanography in 1926; may seem to have become inappropriate to-day. However, it may help to remind us of the real magnitude of the field if I reiterate my statement of that time, the recruits to the number of exe plorers being relatively few, though actually numerous. So, “As to present condition, the field is practically unexplored. The work done about the shores of Europe, America, Asia and Australia, covers only part of the ground, at best, and it is not in any respect complete for any locality. The combined total of investigation throughout the world approximates only a negligible fraction of the investigation yet to be done.” Perhaps this statement appears too extreme without a little explanation or illustration, for which the history of marine biological investiga- tions affords excellent material. Unless I have overlooked some important information, the general region of Woods Hole has been the seat of investigations in marine biology second to none in the world. From the days of Louis Agassiz, Hyatt, Peck, and Verrill to the present time, a long line of biologists of outstanding ability has contributed single and collective studies of re- markable quality and extent. So far as the field of marine biology is concerned, it is evident that it has been largely explored, although it may be that no one would care to say that it has been ex- hausted. It may be supposed that there is still abundant opportunity for discoveries in anatomy, embryology, physiology, ecology, behavior, life cycles, genetics, and even taxonomy of many species of plants and animals in the region. Nor has biological oceanography been neglected. There has been more or less frequent scrutiny of the interrelationships between characteristics of marine organisms in the Woods Hole region and the Gulf Stream for at least fifty years. Tidal currents, inshore currents of various types, modi- fications of land drainage, bottom topography, and even meteorologic or climatic influences have been recognized and considered from time to time in relation to observed biological conditions. Yet it is my impression that even in the highly favored Woods Hole region, biological oceanography is still a virgin field. For example, perhaps the common barnacles are the most familiar and longest known of marine animals in the vicinity, but there is little information as to how their dif- ferences in vigor and abundance from day to day, season to season, and year to year, are related to differences in the characteristics of the sea water Jury 14, 1934 ] at any given time, or as to how its characteristics are influenced by differences in vigor and growth of the barnacles. It may be objected that the complexities of oceanic circulation near Woods Hole are un- favorable to exact determination of these in- fluences and relationships, but that does not alter the fact that opportunities for research are practically unlimited in this field of biological oceanography. Even in regions like those, oc- cupied by the oyster fisheries where highly valu- able biological information has been accumulated, it appears that there is much to be learned con- cerning the interrelationships of those organisms and the sea. It is extremely probable that the more satisfactory conditions of the fishery in one year, and the less satisfactory conditions in an- other, may be due to a considerable extent to local oceanic conditions at a time preceding the season of most intensive study. That is to say, even where the investigations of marine inter- relationships have been carried on most inten- sively, there is still ample opportunity for tracing them in greater detail, as well as in more remote contacts or connections. It is still more evident that such things are true of investigations of cod and mackerel relationships to surrounding waters, and of the causes of oscillations in abundance of such organisms. For some problemis of this type, the strictly biological researches have reached an advanced stage beyond which they can make little progress until investigations of the oceanic com- plex as a whole are pushed forward. In marine biology, one becomes accustomed to drawing his conclusions from observable condi- tions immediately at hand, or even subject to his control. In biological oceanography, according to my view, it is necessary for one to consider conditions and influences remote, as well as im- mediate. The key to the explanation of some local abundance of barnacles in 1934 may lie in a condition developed in the sea several hundred miles away, or in one which has been exhibited in local waters months or years before. I do not say that we are remiss for not knowing such things, but I do think that biological oceanography is the field to which they belong, and in which indefinite opportunities exist. NEEDS IN THE FIELD In 1926 I stressed the idea of need for better general organization. While I still think that such a need exists, I am inclined to abandon that position for two reasons; first, because organiza- tion of the type suggested then is not likely to be possible in practice ; second, because I believe that it ought to be approached or approximated as the result of development from local enterprises, after the latter have ascertained the basic require- ments of organization under differing conditions of operation. THE COLLECTING NET 65 For reasons which I assume to be obvious, I have not changed my views that biological oceano- graphy needs more workers, and more funds to support them and their activities. Directly con- nected with these needs of workers and resources are two others which seem to me of primary im- portance at present. They are the need for more effective and prompt distribution of information as it is accumulated, and the need for more ac- curate correlation of physical and chemical ob- servations with the biological observations in bio- logical oceanography. So far as I can seen, deficiencies in distribution of oceanographic information are due mainly to two things; first, to lack of assistance sufficient to assemble, classify, and otherwise prepare data and materials for reports, rapidly and promptly ; second, to lack of adequate provision for publica- tion and distribution of reports. A great deal of valuable oceanographic information does not get into print until ten or twenty years after the in- vestigations are made, and even then it may not be readily accessible to all in need of it. Rela- tively little of it reaches publication in less than five years, although it is true that much of it could be published in shorter time if investigators were supplied with proper clerical and technical assistance. However, it is true that even after a report is prepared it may have to wait from one to ten years because of lack of space or lack of funds to cover costs of printing. For the same reasons, a report may appear in an abbreviated form which fails to present certain results of a series of investigations clearly. The need for sufficiently close correlation of observations of other aspects of Oceanography with those of biological oceanography is not likely to be satisfied, largely because the demands for direct application of physical and chemical data‘ in non-biological lines are too great. A hydrogra- pher who has the training and ability to carry studies sufficiently illuminating for many bio- logical problems, is not likely to have the spirit oi sacrifice necessary to maintain his contacts with the biological work, or the patience to ac- cumulate physical data fundamental to the bio- logist while being insignificant to him. There are many times when a biologist would need to have continuous, or highly frequent ob- servations made in a limited area and at different depth levels, by physicists, chemists, and biolo- gists, to trace the relationships of certain features to observable biological conditions or occurrences. For example, a case of “red water” appearing in the plankton of a favorable locality at any time may seem to be highly important to a_ biologist (rather biological oceanographer), because of the opportunity to determine its extent, and to study it constantly throughout its existence in an effort to learn fundamental facts about its natural his- 66 THE COLLECTING NET [ Vor. IX. No. 74 tory and relationships. As I have already sug- gested, such a study could not be complete unless detailed physical and chemical observations were carried with a continuity similar to and in a man- ner supplementary to the biological studies. Yet, every experienced marine worker knows: first, that a physicist or hydrographer may not be in- terested in running his researches exactly to suit the biologist, even if run parallel; and second, that a competent physicist or hydrographer is sure to have a program, or series of problems of his own, which he is unwilling to interrupt or neglect to suit the needs of the biologist. Still, the diffi- culties opposing such correlations cannot alter the fact that the need for them exists in the field of biological oceanography. After all, it must be admitted that biologists are sadly deficient in detailed knowledge of important biological conditions, as necessary to full under- standing of problems in biological oceanography as are physical and chemical details. The detailed life history is known for surprisingly few marine plants and animals, and one can only guess at the effects of certain physical and chemical influences upon different phases of the life history of any particular organism. Knowledge of food chains is no better, although we know that some physical and chemical influences are exerted on or through the food or food chain. Exact information con- cerning details of geographical and seasonal dif- ferences in characteristics of particular species, or whole populations, is lacking or unreliable in most marine localities. Much valuable information in these lines can be obtained by biologists without close dependence on physical and chemical inves- tigations. And, it may be urged with propriety that biologists need not falter in attack upon prob- lems of biological oceanography, merely because correlative work in physical and chemical ocean- ography may be less than biologists may desire. Furthermore, there seems to be increasing recog- nition of the need for correlated work in all lines, and there is definite need to advance biological knowledge to better foundations for such corre- lated effort. THE CHEMICAL NATURE OF THE AMPHIBIAN ORGANIZER Dr. L. G. BartH Department of Zooloyy, Columbia University Il. THe Usr oF THE CEPHALIN FRACTION OF MAMMALIAN BRAIN AS AN INDUCING AGENT A number of recent papers have thrown some light upon the chemical nature of the substances which induce a neural plate in the amphibian em- bryo. The work of Holtfreter (1933) has shown that the dead organizer functions after treatment with xylol or 100% alcohol but loses its ability to induce a secondary neural plate in Triton after treatment with ether. Further, Spemann, Fischer and Wehmeier (1933) showed that the organizer is not destroyed by acetone. Finally Needham, Waddington and Needham (1934) were able to obtain induction in Triton with an etheral extract of neurulae. These researches indicate that some substance or substances soluble in ether but prob- ably insoluble in alcohol, xylol or acetone are re- sponsible for the early embryonic induction of the neural plate. Since the chemistry of amphibian neurulae is not well known, it was thought that extracts of the mammalian brain, a structure which Holtfre- ter (1933) has shown to have powers of induc- tion, would give more information about the chemical nature of the organizer. The chemistry of the mammalian brain has been extensively studied, and the cephalins have been analysed by Levene and Rolf (1922) and Page and Btilow (1931). ther extraction of the brain gives chiefly sterols, lecithins and cephalins. The sterols are soluble in acetone, which leaves the lecithin and cephalin. The lecithin may be separated from the cephalin since it is soluble in alcohol, which pre- cipitates cephalin. Bearing in mind that the or- ganizer is an ether-soluble substance and probably insoluble in alcohol and acetone, we might expect that the cephalin fraction of the ether-soluble sub- stances would act as an organizer. One half of a freshly killed calf’s brain was chopped fine and extracted for twenty-four hours at 5° C. with 500 ce. of ether. The suspension centrifuged and the clear ether extract evaporated to about 25 cc. at room temperature. Then 250 ce. of 100% alcohol were added with the formation of a precipitate, and the flask was placed at 5° C. for 24 hours. The precipitate was separated from the solution containing lecithin by centrifuging and was redissolved in ether. Not all of the pre- cipitate dissolved, and the residue after 24 hours in ether at 5° C. was discarded. The cephalin was reprecipitated from the ether by addition of acetone to remove cholesterol. The precipitate, consisting chiefly of cephalin, was redissolved in ether and once more precipitated with 100% al- cohol to remove lecithin and cholesterol. The pre- cipitate was again dissolved in ether and repreci- pitated with acetone to remove cholesterol. Fin- Jury 14, 1934 } THE COLLECTING NET 67 ally the precipitate was redissolved in ether and allowed to evaporate to dryness at room tempera- ture. The substance obtained was a white flaky compound which readily formed an emulsion with water. The cephalin obtained is obviously impure and further experiments are planned with better preparations. In the experiments described here, the cephalin was used 30-40 days after prepara- tion, and during this time must have taken up oxy- gen. The extract was placed in the blastocoel of the early gastrula of the black Mexican Axolotl and was found to induce a neural plate. The experiments show that cephalin, or some impurity in it, will induce a neural plate in axo- loti. It is possible that lecithin or cholesterol is present along with the cephalin, and these sub- stances have not been tested on axolotl. How- ever, Needham, Waddington and Needham (1934) have implanted cholesterol in Triton and obtained no signs of induction. In regard to lecithin, Spemann, Fischer and Wehmeier (1933) have implanted the yolk of the hen’s egg with negative results. These results of course do not conclusively rule out the action of specific brain sterols or lecithins. According to Page and Bulow (1931), cere- brosides and sphingomyelin may be present in cephalin if the alcohol is not carefully removed from the cephalin precipitate. Since these sub- stances are described as insoluble in ether, they could he present in only small amounts. How- ever, the possibility should not be discarded that one of these compounds may be the organizer, especially since Fischer and Wehmeier (1933) re- ported induction of neural plates with glycogen. Here, according to Needham, Waddington and Needham (1934) and Holtfreter (1933), the in- duction was probably due to an impurity in the glycogen. This is evidence that the organizer can function in very low concentrations. Considering cephalin itself, it is found to pos- sess some of the properties of the amphibian or- ganizer as indicated in the following summary : Property Cephalin Organizer occurrence brain and most brain and most of of tissues of tissues of mammals mammals Solubility in ether sol. sol. alcohol insol. insol. acetone insol, insol, xylol sol. insol. HCl insol. insol, H,O fine emulsion diffuses which diffuses The properties of the organizer are obtained from the papers of Mangold (1933), Holtfreter (1933), Spemann, Fischer and Wehmeier (1933) and Needham, Waddington and Needham (1934). It should be pointed out that “insolu- bility” in any solvent means merely that the or- ganizer will induce a neural plate after it has been treated with the solvent. This does not necessar- ily mean that the organizer is insoluble in the solvent used, but may mean that the organizer is in combination with some substance within the cell. On the whole it is not possible at present to dis- tinguish between cephalin and possible impurities in it as causing the induction in axolotl. I merely wish to point out here that cephalin possesses many of the properties of the amphibian organi- zer. WATER CHANGES IN TROUT EGGS AT THE TIME OF LAYING JEANNE F. MANERY AND Dr. LAURENCE IRVING Department of Physiology, University of Toronto It is a matter of common knowledge that fer- tilized eggs of many marine and fresh water or- ganisms have a higher water content than unfer- tilized eggs from the oviduct. Many investiga- tors have studied this dilution of the egg contents in fresh water forms by measuring the depression of the freezing point. Backmann and Runnstrom have found that frog oviduct eggs have a lower freezing point depression than fertilized eggs, while Runnstrom in 1920 and Svetloy in 1929 have demonstrated a similar difference between oviduct and fertilized eggs of trout. By directly weighing speckled trout eggs we have observed that fertilized eggs contain about 25% more water than unfertilized oviduct eggs. That the difference is due to water intake is shown by the fact that the dry weight, when ex- pressed in percentage of the wet weight, decreased about 25%. The chloride concentration, which was used to indicate any change in electrolytes, was also found to be 25% less in fertilized than in unfertilized oviduct eggs. This decrease, which is the same order of magnitude as the increase in wet weight, indicates merely a dilution of the electrolytes with-water. Many workers in other fields refer the water intake to the process of fertilization. Although such a reference is not erroneous in a sense, it nevertheless creates a wrong impression. In order to correct our impression we followed the changes in water content of fertilized and unfertilized rainbow trout eggs after laying. (Continued on Page 74) 68 THE COLLECTING NET [ Vor. IX. No. 74 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel W. Parker, and An- naleida van’t Hoff Snyder Cattell. Printed by The Darwin Press, New Bedford STANDARDS OF CONDUCT I. Two men representing a national encyclopedia came to Woods Hole last week in the interests of their series of expensive volumes and called on several investigators. They came into the office of Tue CottectinG Net in the afternoon and told us that Professor Conklin thought it would be desirable for us to print a review of their vol- umes. [Professor Conklin has asked us to mention the fact that he did not name a single person or or- ganization in Woods Hole or elsewhere to the agents of the National Encyclopedia as possible purchasers of their series. | Providing a favorable review was printed, they offered to give us a set of their encyclopedia which they said was worth $364.00. Naturally we informed them that our magazine was not con- ducted on this basis and reminded them that if they wanted to praise their series in THE CoL- LECTING Net they could do so at the rate of $22.00 a page in the advertising section. They apologized and agreed that our editorial policy was a wise one! Then their second plan was presented: would we write a letter recommending their encyclopedia if they presented the set of volumes to us with their compliments? To justify their proposal, the men produced a letter signed by a very dis- tinguished physiologist singing words of praise about their volumes. We were given to under- stand that their usual technique had been applied in obtaining it. One of the men, formerly pub- licity manager for a prominent radio broadcasting company, was asked what he thought of conduct- ing business on this plane. He did not like it, he said, but inasmuch as his organization had found its competitors resorting to the same questionable tactics they, in self defense, were forced to follow suit. It is no worse, he continued, than the pro- cedure used by the cigarette and other manufac- turers who pay handsomely for their testimonials. Not long ago the editor of a national scientific magazine was offered an expensive series of bio- graphical volumes if he would write a three-line letter commending them. This he of course re- fused to do. In proposing this arrangement a circular was exhibited in which were printed brief testimonials by distinguished men. It was imme- diately evident that their comments meant little or nothing. The same editor makes it his policy not to talk with authors concerning the manuscripts that are submitted to him in order that his judg- ment will not be influenced by their arguments and personality. It is difficult to make an absolutely impartial judgment if one is exposed to friendship, personal contact or gifts. If the distinguished physiologist in question was offered a set of the encyclopedia provided he would consent to give his opinion of the series, it is likely that his letter was couched in more favorable words than it otherwise would have been. Obtaining recommendations through bribery is a condemnable practice; it is unfortunate that commercial establishments can persuade even a limited number of scientific men to join them in such deceit. Introducing Dr. Borts Epurusst, who is spending the earlier part of this summer at the Marine Biological Laboratory, was born in Moscow, Russia, and at- tended the University there. After the Revolu- tion in 1919 he went to Paris and received his doctor’s degree at the Sorbonne. He lived in Paris until last January, when he came to America as a Rockefeller Foundation Fellow. Dr. EpHrusst spent the earlier part of the year in Pasadena at Dr. MorGan’s laboratory; though he likes the east, he considers California one of the most beautiful spots in the country. In Paris Dr. EpHrusst is in charge of the tis- sue culture laboratory at the Institute for physico- chemical biology. His earlier investigations there were on the development of the sea-urchin egg, its chemical composition and variations during the course of development, and especially its seusibil- ity or resistance to heat, and the temperature co- efficients of mitosis. During the last two years his work has been with tissue culture and is chiefly concerned with the laws of growth and regenera- tion in vitro. At the. Marine Biological Lahora- tory he is conducting investigations on regenera- tion in planarians. Dr. Epnrussi’s primary pur- _ pose in coming to this coyntry was for orientation in genetics, and to apply tissue culture methods to genetical problems. He will leave America for Europe on August 16. Jury 14, 1934 ] THE COLLECTING NET 69 FEEMS OF Dr. CLARENCE E. McCLunG, professor of zo- ology and director of the laboratory at the Uni- versity of Pennsylvania, has completed his year as Rockefeller Visiting Professor at the Keio Medical School in the University of Tokio. Dr. McCLunG was a member of the embryology staff at Woods Hole last year. He and Mrs. Mc- CLUNG are returning to America by way of Java and Ceylon. They will be in Cairo during August and will also visit the Zoological Station in Naples. Before sailing from Glasgow they will attend the meetings of the British Association in Aberdeen. Dr. AND Mrs. LAurENCE IRviING of Toronto are leaving Woods Hole with their two children on July 15 for the Atlantic Biological Laboratory at St. Andrew’s, New Brunswick. Dr. IRVING is an instructor in the physiology course at Woods Hole, and is associate professor of physiology at the University of Toronto Medical School. The dedication of the buildings of the Moun- tain Lake Biological Station of the University of Virginia will be held on July 21 in the laboratory building at 2:30 P. M. The principal speaker will be DEAN Harvey L. Price of the Virginia Poly- technic Institute. The exercises will be followed by a trip to Butt Mountain, after which a picnic supper will be held at the Cascades. Dr. JAMES LAWRENCE KeELLoGG has retired after thirty-five years of work in the department of biology at Williams College. He spent several summers in Woods Hole as an investigator at the U. S. Bureau of Fisheries and as an instructor at the Marine Biological Laboratory. ALLyNn J. WaterMAN from Brooklyn College has been ap- pointed assistant professor and will take over some of the work that Professor Kellogg pre- viously did. The Bureau of Fisheries is being conducted on a much reduced scale this year owing to a reduc- tion in appropriations. The main investigation is being done on the physiology and ecology of the oyster, under the direction of Dr. PauL S. Gatt- sorr of the U. S. Bureau of Fisheries at Wash- ington. His asistants are Dr. VERA KOEHRING, aquatic biologist of the Bureau of Fisheries, who is working on narcosis, and GEORGE REPPUN. STANLEY REED, JR., is aquarium attendant, and guide in the museum during the afternoon and evening. ‘year. INTEREST Dr. Norrts RAKESTRAW of Brown University and the Woods Hole Oceanographic Institution is traveling in Europe this summer. He left in April for a Mediterranean trip and then traveled north through Central Europe, Norway and Iceland. He is visiting all the principal oceanographic labora- tories in Europe and will study at several of them. During the midsummer Dr. RAKESTRAW will be at Kiel, Germany and Plymouth, England ; he plans to return to the United States early in September. Dr. H. B. Sternsacu, who has been working on a National Research Council Fellowship under Dr. Ratpu S. Litre at the University of Chica- go, has an extension of the Fellowship for next He will work at the University of Roches- ter School of Medicine with Dr. Epwarp F. ADOLPH, associate professor of physiology, and formerly an instructor in the physiology course at the Woods Hole laboratory. Dr. KatsumMaA Dan of the department-of phys- iology in the University of Pennsylvania, is ex- pecting to drive to California later in the season. He will be accompanied by Dr. DonaLp P, Cos- TELLO who is going to work at the Hopkins Ma- rine Station at Pacific Grove, California. Dr. Dan is leaving the United States for Japan where he will stay for some time. Dr, FRANK A. Brown who has been an inves- tigator at the Oceanographic Institution for the last two years, was married in June to Miss JEN- NigE PetticRove. The wedding took place in Ber- muda. Dr. Puitrp WHEELER and his wife, formerly Miss Betty Parsons, both of whom worked at the laboratory in 1926, 1927 and 1928, visited at Woods Hole over the week-end of July 7. Dr. Wheeler is now a surgeon in Brattleboro, Ver- mont. Miss Loutse Mast has announced her engage- ment to Dr. Heinz Specht, who has been working with Professor S. O. Mast in the Department of Zoology at Johns Hopkins University. Miss Mast ‘received her B. A. from Oberlin College and spent last winter doing graduate work at Johns Hopkins University. Dr. Epmunp S. Nasset of the University of Rochester, formerly instructor of physiology, has been promoted to the position of assistant profes- sor of physiology at the University. 70 THE COLLECIING NED [ Vor. IX. No. 74 ITEMS OF A COMPENDIUM OF CULTURE METHODS At the Atlantic City meeting of the A.A.A.S. a committee was appointed to consider the ques- tion of preparation and publication of a compend- ium of culture methods for invertebrate animals. The committee has found a very general feeling of need for a reference bcok containing reliable information that is needed by any one who is try- ing to rear and maintain cultures, and is proceed- ing with plans for it. It is proposed to have a short introductory chapter on general principles of management, with the main body of the book made up of signed articles, volunteered by indi- vidual workers and based on their own practical experience. The committee reserves the right to condense and to combine where necessary to avoid duplication. An effort will be made to include bibliography, summaries and cross-references to aid in finding things. The scope proposed is North American terres- trial, fresh water and marine invertebrates. teas hoped that the American zoologists will cooper- ate in this undertaking. Let any one who has tested out a reliable method of culture mainte- nance or any device that he has found to be par- ticularly useful to that end, write it up and submit it for a place in this book to any member of the committee. The following persons were appointed to serve on the committee: Dr. James G. NEEDHAM, Cor- nell University, chairman; Dr. PAUL Se GAmiss orr, U. S. Bureau of Fisheries; Dr. FRANK E. Lutz, American Museum of Natural History + and Dr. Pau S. Wetcu, University of Michi- gan. Detailed information regarding the work of the committee may be obtained at Woods Hole from Dr. Pau S. Gattsorr, U. 5S. Bureau of Fisheries Laboratory, Room 122. On Saturday evening, July 14, a small rowboat returning from a beach party at Nonamesset, was caught in the Hole and the passengers were for- tunate to escape a possible tragedy. The boat was swept between rocks off of Pine Island, and was overturned and wedged fast. The four people in the boat were Mr. anp Mrs. P. D. ApAms, Mrs. Wittiam Rosertson, and Dr. R. C. MACCARDLE of Temple University who arrived at Woods Hole on Saturday morning. The Coast Guard heard the cries for help and went to the rescue, but they failed to discover the party and returned report- ing that the boat was safe. Half and hour later Oscar Hiiton’ picked up the group in his motor boat: no one was injured. The rowboat was loosened from the rocks and towed in later by a motor boat, with no damage done but one broken seat. INTEREST THE M. B. L. CLUB The M. B. L. Club is now in full swing for the season’s activities. The club is greatly indebt- ed to the house committee, and in particular to Mrs. Moser, its chairman, for its zeal and con- tinued successful efforts in decorating the inter- ‘or of the clubhouse. This committee hopes to see the clubhouse and its facilities well used by the members. Dances will be held every Saturday evening, during the next few weeks, at least, contrary toa previous decision. There are plans to have one or two smokers during the season, to be held after the Friday evening lectures. It is hoped that these will serve the primary purpose of the club in bringing the various members of the scientific community, old and young, into social contact. Forums and informal discussion groups are heing organized with the assistance of Dr. DANTELLI. The ping-pong committee, under the chairman- ship of Dr. CosTELLo is planning a tournament, it is hoped that more timid players who do not feel up to tournament play will also avail them- selves of the ping-pong equipment. The library committee reports that their circulating library 1s in use. They have also subscribed to several New York and Boston papers and to current period- icals. Several persons have found the clubhouse a convenient place for letter writing and for read- ing during the day. "The music committee is particularly active with programs for Wednesday and Sunday evenings. The piano is available for practice during restrict- ed hours by arrangement with Mrs. SCHWEITZER of the music committee. The membership list of the club has shown a large increase during the past two weeks. There are still, however, several of its past members, and many persons in Woods Hole for the first time, who have not yet paid their dues. The club depends on these to inaintain its activities. Dues should be paid as soon as possible to Miss Cro- WELL in the M. B. L. office, or in Room 340 in the Brick Building. F. J. M. SICHEL. Dr. Henry H. Donaldson, member of the Wis- tar Institute of Anatomy and Biology and a mem- ber of the Committee on the Public Welfare Medal award, made the presentation address for the National Academy of Sciences when the medal was awarded to Dr. David G. Fairchild, tormerly of the U. S. Department of Agriculture, Washington, D. C., at the annual meeting. Dr. Fairchild received the award for his “exceptional accomplishments in the development and promo- tion of plant exploration and the introduction of new plants, shrubs and_ trees in the United States.’ Dr. Donaldson has just arrived in Woods Hole. Jury 14, 1934 ] THE COLLECTING NET 7\ 0 fer 4 oe The laboratory opened its 36th season on June 20th. The following biologists are working at the laboratory. Drs. WARREN H. ano MarG.ret Rk, Lewis, Carnegie Institution of Washington, are continuing their studies on tissue cultures of nor- mal and cancerous cells. They are being assisted by Perry S. MacNeav from the Michigan Med- ical School. Dr. Horr Hresarp, of Oberling Col- lege, is culturing various invertebrate tissues un- der the direction of Mrs. Lewis. Pror. ULrRic DAHLGREN AND CorNELIUS T. Kaytor, of Princeton University, are making a survey of the embryological forms found in Frenchman’s Bay. The large planktonic population and its daily fluctuations in species make such a survey a long and laborious task. Several summers will be nec- essary to complete it. Dr. AND Mrs. JAmeEs A. SHANNON of New York University Medical School, are investigating glomerular function of the Elasmobranch kidney. Later in the summer Mr. Hernz Specut, also of New York Univer- sity, will assist in the work. Drs. Witttam H. CoLe AND J. B, ALLISON are again using the barnacle, Balanus balanoides, as a test animal for determining the stimulating efficiencies of various organic and inorganic salts, certain non-electro- lytes and mixtures of the same. The Mt. Desert laboratory is one of the few places where barnacle reactions can be satisfactorily studied for long periods of time, because of the cold, uncontam- inated sea water at its dosposal. Mr. I. W. Sizer, of Rutgers, is determining the temperature char- acteristics of the response of Fundulus to the di- carboxylic acids in sea water to compare with the values found for fresh water solutions. Later in the summer Drs. D. S. Jounson Anp J. N. Coucu of Johns Hopkins University with two student assistants are expected to work on botan- ical problems, and Mr. F. G. Watton Smiru of the Royal College of Science, London, to study the embryology of Acmaea. WititrAm Crapstree of Bar Harbor is in charge of boats, motors and all collecting this year. He reports an abundance of dogfish. WAt- TER RusseLtt of Salisbury Cove continues his caretaking and general repairs. Only constant vig- ilance on his part this past winter saved the run- way and pier from the ice. As late as Easter the ice was 12 feet thick along the shore. In view of the fact that many piers were completely wrecked around Bar Harbor, the laboratory feels fortu- nate to have escaped with only minor injuries, The Mt. Desert Island Biological Laboratory Insulation of the 2100-gallon salt water tank by a six-inch air space has prevented a rise in temp- erature of the water of more than | degree C., even on the hottest days of this unusually hot summer so far. The freshly pumped sea water this year has not yet exceeded 12.0”, being usual- ly around 11.0°, so that the runing salt water in the laboratory aquaria has not exceeded 13.0°, Unless the temperature of the sea shows a phen- omenal rise this year, the maximum temperature of running sea water should not exceed 16°. The tank is pumped full twice daily. The uncorrected pH by phenol red is 8.32 + 0.03. Informal races are being planned for the mixed sailing craft of the community. Possible entries include the Friendship sloops of Drs. LEwis and SLACK, the Wee-Scotts of Mrs. M. M. Hoskins and Dr. E. K. MARsHALL, Jr.; the cat boat, Baca, of Dr. CoLr and the O-boat of Muss MIRIAM SLACK. At the annual Fourth of July picnic on the laboratory shore-front large quantities of steamed clams, salad, coffee and ice cream were con- sumed by the 44 persons attending. In the evening the SLACKs entertained the laboratory group at a fireworks party. Mr. A. S. JoHNsSoN, comptroller of Rutgers University, and his family are summering again in Salsbury Cove. The residences of Drs. NEAL, HeEGNER and H. W. Smiru, who are vacationing elsewhere, are being rented by Mrs. WILLIAM McCormick Brrr, the Misses DaBNey and GRUNOow, and the Henry SLACKS respectively. The laboratory property along the state high- way has been greatly improved by CCC workers engaged in beautifying the approach to Acadia National Park. The annual meetings of the Corporation and of the Board of Trustees will be held on Thursday, August 9th, at 3. o'clock. Election of officers and five trustees will be the chief items of business. Excellent board is being served at the dining hall by the Misses Martua AND SARAH Hayes of New Brunswick, N. J., and Miss MARGARET TALLMAN of Bridgeport, Conn. Workers at the laboratory are enjoying the new shop building where facilities are available for carpentering, painting and simple machine work, 72 THE COLLECTING NET [ Vor. IX. No. 74 THE BIOLOGICAL LABORATORG COLD SPRING HARBOR Among recent visitors to the Laboratory are Dr. AND Mrs. RatpH Wyckorr, Dr. DUNCAN MacInnes, Dr. L. G. Loncwortu and Dr. T. SHEDLOVSKY of the Rockefeller Institute; Dr. and Mrs. THero L. Jaun, Dr. Oscar W. RIcH- ARDS, Dr. AND Mrs. Harotp Mestre, and others from Yale University; Dr. Harry A. CHANNI- PER and members of his staff from New York University ; Dr. FELIX BERNSTEIN of Columbia; Dr. Stuart Mupp of the University of Pennsyl- vania School of Medicine; Dr. H. K. HARTLINE of the Johnson Foundation; Dr. ALEXANDER HoLvLeNDER of the Rockefeller Foundation; Dr. Hucu H. Darsy of Swarthmore; Mr. anp Mrs. Gus RyspacHok and Dr. AND Mrs. CHARLES Hopce of Temple University; Dr. anp Mrs. SWINGLE of Princeton; and Dr. P. S. HENSHAW of Memorial Hospital. Some of these have visit- ed the Laboratory several times as participants in discussions concerned with the symposia. Dr. Mupp and Dr. SHEDLOvsKyY will be in residence for a few weeks beginning August first. Dr. CHarctes B. Davenport retired recently from the directorship of the Department of Gene- tics of the Carnegie Institution of Washington at Cold Spring Harbor. Previous to his retirement he delivered a paper at the Institution covering the early history of the founding of the various laboratories at Cold Spring Harbor, and the de- velopment of the Department of Genetics of the Carnegie Institution. Following his paper a state- ment of appreciation was made by Dr. Oscar Rippte of the Department. Dr. E. CARLTON MacDowELt, on behalf of workers in the Depart- ment, presented Mrs. Davenport with a bouquet of roses, and Dr. Davenport with a fund for the purchase of books for his private library, together with bookplates having the signatures of all the workers in the Department. Dr. DAveNPort has been appointed Research Associate in the Institu- tion and is conducting his work at the Eugenics Record Office. Dr. A. E, BLAKESLEE has been named Acting Director. Of the students in the course in surgical meth- ods in experimental biology, four hold Ph.D. de- grees, three are undergraduates, the others are graduate students. The course is normally limited to twelve students. This year due to a misunder- standing, fifteen were admitted. And even so some desirable candidates had to be rejected. Miss CATHERINE R. Brown, secretary at the Laboratory for ten years, recently married Mr. John MacLeod, assistant to Dr. Ponder. Mr. and Mrs. MacLeod have just returned from their honeymoon in Canada. Dr. Ertc PonbER’s monograph on “The Mam- malian Red Cell and the Properties of Haemoly- tic Systems” has recently appeared as Volume 6 of Protoplasma-M onographen. Dr. Harotp A. AspRAMSON’s new book, “Elec- trokinetic Phenomena and Their Application to Biology and Medicine’, published by The Chemi- cal Catalog Company, became available for distri- bution early in the month. Three members of last year’s conference-sym- posia have joined the staff of the Laboratory and one has been elected to the Board of Directors. Dr. Stuart Mupp, professor of bacteriology at the University of Pennsylvania School of Medi- cine, is on the Board. Dr. Eric PoNnpEr has joined the all-year staff, while Dr. Harotp AprRAMSON and Dr. KENNETH S. COLE are mem- bers of the summer staff. A party was given for the Abramsons to cele- brate their first wedding anniversary. On Monday evening July 9, Dr. GEorGE W. CoRNER gave a historical lecture to which lay- members of the Association were invited. His subject was “Medicine in the Poems of Chaucer.” Regular evening lectures delivered thus far are as follows: Wednesday, June 27—Dr. GEORGE W. CORNER, “The Present State of the Corpus Luteum Question.” Friday, July 6—Dr. Hans MUtteEr, “Cooperative Phenomena in Physics.” Wednesday, July 11—Dr. Grorce L. Crark, “X- rays as a Research Tool in Biology.” Friday, July 13—Dr. Eric Ponper, “The Factors Controlling the White Cell Count.” The paper of Proressor W. AstBury to be read by Dr. Clark in one of the symposia did not arrive from England in time to be given on the day originally scheduled. It will be presented to- gether with the papers already announced for Monday, July 23rd. Jury 14, 1934 ] THE COLLECTING NET 73 The Biological Laboratory Directory for 1934 INVESTIGATORS, ASSISTANTS and TECHNICIANS Abramson, H. A. research, assoc. bacteriol., Cornell Medical School. Bacon, Annette L. ass’t, Temple. Blanchard, Ernest W. inst., assoc. biol., Bryn Mawr. Bozler, E. research, fellow med. physics, Johnson Foundation School of Medicine. Brink, Frank Jr. research, Pennsylvania State Col- lege. Cain, Stanley research, ass’t prof. bot., Indiana. Chouteau, Ellen M. librarian, Biol. Lab. Clark, George L. research, prof. chem., Univ. of Il- linois. Climenko, Robert research, lect. pharmacol., Cornell. Cole, Kenneth S. research and inst., ass’t prof. physiol., College of P. and S., Columbia. Cole, Robert H. research, undergr., Oberlin. Conard, H. S. inst. and research, prof. bot., Grinnell. Corner, George W. inst., prof. anatomy, Univ. of Rochester School of Medicine and Dentistry. Cunningham, Bert inst. and research, prof. biol., Duke. Curtis, Howard J. research, physicist, Biol. Lab. Deery, Edward glassblower, Bell Telephone Labs., Fricke, Hugo research, in charge biophysics, Biol. Lab Fulton, MacDonald, research, dept. biol., Brown. Gallagher, D. M. research, radio engineer, Biol. Lab. Galligar, Gladys research, dept. bot., Univ. of Illi- nois. Gaunt, Josephine research. Gaunt, Robert research, prof. biol., Charleston. Gold, Dorothy secretary, Biol. Lab. Harris, R. G. director, Biol. Lab. Hart, Edward research, chemist, Biol. Lab. Haterius, H. O. research, ass’t prof. biol., Washing- ton Square, New York Univ. Hinchey, M. Catherine dept. biol., New Jersey State Normal School. Keen, Maurice F. research, inst. biol., Temple. Kornhauser, S. I., inst. and research, prof. embry., Univ. of Louisville Medical School. Leitch, Maurice L. research, inst. biol., Temple. Marsh, Phronsie secretary, Biol. Lab. MacLeod, Catherine secretary, Biol. Lab. MacLeod, John research, research ass’t general physiol., Biol. Lab. McAfee, Janet Brokaw research, ass’t, College of P. and S., Columbia. McCurdy, Harriet (Mrs. Blanchard) research. Muller, Hans research, prof. physics, Mass. Inst. Tech. Oltman, Clara research, inst., Brooklyn College. Parkins, Phyllis. Parkins. W. M. research assoc., Princeton. Parks, Mark E. research, inst. biol., Washington Square, New York Univ. Peterman, Robert research, Gettysburg College. Ponder, Eric investigator general physiol., Biol. Lab. Otto research, prof. bacteriol., Cornell. Rashevsky, Nicolas research, dept. of physiol., Univ. of Chicago. College of Robinson, Ellis J. research, ass’t biol., Washington Square, New York Univ. Sargent, Louisa M. inst., ass’t prof. bot., Grinnell. Schaeffer, A. A. research, prof. biol., Temple. Spieth, Herman T. inst., inst. zool., College of City of New York. Stoudt, Harry N. research, inst. biol., Temple. Taylor, I. R. inst. and research, ass’t prof. physiol., Brown. Twitty, Victor C. research, ass’t. prof. zool., Stan- ford. Victoreen, Ernest research, technical ass’t, Biol. Lab. Walp, Russell Lee, research, prof. bot., Marietta College. Walzl, Edward research, fellow biol., Johns Hopkins Univ. Winsor, C. P. research, dept. physiol., Harvard. STUDENTS Ballard, W. W. inst. zoology, Dartmouth. Bifoss, Rev. Callistus G. prof. biol., Quincy College. Boettiger, Edward grad. stud., Brown Univ. Bookman, John J. undergr., Brown. Booth, Lois grad. stud., Pittsburgh. Brambora, Erna C. grad. stud., Brooklyn College. Cone, Avis Wood, Montreal. Corner, George. Curtis, Brian C. grad. stud., Stanford Univ. Downes, Nancy undergr., Barnard. Finkelstein, Allen, grad. stud., Bellevue Hospital Medical College. Fraad, Daniel J. undergr., Brown. Fuller, Caleb A. grad. stud., Brown. Harrison, James S. undergr., Brown. Harwood, Paul H. Jr. undergr., Princeton. Himmelright, Mary Eleanor grad. stud., Maryville College. ry Hunt, Thelma, ags’t prof. psychology, George Wash- ington University. Hunt, Thomas E. prof. histology and embryology, Alabama, School of Medicine. Jones, Herman D. assoc. prof. organic and bio-chem- istry, Alabama Polytechnic Inst. Kleinberg, William grad. stud., New York Univer- sity. Laity, Elsie M. grad. stud., Grinnell. Lytle, Theodore L. undergr., Princeton. Maynard, Francis L. grad. stud., Brown. Ochs, I. L. undergr., Wesleyan. Ross, Donald grad. stud., Grinnell. Rouse, Sylvia B. grad. stud., Brown. Rubin, Morton A. grad. stud. and ass’t general physiol., Clark. Taylor, C. R. inst. biol., Shaw. Tobin, Charles E. grad. stud., College of Charles- ton. tum Suden, Caroline, research fellow, Evans Mem- orial hospital and Boston Univ. School of Medi- cine. Vandam, Leroy K. grad. stud., Brown. Wahlers, Alice M. grad. stud., Adelphi College. Wawro, N. W. grad. stud., Brown. Whitaker, Elizabeth A. grad. stud., Brown. Young, Lawrence E. undergr., Ohio Wesleyan. 74 THE COLLECTING NET { Vor. TX. No. 74 WATER CHANGES IN TROUT EGGS AT THE TIME OF LAYING (Continued from Page 67) The great variation which we observed to exist in the sizes of eggs from different speckled trout females necessitated following water changes in the eggs of a single rainbow trout. A sample of eggs was procured immediately after extrusion from the oviducts, and the remainder was divided into two lots. One lot was then fertilized and both were covered with stream water and left un- disturbed for about an hour to “harden up”. Samples were taken at certain intervals of time and their wet weights and dry weights (after dry- ing for 18 hours at 100°) were found. It was observed that at the end of the first hour after laying, the wet weight of both fertilized and unfertilized eggs had increased from 65 to 77 mgm. per egg. No further increase occurred up to 40 hours and other data show no subsequent changes in wet weight up to a period just prior to hatching. The dry weight of both lots at all in- tervals of time studied was identical with that of oviduct eggs. Using the decrease in percentage of dry substance and the decrease in chloride con- centration as indicators we were able again to conclude that only an intake of water had oc- curred. Similar results were obtained on the eggs of three separate females. We concluded, then, that trout eggs, fertilized or unfertilized, take in the same quantity! of water during the first hour after laying; and we would in the future refer such water intake not to the process of fertiliza- tion but to the process of laying. The mechanism involved in this water intake presents a problem. Eggs with depressions of the freezing point of about —0.5 to —0.6° are placed in fresh water, which results in a high osmotic gradient being set up between the interior of the egg and its external environment. If, however, one postulates osmotic forces to be involved in the entrance of water into the egg, one finds it difficult to explain the arrest of its entrance at the end of one hour. Although we do not propose to attempt an explanation, nevertheless certain facts are clearly established by a simple study of this kind. In the first place oviduct eggs are sur- rounded by a delicate membrane. They are soft and lack a perivitelline space. When placed in fresh water the membrane thickens, becomes firm, tough and elastic, a perivitelline space forms, and at the same time the yolk becomes surrounded by a cytoplasmic layer. In a relatively short time two membranes are formed. Runnstrom_ has shown that their formation depends on a fresh water environment and will not occur in sea water. The rapidity with which the membranes form and the fact that the chorion seems to be the result of a condensation or flocculation of col- loid-like substances suggests the occurrence of some physical reaction at the surface of the egg which depends on its contact with fresh water. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 10). A METHOD FOR THE ESTIMATION OF CHANGES IN THE RATE OF INTESTINAL SECRETION Dr. E. S. NASSET Assistant Professor of Physiology, University of Rochester In the search for the humoral agent concerned in the secretion of succus entericus it was desir- able to have an acute animal preparation similar to those ordinarily used in the study of pancrea- tic and salivary secretion. In the dog there are no intestinal ducts which can be cannulated and, therefore, it was necessary to use a portion of the mucosa. Since the mucosa performs the double function of absorption and secretion some differ- ential method was required. The Wells modification of the Hamburger technic for the study of fluid exchange in the in- testine was found to be satisfactory for the pur- pose. The method is based on the fact that at an intraintestinal pressure of about —15 cm. of water the apparent absorption or secretion ceases. The loop of intestine is kept at constant volume by a helical wire coil in the lumen. Appropriate chemical analyses and enzyme studies of the in- testinal contents were made at different pres- sures, both above and below the apparent zero point. It was shown that, within the range of pressures studied (—5 to —25 cm. water), it is the secretion process which changes with changes in pressure. The absorption of glucose was in- dependent of pressure within the same range. It is felt, therefore, that changes in the volume of the contents of such a preparation can be said to represent changes in the volume of the secretion. (This article is based on a seminar report presented at the Marine Biological Laboratory on July 10.) Jury 14, 1934 } THE COLLECTING NET hee MeUSe SSeS SE SES SUSL StS SE Se 0%. 8,08, 00 897,00. 80,8°0,04, 00.00.06, 06,04,00,04,06, ES padscetisteetetatehaPatiOetatvatvetvetvatvaDvatvatvetvetvatvetvatvetvatvetostvaty ‘oe’ ‘ev'es'ov'os'oe' ‘ov'evos'os'es'os'es'os'os Tavlevevies's sas'esasiesiasasteseeesesesesiesesiesiesesasesaseses esos esesecesesaesecesevesesesesieceseciesieses SCOMPLETE Relaxation for Sustained OBSERVATION @ ONE of the primary purposes in designing the Spencer Lens series of research microscopes was to make the phy- sical part of the scientist's work more comfortable. N lun ait 208, =, ‘ee'oe ECC eesales eases eased ev edevedodovevedevedededededevededetevevelecede SUSUAEREARE AERA AS PEPE EE AE OO 808088 00,09 8008089188 SU 8980900000 00.0000 CeCe eee es eee er ee ee er ee eS ee ee ee ee veka hekaketitetiietetete ee ee ee ee ee er et ero’ ve esires The man who sits down to operate one of these instru- ments is at once aware of the velvet-like smoothness of the operation of all adjustments. He is well aware of the superiority of Spencer optics. But suddenly, after continued observation, he realizes that all physical strain has been eliminated. $4,299 0,0 992,928,009 699,90,20,99, Voe'ee'eeoe'es esos on ee ee ee ee" & "exes" In reading, one’s attention is concentrated on the mean- ing of the words. In using a Spencer research microscope, BY his attention is concentrated on interpretation of the is Spencer Research Micro- specimen. i pecreaNo-as JECecisely 1. Eyes are used as naturally as in reading. x built for the practical re- 3% search worker. Low-type 2. Head is held in normal reading position. it fine adjustment, Inter- ghangeablabincculcrand 3. Body is as relaxed as when reading in an arm chair. i] monocular body tubes. : 5 3 Write for Folder M-56-C. All adjustments (except the coarse focusing) are placed 33 so they can be manipulated by the hands resting comfort- 3 ably and relaxed on the table. The designers of Spencer £Y3 research microscopes are aware of the trying conditions of 3% research work. 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At Junior and Senior High School Levels (Just Published) 1 By GeorGE W. Hunter, Pu.D., Lecturer in Methods of Education in Science, The Claremont Colleges and University of Southern California, Los Angeles, Cal. 560 pages. List Price $2.50 A COMPREHENSIVE survey of the latest and most approved methods of the teaching of science in junior and senior high schools. Throughout, the book is vitalized by the author's rich philosophy evolved from his many years of successful teaching of science in both secondary schools and colleges. Tue newest methods and concepts are here explained clearly and fully. Emphasis is placed on the need for making science teaching of direct value in training pupils for citizenship. Chief attention, how- ever, is focused on the ways by which teachers may deal with classroom problems in a sane, sympathetic, and realistic fashion without yielding to hasty fads or generalizations. Eacu chapter begins with a preview and closes with a comprehen- sive series of study questions and a helpful list of reading references. Generous use is made of citations and quotations from other publica- tions. Charts, diagrams, score cards, etc., give graphic aid to the text. AMERICAN BOOK COMPANY New York Cincinnati Chicago Boston Altanta San Francisco 0 0 wc cece oc ee 00 0 0 ee eo. Jury 14, 1934 ] THE COLLECTING NET 77 Biological Specimen Dishes Now in Two Sizes The small Dish has a capacity to the brim of 350 cc., inside height 45 mm., inside diameter 100 mm., height overall 50 mm. The large Dish has a capacity of 1750 cc., inside height 70 mm., inside diameter 175 mm., height overall 80 mm. Both Dishes are made from clear heavy glass. The bottoms are flat and the Dishes will stack perfectly. It is applicable to work in embryology, especially with chick embryos; to small aquatic organisms, living or preserved; to the development of Echinoderms and other eggs. 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EIMER & AMEND Est. 1851 Inc. 1897 Headquarters for Laboratory Apparatus and Chemical Reagents Third Ave., 18th to 19th St. NEW YORK, N. Y. THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and 37 plates, published January, 1934 This guide for authors, in preparing manu- scripts and drawings for the most effective and economical method of publishing biologi- cal research, has been prepared by the Staff of The Wistar Institute Press and the codper- ative efforts of more than fifty editors con- cerned in the editing of journals published by The Wistar Institute, and presents the con- sensus of opinion on many points relating to the mechanical preparation of manuscripts and drawings for the printer and engraver. Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. 73 Be er! THE COLLECTING NET [ Vo. IX. No. 74 drum Fixed stage, 12 cm. square Illuminating apparatus with rack and pinion Condenser 1.2 with iris Triple revolving nosepiece Fine adjustment with graduated CARL ZEISS PHYSICIANS’ AND LABORATORY MICROSCOPE E S C-106 Magnifications 56-900x Achromatic objectives: 8 na. 0.20 40 n.a. 0.65 90 n.a. 1.25 oil imm. Huygens oculars 7x and 10x Price, $152.00, f. 0. b. N. Y. A good dark field outfit is obtained by adding: Cardioid con- denser $26, extra for oil im., with iris $4. Compensating ocu- lar 15x: $11. ; A satchel type of carrying case can be supplied instead of the standard cabinet at an additional cost of $4.00. CARL ZEISSoclNiee 5eabaietirtihne Xivae couse E OR (Oye. 1S Dor 0 0: Electrokinetic Phenomena and Their Application to Biology and Medicine By HAROLD A. ABRAMSON, M.D. A.C.S. Monograph No. 66 Tus monograph deals with singular uni- formity of purpose, with the chemistry of the surfaces of proteins, organic and inorganic substances, blood cells, spermatozoa, bacteria, immune substances and tissues as determined by electrokinetic methods. It is unusual in its treatment because the available experimental data have been clearly related by suitable cal- culations to the modern theory of electrolytes, and it provides the reader with an invaluable means of examining experimental results in the light of recent theory. For those interested in industrial applications there is an appendix with a rather complete list of references to the patent literature. 332 Pages Illustrated 728 So. Hill Street LOS A N’'‘G EVESESS CHAPTERS Historical Early Theory and related Experi- ments: Part I, Classical Theory of the Rigid Double Layer—Part II, Early Confirmation of Theory. Methods—Recent Theory and Related Experiments—Proteins and Some Related Com- pounds—General Effects of Salts on “Inert” Sur- faces—Inorganic Surfaces—Organic Surfaces — Gases—Blood Cells, Spermatozoa, Tissues, etc.— Bacteria, Antibodies, Viruses and Related Sys- tems—Appendix I. Notation—Appendix II. Con- stants and Conversion Factors—Appendix III. Patent Literature—Author Index—Subject Index, $7.50 The Chemical Catalog Company, Ine. 330 WEST 42nd STREET Nemec oem, NEW YORK, N. Y., U.S.A. Jury 14, 1934 ] THE COLLECTING NET wre 79 Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, etc. Catalogs will gladly be sent on request. Please mention name of school and Spalteholz subjects taught, to enable us to : = Transparent send the appropriate catalog. Fite tery Preparations : os oe nates Human ~U as A (ee i Cusv-Apams Company Zoological 25 EAST 26th STREET NEW YORK Model of Human Heart Visit our display rooms and museum. High Speed for Greater R. C. F. Relative Centrifugal Force, about nine times greater than that of the ordinary centrifuge, can be obtained with the Multispeed Attachment for samples of material up to 40 ml. capacity. Increasing speed from 3,700 R.P.M. to 18,000 R.P.M. increases the relative centrifugal force at the tip of the tubes from 2,800 x gravity to 25,000 x gravity. The International Multispeed Attachment with a maximum speed of 18,000 R.P.M., can be used in Size 1, Type SB and Size 2 Centrifuges, when equipped with welded steel guard. The Attachment consists of a duplex support for a secondary high speed roller bearing spindle and pulley, a belt tightener spindle and pulley and a bronze driving pulley for the centrifuge shaft. The great centrifugal force calls for the best available material (Duralumin) in the No. 295 head which holds six tubes of 7 ml. capacity at an angle to conserve space. The head is coni- cal in shape and highly polished to reduce air friction. Send for 1934 Bulletin MS INTERNATIONAL EQUIPMENT CO. 352 WESTERN AVENUE BOSTON, MASS. Size 1, Type SB with Multispeed Attachment = 7S Makers of Fine Centrifuges _THE COLLECTING NET [ Vor. IX. No. 74 —— VISIT THE BAUSCH & LOMB EXHIBIT j JULY 23 to 31 Inclusive IN THE OLD LECTURE HALL | B-& L Centrifuge Microscope (after Harvey Loomis) F the excellence of microscope optics is to be fully utilized, precise construction of mechanical parts is absolutely necessary. The B & L Centrifuge Microscope is evidence of the Bausch & Lomb mechanical ability. With this instrument you see a continuous picture of micro-organisms being centrifuged at the rate of 10,000 r. p. m. The specimen is traveling four miles per minute and is sub- jected to a centrifugal force equal to 12,000 times gravity! The Centrifuge Microscope is a mechanical as well as an opti- 5S cal achievement — and the same mechanical excellence is built into every B & L instrument. For information on the Centrifuge Microscope or other B & L Optical Instruments, write to Bausch & Lomb Optical Company, HO UEIEN LGR cere) or 670 St. Paul Street, Rochester, New York. above is one of the new H series. Binocular vision, divisible body tube, and 115 x 130 mm stage are characteristics of this instru- ment, It is built with the same mechanical precision as the Cen- OPTICAL INSTRUMENTS FOR THE SCIENCES Vol. IX. No. 4 SATURDAY, JULY 21, 1934 Annual Subseription, $2.00 Single Copies, 25 Cents. EXPERIMENTAL STUDY OF SEX-DIFFER- ENTIATION IN THE CHICK EMBRYO Dr. B. H. WILvieR Professor of Zoology, The University of Rochester The talk tonight is a sort of comprehensive re- view of the experiments we have been doing on the subject of sex-differentiation in the chick em- bryo. The principal phases of the subject to be considered are largely an out- PHYSIOLOGY OF DETERMINATION IN THE SEA-URCHIN DEVELOPMENT Dr. JouNn RUNNSTROM Professor of Experimental Zoology, University of Stockholin In the following paper I shall present outlines of the studies on the mechanics of development in the sea-urchin, which were carried out by mem- bers of the staff of the department of zoology in Stockholm. growth of experiments, de- signed primarily to analyze the action of the sex-hormones =! 2 i. Fig. 2 shows a scheme of alendar ain - A - Calendar the division of the egg of the TUESDAY, July 24. 8:00 P. M. sea-urchin, Paracentrotus. In in the embryo. We set out es : ; the 16-cell stage (d) three originally to duplicate in the Se ae meer ene aa layers of cells can be recog- chick embryo natures exper- they change without chromos- nized, the mesomeres, the iment in cattle twins, where omal inversions or transloca- macromeres and the micro- you will recall from Dr. tions?” meres. In the next stage (e) Lillie’s studies, the female Dr. Roberts Rugh: “Ovulation and ‘Ai RRS ONTASTROS Emre Chala imi twin is modified: in the male direction apparently by the action of sex hormones from the male twin. By combining a gonad rud- iment as a graft with a sex- ually differentiated host em- bryo a relationship is set up which simulates rather closely egg transport in the frog.” Dr. John W. Everett: ‘Certain un- usual cytoplasmic elements in the yolk-sac epithelium of the white rat.” Dr. Henry J. Fry: ‘Notes on the behavior of asters.” FRIDAY, July 27, 8:00 P. M. Lecture: Prof. Rudolf Héber: “Ex- two layers called by Horsta- dius an; and ans. The macro- mere material is separated into two layers designated by Horstadius as veg; and vegs in the 64-cell stage (f). The limit between the future ecto- derm and entoderm coincides that existing between cattle twins in utero. That is, the signiicant means of communication between graft and host is estab- (Continued on Page 85) 1 = rantl , ree perinedtated theloamnticke ras with the limit between veg: erties of glands.” and vegs, as established by Horstadius against the view of Boveri. If in the 16-cell stage the mesomere material is separated from the macro- and micromere material (Fig. 3), the TABLE OF Experimental Study of Sex-Differentiation in the Chick Embryo, Dr. B. H. Willier........ 81 Physiology of Determination in the Sea-ur- chin Development, Dr. John Runnstr6m....81 Effect of 1934 Winter in Woods Hole........ 82 A Potentiometric Study of Phthiocol, Dr. Dake Cy IRN Secor onpacennnocpaGneerbeouac™ 88 Relationship Between the Water Content and Oxygen Consumption of the Organism, Dr. SVMS UNV ES) 2). oye ici a teleteue eevee wets enn 89 CONTENTS The Chlorophyll Unit in Photosynthesis, femny. Vy EXON «2. oz tye! Wereievctarsestoendrolakertel ares 90 Solvent Water in the Erythrocyte, Dr. A. K. d St: of OF: 1 ol Oe ee eR RE eee ae ener ickeer Ars chen here cur er 90 | Function of The Collecting Net—Letter from DryiG. Regal Ruew.yrae slecicietctetc ieee iets 91 ML GHTOTI al RAS OL ey tate tayokere aie hadciokenets easreleneneree sicher: 92 WTA OP WMO Sono ohcn mo oneaweoseceuceane 93 Critique of Curves of Growth and of Rela- tive Growth, Dr. C. B. Davenport ........ 95 82 THE COLLECTING NET [ VoL. IX. No.7 former develops into a blastula with a great area of stiff cilia (to the left of Fig. 3). In the normal larva these stiff cilia cover a very small re- gion of the animal or an- terior part of the larva. This is the “ciliary tuft.” According to our views this tuft has the tendency to ex- tend over a greater area, but this tendency is checked by the inhibiting activity of the vegetative or posterior part of the larva. In a very elaborate study Horstadius isolated the different layers of cells already referred to. Some of the results are shown in the first vertical column of Fig. 4. An, de- velops to a blastula covered with stiff cilia. When these peculiar larvae are kept, the Seay Fig. 1. stiff cilia are replaced by Effect of lithium on the sea- urchin larva. (After Herbst.) mobile cilia, but no further development takes place (cf. Fig.). The layer ang forms a blastula in which the ciliary tuft does not cover the whole surface of the larva. In the vegetative part mobile cilia are present. The fate of veg, which normally forms the more vegetative region of the ectoderm, is variable. Larvae with a ciliary tuft can be formed; but this is not always the case. The tuft can be absent and in this case a rudimentary entoderm is formed. The isolated vegy material, nor- mally forming ento-meso- derm, develops into a small larva with ecto- and meso- derm. The next three vertical columns of the figure dem- onstrate how the develop- THE WINTER OF 1934 AND ITS EFFECT UPON THE FAUNA AND FLORA OF WOODS HOLE The winter of 1934 was of such unusual sever- ity in Woods Hole and the Cape Cod region that it has been classed with 1918 as one of the worst seasons 1n history. The salt water froze in great sheets across Buzzards Bay, through the “hole,” and throughout the length of Vineyard Sound. Changing tides broke the ice into huge blocks which swept back and forth doing uncountable damage. At Chappaquoit, Silver Beach, Pen- zance and elsewhere along the shore the ice cakes piled up in immense heaps, and the unusual arctic beauty of the scene attracted sightseers from Bos- ton as well as from neighboring towns. (Continued on page 99) Photographed by Edith Billings ICE BLOCKS IN FEBRUARY AT CHAPPAQUOIT ] Jury 21, 1934 ] THE COLLECTING NET 83 Mikromeren a Fig. 2. Segmentation of the Paracentrotus egg. (After Horstadius. ) ment changes if one, two or four micromeres are implanted into the different cell layers referred to; the micromeres normally form the primary mes- enchyme. Already one micromere is able to in- hibit considerably the extension of the ciliary tuft in larvae developed from an; or ans. Veg; plus one micromere develops into larvae which are de- prived of the ciliary tuft. Also material from the macromeres is able to stop the extension of the tuft area, but two to four times as much macro- mere material is necessary to produce the same effect as is given by the micromere material. The inhibiting effect on the extension of the tuft area decreases rapidly from the vegetative toward the animal pole. The experiments of Horstadius sug- gest the presence of two opposite overlapping gradients in the sea-urchin egg, (cf. the scheme Fig. 5a). We imagine that certain types of meta- bolic activities decrease from the animal towards the vegetative pole, others from the vegetative toward the animal pole. The development is de- cided by the balance between the animal and the vegetative activities. If one of the gradients of activity predominates, such abnormal formations can develop as the blastula covered with stiff cilia from an isolated an, layer, or a larva consist- ing almost exclusively of entoderm as from veg» plus four micromeres. On the other hand almost normal plutei can develop from an, plus four micromeres or ans plus two micromeres. In these cases the greater part of the egg material is elim- inated but the balance between the animal and the vegetative activities resembles that in the normal egg. The same conclusions can be drawn from maay more of the experiments carried out by Horstadius. Herbst discovered the interesting fact that com- paratively small amounts of lithium-salts added to the sea-water profoundly modify the devel- opment of the sea-urchin larvae. The limit be- tween the entoderm and the ectoderm is trans- located in the animal direction. The ectoderm con- sequently is reduced in size and the ento-meso- derm enlarged. The differentiation of the ecto- derm is more or less simplified. The parts which are most animal disappear first. In extreme cases the whole egg develops into an ento-mesoderm. Different steps of these remarkable transforma- tions are seen in Fig. 1. Comparing Fig. 1 with Fig. 4 it is striking how much the lithium larvae resemble the result of such combinations as veg; plus four micromeres, veg» plus one micromere, veg» plus two micromeres, vege plus four micro- meres. The resulting larvae in the order named could indeed be chosen as illustrating an increas- ing lithium effect on the sea-urchin egg. Indeed the combinations in question illustrate the influ- ence of an increasing predominance of the vegeta- tive over the animal activities as an examina- tion of Fig. 4 makes clear. It seems legitimate to conclude that lithium impairs the animal activities in the developing eggs and thus produces an un- balance between animal and vegetative gradients as roughly illustrated by the scheme Fig. 5b. In a series of experiments it has been shown that carbon-monoxide sensitizes the egg to the influence of lithium. The eggs were brought into solutions of lithium in sea-water so weak that they produced no effect or only a very slight one on the larvae. The suspensions of eggs were brought into closed flasks under an atmosphere of 5% oxygen and 95% carbon-monoxide. In the dark- ness a strong lithium effect is now produced re- sulting in larvae with a small ectoderm and a large entoderm region. Carbon-monoxide depresses the respiration. This effect can be reversed by illum- ination. The sensitizing effect of carbon-monoxide on the action of lithium decreases on illumina- tion, and this to a degree corresponding to the intensity of the light. Thus it is evident that the sensitizing effect of carbon-monoxide depends on its inhibiting influence on respiration. The question then arose whether lithium in itselt acts as an in- hibitor of respiration; this has been proved by Lindahl to be the case. Warburg has shown that after the sudden increase in respiration follow- Fig. 3. ated animal and vegetative parts, Scheme of the development of the separ- 84 THE COLLECTING NET [ Vor. IX. No. 75 +2 Mikr Fig. 4. Separation of the different layers of the segmenting sea-urchin egg (cf. text). (After Horstadius). ing the fertilization of the sea-urchin egg, a more gradual increase ensues during the development. Lindahl was able to show that lithium has almost no influence on the respiration of the newly fer- tilized egg, but the decrease of oxygen consump- tion due to the presence of lithium increases dur- ing the development. Lindahl concludes from this that lithium has a specific influence on the in- creasing part of the respiration. The inhibition obeys a simple law indicating perhaps that a mono-molecular reaction is involved. Potassium antagonizes the lithium effect. By adding a suitable amount of an isotonic solution of KC1 to the lithium sea-water an almost normal development can be restored. It is then significant that the addition of potassium also removes the inhibiting action of lithium on the respiration. In a series of experiments Lindahl has treated the unfertilized Paracentrotus eggs during 8-20 hours with a mixture of 10 parts calcium- free sea-water and 1 part isotonic sodium sulphocya- nate. After the treatment the eggs are washed with normal sea-water and fertilized. The whole eggs now develop as isolated animal parts of the egg, cf. Fig. 3. The blastula has a very en- larged ciliary tuft. In other cases a very small intestine can appear. In these experiments the vegetative activities have been suppressed as in- dicated in the scheme Fig. 5c. In this way an un- balance is produced just as in the experiments with isolation of animal parts not counterbal- anced by implantation of micromeres. Lindahl found that the treatment by the cal- cium-free sea-water plus sodium sulphocyanate is quite ineffective, if the respiration is suppressed during the treatment. The air was replaced by ni- trogen or by a mixture of 5% oxygen plus 95% carbon-monoxide. The normal. Lindahl tried also to balance the effect of the calcium-free sea-water mixture by adding lithium after the fertilization of the egg. In this way he succeeded in bringing about an almost normal development. The balance between the gradients of activities as schematized in Fig. 5a is restored. Very often the effect of lithium predominates somewhat. Comparison with a culture of the same material with pure lithium effect reveals, at once, how much more normal the larvae are which de- velop from eggs treated with the calcium-free nuxture before fertilization. development was now In many cultures of the eggs from the calcium- free sea-water mixture Lindahl found blastulae with two ciliary tufts (cf. the rough sketch Fig. 6a). The impression is gained that two animal centers have been formed through a sort of “heteromorphosis”. Eggs from the same culture had been treated with lithium. Among the larvae developing in the lithium sea-water a number are formed with two ectoderms and two correspond- ing entoderm systems, Fig. 6b. These larvae prove that two animal centers have been formed. These correspond in all probability to the original ant- mal and vegetative parts of the egg; a consider- able rearrangement of the original polarity has taken place in these larvae. Sometimes larvae de- velop in these cultures in which a number of smaller intestines are formed at the equator of the larva, cf. Fig. 6c. The differentiation of the sea-urchin egg has been interpreted above as determined by a bal- ance between two types of metabolic activities de- creasing along two opposite gradients, cf. Fig. 5a. The animal activities are dependent on normal respiration of the egg. Already a general decrease in the respiration of the egg has an inhibiting ef- fect on the normal development of the animal part of the larva. The sensitivity decreases in an ani- mal-vegetative direction. Lithium has a specific inhibiting action on the animal part of the larva. It has also been shown that the influence of lith- ium on the respiration is not a general but a spec- Fig. 5. Scheme of the gradient systems in the sea-urchin egg, Jury 21, 1934 } THE COLLECTING NET 85 Fig. 6. Larvae treated with calcium free sea- water 2 NaSCN, without (a) or with (b, c) sub- sequent treatment with lithium. ific one. It must be admitted as a possibility that lithium inhibits oxidations characteristic of the animal activities. It has also been shown that the treatment of the eggs with the calcium-free sea- water plus sulphocyanate is ineffective if the res- piration is decreased or suppressed. The animal activities have a decidedly aerobic character. They may be based on the presence of certain sub- stances in an oxidized state or dependent for their formation on oxidation processes. The vegetative activities on the other hand are tolerant of a lower level of respiration. Processes of a more anaerobic character or substances in a reduced form are perhaps involved in the activities along the vegeta- tive gradient. EXPERIMENTAL STUDY OF SEX-DIFFERENTIATION IN THE CHICK EMBRYO (Continued from Page 81) ished via the blood circulation of the embryonic membranes. The special topics to be discussed are: (1) The nature of the organization of the gonad-forming area at various stages of its early development—primarily the time that this area ac- quires an organization which is fixed specifically as to sex. (2) The role of sex-hormones in sex-differ- entiation of the embryo. (3) The significance of the so-called primor- dial germ-cells to the origination and differentia- tion of the gonad. (4) The differentiation of the germ-cell itself. ie Evidence on the nature of the organization of the gonad-forming area of the chick has come from a study of the differentiation of this area in grafts to the chorio-allantoic membrane. It was discovered in such grafts in 1925 that the gonad rudiment of genital ridge stage, although mor- phologically indifferent as to sex, is capable of self-differentiation into a gonad of specific sex. The right genital ridge invariably forms a testis or a right ovary while the left genital ridge invar- iably develops into a testis or a left ovary. The rudiment is thus found to be specifically organized as to sex and in the female as to laterality also. The interpretation with respect to laterality is made possible by differences in structure of the right and left ovaries in birds. The right ovary possesses only primary sex-cords while the left has both primary and secondary (or cortical) sex- cords. The right and left testes also exhibit cer- tain differences in structure but they are not suffi- ciently striking as a rule in the grafts to be de- tected. Irom these results the conclusion is reached that the gonad rudiment of the genital ridge stage is not physiologically but only morphologically in- different as to sex. These findings led quite naturally to an analysis of potentialities of gonad-forming areas of stages earlier than the genital ridge. These include: (a) stages prior to the formation of the germinal epi- thelium-donors 29-34 Somites; (b) stages at which a germinal epithelium is visibly differen- tiated—donors 35-41 Somites. In these stages germ cells are present in the gonad-forming area. (At this point a brief description of the method of isolating the gonad-forming areas and their transplantation to the chorio-allantoic membrane of the host embryo was given.) With respect to differentiation of gonad the re- sults of this analysis are as follows: (1) The gonad-forming anea just prior to as well as during the formation of germinal epithe- lium has the power to form a gonad of specific sex but with distinctly less frequency than the genital ridge. A gonad of specific sex differen- tiates from the gonad-forming area of. a stage prior to the origin of the germinal epithelium in 23% of the grafts, whereas after its formation it is 48% of the grafts. Thus it is evident that a gradual increase in frequency of differentiation of a gonad of specific sex occurs as the morphogene- sis of the gonad rudiment takes place. (2) A gonad-like body of undistinguished sex forms commonly (56%) in grafts of the gonad- 86 THE COLLECTING NET [ Vor. TX. No. 75 forming area whereas it rarely arises in grafts of the genital ridge. Prior to 31-Somite stage this is the only type of gonad thus far formed in eight of the grafts examined. Several grades of organ- ization of these bodies are recognized, ranging from a mass of stroma and germ cells to a defi- nitely circumscribed body of stroma with sex- cords of germinal and non-germinal cells. (3) The gonad-forming area yields frequent- ly multiple gonads whereas a well-defined genital ridge gives a single gonad. (4) The size of the gonad of specific sex which arises from the gonad-forming area is smaller than one from the genital ridge. Thus we see a progressive increase in develop- mental potentialities of the gonad-forming area occurring as it transforms into a genital ridge. What does this signify? This is interpreted as indicating a progressive change of some sort in the organization of the gonad- forming area, 1. e. it is an index of an ascending organization. Prior to the 31-Somite stage the gonad-forming area is apparently wholly dependent upon the neighboring regions of the embryo, since it has failed in eight cases to differentiate into a gonad of specific sex. Evidence not fully analyzed yet shows the importance of adjacent regions of the embryo for its differentiation. Beginning with the 31-Somite stage there is exhibited less and less dependence, finally culminating at the 50-Somite stage in a condition which is wholly independent of its surrounding parts. The effect of the neigh- boring regions of the embryo upon the gonad- forming area seems therefore to be of a cumula- tive nature. The conclusion is reached that the gonad-forming area during its initial development exhibits an ascending organization which attains at the genital ridge stage a fully “determined” con- dition—a_self-differentiating system in a strict sense. Recently we have been able to carry our analy- sis to far earlier stages than those just examined, by means of which very interesting results on lo- calization of the gonad-forming potencies are re- vealed. By making a study of the developmental powers of the various regions of the blastoderm of the head-process stage the blastoderm is found to be a mosaic of specific organ-forming areas, such as the eye-forming area, thyroid-forming area, and the mesonephros-forming area. [At this time-maps (prepared by Miss Mary E. Rawles and myself) illustrating the developmental potencies of the eighteen pieces into which the blas- toderm was cut, were shown. ] Of particular interest here is the mesonephros- forming area situated just behind the primitive pit. Centrally situated within this larger meson- ephros-forming area is a much smaller area hay- ing gonad-adrenal-forming potencies. The meson- ephros-forming area exhibits certain properties of unusual significance. First, it is a gradient in de- velopmental powers, with greatest power in the center gradually diminishing peripherally. On the basis of a study made on the eye-forming area the contours and potencies of this area will in all probability change with advance in development, i. €., as it assumes a distinct bilateral organiza- tion. Second, it possesses an asymmetrical organ- ization. The median part of the area has the great- est power for producing mesonephros, the left side less and the right side least of all. Also the amount of growth and the degree of differentia- tion attained follows the same order. Thus the asymmetry of the urino-genital system of birds is traced to the blastoderm “of an early stage. Il. We come back now to the original problem, that of sex-hormones and sex-differentiation. The gonad-forming area or primordium differentiates according to its specific organization, acquired be- fore its isolatton from the donor, and is not modi- fied by the sex of the host embryo. Furthermore, a graft of sexually differentiated gonad tissue, even in large amounts and from juvenile fowls, known to be producing sex-hormones, fails to bring about any signs of sex-reversal. One ex- - ceptional case of an ovotestis appeared recently in our grafts. It was obtained from the gonad- forming areas of a 3l-Somite donor and grown for nine days on a female host. In the light of recent studies of the action of the female sex-hor- mone preparations when injected into the white of the hen’s egg, this is probably a left testis in which germinal epithelium or incipient cortex was present and which responded to the female sex- hormones of the host. This interpretation is based upon asymmetrical structural differences discovered in the testes of duck (Burwell 31), and of chick embryos. In the left testis a dis- tinct germinal epithelium (continuous covering in the duck; intermittent in the chick) is found, whereas in the right it is completely absent. (These structural differences were illustrated with photomicrographs.) It might be expected that such testes would exhibit fundamentally different types of response to female sex-hormones, the left forming cortex whereas the right would not. The senetal lack of cases of sex-modification in the chick is very puzzling, particularly since in the amphibian embryo sex-reversal may be readily induced by similar methods of grafting. It is only recently, as has just been indicated, that a possible clue to this baffling problem has become appar- ent. LU The nature of the problem to be considered here may be pictured in the form of a question. Are the primordial germ-cells, which according to current theory segregate early and outside the gonad-forming area with subsequent migration Jury 21, 1934 } THE COLLECTING NE ” into it, essential for gonad-origin and differentia- tion? Various attempts have been made to show that the “germ-cell crescent” of Swift is the source of the germ cells of a gonad rudiment. By excision of the germ-cell crescent (Reagan, 1916) and by irradiation with ultra-violet (Benoit, 1930), or with X-rays, (Dantschakoff, 1932) it has been shown that the prospective germinal epithelium may be freed of germ-cells. That such a germinal epithelium has the power to differentiate into a gonad has not been ana- lyzed. This we have recently accomplished in two different ways. One, by grafting the whole blas- toderm of early somite stages following the re- moval of the germ-cell crescent. Second, by grafting from the blastoderm of the head-process stage a small median piece situated just behind the primitive pit. Each method has given a sterile gonad with typical male-like sex-cords in a total of ten cases.. In no case was ovarian cortex formed although this is to be expected in half of the cases on the basis of a normal sex ratio. Ap- parently only the first set of sex-cords—known to possess male potentialities in both sexes in birds —are capable of formation under the conditions of the experiment. Besides giving support to the theory of extra-gonadal origin of germ-cells, the results are in themselves very interesting, and [| think significant for an understanding of the com- plicated story of gonad origin and differentiation. Ve In chorio-allantoic grafts of the gonad-forming area two kinds of germ-cells, differentiated and undifferentiated are distinguishable. The former occur singly or in masses in the mesenchyme. The latter is found in the male sex-cords and in the female or cortical cords. The undifferentiated germ-cell is characterized by dark-staining gran- ules uniformly distributed in the cytoplasm; the differentiated, by a grouping of the granules to one side of the nucleus. This difference in gran- ule distribution in the cytoplasm appears to be an indicator of differentiation. Differences in cellular differentiation of germ cells are seen in the two sexes. In the male the germ-cells occur singly in the sex cords and the granules are usually finer and lighter staining than those found in the cortex of an ovary. In the cortex of the ovary the germ-cells are smaller and occur in groups. The fact that the germ-cells in the mesenchyme remain unchanged, like pri- mordial germ-cells, while those which come to be situated within sexual cords (male or female) undergo structural changes, furnishes convincing evidence that they are dependent upon a specific tissue environment for their differentiation. The germ-cell, although specialized in a general way as a sexual cell and thus differing from non- germinal cells, is really sexually indifferent and remains in this condition in such non-sexual tissues as mesenchyme. It undergoes speci- fic cellular transformation as to sex only in the specific tissue environment of the sexual cord: What is to say, in a male sex-cord the germ cell transforms in the male di- rection; in a female sex-cord it transforms in the female direction. The capacity of the indifferent germ-cell to dif- ferentiate in either the male or female direction in a graft indicates that it possesses bisexual poten- tialities. This has been beautifully demonstrated in the female fowl by an experimental study of the right ovary (Domm et al). It has been found that if the left ovary is removed during the per- iod when the germ-cells are still present in the right ovary (hatching to the third week) it trans- forms into a testis containing male germ-cells; if removed after they have disappeared a sterile tes- tis differentiates. It would appear from these ex- periments that, (1) germ-cells remain indifferent in the medullary cords of the normal right ovary, (2) when the inhibiting influence of the cortex of the left ovary is removed, the latent male poten- cies of the medullary cords become expressed in the form of seminiferous tubules, the germ-cells therein undergoing a specific differentiation into male sex-cells, and even into their definitive form, the spermatozoa. Similarly from a study of sex-reversal in frogs Witschi has brought forward evidence of consi- derable weight that the germ-cell is dependent upon a specific tissue for its differentiation in the male or female direction. According to him the “sex-differentiating factor” is localized in the cor- tex and medulla. The cortex is a female differ- entiating system while the medulla is a male dif- ferentiating system. Thus in an ovary undergoing sex-transformation some of the germ-cells of the degenerating cortex wander into the medulla where they differentiate into spermatogonia. On this question Humphrey (1933) has given us a most convincing demonstration in frog embryos. By combining the prospective gonad area of one sex with the germ-cells of an embryo of the op- posite sex in an ingenious grafting experiment, he was able to prove that the germ-cells differen- tiate in accordance with the sex of the gonad into which they enter, and not according to their ori- ginal sex-determination. That is, the germ-cells from a male embryo differentiate within an ovary into oocytes, whereas germ-cells from a female embryo differentiate within a testis into sperma- togonia. Tn conclusion, I should like to make a summary of the principle generalizations brought out. (1) The gonad-forming area exhibits an as- cending organization finally attaining a fully “de- termined” condition just prior to the genital ridge stage. This is probably the stage of embryonic 8¢ hb HE COLLECTING NEWT [ Vor. TX. No. 75 segregation. Lateral differences in the organiza- tion of the gonads of the domestic fowl has been traced to a fundamental asymmetrical organiza- tion of organ-forming germs of the early blasto- derm. A knowledge of these inherent differences in organization is essential to an understanding of the nature of the response to the action of sex- hormones. (2) A gonad may originate and differentiate independently of the primitive germ-cells, thus supporting the theory of their independent origin in the chick embryo. (3) Germ-cells exhibit no capacity for inde- pendent differentiation. They merely have the ca- pacity of repeated multiplication. The germ-cell also has bisexual potentialities; in other words, the differentiation of the germ-cell is the realiza- tion of alternate, male or female, forms of re- sponse. The direction of the response is mediated by a specific tissue environment. A POTENTIOMETRIC STUDY OF PHTHIOCOL, THE PIGMENT OF THE HUMAN TUBERCLE BACILLUS Dr. Eric G. 3ALL Associate in Physiological Chemistry, Johns Hopkins Uniwersity Medical School More data are needed on those constituents of living tissue which are components of definite oxidation-reduction systems. Such information will aid in defining the intensity level of energy changes in living cells, a level essential for the activation of certain enzymes and for the protec- tion from destruction of compounds like epine- phrine. Phthiocol, a pigment isolated from the tubercle bacillus by Anderson and Newman, has been shown by these workers to be 2-methyl-3 hydroxy-1, 4-naphthoquinone. On the basis of its structure this compound should be amenable to potentiometric study. It has been found to be the oxidant of a reversible system whose potential in the more usual pH range is among the lowest re- ported for systems of biological origin. The phthiocol used in this investigation was a synthetic sample kindly supplied by Dr. R. J. An- derson. This compound is a weak acid, poorly soluble in aqueous solutions of pH lower than its pK value (5.08), more readily soluble on the al- kaline side. In aqueous solutions the undissoci- ated form has a pale yellow color, the anion a dark red. The reduced form of each species is colorless. Since both components of the system were ent’rely stable throughout the usual pH range, the procedure for potentiometric measurement was the usual one. Two electrons are involved in the reaction. The value of n=2 was found to hold at all pH values and no indication of a two- step oxidation was encountered as described for the bacterial pigments, pyocyanine and chlorora- phine. For 30° C., the normal potential of the system is 0.2987 volt, the apparent dissociation constant of the hydroxyl group of the oxidant is 8.32 & 10°, and the first and second apparent dissociation constants of the reductant are 1.26 « 10°° and 2.88 * 107°. The dissociation of the third hydroxyl group of the reductant was not ef- fective in the pH range studied. Other hydroxynaphthoquinones have been iso- lated from natural sources. All, however, are of plant origin. Four of these, lapachol, lo- matiol, lawsone, and juglone have been submit- ted to a potentiometric study by various workers. The first three resemble phthiocol most closely in structure and consequently their systems exhibit normal potentials of the same magnitude. The oxidant of these systems is the naturally occurring form. The juglone system is the most positive ot the group. The occurrence of the reduced form of juglone in living cells is in keeping with the higher potential of this system. Nearly a dozen pigments from natural sources have been shown to form reversible oxidation-re- duction systems. Most of these systems lie be- tween those regions of potential shown to repre- sent the state of reducing intensity of ameba in aerobic and anaerobic conditions. This interest- ing fact, coupled with the rapidity with which the reductants of these systems are oxidized by air, has led to the assignment of the role of accessory respiratory ferments to these pigments. Such in- deed may be their role, though the increased oxy- gen consumption caused by their addition to tissue to which they are foreign is not a convincing test of their physiological importance in normal res- piration, especially since increases in quantity of oxygen consumed give no indication as to the use- fulness of the energy released to the normal pro- cesses of the cell. Most of these compounds oc- cur in lower organisms or plants in which the chief method of respiration is still unknown so that the assignment of even a minor role in this process, even in their native tissue, seems at pres- ent premature. Phthiocol should form a valuable addition to the series of oxidation-reduction indicators at pH values greater than 6.0. Its solubility and color intensity in this range are satisfactory. At pH 7.4 the system lies intermediate to the indigo- monosulfonate and phenosafranine systems and slightly negative to the less desirable brilliant alizarin blue system which heretofore was the only one available in this region. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 17.) —E—— Jury 21, 1934 ] THE COLLECTING NET 89 THE RELATIONSHIP BETWEEN THE WATER CONTENT AND OXYGEN CONSUMPTION OF THE ORGANISM Dr. WiLL1AM RANKIN DURYEE Department of Zoology, Northwestern University The role of water in active protoplasm has not yet been fully comprehended. By far the most abundant of the constituents of the living cell, it is particularly important as a milieu or theatre of action for the complex series of metabolic reac- tions. This paper presents a new point of view bearing on two rather old problems—namely, that water content changes and the rate of cell oxida- tions may be inversely related one to another on the basis of the Mass Action Law and from the standpoint of LeChatelier’s Principle. Stated more specifically the problem was to see whether reversibly hydrating and dehydrating certain fresh-water organisms would cause changes in their rates of oxygen consumption. The animals used were: Euplanaria dorotocephala, Procotyla fluviatilis (sometimes mis-called Dendrocoelum lacteum), Planaria maculata, and Amblystoma punctatum embryos. It is a pleasure to acknowl- edge my indebtedness to Dr. J. W. Buchanan of Northwestern University, who suggested and di- rected this work. Water content changes were brought about by exposure to different anisotonic media made non- injurious as far as possible by balanced cations. The general picture of hydration and dehydration in planarian tissues shows changes that are both inter- and intracellular and which vary markedly both with time and concentration. Oxygen consumption was measured by two widely different techniques: first by the Winkler method in which relatively large volumes (about 500 cc.) of media were used, and second by a microrespirometer using very small volumes. This respirometer, which was designed to take the place of a Warburg manometer, is more simple and reliable to operate than any other apparatus that I know of—readings are accurate to within half a cubic millimeter of gas. Exposure to distilled water—a hydrating agent —depressed the average rates of oxygen con- sumption in Euplanaria dorotocephala about 60 per cent. Conversely a dehydrating Ringer-Mur- ray solution stimulated the same sets of individ- uals from 29 to 58 per cent. above normal de- pending on the number of hours of exposure. Each period in a test solution was preceded by one in tap water to establish a norm, thus form- ing a series of interlocking control values from successive tests on the same individuals, and show- ing that the effects were completely reversible. It is to be emphasized that the same sets of worms could be hydrated and dehydrated repeatedly— one pair of Procotyla were observed through 15 transfers. On the other hand, developmental changes in Amblystoma embryos made such suc- cessive experiments impractical. Oxygen uptake was not materially affected un- til the A value of the medium reached about —0.4° C. (the strength of the Ringer-Murray solution) showing the tolerance limits of the worms. With Ringer + 1/6 to Ringer + 1/3 there were marked increases. In higher salinities a falling off was noted. Winkler determinations yielded more extreme values than did respirometer ones, thus clearly in- dicating the effect of the larger volumes of media employed in the former method, since these ani- mals could osmotically condition their environ- ments. Recent experiments indicate .that non- electrolytes, such as manitol, cause similar in- creases when used in the same osmotic concentra- tions. The following tentative explanation is offered for these observations. A consideration of four major cell oxidation mechanisms, 1. e. Bach and Engler’s, Warburg’s Respiratory Enzyme, Wie- land’s Hydrogen Activation, and Keilin’s Cyto- chrome-Oxidase theories, shows that the last three have this fact in common—namely, that water molecules are formed as end products of these enzyme-controlled reversible reactions. From this there follow two important theoretical reasons why water content should be inversely related to the rate of oxygen utilization: I. “The course of enzyme reactions can be ex- pressed with fair accuracy on the basis of the Mass Action Law.’—Waldschmidt-Leitz. II. An accumulation of water in the cell tends to displace these equilibria toward the left to re- tard these processes, and conversely a diminution in the amount of water tends to accelerate them according to the Law of Mass Action and to Le Chatelier’s Principle, which states that a reversi- ble system (or reaction) will proceed in the direc- tion of least stress. So, inasmuch as more than twenty other inves- tigators—notably Harden and Paine, Caldwell, and Kidd and West—have observed evidence of a similar relationship (in barnacles, echinoderm eggs, planarians, frog tissues, teleosts and certain mammals and also in yeast and other plant tis- sues) and in spite of objections raised by the ex- periments of Beadle, Schlieper, and others, it may be concluded that over short periods of time, and within the definitely restricted limits of water content change that organisms may withstand without injury—-one of the primary effects of hy- dration is to retard cell oxidations, and converse- ly, one of the primary effects of dehydration is to accelerate them. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 17.) 90 THE COLLECTING NET [ Vor. TX. No. 75 SOLVENT WATER IN THE ERYTHROCYTE Dr. A. K. PARPART Assistant Professor of Physiology, Princeton University Studies on the distribution of substances be- tween the erythrocyte and its environment have necessitated a knowledge of the extent to which the water within the erythrocyte is capable of act- ing as a solvent for the substance in question. On the basis of dry weight determinations the mam- malian erythrocyte appears to have about 69% of its volume occupied by water. The amount of this total water content which is capable of acting as solvent water according to one group of inves- tigators is 50% ; according to another group it is 70-75% ; and a third group believes that all of it is capable of acting as solvent. In the present work studies were made on the distribution of ethylene glycol, glycerol and urea between the erythrocytes of several species of mamuials and their environment of either Ringer- Locke solution or plasma. In the case of both ethylene glycol and glycerol the data secured in- dicate that about 70% of the total water acts as solvent for these substances. Urea distribution data on the other hand indicate that there is more water in the cell than can actually be found there. The latter result may be due to a variety of fac- tors, chief of which are the effects of urea on pro- teins and its solubility in lipoid materials. By a repetition of Hamburger’s experiments on the degree of swelling and shrinking of the eryth- rocyte in hypo- and hypertonic Ringer-Locke solu- tions and in hypotonic plasma it was found that about 70% of the water is capable of being trans- ferred osmotically, i. e. capable of solvent action. (This article is based on a seminar report given at the Marine Biological Laboratory Auditorium on July 17.) THE CHLOROPHYLL UNIT IN PHOTOSYNTHESIS By Henry I. Koun Assistant in Physiology, Harvard University From the point of view of energetics, the pro- cess of photosynthesis is a mechanism whereby the energy of light can be converted into the po- tential energy of chemical structure, and since chlorophyll absorbs the light utilized, the fun- damental problem is to describe the chemical re- actions into which this pigment enters. Little is known about these reactions, but it is assumed generally that the first of these must be the union of COs, probably in the form of HsCOs, with chlorophyll. This hypothesis raises the question wtih which we shall deal: How many molecules of COs combine with how many molecules of chlorophyll to form one molecule of the COz- chlorophyll compound? Since photosynthesis does not occur i vitro, we must seek our answer in the living green plant. The question we have raised is that which Em- erson and Arnold? sought to answer several years ago, and since we have used their methods, a brief recapitulation of their work is appropriate. As is well known, the process of photosynthesis may be divided into two parts, the photochemical and Blackman reactions, the latter of which is independent of light. The Blackman reaction, which is sensitive to cyanide and temperature, may be considered as a recovery process, and any device which allows it to run to completion will exclude it as a limiting factor. To accomplish this, Emerson and Arnold illuminated their plants with very brief, intense flashes of light, separ- ated by adequate periods of darkness. These dark periods were of such a duration that lengthening them did not further increase the amount of photosynthesis per flash. Since the plants were saturated with respect to light intensity and COs tension, and since the flashes were so brief (1 x 10+ secs. or less) compared to the life time of the Blackman reaction (about 1.5 x 10% secs. in Chlorella pyrenoidosa at 25°C.), it was argued that the amount of photosynthesis per flash rep- resented the maximum yield per cycle of which the photosynthetic mechanism was capable. Under these conditions one may ask the ques- tion of how many molecules of chlorophyll are present for each molecule of COz reduced. Emer- son and Arnold extracted the chlorophyll from their plants and determined the value of the ratio, mols of chlorophyll/mols of CO. reduced per flash. In the green alga, Chlorella pyrenoidosa, they obtained values lying between 2,000 and 3,000, which are independent of the chlorophyll content of the cells. Such a result is surprising—to say the least— and in order to subject it to a severe test Arnold and the writer? have investigated a half-dozen other plants of varied systematic position. These included species of Nicotiana, Bryophylhun, Lem- na, Selaginella, Sticchococcus, and Chlorella. In all cases the value of the ratio was approximate- ee Jury 21, 1934 | THE COLLECTING NET 91 ly that originally obtained, and the fact became manifest that though the value of the ratio might be increased (for example, by brief exposures of the plants to ultra violet light), in no case does it fall appreciably below 2,000. And so we are confronted with the problem of interpretation. Are these results due to a sys- tematic mistake in analysis? If not, what is the physical and chemical significance of this “chloro- phyll unit” for the process of photosynthesis ? 1. Emerson, R., and Arnold, W. A., J. Gen. Physiol., 16, 191, 1932. 2. Arnold, W. A., and Kohn, H. I., J. Gen. Physiol., 1934, in press. (This article is based on a seminar report presented at the Marine Biological Laboratory on July 17.) THE NAMING OF WOODS HOLE There is considerable uncertainty as to the ori- gin of the name “Woods Hole” which is the pres- ent approved spelling for the name of this village, situated in the township of Falmouth, Barnstable County, Massachusetts. At one time the village was called “Woods Holl.” In the tenth century, this continent was visited by the Norsemen. In fact, they are said to have landed at Woods Hole in 1000 A. D. Among the Norse words which were thought to have been adopted by the natives of the Falmouth district was the noun “holl,’” which in the lan- guage of the sea king means “hill,” but which to- day is spelled “hole,” and has quite a different meaning. Webster’s Dictionary places the following defi- nition sixth, in the meaning of “hole”: Local U. S. (a) a small bay; a cove. (b) a narrow water- way. To be sure, Woods Hole is located where a narrow waterway of strong tides connects Buz- zards Bay and Vineyard Sound. It is the south- western extremity of Cape Cod, where the land meets the sea in gracefully rounded /ills. In this connection it should be noted that “Woods Hole,” in common with Robinson’s Hole, Ouick’s Hole, and Butler’s Hole, is the name applied to a body of water near what is today, or what was in years past, a well-defined /ill. While there seem to be good grounds for think- ing the name “Woods Hole” originated with the Norse word “holl’’ and its spelling to have heen corrupted into “hole,”’ there appears to be con- siderable doubt of this on the part of most of the old-timers around Woods Hole. By the same token that Quick’s Hole and Robinson’s Hole have always been known as “Hole” and not “Holl,” Mr. Gifford feels that he voices the opin- ion of the majority of the early settlers of Woods Hole in stating that the correct name for this village, and for the waterway on which it is lo- cated is Woods Hole, its origin being the defini- tion given as “a narrow waterway.” THE FUNCTION OF THE COLLECTING NET To THE EpirTor, I regret that it was impossible to prepare the article requested on the Michigan Biological Sta- tion for use in your issue of June 30. Our Sta- tion has just gotten under way and it will soon be possible to prepare a report covering the re- search in progress. I think that the report on the Michigan Station in the CotLtectinc Net last year covered our research too well to justify a repetition. Concerning the further development of the CotLectinG Net, I am inclined to believe that it might well develop into the unofficial and infor- mal organ of the marine and freshwater labora- tories of North America. There are now quite a number of such laboratories, most of them alto- gether too small to support a journal of their own. Your modest journal might very well meet the needs of this group. If this journal could be used as a medium for the publication of brief pre- liminary articles based upon research at the vari- ous laboratories, it would help these laboratories very materially. This need not stand in the way of the publication of some of the longer articles which you have been carrying in the past. In your letter you indicate the possibility of ob- taining funds for the creation of scholarships for promising young investigators, without indicating the manner in which such funds have been col- lected. I have always had a feeling that profits accruing to a publication ought to be plowed back into the publication, but if your journal can serve as an agency for the collection of funds from outside sources for the establishment of scholar- ships, such collection is desirable and perhaps should be pushed even further. Concerning the publication of lectures and sem- inars presented at our Biological Station, I have as yet given little thought. We have fewer in- dependent investigators than many of the larger laboratories although we have usually a good group of younger people working under direction. Thus far only one group, the parasitologists, have carried out a definite program of weekly research conferences. The reports given at these confer- ences are usually on research already accom- plished or upon research well under way and with fairly definite idea as to place of publica- tion. It is possible, however, that this group and certain others might welcome a place for the pub- lication of reports of a less extensive character than those published for the Woods Hole Semi- nars and the Cold Spring Harbor Lectures and Seminars. You may expect shortly some notes on the Sta- tion and a little bit later a report on the research activities. > Very truly yours, (Signed) Grorce R. La Rur, Director. 92 THE COLLECTING NET [ Vor. IX. No. 75 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel W. Parker, Marjorie Mitchell, and Annaleida Snyder Cattell. Printed by The Darwin Press, New Bedford STANDARDS OF CONDUCT. IL. Beach Ethics We have intentionally avoided commenting on the beach situation in Woods Hole. That is not an indication that bathing conditions are better or that we have altered our ideas concerning them. We planned to continue the discussion last sum- mer, but high authority warned us that it would be detrimental to the best interests of the laboratory, the situation in question as well as ourselves to open it again. We bowed reluctantly to their wishes, partially agreeing with them that if the matter were dropped the fence on the Bay Shore Beach would be voluntarily removed. The fence is there—part of it contrary to law—the “public” portion is crowded and rocky ; the “private” beach is vacant and sandy. We may be criticised severely for bringing the matter up again but we sincerely believe that it is in the interests of the laboratory community that the attention of its members be directed to the unfortunate situation. We take it for granted that the lapse of two years permits of a calm and careful analysis of the situation. In any case, being an independent publication, no objection can properly be made to any facts or opinions that Tue CoLLectinG Net may print. Certainly the facts of the case should be presented to new workers at the laboratory and be recalled for the people who worked here in 1932. In our opinion it is an extraordinary situation that five members of the Corporation of the Ma- rine Biological Laboratory, three of whom are trustees of the institution, should act together to exclude other biological workers, their wives and their children from a sandy area upon which everyone has always been free to bathe. To some the action would be forgivable if the open beach would cause serious discomfort to the property owners. The facts are, however, that few or none of the several property owners are living on their beach property, and that two of them have given their houses, presumably for a consideration, to people who are not working in biology. Some of them, then, have taken for private gain a sandy stretch on the shore which has always been con- sidered public property. We have been given to understand that the property owners in question purchased their beach lots at a comparatively low sum because of their adjacency to a public beach. Naturally there are two sides to the question, although these remarks present only one of them. The property owners believe that they are within their legal and their moral rights by closing the beach in front of their property; they would not admit that “ethics” enters into the question. Our readers should not, therefore, form opinions of their own from our statements until we have the opportunity of presenting the views of the prop- erty owners; they feel just as strongly about the matter as the people on the opposite side of the fence. The Effingham B. Morris Biological Farm of the Wistar Institute of Anatomy and Experi- mental Biology, which is located at Emilie, Penn- - sylvania, has in residence this summer the great- est number of active members in its history. For- merly there were only one or two workers to make use of the very excellent equipment of the laboratory, but at present there are eight investi- gators. The farm has opened a second new build- ing for the study of amphibia. CURRENTS IN THE HOLE At the following hours (WVaylight Saving Time) the current in the Hole turns to run from Buzzards Bay to Vineyard Sound: Date A.M Ee July 26 4:35 4:47 July 27 9:27 \ Sil July 28 6:17 6:34 July 29 7:06 7528 July 30 7:00) Snell July 31 845 9:16 August 1 9:37 10/4 NBEBR A By eccesc. 10:30 11:14 INGER 8) eapotat UiAey August 4 ........ Se ABA il 25) In each case the current changes approxi- mately six hours later and runs from the Sound to the Bay. It must be remembered that the schedule printed above is altered somewhat by wind conditions. Prolonged winds sometimes cause the turning of the cur- rent to occur a half an hour earlier or later than the times given above. The average maxi- mum velocity of the current is five knots. Jury 21, 1934 ] THE COLLECTING NET 93 REVS: Or Proressor RrcHARD GoLDscH mip? of the Kai- ser Wilhelm Institute for Biology, Berlin-Dahlem, is expected to visit several universities in Spain during the month of August to give lectures on genetics. He will return to Berlin in September. His daughter, Dr. Ruth Goldschmidt, is planning to be in New York late in August and will also visit Woods Hole. Dr. GrorcE S. DEReENy1, of the University of Pennsylvania Medical School, arrived on July 18 at the Marine Biological Laboratory, coming from the Tortugas Islands where he spent the earlier part of the summer working on the nery- ous systems of various invertebrates, especially crustaceans. Dr. JosepH M. Murray has become head of the Department of Zoology at the University of Maine, haying been appointed in July. Dr. Mur- ray was formerly associated with Dr. C. C. Lir- TLE in cancer research at the Roscoe B. Jackson Memorial Laboratory at Bar Harbor. Dr. Frank A. Hartman, for many years Pro- fessor of Physiology at the University of Buffalo, and distinguished for his work on cortin, has been appointed, Professor of Physiology at the Ohio State University. Miss Exsa M. Kerr, formerly instructor of zoology at the New Jersey College for Women of Rutgers University, has recently been pro- moted to the position of assistant professor of zoology. Miss Keil has been carrying on re- search work at the Marine Biological Laboratory as well as assisting in the Chemical Room for a number of summers. Dr. Lestie C. Dunn, professor of zoology at Columbia University since 1928, is on a year’s leave of absence during 1934-1935. In August he and his family will leave the United States for Oslo, Norway, where Dr. Dunn will continue his work on genetics. His children will go to school in Oslo. Dr. FernanpDus Payne, professor of zoology at the University of Indiana, recently came to Woods Hole to attend the first meeting of the Board of Review of the Marine Biological Lab- oratory. INTERES? Dr. Ivey F. Lewts, professor of biology at the University of Virginia since 1931, replaces Pro- FESSOR KERNANDUS PAYNE as chairman of the Division of Biology and Agriculture of the Na- tional Research Council. Dr. Lewis is working in Virginia this summer where he is director of the Mountain Lake Biological Station of the Univer- sity of Virginia. Dr. W. RK. BRENNEMAN of the University of Indiana will work as a National Research Council fellow in zoology with Dr. FRepERIcK L. Htsaw, professor of zoology at the University of Wis- consin, Dr. Brenneman is interested in the effect of endocrines on the early development of the chick. Messrs. Francis R. Hunter of California Insti- tute of Technology and Cornelius T. Taylor of Rutgers University have been appointed part- time assistants in the Department of Biology, Princeton University. Dr. Loutsre H. Grecory, associate professor of zoology and associate dean at Barnard College, recently spent ten days at the Marine Biological Laboratory and expects to return again in Au- gust. Dr. T. T. CHEN, of the Department of zoology at the University of Pennsylvania, received his Ph.D. in June. Dr. Chen has also received a Sterling Fellowship to carry on work at Yale University during 1934-1935. Dr. Ettice McDonatp, Director of the Can- cer Research Institute at the University of Penn- sylvania Medical School, visited Dr. Clowes and Dr. Chambers at the Marine Biological Labora- tory on July 19. Dr. Donatp Y. Sotanpt and Omonp M. So- LANDT who have been working at the Laboratory this summer on a CoLtLtectinG Net Scholarship, are leaving Woods Hole on July 23 for the At- lantic Biological Laboratory at St. Andrew’s, New Brunswick, where they will work with Dr. Lau- RENCE [RVING on the respiratory mechanisms of seals. In September Dr. Solandt is planning to go to England where he will study under Dr. A. V. Hirt, Foulerton Professor of Physiology at University College, University of London. 94 THE, COLLECTING NET [ Vor. IX. No. 75 ITEMS. Oe The Board of Scientific Directors of The Rockefeller Institute for Medical Research an- nounces the election of Dr. WARFIELD THEOBALD LoNGcorE as a member of the Board of Scientific Directors to succeed Dr. William H. Welch, de- ceased. The following promotions and appointments are announced: PROMOTIONS Associate Member to Member, Dr. Leslie T. Webster. Associate to Associate Member, Dr. Richard E, Shope. Assistant to Associate, Dr. Francisco Duran-Rey- nals, Dr. Kenneth Goodner, Dr. Geoffrey W. Rake. Fellow to Assistant, Dr. Kenneth S. Chester, Dr. Erich Traub, Dr. Philip R. White. NEw APPOINTMENTS Associate Member, Dr. Max Bergmann. Assistants, Dr. Donald C. Boughton, Dr. Jack Compton, Dr. James R. Dawson, Jr., Dr. Lee E. Farr, Dr. Delavan V. Holman, Dr. John G. Kidd, Dr. Colin M. MacLeod, Mr. William F. Ross, Dr. Thomas F. M. Scott, Dr. Joseph Ix. Smadel, Dr. Carl V. Smythe. Fellows, Dr. Bacon F. Chow, Mr. Joseph S. Fru- ton, Dr. George I. Lavin, Dr. Charles V. Sea- stone, Jr., Dr. William Trager. NOTES FROM THE BIOLOGICAL LABORATORY AT COLD SPRING HARBOR Among the women who are working at the Laboratory there are: Mrs. ErNest BLANCHARD (Dr. Harriet McCurpy) who is carrying on in- dependent research; Mrs. Rogpert Gaunt (Jo HowLanpD) who is conducting research with her husband; Mrs. Howarp Curtis who took over Miss Brown’s work during her absence; Mrs. Joun MacLrop (Miss Brown) who continues her duties for the remainder of the summer; and Mrs. WiLttAM PARKINS (PHYLLIS PLyER) who is in charge of living quarters. New boat owners at the Laboratory this year include: ANNETTE Bacon, sailboat; Dr. aNnp Mrs. Curtis, sailboat; D. M. GaLLaGuer, out- board motor speed boat; Mr. AND Mrs. WILLIAM PaRKINS and Prorrssor and Mrs. Gaunt, in- board motor speed boat; ProressorR THOMAS Hunt and Dr. CARoLINE TUM SUDEN, canoes. Dr. A, A. SCHAFFER has won an all time record by making his sailboat again “calm-seaworthy”’. Dr. AND Mrs. Harris’ sailboat is among those present. There are a few boats whose type and ownership is still in doubt. INTEREST Dr. H. B. Sternsacu, National Research Council fellow in biology at the University of Chicago, was married early in June in Chicago to Mary Eleanor Parsons ot Louisville, Kentucky. Dr. and Mrs. Steinbach arrived at Woods Hole last week, and are living in the Edwards House on School Street. Dr. Oscar Sette, Director of the Woods Hole station of the Bureau of Fisheries, is expected to arrive next week with Mrs. Sette. The out- door pool is being stocked with mackerel awaiting his arrival. Dr. Sette will continue his research on mackerel migration for the remainder of the Summer. About 200 mackerel are in the pool at present, and more will be added before the stock- ing is complete. The engagement of Miss MArcrery J. GREENE to Mr. Lorus J. MILne has been announced to their friends. Both are members of the Proto- zoology Class at the M. B. L. Miss Greene is from Hunter College of New York City, and is now a graduate student at Columbus University. She will teach in the New York City public high schools during the coming year. Mr. Milne is a graduate of the University of Toronto. He is in the United States as a research worker in En- tomology at Harvard University, where he is studying for his PhD. THE STRING TRIO CONCERTS A string trio of New York artists will give the first of two chamber music concerts here on Sun- day evening at Community Hall, starting at 8:30. Nancy Wilson, ‘cellist of the group, is already well known to Woods Hole audiences; Wolfe Wolfinsohn, violinist, and Bernard Wagenaar, pianist and composer, are the other members. The first number on the program is the Haydn Trio in G Major, of which the finale, Rondo all” Ongarese, is perhaps the most popular movement from any of the composer's chamber music works. Following this, Miss Wilson, accompanied by Mr. Wagenaar, takes the stage with a Handel Sonata for ‘cello and piano and another Haydn number, this time a tempo di Minuetto. Mr. Wolfinsohn’s portion of the program comes next, and consists of three characteristic national dances, Hungar- ian, by Brahms, Spanish, by De Falla, and Sla- vonic, by Dyorak. Dvorak is also the composer of the final number, the “Dumky” trio in E. Tickets are 75 cents and one dollar, with re- duced rates for subscribers to both concerts. LS eee el rr CrrtCC rr i Jury 21, 1934 ] THE COLLECTING NET 95 Wa BIOLOGICAL EABORATORY COLD SPRING HARBOR CRITIQUE OF CURVES OF GROWTH AND OF RELATIVE GROWTH C. B. DAVENPORT Department of Genetics, Carnegie Institution of Washington, Cold Spring Harbor, L. I., N. Y Cr ecm ane eG Te Ay eS oy MN te ay 4 1S Yrs. Figure 1. Development curve of human stature, conception to maturity, based upon measure- ments made upon about 100,000 boys, mostly Nordics. 1. GrowrTu CuRVES Typically, any animal which develops from the egg starts with a weight near zero and increases at first by small absolute increments in a unit of time; by gradually increasing increments it reaches a maximum velocity of growth; then slowly the age-weight curve of absolute growth flattens out until it is practically horizontal. The result is that the age-weight curve of absolute growth forms (disregarding certain irregulari- ties) an S-shaped, or logistic curve. (Figs. 1, 2). Thal > 4 we 1 16 4 Figure 2. Development curve of body weight, birth to maturity, based on measurements made of about 100,000 boys, mostly of Nordic origin. PPTs cts 6) tT 8 So Ny If, now, instead of this age-weight curve of absolute growth of the child we plot the incre- ment in a unit of time from the start of the fer- tilized egg onward—with age as abscissa and in- crement in weight as ordinates, we get quite a different shaped curve—one which shows the average velocity of growth of total body weight. This is, indeed, obviously not a simple curve but is made up of several components, two of which have roughly the properties of a Gaussian distri- bution curve. Especially if we examine the part of the increment curve centered (in males) at about 15 years, we find very nearly a typical Gaussian curve of error. (Figs. 3, 4). Figure 3 Unbroken line: Curve of varying veloci- ties of growth in weight, at yearly rate, of approximately 100,000 boys mostly of Nordic stock. Broken line: Two curves computed for the cir- cumnatal and adolescent spurts respectively, following Robertson’s hypothesis. Dot-and-dash line: Computed “residual growth”. Now such curves have long been plotted and considered to show the law of growth. The late T. Braisford Robertson based upon such a curve his conclusion that there are in humans three spurts of growth, one intrauterine, one at about 5 to 6 years of age, and one adolescent, and that each one corresponds to an autocatalytic reaction. In 1926 I had occasion to analyze this summation curve of growth and weight and reached the con- clusion that besides two main spurts of growth was a residum of real importance which was not due merely to an overlapping of the main spurts. THE COLLECTING NET [ Vor. IX. No. 75 cm. B [ mr | | to A Le ©) 8 7 A © ic s t—- 4 | A A i 7 if > 5 : c i e i] B ' | 1 a 615 LT Bio loan DI Ne SST ae 1B Sm rzom alice Figure 4. Light line: Curve of varying velocities of growth in stature, at yearly rate, based upon about 100,000 boys, mostly Nordic stock. Heavy lines: Three curves of velocities of growth at different ages of 3 boys: A-A, B-B, C-C. These three curves show typical variations in age at spurt and in intensity of the spurt. Juvenile and adolescent spurts are revealed. (Fig. 3). I then called attention to the fact that the curve of the adolescent spurt is essentially a Gaussian curve. Also that since the various organs of the body had each its own law of growth it could hardly be expected that the growth of the body as a whole, at adolescence, could correspond to one autocatalytic reaction. In 1931 I called attention to the fact that the in- dividual increment curve of growths, such as the weight or stature of an individual, are never like the summation curve (Fig. 4). Since the hypo- thesis of autocatalysis relates to individual growth and not merely to mass statistics, and since no individual grows the way shown by the mass growth curves, Robertson’s conclusion was based on incorrect premises, and had no validity. In this paper I suggested that the curve of adole- scent spurt, at least, depended for its significance upon the law of variation, and was due to the fact that the spurts of growth in boys tended to group themselves about 1414 years, but might occur three years earlier or later. To test this hypothesis I have had the distri- bution of spurts and the quantitative part played by these spurts by a number of cases plotted for 75 boys during the period from 12 to 18 years (Figs. 5, 6). The conclusion to which this table points is that the adolescent spurt is not at all evidence of a monomolecular catalytic reaction, but is an evidence of the method of distribution of the growth spurts. It has really nothing to do with the way a child, or other young animal, grows. This point I made in a paper published by the American Philosophical Society in 1931. Yet al- most till the present time analyses of the method of human growth are made from mass statistics, and conclusions concerning the energy of synthesis in growth are computed from these mass curves, some of which are really distribution curves. * In view of the ease of misinterpreting the law of growth from mass data, doubt is thrown upon all laws of growth based on mass statistics. The only way to learn how an organism grows is to study its growth. It is misleading to analyze merely mass data, and this holds for man and the lower mammals. Whether or not it holds for invertebrates and plants has yet to be determined. During many years investigators in the field of growth have tried their hands at devising general formulae for growth of the organism as a whole. Analysis of these curves has been made by W. Ludwig, who concludes that in all biologic growth the ratio V1/V = ¢ is decreasing; i. e. the pro- portional value to V1 to V is diminishing as V increases. Moreover, there are only four simple functions ¢, namely a linear decrease or reversed proportionate growth, and, indeed, either in re- lation to time (t) or mass or volume (v). Of these only two are important. These are: 1. ¢ =K (C—y). In this formula the speci- fic velocity of growth diminished with the body volume, V, reached is the smaller the nearer V comes to that final specific body weight (c). This is the logistic curve of Robertson and others. 2. ¢=K/t. In this formula the specific velocity of growth is inversely proportional to time; or, the specific velocity of growth is the smaller the longer the body has been growing. To this class belong the parabolic growth curves of Schmalhausen. But Saller (1927, p. 585) has concluded that a mathematical fermulation of external cause of growth (weight) is not today possible 1f such fomulation is required to come near to the essence of the growth process. A comparative consideration of the laws of growth leads to the result that the logistic and parabolic laws are on the one hand too simple for a treatment of growth by formula, on the other hand too complicated in order to be physiologi- cally founded. Also, Ludwig (1929, p. 758) concludes that treatment of growth by formula appears at pres- ent to be “played out” and ought appropriately to be relieved by a more descriptive method. Were there a growth activating factor that acted on all parts of the body at the same time we might lay more stress upon the curve of body erowth. But we know that the different organs grow at different rates and their maximum per- iods of growth occur at different times. I may be permitted to repeat a summary of this subject that I published eight years ago: “Thus Starkel and Wegrzynowski (1910) and E. Thomas (1911) find that the suprarenals grow * Cf. Wetzel, ’32, 33. Jury 21, 1934 } rapidly in the fetus, attaining, at or about birth, a weight of 3 gm. After birth the weight falls, absolutely, to about 1.5 gm. at about 12 months of postnatal life. It then increases very slowly to about 3 gm. at about the end of 5 years. Thomas shows that the degeneration after birth affects, especially, the deeper layers of the cortex. Scammon (1926,b)* shows, in addition, that in the suprarenals there is no extraordinary prenatal acceleration of growth but only a postnatal in- volution. A similar postnatal retardation of growth-velocity occurs in the cerebellum (Scam- mon and Dunn, 1924). “The length of the uterus in the fetus under- goes extraordinary changes that have been worked out by Scammon (1926, a)®. Thus in the 7th fetal (lunar) month the uterus begins to show an extraordinary spurt in growth, as compared with the body as a whole. At birth the length of the uterus is 35 mm. while, had the spurt not occurred, it would have been only about 23 mm. Within 3 weeks after birth the length of the uterus has fallen to 24 mm.; and then increases slightly during the following 5 months. “This suggests, says Scammon, ‘that the growth of the uterus in the latter fetal months consists of a substrate of typical fetal growth plus a secondary growth increment, which, presumably, is due to an extra stimulus furnished by a hormone of placental or possibly ovarian origin. After birth the organ loses this secondary increment but retains that resulting from the early, fetal growth rate.’ Again, refer- ence may be made to the well known case of the thymus, which, according to Hammar (1921)° undergoes a rapid reduction of size and function as adolescence sets in at 11 to 15 years. This involution seems to be determined and controlled by the development of the gonads. “The foregoing interesting studies on varia- tions in the velocity of growth of human organs justify the conclusion that the development of weight in man is the resultant of many, more or less elementary growth processes.” II. Curves or RELATIVE GRowTH It is clear that in the development of one of the higher animals we can distinguish between growth of the body as a whole and that of one of its ap- pendages, or any other part that grows differenti- ally. This is a point that Julian S. Huxley has stressed in his work on relative growth. For ex- -ample, he finds that the weight of the large chela of male fiddler crabs bears a constant growth ratio (from crabs of 60 mg. to 3144 gm.) to the weight of the rest of the body except that at 1.1 em., total weight, the ratio, known as K, abruptly changes to about 80 per cent. of its former value. The relation of growth of the smaller fraction of total body weight to the larger fraction is given THE COLLECTING NET 97 by the formula: y = b.x* or log. y = log. b + k log x. The theoretical justification for this formula is stated by Huxley thus: “One essential fact about growth is that it is a process of self multi- plication of living substance i. e., that the rate of] growth of an organism growing equally in all its parts is at any moment proportional to the size of the organism.” Now it is to this general thesis that I will first draw attention. Growth is not simply a pro- cess of self multiplication of living substance, if this phrase is used in the usual sense of micellor substance (protein particles). Studies that I made long ago on tadpoles indicate that the growth process is for a considerable time that of imbibition of water. In my study of frog de- velopment the total dry material diminished dur- ing 10 days from .80 mg. to .72 mg. per tadpole, whilst the weight increased 91% times. The abso- lute amount of water added per day increased slightly during this period, but it is not because the protein molecules increased, but possibly be- cause the lyophilic properties of these molecules became more efficient, perhaps just through their ets > % distribution of seu ° (UNAS uA TLS RIG NRITINT st ce 3) 14D IS Ie IT Figure 5. Unbroken line: Curve of total annual increments in stature of boys measured at Letchworth Village who had maximum spurts of growth in stature between the ages of 11% an 17% years. Broken line: The percentage of these individuals who had their maximum growth within the same age limits. Figure 6. Unbroken line: Total increments in weight of the same boys represented in Fig. 5 who had maximum spurts of growth in weight between the ages of 121%, and 18% years. Broken line: The percentage of individuals who had their maximum growth within the same limits. Figures 5 and 6 illustrate the point that the ado- lescent spurt of Fig. 3 is not primarily a biological but rather a statistical phenome- non. No child grows in the fashion of the adolescent spurt of Fig. 8, but rather as shown in Fig. 4. 68 THE COLLECTING NET f Vor. IX. No. 75 teal il ie! 102.5, = +— 100.0) + t— mS a 95.0) eek ae eee =| 925 a Bl | a 615 } = sa B50 =| 25 (eas ea ti B00 | mf | ed Te Yee Figure 7. Development curve of growth in length of the distal arm segment relative to that of the proximal segment, in Homo, Chimpanzee, Gorilla and Gibbon. There is a clear hetero- gony in the Homo curve, but it bears little relation to the formula: y = b.xk. The curve represents well the complex of factors in- volved in heterogony; phylo-ontogenetic and others. dispersal by the imbibed water. This is a quali- tative rather than a quantitative change. To reduce this case of the growth of the frog embryo, already cited, to one of relative growth, I have plotted the growth of water content to total weight, corresponding to Huxley's (1932) Fig. 20. Despite the different form of growth of the weight of the body as a whole during the days before 9 and after, the points actually found do not diverge widely from the straight line whose k value is 1.00. Apparently, the small differences in rate and method of growth, though significant, are swallowed up in the general trend. With the results found in the frog may be compared those found by Scammon and Calkins. According to their careful measurements the rate of growth of all external dimensions of the fetus from 5 to 35 cm. total body length (stature) is the same. Hence always k = 1. Another illustration may be taken from the growing plant. Reed, (Am. J. Bot. 8:377, 1920) measured a growing Bartlett pear shoot during 126 days. From 7 days to 14 days of age the seedling grew from 5 cm. tall to 11 cm. tall, an increase of 6 cm. When the organism was of size 112 cm. tall it grew in one week 1 cm. When it was half that size, or 55 cm. to 63 cm. tall, it increased about 8.5 cm. per week. There is thus no obvious relation between size of organism and the velocity of its growth.* Of course, in both illustrations the reason is that most of the in- *Tf it be objected that the growth of a seedling is not “equal in all of its parts” it may be pointed out that the same thing is true for practically all of the cases cited by Huxley as illustrating his fundamental equation. crease in weight is not self-multiplying living substance, but water in one instance and cellulose in the other. Thus it appears that Huxley’s “Essential fact about growth” is conceived too narrowly, and hence is not generally applicable. Whatever may be the objection to the theore- tical basis of Huxley’s fundamental principle, we should nevertheless examine his formula y = b.x* to see how far it may be useful in the analysis of relative growth. That it does express the rela- tion between the increase of chela weight to body weight in male fiddler crab between the limits of 1.7 and 28 mm. is clear. Various investigators have found the formula useful. In some curves of relative growth, as of the lower, in relation to upper, arm segment (brachial segment), the relation is so complex (as shown in Fig. 7) that the k value is quite diverse for the various ages. One can, of course, compute k for each age period, but it is questionable if the series of k values are easier to compare ana 1n-— terpret than the curves of varying proportions. Where the growth in length of the arm as a whole is compared with that of the chest diame- ter from 3 months after conception, the value of k is found to change from 1.00 up to a length 250 mm; to 2.00 from 250 to 450 mm; to 1.8 from 500 to 700 mm. (or about the adult length). The attempt to fit 3 straight lines to those points when plotted on double logarithmic paper is only partially successful. One feels that many more breaks are required to fit the points satisfactorily. Admitting that it is useful may times to ex- press quantitatively, by the differential coefficient, the rate of growth of different dimensions, one may look for a simpler qualitative expression than that of Huxley’s with its unjustified theo- retical basis. The suggestion has arisen that if the curves of growth of the two dimensions to be compared be plotted on ordinary graph paper, y, size, being plotted against x, time, the comparison of the tan- gents of their slopes at any age would give the relative growth ratio at that time. Or, the rela- tive growth of the two curves may be expressed by comparing’ their differential equations for any desired value of t. REFERENCES C. B. Davenport, 1926, J. Gen. Physiol., 10: 205-216. Cc. B. Davenport, 1930, Mem. Vol. 60th Birthd. Prof. V. Riizicka, Prag. C. B. Davenport, 1931, Proc. Am. Phil. Soc., 70: 381- 38 9. C. B. Davenport, 1934, Proc. Nat. Acad. Sc., 20: 359- 364. J. S. Huxley, 1932, Problems of Relative Growth, London. W. Ludwig, 1929, Biol. Centr., 49: 735-758. T. B. Robertson, 1923, Chemical basis of growth and senescence. Phila. K. Saller, 1927, Roux’s Archiv., 111: 453-592. ; } : | menus, are no longer to be found. Jury 21, 1934 J THE COLLECTING NET 99 THE WINTER OF 1934 AND ITS EFFECT UPON THE FAUNA AND FLORA OF WOODS HOLE (Continued from page 82) The abundance of certain biological material was seriously impaired by ice and exposure, while a few perennial garden shrubs were killed. To the delight of the younger generation and the dis- may of boat owners the Eel pond was frozen over for the first time since 1918, and afforded a per- fect surface for ice-boating and skating. How- ever, continued sub-zero weather together with the rise and fall of tides, ripped up piles and wharves which had to be anchored with ropes to prevent their being lost completely. Steamer service between New Bedford and the islands suffered many disruptions. Because of the floes packing the “hole” it was necessary to de- tour through the Quick’s Hole passage, which de- layed the schedule forty-five minutes. For ten days in February Nantucket was ice bound and cut off from all boat service. It was reported from the observation tower there that unbroken ice could be seen extending in every direction. Since transportation of mail and supplies by boat was impossible, the island was served for a week by planes from Woods Hole and Boston. By February 15 a Coast Guard cutter, with 3450 pounds of mail was able to detour through Vine- yard Sound and around Gay Head, approaching Nantucket from the east. The drifting floes did damage to wharves, spars, buoys and*piles. Oscar Hilton and James MelInnis report that for several days they had to chop out the ice from around their boats in the Eel pond each morning. The piles which were not completely lost all had to be redriven in the spring. The Supply Department of the Laboratory has been inconvenienced by the decrease of several formerly numerous species of invertebrate mater- jal. Cirratulus and arenicola were killed by being exposed to frost when the tide was low, for strong gales of wind swept over the bare flats causing unusually severe conditions. The plentiful beds of mytilus and other mussels, which in for- mer summers were the stock item on beach-party Hundreds of thousands of tons of ice were swept clean of mus- sel beds. Through the cooperation of the game warden and Falmouth sportsmen many waterfowl and upland game were protected from the hard con- ditions of the winter. Feeding stations were maintained along the shore and in the woods, and 1,600 pounds of grain were distributed. Even with this provision it is interesting to hear house- holders’ accounts of hundreds of gulls and terns which came to their backdoors far from the shore, in search of food. FRWue: THE WOODS HOLE CAMERA CLUB The Woods Hole Camera Club will hold its third meeting on Monday evening, July 30, at 7:30 o'clock in the Old Lecture Hall. At this meeting, the club will have as its speaker Mr. Er- nest H. Anthes of the Bausch & Lomb Optical Company, who will talk on “The Use of the Mi- croscope and Its Manipulation in Photomicro- graphy”. The meeting will be open to all who are interested in this subject. There will be a period of discussion following the talk. The Camera Club has been organized by a group of about twenty camera enthusiasts, both summer workers and all year residents of Woods Hole. It is planned to hold meetings each Mon- day evening at 7:30, at which some topic of in- terest to photographers will be presented by a competent speaker. These meetings will be open to other than members of the club. The talks for the remainder of this season will consider topics such as pictorial photography, portrait work, plates and films, the use of filters and color photography. During the latter part of August a salon will be held, at which will be exhibited work by members of the organization. Officers elected for this year are E. P. Little, president, and Thomas Goffin, secretary-treas- urer. The program committee consists of C. G. Grand, chairman, George Striker and R. L. Car- penter. An invitation is extended for anyone in- clined to photography as a hobby to become ac- tively associated with the Camera Club. M. B. L. CLUB NOTES A “smoker” is to be held at the M. B. L. Club on Friday, August 3, after the evening lecture. This occasion will be in honor of the foreign visitors at the laboratory and the Club extends a cordial invitation to everyone to be present and to meet them. Cigarettes will be provided and coffee served. The ping-pong tournament is well under way and is being handled very efficiently by Dr. Cos- tello. There are about twenty entrants. Several games have attracted large audiences. The whaleboat which was given to the Club through the generosity of the Supply Department is being used constantly and is much appreciated. Those wishing to use it should arrange in ad- vance with Mr. Bosworth who can be found in the botany building or the mess or with Dr. Sichel, Room 340, in order to obtain the key. There is a charge of fifty cents for each time the boat is used, and an additional charge of ten ceuts for each person using the boat who is not a member of the club. 100 THE COLLECTING NET [ Vor. IX. No. 75 Leitz Photometer “Leifo” A new Universal Instrument for Colori- metry, Nephelometry and Simplified Spec- tro Photometry will be displayed at our Exhibit in August 1934 at R. G. Thomp- son’s, Main Street, Woods Hole, Mass. With this instrument, which is based on a new principle, the following determinations can be made with highest accuracy: 1. Determinations of concentration of colored solutions. 2. Determinations of Nephelometric and Luminotric Intensities of solutions. 3. Determinations of Hydrogen-Ion con- centration. 4. Measurements of surface glare of solids. 5. Densitometric measurements of films, plates and other solid objects. 6. Measurements of color by the « {dditive Method. And many others It opens up many new fields for Colorimetry and Nephelometry. LER linc. DEPT ..562 | 60 EAST 10TH STREET NEW YORK CITY BRANCHES: Washington, D. C., Chicago, Ty San Francisco, Calif. Los Angeles, Calif. THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and 37 plates, published January, 1934 This guide for authors, in preparing manu- scripts and drawings for the most effective and economical method of publishing biologi- cal research, has been prepared by the Staff of The Wistar Institute Press and the codper- ative efforts of more than fifty editors con- cerned in the editing of journals published by The Wistar Institute, and presents the con- sensus of opinion on many points relating to the mechanical preparation of manuscripts and drawings for the printer and engraver. Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. Books Reduced 1-3 to 2-3 OFF Call at our Office on Water Street THE COLLECTING NE WOODS HOLE, MASS. \ EO 25 © ¢ MICRO SLIDES COVER GLASSES DO NOT FOG Ask your dealer —or write (giving dealers name) to AN “Ciay-Apams COMPANY S 25 East 26th Street NEW YORK /, wi bo oun ant NS Jury 21, 1934 ] THE COLLECTING NET as 101 Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, etc. Catalogs will gladly be sent on request. Please mention name of school and Spalteholz subjects taught, to enable us to 3 z Transparent send the appropriate catalog. rite Pie Mry, Preparations 5 : of Chic Human | (A : | NE ey Fa Tp. a me Cusv-Apams Company reac 25 EAST 26th STREET NEW YORK Model of Human Heart Visit our display rooms and museum. GRUEBEER Microscopical Stains Staining Solutions Physiological Preparations Sole Distributors AKATOS, Inc. 55 VAN DAM STREET NEW 'YORK CITY 102 _ THE COLLECTING NET [ Vor. IX. No. 75 BINOCULAR MICROSCOPE XIIlA One of the new series of low power, wide field binocular micro- scopes. Characterized by good definition, brilliant illumination, and ease in operation. Table of Magnifications, Free Working Distances, and Diameters of the Field of View | | Pair of objectives | % 144 214 Free Working | Distance in cm. | 14 12 8 2 Diameter Diameter Diameter Pair of Eyepieces Magnifi- | of field Magnifi- of field Magnifi- of field | cation ofwiewi |) cation CERES cation Ofralewe 8x 4 40 mm. 10 14.9 mm. 20 7.7 mm. 12% | 6% 36 mm. 15% 13.5 mm. 31 6.9 mm. Microscope XII A, as illustrated, including pair of objectives 14 x and pair) Of oCulars S26. one eee aeae $90.00 f.o.b. New York A copy of catalog’ Micro 464 will be supplied on request. CAR Ly Z ET spol N ee Al ty ey 1 ie ie lave VEG Oy Wy 728 So. Hal Sitwesce NE EW. Yr JOMRe ks L © S A NG Ege COLD SPRING HARBOR SYMPOSIA » LON QUANTITATIVE BIOEOG Volume I (resulting from conference-symposia of 1933 and dealing largely with surface phenomena) contains papers by Harold A. Abramson, D. R. Briggs, Robert Chambers, Barnett Cohen, Kenneth S. Cole, Hugo Fricke, Herbert S. Gasser, A. V. Hill, Duncan MacInnes, L. Michaelis, Stuart Mudd, Hans Mueller, W. J. V. Osterhout, Eric Ponder, Theodor Svedberg, D. D. Van Slyke. From a book review: “Jf this initial high standard (Volume 1) is maintained, it is diffi- I cult to see how a worker in this field can do without these volumes .. Volume II will appear in the autumn of this year. It will contain papers and edited dis- cussions resulting from the conference-symposia of 1934 concerning some aspects of growth. Authors include: W. T. Astbury, Felix Bernstein, H. W. Chalkley, George L. Clark, Charles B. Davenport, M. Demerec, Hugo Fricke, J. WW. Gowen, F’, Gudernatsch, F. S. Hammett, Theo. L. Jahn, L. G. Longsworth, Hans Mueller, Charles Packard, Otto Rahn, Nicolas Rashevsky, Oscar W. Richards, Charles R. Stockard, Victor C. Twitty, Harold C. Urey, C. Voegtlin, C. P. Winsor, Sewall Wright, Ralph W. G. Wyckoff. The prepublication price of Volume II, bound in cloth, is $2.90, cash with order. A fter publication, the price will be $3.35. The price of Volume I is $3.35. Persons purchasing Vol- ume I] may obtain Volume I for $3.00, Address the Biological Laboratory, Cold Spring Har- Donde. We N.Y Jury 21, 1934 | JHE COLLECUINGSNEU se a a 103 B & L Micro-Manipulator after the design of Dr, G. W. Fitz VISIT THE B & L EXHIBIT JULY 23 to 31 INCLUSIVE IN THE OLD LECTURE HALL l The GSET Microscope shown above has an inclined eyepiece and divisible body tube mounted on the stable, rigid, well balanced G type stand. OPTICAL IN AN IMPORTANT FACTOR IN AN OPTICAL INSTRUMENT Working with minute organisms the scientist receives real sat- isfaction from the amazing precision attained in the B & L Micro- Manipulator — evolved from 75 years of experience in the pro- duction of precision instruments. Men who have made the production of such instruments their life work* are responsible for the mechanical excellence of this instrument — designed for efficient micro-manipulation in its widest range. Smooth mechanical action, resistance to wear, absence of lost motion, are all the result of the ability of highly skilled workmen in holding to the close tolerances set forth in the specifications. The men who are responsible for the mechanical precision of the Micro-Manipulator put the same fine workmanship into all B & L Optical Instruments. For complete details on the Micro- Manipulator or the G line of Microscopes, write to Bausch & Lomb Optical Co., 671 St. Paul Street, Rochester, N. Y. *Over 220 B&L employees have been engaged in making B & L instruments for more than 25 years. Bausch s Lomb RUMENTS FOR THE.SCIENCES 104 __ THE COLLECTING NET { Vor. INS Now Phcto courtesy of Botany Dept. University of Toronto SPENCER Laboratory Lamps ERE is a typical university installation of the new Spencer Labo- ratory Lamp No. 372. Seventy of these lamps were installed in this college laboratory. Their merit in such laboratories is due to: 1. A single lamp provides efficient illumination for at least four microscopes. 2. No direct light can reach the eyes of the observers. 3. No additional illumination is needed in the laboratory. 4. The illumination approximates daylight... micro-spec- imens are observed in their true color values. 5. The beautiful crystal black enamel finish is alcohol and reagent proof. 6. Size is 17’’ high, shade 17” wide, globe 10”’ diameter. Spencer No. 372 Laboratory Lamp... complete, heavy stand, special inside frosted globe, 150 watt daylight bulb, metal shade and 8 feet of silk cord with switch . $22.50 Spencer Bakelite Substage Lamp An improved substage lamp combining low price with Spencer Quality. Made of bakelite which does not heat up as does metal. Construction gives im- proved ventilation. Spencer Lamp No. 385-A with the blue ees ground ononeside . . . . « $3.00 Spencer Lamp No. 385-B with doylion ‘los substi- tuted for blue glass . . Boo 6 fee No. 386 110 Volt Bulb . . . . 25¢ LAMP PRICES SUBJECT TO QUANTITY DISCOUNTS, OF COURSE Vol. IX. No. 5 S..TURDAY, JULY 28. 1934 Annual Subscription, $2.0 Single Copies, 25 Cents. THE COOPERATION OF MARINE BIOLOGICAL LABORATORIES Dr. WitttAm H. Cote Director, Mount Desert Island Biological Laboratory SOME EXPERIMENTS ON THE OSMOTIC PROPERTIES OF GLANDS ProF. RupOLF HOBER Professor of Physiology, University of Pennsylvania In the latter part of Dr. Harris’s article in the CoLtLectTinG Net for June 30th, he briefly sug- gested that cooperation between the marine bio- logical laboratories on the Atlantic seaboard might result in a real gain for biology. This suggestion is re- markable only because it has never been adopted. Coopera- tion between research organ- izdtions carrying on similar programs can not help but he beneficial if it involves no dic- tatorial concomitants. Amer- ican investigators are right- fully jealous of their freedom I propose to give you a review of a series of experiments on the osmotic properties of glands. My own interest in adhering to this object is not because I want to explain the specific function of this or that gland, especial- ly its role and position in the household of the human body, but because I feel attached to the general problem of the gland-like activity which we meet everywhere in a single cell as well as in the whole organism of plants or animals. In this connection, what is the M. B. E. Calendar TUESDAY, July 31, 8:00 P. M. Seminar: Dr. Coleen Fowler: “Per- meability of Amoeba proteus to water.” Dr. J. F. Danielli: “‘The tension at the surface of mackerel oil.” Dr. H. Burr Steinbach: “Injury po- tentials in scallop muscles.” Dr. Charlotte Haywood, Misses T. to conduct their research ac- cording to their plans, and justifiably refuse to enter into agreements with other investi- gators (even in the same field) for fear of losing that freedom. But if a plan of co- operation between laboratories can be devised which will not fetter in any way the scientific freedom of the respective in- vestigators and will not commit the governing boards of those labora- (Continued on page 115) Stevens, H. TeWinkel and M. Schott: “The relative effects of increased carbon dioxide and diminished oxygen upon _ the heart rate of young trout.” Dr. Robert Chambers: “The hy- aline plasma membrane of the echinoderm egg.” FRIDAY, August 3, 8:00 P. M. Lecture: Professor Gary N. Calk- ins: “Factors controlling proto- plasmic longevity in Protozoa.” meaning of gland-like activity ? In every organ we have to deal with its specific metabol- ism; in the muscle, with the building up of—let me say— myosin or of creatin-phos- phoric acid; in the brain with cerebrosides; in the connec- tive tissue with collagen. In the same way, if you review the typical glands, you will find that, like all the other organs, they fabricate characteristic substances, appearing in the glandular secretions; the salivary Hober Some Experiments on the Osmotic Prop- erties of Glands, Professor TABLE OF CONTENTS Rudolf 1929 105 Lhe Biological Laboratory at Cold Spring Harbor Editorial Page Airplane View of Woods Hole Taken in Items of Interest AIRPLANE VIEW OF WOODS HOLE TAKEN IN 1929 | Showing in the foreground at the left, the Church of the Messiah and the Falmouth Road; in the » background (from left to right) Little Harbor and the U. S. Lighthouse Service, the steamboat wharf, the drawbridge, the Bureau of Fisheries buildings, the Fel Pond, main building of the Marine Biologi- cal Laboratory, the Brick Dormitory and Penzance Point. THE BIOLOGICAL LABORATORY AT COLD SPRING HARBOR Showing (from left to right) the main building of the Carnegie Institution, summer laboratory buildings, Blackford Hall, dormitories, main building of The Biological Laboratory (in the back- ground), and several summer residences. . —= SS eee en SS ee, Jury 28, 1934 ] THE COLLECTING NET 107 gland produces mucin or special enzymes, the mammillary gland lactose, the liver glychocholic and taurocholic acid, and so on. But beside this specific metabolic activity we observe another sort of activity which is common to all glands, that is their osmotic activity. The gland produces its secretion out of the blood or out of the lymph. The composition of the secre- tion is, more or less, comparable to that of the issuing solution in regard to several constituents, but unequal in the concentrations of the con- stituents so that an osmotic “unequilibrium” has been established. This means: a dissolved sub- stance present in the issuing solution, viz., blood plasma or lymph, is brought to a higher or lower concentration in the secretion; therefore water is separated more or less from its solutes. In any case, work is done against the affinities between solvent and solute, and the gland has to obtain the energy supply for this osmotic work. Now it is evident: if we want:to analyze gland activity, it is necessary to compare the solutioms on both sides of the gland tissue; it means at first to become acquainted with the singular con- centrations: of every compound on both sides of the gland. It is not easy to work out the experi- mental conditions for this purpose in every case. For instance, Starling’s marvellous heart-lung- kidney preparation seems to me not to be an ideal method for analyzing kidney work because of the highly complicated composition of the perfusing blood. In comparison with this, the perfusion with a hemoglobin-Ringer solution, which Dr. Amber- son and I have used in our experiments on the osmotic properties of the submaxillary gland of the cat, seems obviously to be an improvement. Much closer to an ideal solution of the experi- mental problem is the procedure of Richards in his work on the frog’s kidney as he compares the composition of the perfusing Ringer solution with the content of a single convoluted tubule, ob- tained by puncturing it. This method is very sim- ilar to the experimental treatment of plant cells like Nitella, where the relatively high amount of cell sap allows an exact chemical analysis, so that you are enabled to follow the transport of chloride-ions from the outside medium across the thin living protoplasmic layer into the vacuole, which, according to Hoagland and Davis, is brought about by light as the energy supply. In the same way, according to F. C. Steward, the cells of potato slices are able to concentrate in their sap K and Br ions about 1,000 times, the energy for this storage deriving presumably from their high respiratory metabolism. These last examples demonstrate fairly well that we have to do here with a general and funda- mental problem of general physiology. The osmotic work of the plant cells, their transport of substance against the concentration decrease, is not at all different from the characteristic gland work. Here and there we are meeting the same problem of transport, necessary in cells either for nutrition or for elimination of waste prod- ucts, and in glands for different purposes. In many cases of this sort it is not difficult to demonstrate the activity of the living cell in these transport effects. The general property of all liv- ing material, viz., to be narcotized, enables! us every time to switch off the living machine, and by removing the narcotic, to switch it on once more. This method of procedure, to abolish the gland activity in a reversible manner, is often a great help in analyzing the function of a gland, for it would be misleading to argue that every substance which you find in a secretion has been worked out by the gland. In a detailed analysis of glandular function you must discover how far the organ behaves like a passive dead filter and how far like an active living transporting mech- anism, or in other words, how far you may speak of “passive physical permeability,” and how far of “active physiological permeability.” After this long preamble let me begin now with the reference to special glands. I propose at first to speak about the passive filter-like per- meability; later on about the active permeability and finally about the energy supply and the un- derlying chemical reactions. Suppose you make experiments on the isolated frog kidney perfused with Ringer solution, then you must differentiate mainly between the two types of walls separating the perfusing solution and the secretion product; one is the wall of the glomeruli, the other, the epithelial layer of the tubuli. The special circulatory conditions in the frog kidney enable us to offer any dissolved sub- stance on the one hand only to the outside of the proximal convoluted tubules, perfusing through the renal portal vein, or on the other hand only to the glomeruli, perfusing through the aorta. If you do perfuse through the renal portal only, you will find that when you add to the perfusing Ringer solution several lipoid insoluble non-elec- trolytes with different molecular sizes the small sized molecules like ethylenglycol or acetamide pass through the tubular wall, and the larger sized like mannite or asparagin will not pass through. In order to maintain as nearly as possible normal conditions and to obtain a copious urine the aorta is simultaneously supplied with normal Ringer solution only. The following table gives a sample of this type of our experiments : A series of experiments of this kind showed that the threshold value of molecular size is likely to lie between that of erythritol and that of ara- binose. We conclude that the epithelial wall of the tubules is comparable to a sievelike membrane with limited pore diameter. This conclusion is in best agreement with the fact that, as can be seen in the table, narcotization has no influence on the amount of penetration, The penetration is a 108 THE COLLECTING NET [ Vou. IX. No. 76 TABLE I. KIDNEY. Passive Penetration of Ethylenglycol m/100 ethylenglycol only from the venous side. 0.075% Phenylurethan the same. 0.0005% Phenolred from the art. and the venous side. ne Urine Time Addition to G = rfus. solution es onc. ot Perrus.Som Cone. of eth. el. phedolred 10.40 — 11.50 10 11.57 — 12.57 ethyl. glyc. 2.12 = 92% 9 1.00 — 1.30 eth. gl. + ureth. 1.80 = 80% 1.5 1.40 — 3.50 eth. gl. 1.56 = 69% 5 3.45 — 4.35 eth. gl. + ureth. 1.68 = 73% 125) purely physical process, where diffusion operates alone. But there is a remarkable deviation from this rule—here as well as in other cells and tissues,— if you take into account the behavior of amino acids; you find that they pass much more slowly than is to be expected or rather that they do not pass at all. We have found an explanation by experiments on collodion membranes with sufh- ciently small pores; the amino acids behave in re- spect to these artificial membranes like molecules with a higher molecular volume than follows from physico-chemical data. The reason is that, according to Bjerrum, the molecules of the amino acids are present nearly completely in the form of “Zwitterions,’” -+-NH3.R.COO_, which assumes the character of strong electric dipoles and there- fore are likely to be enveloped by a fairly ex- tended shell of water dipoles. Comparable to the lipoid insoluble nonelectro- lytes are several inorganic ions. For instance, the wall of the tubules appears to be totally imper- meable to chlorine, although Cl is washing con- stantly the outside of the tubules in the relatively high concentration of the Ringer solution. If you send through the glomeruli a Ringer solution in which the Cl-ions are replaced by SO,-ions, and if at the same time normal R is sent through the renal portal vein, you will find eventually a Cl-free urine. This is the more astonishing as from the inside of the tubules Cl-ions penetrate easily. Also a number of acid dyestuffs, charac- terized by a relatively high diffusion rate, have been demonstrated not to permeate the wall. In this way we learn that a tissue, which is a layer of cells plus intercellular substance, as is the simple layer of epithelia forming the walls of the tubules, can behave like a single cell with its selective impermeable membrane. What we find here, represents a remarkable behavior as for ex- ample the intestinal epithelium shows a much greater permeability. There exists still another way to study the passive physical permeability—as we have called it—on the isolated kidney. According to a great number of well-known observations on animal and plant cells you may assume that organic non- electrolytes can pass through the cells independ- ently of their molecular size, provided that they are lipoid soluble. As a matter of fact, we have. found that pinaconhydrate—comparable by its molecular size to the nonpenetrating arabinose— penetrates easily; even triethyl citrate of anti- pyrin, the molecular size of which equals nearly that of saccharose, goes through. Quite another aspect you get from the glom- eruli, if you regard their passive permeability. We are well informed about it, since Richards has succeeded in analyzing the punctate of the Bowman capsule, and since he has found out that apparently all crystalloidal substances which are present in the perfusing solution, are fit to pass through the wall of the glomerular tuft; sugars, amino acids, Cl, phospates and others appear in the capsule in the same concentration as they have in the perfusing solution. Only if you ap- proach more and more the field of colloidal dis- persion, you will find limits in the penetration. Many years ago I found out, that if a dye-stuff is retained by the glomeruli, it appears to be high- ly colloidal. More recently it has been shown by L. E. Bayliss and Miss Kerridge that only the relatively small sized protein-bodies, like oval- TABLE IT. Series of Molecular Series of Diffusion Volumes Rates ethylenglycol 65.5 | ethylenglycol 65.5 elycocoll 76.5 | glycerol 87.8 glycerol 87.8 elycocoll 70.5 alanin 98.5 erythritol 130.2 erythritol 130.2 alanin 98.5 asparagin 134.7 arabinose 153.4 arabinose 153.4 asparagin 134.7 lysin 177.0 | mannite 189.2 mannite 189.2 lysin 177.0 — Jury 28, 1934 } THE COLLECTING NET 109 TABLe ITI. Lipoid insoluble Lipoid soluble MV. M.V. ethylenglycol_ 65.5 acetamide 68.7 propionamide — 90.7 dioxyaceton_ 93.8 diethylurea 103.2 butyramide AZ, erythritol 130.2 asparagin 134.2 arabinose 153.4 pinaconhydrate 158.6 mannite 189.2 antipyrin 188.1 triethylcitrate 309 saccharose 345.6 The underlined substances are going passively through the tubular wall. bumin, Bence-Jones-protein and hemoglobin, pass through the glomerular wall. After all, we arrive at the conclusion that the wall of the kidney gland resembles at the same time a sieve-like membrane with pores of differ- ent diameter and membranes of lipoid material; the glomeruli especially resemble an ultrafilter insofar as they are impermeable to a great num- ber of colloids. Now, let us take into account the passive per- meability of another gland, the submaxillary gland of the cat. As you know, saliva is a strong- ly hypotonic secretion, the mean freezing point depression is about 0.15. It means that the secre- tion must be performed by the gland against an os- motic pressure of about 5 atmospheres, or in other words, the blood fluid must be pressed through the gland with a pressure of 5 atmospheres, since by far the most substances dissolved in the blood plasma are retained by the intact salivary gland. Is there no limiting, molecular size, beneath which, as in the artificial precipitation membranes, mole- cules of dissolved substances can be filtered through? Dr. Abramson and I tried two years ago to answer this question. As I have mentioned already, we succeeded in securing the survival of the gland for about 3 hours or more by perfus- ing it with hemoglobin-Ringer so that we were enabled to offer to the gland organic electrolytes in well-defined concentrations, uncomplicated by the presence of the more or less permeable and more or less absorbing erythrocytes. The ex- periments have shown, that under our experi- mental conditions, substances usually present in the blood, such as glucose, glycocoll, alanin, Cl, SO4, can be retained—more or less, and this is dependent on the rate of survival of the gland— while smaller molecules as those mentioned— acetamide, propionamide, dioxyacetone or lipoid soluble molecules like butyramide or dimethylurea pass into the saliva. It is hardly necessary to add that the saliva was obtained only by stimulating the chorda tympani. But it is worth mentioning that in the behavior of urea we met one remark- able exception; urea penetrates decidely more slowly than we would expect from its molecular size, compared with ethylenglycol or acetamide. We can explain this in a very general and there- fore unpresuming manner as indicating specific affinities to the gland; specific affinities which we will meet later as doubtless existing in the kid- ney. But there are existing several more obvious- ly comparable observations. According to ex- periments of Jacobs on hemolysis, urea pene- trates mammalian red blood cells with higher speed than acetamide or ethylenglycol; in fishes the opposite is true. Further, the studies of Collander and Barlund on plant cells gave the result that in Chara urea permeates more easily than glycerol; in Rhoeo, glycerol more easily than urea. Similar observations have been published by Wilbrandt, Orskov and myself. It seems to me, that these specific differences are dependent on slight variations in the chemical constitution of the plasma-membrane of the cells, so that there are operating either more basic or more acid va- lences. But this evening I have not time enough to follow this conception in detail. I Jeaye now the phenomenon of passive per- meability and turn your attention to the much more interesting chapter of active physiological permeability, to the appearance of real osmotic glandular work. I start once more with the iso- lated kidney of the frog. It is well known that the urine of the frog is often free from Cl: this seems to be in contrast to the before mentioned fact that Cl passes easily into the Bowman cap- sules. The same is true with glucose. The sim- ple explanation is, as you all know, that both of them are reabsorbed by the tubular epithelia. We have found recently still another substance, which does not appear in the urine, although it passes through the glomeruli, that is glycocoll or alanin, TABLE IV. Submaxillary Gland Perfused with m/30 butyramide = 55.6mg% NHg m/30 malonamide = 111.2mg% NH, Perfused with Sela % NaCl | mg% NHg blood 0.017 7 Hb. R. + butyramide 0.040 65 Hb. R. 0.055 2 Hb. R. + malonamide 0.058 30 Hb. R. 0.099 22 —_ 110 THE COLLECTING NET [ Vot. TX. No. 76 TABLE V. Kipney. Influence of phlorizin (2.10% mg%) on exeretion of glucose (60 mg%), uric acid (1.35 mg%) and sodium chloride (650 mg% ). - 1UF ie ay save Time Addition to % e perfus. solution 30 algeus mg% meg Yo P/M seess uric acid Na Cl 12.25 — 12.50 0) O7 319: 12.53 — 1.23 phlorizin 40 6.7 476 1.28 — 2.28 8 4.7 520 2.30 — 3.30 10 3.8 550 The reabsorption of these different substances has been demonstrated in several ways: it has been shown already twenty years ago by Bainbridge and his co-workers, that, after you have poisoned the proximal convoluted tubule by perfusing from the renal portal vein with mercuric chloride, Cl appears in the urine in the same concentration as it is in the perfusing Ringer solution. More physiological is our procedure, viz., to inhibit the reabsorbing epithelia with narcotic substances or by asphyxiating with cyanide, because these in- toxications are reversible ones. Another way to demonstrate the reabsorption is by analyzing the fluid obtained by puncturing the lumen of the tubules, as Richards and Wearn have done; they find that the Cl-concentration decreases more and more, the farther from the glomeruli the punctur- ing cannula is put into the lumen of the tubule. Now, the only substances the reabsorption of which has been noticed till today in the frog kid- ney are Cl, sugar and amino acid; others, like sul- fate, ethylsulfate, phosphate, ferrocyanide, dye- stuffs, are not reabsorbed. This faculty of the kidney is useful from the standpoint of the frog; for this faculty enables the frog to conserve food materials, like sugar and amino acid, which are on the way to escape the body, and to save chlor- ides, which the frog cannot get in sufficient amount from the fresh water. This interpretation seems justified also by studies relating to the properties of the kidney of elasmobranchs. From the old observations of von Schroeder in 1890 we know that the osmotic pressure of blood plasma in this class of animals is maintained to a great degree by urea. Therefrom we conclude that this metabolic product, urea, has to fulfill a special and important role, and following this it seems only natural and conceivable that, according to R. W. Clark and Homer Smith, urea is reabsorbed nearly completely in the kidney of elasmobranchs, so that the urine is almost ‘free from it. Thus we face the conclusion that there are created special machines in the kidney, fit for: driving the useful substances against the concen- tration gradient to the higher level of the blood fluid, whereas the path in the opposite direction from the blood to the lumina is closed. Is it pos- sible to tell anything about the nature of these machines ? Following old experiments of Hamburger we- have perfused the frog’s kidney not only with glu- cose but also with other hexoses like fructose, mannose, galactose, and with pentoses like xylose and arabinose. The result of our experiments is this: the reabsorption is very different for the dif- ferent sugars, viz., the reabsorption of glucose predominates, the other sugars follow in the series; galactose, mannose, fructose, xylose; the reabsorption is zero with arabinose. It is inter- esting to see that nearly the same series has been observed by Cori and Wilbrandt in the intestinal absorption of rats. What does it signify? It is well known that the sugars underlie metabolic changes in the organs at a very different rate, that they differ very much in their power of supplying energy to the body by their transformation, and therefore we may guess that special conditions are realized in order to fix the different sugars in the TABLE VI. Liver. Perfused by 0.0005% tropeolin in Ringer. Sac mir meyin Time Addition mg/hour concentr. of dye_ 12.05 — 12.50 3.25 430 12.50 — 1.40 +0.025% phen. ur. 4 0) 2.40 — 3.30 250 3.30 — 4.50 -+- m/1000 KCN 5 ; 9 4.50 — 6.15 8 70 Jury 28, 1934 ] THE COLLECTING NET 111 interior of the cells with different strengths for further chemical changes. These special binding faculties we meet also in the kidney. We have known for a long time that there exists an inter- esting poison, phlorizin, which is able to block the power of reabsorption of glucose in the kidney; you all know the phlorizin- or renaldiabetes. Now we have made experiments with phlorizin on the frog’s kidney with the following results: first, if you follow, in the same experiment on an isolated artificially perfused frog kidney, the reabsorption of Cl, of amino acid, and of glucose, you will find that phlorizin is injurious only for the absorption of glucose, not for that of Cl or of amino acid, or for the secretion of uric acid. We learn by these and other experiments that the dif- ferent reabsorbing machines in the walls of the tubules are working independently of each other. Second, the phlorizin prevents the reabsorption of all sugars, the large reabsorption of glucose as well as the small one of xylose. The explanation for this may be found in several observations by Lundsgaard and by Wilbrandt. These authors have shown, that in higher concentration phlori- zin can prevent sugar absorption also in the intes- tine, that phosphate favors the absorption in the intestine, and that phlorizin has a specific power of inhibiting the phosphorylization and dephosphory- lization. of glucose. Therefore they came to the con- clusion that hexosephosphoric acid might play a role as an intermediate compound in the transport of sugar, wherever it is absorbed. It might be possible to arrive through this hypothesis (the in- termediate formation of speci an understanding of the peculiar fact that the epi- thelia of the tubules behave like valves, so that the sugars can enter the epithelia of the tubules only from the inner side, while the path in the opposite direction is closed. Such a valve-like behavior we meet also with the transport of amino acids and of Cl. We shall now turn to the most significant os- motic work of the glands, the accumulation of solutes in the secretion, their transport from a lower concentration level to a higher one, as the gastric glands realize it with the hydrogen ions, the kidney with urea, the liver with the bile dyestuffs. I begin again with the kidney. The old controversy, whether every rise in concentra- tion in the urine, compared with the blood, is the effect of reabsorption of water, or whether it might be also for some compounds the result of a secretory transport, seems to me decided conclu- sively to the latter concept. Secretion can no longer be denied. This has been demonstrated especially by a number of dyestuff-experiments. I mention only the following: 1. One perfuses the isolated frog kidney through all its blood vessels, through the aorta as well as through the renal portal vein, with a faint pink mixture of very little phenol red and very little of the blue acid dyestuff cyanol in Ringer; one gets a urine almost purely deep red, because the phenol red is strongly concentrated, occasion- ally up to twenty times and more, whereas the cyanol is present in the same very low concentra- tion that is contained in the perfusing mixture. Now, if one narcotizes the proximal tubules only from the renal portal vein, there results a urine sometimes identical in its faint pink color with the perfusing mixture. There cannot be any other explanation than that phenol red is secreted by the tubules and that cyanol is not. 2. The second experiment has been made here in Woods Hole five years ago. Following Mar- shall’s studies on aglomerular kidney we inject- ed intravenously in a goosefish a similar mixture of phenol red and cyanol; both of them are car- ried by the blood to the aglomerular kidney of this fish, and we found that the dyestuffs are separated quantitatively from each other, so that only phe- nol red is secreted into the urine, induced by a se- lective affinity of the tubular epithelia to this substance. This experiment seems to me still more convincing than the former one, since there exists only one way from the blood to the lumina, viz., through the secretory tubules. 3. Armstrong and Chambers have made re- cently very delicate and significant experiments on embryonic tissue. Armstrong observed the pro- nephros of Fundulus and found out that after in- jection into the body of the embryo, phenol red appears to be highly concentrated in the lumina of the tubules be fore the glomeruli have developed at all. The experiments of Chambers have been done on tissue cultures of the mesonephros of nine to ten day old chicks. The isolated segments of proximal tubules, when cut into pieces, close their ends and remain alive. Incubated in a phe- nol red solution they begin, at a temperature ot 39°, to take up phenol red and to concentrate it in the lumina against a concentration gradient, till it reaches twenty or thirty times the outside concentration. In all these experiments phenol red has been used, because it has been found earlier by several authors that it can be concentrated by the kidney and that it resembles in this way one of the nor- mal characteristic constituents of the urine, e.g., urea. But as we have seen already, by no means all dyestuffs are fit for secretory concentration and it is worthwhile to ask what properties a dye- stuff must have ‘for being taken up by the epithe- lia of the tubules, stored by them and pushed through them. For if we know the nature of af- finities concerned in this transport we begin to understand the mechanism. Up to the present time we cannot answer this question; as far as I see only one rule holds true, viz., all hpoid soluble dyestuffs, whether basic or acid can be concen~ trated by the frog’s kidney; you supply the kid- ney with such a dy estuff only from the renal por- (Continued on page 113) 112 THE COLLECTING NEG [ Vor. IX. No. 76 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel Warner Parker, Margery Fuller Mitchell, and Annaleida Snyder Cattell. Printed by The Darwin Press, New Bedford PLAN OF PUBLICATION The publication of THE CoLLectinG NET has been delayed recently, and in order to enable us to return to our regular day of publication we are issuing a sixteen-page number; it may also be necessary to put out a small number next week. This scheme will not affect the total num- ber of pages which we print during the summer, for we plan to increase the number of pages in the last three numbers by an amount equivalent to that which we delete now. This arrangement will make it possible for us to accomodate some of the material which will accumulate towards the end of the season, and therefore result in a bet- ter magazine. In abbreviating this number it has been neces- sary to postpone for a week the publication of the following material : “Spindle Fiber Attachments, Do They Change without Chromosomal Incisions or Transloca- tions?” by Dr. E. ELEANOR CAROTHERS; “Ovu- lation and Ege Transport in the Frog’ by Dr. Roperts Rucu; “Certain Unusual Cytoplasmic Elements in the Yolk-sac Epithelium of the White Rat” by Dr. Joun W. Everett; “Com- ments on a Seminar Report by Dr. W. R. Dur- YEE” by Dr. M. E. KRAHL; an article from the Biological Laboratory at Cold Spring Harbor; news notes from the Mt. Desert Island Biological Laboratory and the Scripps Institution of Ocean- ography, as well as most of our “Items of Inter- est ;’’ comments on the “beach question ;” “Stand- ards of Conduct. III;” and the “Currents in the Hole.” COMMENTS ON A LECTURE GIVEN BY PROFESSOR RUDOLF HOBER Dr. RoBERT CHAMBERS Professor of Biology, New York University As was to be expected the lecture of Professor Hober was a most stimulating one especially to cell physiologists, who are legion at Woods Hole. Particularly refreshing was Professor Hober’s treatment of his subject in summarizing his ex- tensive researches and relating them to the funda- mental problem of transport of materials across cells. In a treatment of the subject of osmotic work done by living cells we have to thank Pro- fessor Hober for emphasizing the existence of an active “physiological permeability” which differs from what is usually implied in the term per- meability, not only from the fact that the cell is consuming energy in the process but also in the ability of the cells to drive materials in one di- rection against .a concentration gradient. He points out that a similar reaction obtains in plant cells in which the energy required has been found to be drawn from the presence of light and of respiratory processes. The kidney offers a pecu- larly well-adapted organ especially in the frog for an examination of the two types of permea- bility, since the passive filter-type predominates in the function of the glomerular tuft while the energy consuming type appears to be the main function of the tubules. Professor Hober has shown that the epithelium of the tubules may also react to nonelectrolytes as if it were a sieve-like membrane with limited pore diameter. It should be noted, however, that a'striking feature of the epithelium of the proximal tubule is the promin- ence of an intercellular cement-like substance. It is possible that the sieve type of permeability may be a function of the intercellular material rather than of the cells themselves. Of considerable interest in reference to the limit of size in penetration are his remarks on the passage of aminoacids which, by virtue of their molecules existing as Zwitterions, may have their molecular dimensions increased by a shell of water. Professor Hober has also shown that organic nonelectrolytes, which are lipoid-soluble, penetrate cells far more readily than lipoid- insoluble nonelectrolytes of corresponding size. However, the mere fact of lipoid solubility does not determine whether a dye will or will not be accumulated within the lumen of the tubule. There are lipoid-soluble dyes which manifestly accumulate within the cells without passing through into the lumen. Tissue culture exper- iments indicate that lipoid-soluble, basic dyes ac- cumulate within kidney cells and will not pass into the lumen unless the lumen is more acid than the pH of the cytoplasm of the surrounding cells. Even in the so-called passive type of permea- bility Professor Hober has pointed out, from cases in literature in addition to his own exper- iments, that specific differences occur between closely allied tissues. It is therefore impossible te make generalizations from intensive studies on only a few tissues. The other type of transport which Professor Hober happily has called “physiological permea- bility” and in the performance of which osmotic work is done must not be confused with cases of difference in partition coefficients due to solu- bility alone. For example, Kempton and I have shown that phenol red is transported across the (Continued on Page 113) 0 Ee Jury 28, 1934 ] THE COLLECTING NET tS ies “Or Introducing Dr. W. L. Francis of Cambridge, England, who is spending his first season at the Marine Biologi- eal Laboratory. He is working on the effects of electrolytes on the nerve of the crab, and previ- ously has published work which includes studies of protein membranes, and the electrical proper- ties and oxygen consumption of frog skin. Dr. Francis received his M. A. and Ph. D. de- grees at Cambridge University, and his B. Sc. at the University of London. In November 1933 he came to America on a Rockefeller Foundation Fellowship and has been working since then with Dr. W. J. V. Osterhout, at the Rockefeller Insti- tute for Medical Research. Dr. Emit Witscut, professor of zoology and experimental embryology at the University of Iowa, has begun his work at the Marine Biologi- cal Laboratory. Dr. Witschi and his son Hans motored to Woods Hole from Iowa by way of Canada and through Boston. Mrs. Witschi and their daughter Maryanne are spending the sum- mer in Switzerland. Dr. Witschi has done exten- sive work in the field of sex physiology especially on the subject of sex reversal in the amphibia. INTEREST M. B. L. CLUB NOTES There will be an informal reception at the M. B. L. Club in honor of the foreign investigators at the Laboratory following the Friday evening lecture, August 3. The scientific workers in Woods Hole are cordially invited to be present and to meet them. On Saturday evening, August 4th, the second “Mixer” of the season will be held in the Club- house at 8:30 P. M. Everyone who is connected with the scientific institutions of Woods Hole is invited to come. PROFESSOR CHAMBERS’ COMMENTS (Continued from Page 112) tubular wall independently of pH differences be- tween the lumen and the blood plasma. At the same time physiological permeability must not be interpreted as a convenient way of relegating the phenomenon to causes unknown. Professor Hober’s experiments indicate clearly that an ox- idation mechanism of some sort is undoubtedly involved. It is significant that in tissue-culture work the kidney tubules when narcotized have been found to be freely permeable to phenol red in both directions. This indicates that the ox- idative mechanism, when in full action, is respon- sible for the directed movements in the so-called “one way permeability.” SOME EXPERIMENTS ON THE OSMOTIC PROPERTIES OF GLANDS (Continued from page 111) tal vein and it will appear in the urine in a higher concentration. Such lipoid soluble dyestuffs are for instance, most of the basic dyes, the tropeolins, orange R, brilliant orange and others. This same rule holds as I have seen here in the last days, in the aglomerular kidney of the toad- fish; for instance, after intramuscular injection of orange R you may see under the microscope the lumina very distinctly filled with an orange con- tent, while the surrounding transporting epithelia of the tubules appear to be colorless. But beside this one rule for the dyestuff excretion there must be others, because a number of dyestuffs which are ordinarily lipoid insoluble, for instance phenol red, are also secreted. Nevertheless, the above- mentioned rule permits us to assume, that the en- trance of the dyestuff into the cell, by the passive means of dissolution in the lipoids of the cell, is in any case one of the essential steps in the scale of processes, effecting the secretory concentration. Very different from the behavior of the frog kidney in relation to dyestuffs is that of the frog liver. You perfuse first the isolated surviving liver from the vena abdominalis with Ringer solu- tion until the bile secretion is colorless; this you follow with a very dilute dyestuff-Ringer-solution and after some time you obtain often an amazing- ly deeply colored secretion; the dye has been con- centrated sometimes more than one thousand times,—an effect of the living organ, for as soon as you narcotize the liver, it loses this concentrat- ing faculty in a reversible manner. Still one point seems to me remarkable: even if the dye has been concentrated 1000 times by the liver, you do not see anything of it in the liver cells under the mi- croscope ; moreover, you do not see it in the bile capillaries, as they are too thin; as a matter of fact, it is unfortunately impossible to localize the interior surfacé, where the secretory concentra- tion happens. Now, what is different from the kidney is this: that the liver is able to perform the concentrating work with nearly every dye, basic or acid, diffusible or highly colloidal, lipoid soluble or insoluble. We ask again: what is the chemical or physico-chemical basis, the ruling con- dition of the work? What is the mechanism ? Let us return to the kidney. Naturally a spe- cial interest is attached to the nature of the elimi- 114 THE COLLECTING NET [ Vor. IX. No. 76 nation of the normal constituents of the urine. In common with the reabsorbing power of the kidney its secreting power also is intimately con- nected, or better, specifically adapted to the needs of the whole body. We understand that the epi- thelia of the tubules prevent sugar, amino acid and Cl from being lost to the body by escaping through the kidneys. With the same fitness or adaptability the kidney collects special substances which are useless and removes them from the body. For instance: according to Marshall and Grafflin, marine fishes drink sea water so that, by absorption in the intestine, Mg and SO, enter the blood in large amounts and disturb the normal ionic equilibrium. In the aglomerular kidney of Lophius this is avoided by a specific elimination of Mg and SOx, through the kidney, while other substances, like ferrocyanide or glucose which are foreign to the body, when injected into the blood, are not excreted at all. Another similar case is the following: as we have seen already, the molecular volume of urea is a very small one in comparison to many other organic nonelectrolytes. Because of this it can penetrate many cells more or less easily by passive diffusion. But this pas- sive manner of elimination seems not to be suffi- cient for the normal life of the body. As Mar- shall and Crane first have observed, urea is ac- cumulated in the frog kidney, especially on the dorsal side, where the proximal convoluted tu- bules lie. Now, if you perfuse the kidney with urea-Ringer-solution only from the renal portal vein, such an accumulation can be demonstrated. No accumulation occurs if you previously narcot- ize the cells. However, the accumulation occurs only when the tubules receive the supply of urea by way of the blood vessels and not by way of the lumina of the tubules. We meet once more the strange valve-like one-way permeability of the cells, this time in the opposite direction, i.e., pre- venting passage from the inside to the outside rather than in the opposite direction as with sugar, amino acid, and Cl. These and several other rea- sons lend support to the opinion that the accumu- lation of urea in the epithelia of the proximal con- voluted tubule is one step preparatory to the out- put of the urea into the lumen. The supposition of Rehberg seems to me unlikely, that from the stored urea is derived the ammonia production in the kidney. One question remains finally to be raised. Let us compare for a moment the various studies on the working muscle with studies on the working gland. In studying the muscle we are dealing with a great number of physical problems and we have to look for the histological background in the working machinery, but beside this we have also to investigate the chemical reactions deliver- ing the required free energy, which are involved in the contraction process. Likewise we must ask which sort of metabolic reaction is to be regarded as the source of energy for the osmotic machine of a gland. You may remember that in the beginning of this lecture I have compared the active gland with the cells of Nitella which, according to Hoagland and Davis, are transporting Cl ions against a diffusion gradient in the presence of light, and with potato slices which, according to Steward, are accumulating K and Br ions a thou- sand times in comparison to the outside solution as long as a strong respiration of the cells is maintained. Ina similar way the work of a gland is connected with metabolic reactions, and they remind us strikingly of the chemical changes which occur in contracting muscle. It is well known that glandular secretion is very much dependent on a sufficient supply of oxygen. Therefore, blocking the oxidation processes by cyanide inhibits more or less the characteristic glandular activity, as well as it does muscular ac- tivity. But also, as in the muscle, the gland work apparently depends to a good extent upon anaero- bic reactions, partially upon the transformation of carbohydrates into lactic acid and partially upon the splitting of organic phosphoric compounds. Since the observations of Lundsgaard on muscle, we know that the formation of lactic acid can be upset by iodoacetic acid, and that the working ma- chine is interrupted by this defect. The impor- tance of a similar effect on glandular work fol- lows from our observation that the activity of the kidney, of the liver and of the submaxillary gland can be abolished in the same way by iodoacetate. Further, it is well known from experiments of Meyerhof and of Feng that the activity of muscle and nerve, after being suppressed more or less by the 1odoacetate, can be restored by addition of lactic acid. In the same way the secretion of dye- stuff by the liver, which is stopped by perfusing with iodoacetate, is established in the presence of lactic acid. As a matter of fact, there can be no doubt that as in muscles, the formation of lactic acid is one link in the chain of processes, intimate- ly connected with the working metabolism. Fin- ally, according to Bergonzi and Bolcato, stimula- tion of the chorda-tympani sets free in the sub- maxillary gland phosphoric acid and lactic acid, and following this we have found out that after poisoning the gland with iodoacetate the phos- phoriec acid production increases while the produc- tion of lactic acid decreases. In conclusion: I have tried to present to you a survey of some progress in a field of physiological investigation where we encounter the same funda- mental problem of general physiology, the prob- lem of transport through the living cell. This is a problem of intake and output, to which our attention is focused everywhere in our wide field, in a consideration of every living anabolyzing and katabolyzing cell, in the nature of absorption or the nature of lymph formation and in many for 28, 1034} THE COLLECTING NET 115 other topics. I must apologize that, in order to propitiate this survey I had to remind you of a number of well known facts; but the main inten- tion was to show you that in this field of glandu- lar activity, opening more and more, we find highly inviting conditions to enable us to approach the sphinx of transport, this main problem of ap- plied physical chemistry, since the word permea- bility has its fascinating appeal to everybody who is attached to general physiology and nowhere more than in Woods Hole, this beautiful home of science. THE COOPERATION OF MARINE BIOLOGICAL LABORATORIES (Continued from page 105) tories to any inflexible procedure, then the labora- _ tories and their workers need have no fear of co- - operation. If this sort of cooperation is what Dr. Harris has in mind it should appeal to all con- cerned. The questions to be answered then are how can laboratories cooperate in such a way, and what are the steps that might be taken to consummate it? The replies to these questions must be intro- duced by a consideration of the history of biology during the past thirty-five years. At the turn of the century biology was just beginning the period of intensive analysis. Specialists began to appear and delimitation of fields began. This dividing of biology continued until many separated sectors became almost cut off from each other. Persons engaged in each sector became highly skilled tech- nicians and strongly entrenched theorists, with blissful disregard for the need of coordination and sympathetic attention for accomplishments in other sectors. That age of detailed analysis was necessary in the development of modern biology, but the time has come when the discoveries in various special fields must be synthesized, before further progress in any of those fields can be made. Analysis by specialists must continue but simultaneously an active liason between the re- spective fields must be established and maintained. Such coordination and synthesis cannot be limited to biological fields alone, but must include those aspects of mathematics, physics and chemistry that have been demonstrated to be fundamental to biology. There are encouraging signs today that the mathematicians, physicists, Ghernists and bio- logists are becoming more appreciative of the in- terrelations between their subjects, and that they can actually cooperate in solving each other’s problems. For example, at Cold Spring Harbor last summer, distinguished and competent lead- ers in each of those subjects sat around a confer- ence table and discussed with sympathetic consid- eration their mutual problems. It is difficult to imagine how such behavior can result in anything but real benefit to science. Each one of the marine laboratories has certain advantages and facilities peculiar to itself which result from its geographical location and from the past policies of their governing boards. These advantages and facilities should not become un- aac ennate ee ee ee EE EEE eee eee necessarily competitive, but should be so utilized by investigators that the greatest good to the larg- est number would result. In other words, the laboratories should not become scientifically iso- lated and unacquainted with the development in the other laboratories. Although each one might specialize in various branches of biology such specialization should not create a false perspective of biology as a whole. That condition can be pre- vented by informal and flexible agreements be- tween the laboratories so that emphasis can be placed on each laboratory’s peculiar facilities without unnecessary duplication and competition. Included in the policy would be provision for mu- tual criticism, advice and assistance. Personnel will largely determine the direction of each laboratory’s development, as Dr. Harris has point- ed out, but sympathetic consideration of mutual advantages and problems will make for greater efficiency in each one’s program. Furthermore, a united effort to advance biology cooperatively in that way will justify more fully the financial support which must continue to come from individuals and foundations in the form of annual contributions or endowments. Research in biology is becoming more and more expensive as it widens its fields to include quantitative investi- gations along physical and chemical paths. Therefore more and more money will have to be secured, and evidence will have to be presented to those persons and organizations furnishing the funds, that money is not being wasted by short sighted policies, unnecessary duplication and competition, Steps that might be taken to advance such a co- operative program are: (1) conferences between the governing boards and directors of the labora- tories concerned, at which full discussions of the proposition could be had pro and con. Practically unanimous agreement should be attained before any plan is adopted. (2) Informal designation of the CottectinG NE? as the unofficial organ of the laboratories, which would serve as a clearing for presentation of certain research and reports of lectures, for discussions of mutual problems and for news items, resulting in a community of in- terest between the laboratories and their inves- tigators. (3) Public announcement of the pro- posed plan with the invitation for criticism and advice extended to American biologists. 116 THE COLLECTING NET [ Vor. TX. No. 76 Spalteholz Transparent Preparations Human and Zoological Model of Human Heart WHEN YOU ARE IN CHICAGO FOR THE 1934 WORLD’S FAIR You Are CorDIALLY INVITED TO VISIT The Turtox Laboratories and see where many of the biological exhibits were prepared for the Cen- tury of Progress Exposition, GENERAL BIOLOGICAL SUPPLY HOUSE Incorporated 761-763 EAST SIXTY-NINTH PLACE CHICAGO Skeleton of Fish in Case Models, Specimens, for Physiology, Zoology, Botany, Anatomy, Embryology, ete. Catalogs will gladly be sent on Please mention name of school and subjects taught, to enable us to send the appropriate catalog. Charts request. Life History of Chick |Crav-Apams Company 25 EAST 26th STREET Visit our display rooms and museum. NEW YORK THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and 37 plates, published January, 1934 This guide for authors, in preparing manu- scripts and drawings for the most effective and economical method of publishing biologi- cal research, has been prepared by the Staff of The Wistar Institute Press and the co6per- ative efforts of more than fifty editors con- cerned in the editing of journals published by The Wistar Institute, and presents the con- sensus of opinion on many points relating to the mechanical preparation of manuscripts and drawings for the printer and engraver. Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. Jury 28,1934] = THE COLLECTING NET New Laboratory LAL on the Reptilia THE COLLARED LIZARD By BD. DWIGHT DAIS Field Museum of Natural History This is the first general guide to the dissection of a reptilian form to be published in this country. The collared lizard was chosen as the specimen to be studied because of its size and availability, and because it represents a relatively unspecialized terrestrial form which may well serve as a type for the lizards and reptiles in general. Features of particular interest from the standpoint of comparative anatomy have been stressed throughout the manual. Each of the major systems of the body is taken up separately, and the work is so planned that, unless a complete dissec- tion of the circulatory system is undertaken, it may be carried out on a single specimen. In the opinion of the author, it is extraordinary that a vertebrate group as large and important as the Reptilia, which has long held the place it deserves in European institutions, has been neglected to such an extent in America.” His manual will fill a definite need in the study of compara- tive anatomy, vertebrate zoology, etc. To be published this fall. Probable price, $0.90 THE MACMILLAN COMPANY 60 Fifth Avenue New York Pie THE COLLECTING NET [ Vor. IX. No. 76 COLUMBIA : PARAFFIN OVEN A safe, efficient, simple, and inexpensive Oven for individual or small class use, for infiltration, embedding, spreading, and drying. It was designed by Dr. C. E. 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THE NEW SPENCER MEDICAL MICROSCOPE OFFERS THESE ADVANTAGES ® Balanced Optical System ®@ Metal-Mounted Achromatic Oil Immersion Objective ® Conical Eyepiece ® Micrometer Fine Adjustment @ Dual-Cone Nosepiece ® Fork-Type Substage @ Research-Type Stand ® Rhodium Plated Parts WRITE FOR FOLDER M-61-S FOR COMPLETE DESCRIPTION AND PRICES [ Vor. IX. No. 76 increase the range of the ARW Magnifications and. Real Fields for KW Paired Eyepiece Objective Magnification Number _ Magnification No. 445mm Microscope 60X 3.82mm yeaa new development, additional optics for the AKW, is further indication of Bausch & Lomb’s ability to anticipate and meet your needs with the finest in optical quality. The substitution of two new objectives (4X and 7.5X) and addition of one new set of paired eyepieces (20X) adds seven new magnifications* to the AK W Microscope. BOX 292mm Now a range of 7.0 to 150 diameters is available instead of the former 7.0 to 87 diameters! Of great importance is the wide field ob- tained despite the high magnification. The real field in higher powers is much larger than that of ordinary binocular types while in the lower magnifications, it is more than double. *The figures in the black squares in the above table are the new magnifications. Our new catalogue No. D15 describes our Binocular Mi- croscopes completely. It will be gladly sent on request. Bausch & Lomb Optical Co., 671 St. Paul Street, Roches- ter, New York. Bausch s Lomb OR Neot RUMENTS -F OR THE SCIENCES Vol. IX. No. 6 SATURDAY, AUGUST 4, 1934 Annual Subscription, $2.00 Single Copies, 25 Cents. OVULATION AND EGG TRANSPORT IN GROWTH CORRELATIONS IN AMPHIBIA STUDIED BY TRANSPLANTATION * THE FROG Roperts RuGH Instructor of Zoology, Hunter College The sex-stimulating hormone of the anterior pituitary in the frog has as one of its functions the liberation of the egg from its follicle. This is really a dual process, involving first the rupture V. C. Twitty Stanford University The method of transplantation in the study of animal development is essentially a means of in- troducing any given part of the organism into-a new organic environment, and thereby of testing of the follicle and second, the muscular contraction of the follicle wall which forces the egg out through the rupture area. This opening rarely ex- ceeds half the diameter of the egg. Ovulation induced by anter- ior pituitary injection cannot be regarded as an all-or-none reaction, for the degree to which the ovaries of a partic- ular frog are emptied depends upon the amount of this pitui- tary hormone injected. This can be clearly demonstrated in a frog into which only one male pituitary has been inject- ed. Within 40 hours all of the eggs which are going to leave the ovary will have been freed MW. B. UE. Calendar TUESDAY, August 7, 8:00 P. M. Seminar: Dr. L. G. Livingston: “Plasmodesma in Plant Tissue.” Mr. Charles E. Renn: ‘Concerning the Disappearance of the Eel Grass.” Dr. Selman A. Waksman and Dr. Cornelia L. Carey: ‘Origin and Chemical Nature of Organic Matter in the Sea Water and Sea Sea Bottom.” FRIDAY, August 11, 8:00 P. M. Lecture: Professor Henry E. Crampton: “Studies on Evolu- tion in the Islands of the South Seas.”’ Illustrated. from their follicles. Such the effect of altered relation- ships on the subsequent devel- opmental behavior of the part. 3y a comparison of this be- havior in new surroundings with its customary develop- ment, one attempts to deter- mine the extent to which the conduct of this part is due to innate factors, resident in its cells, and to what extent its development is a_ function of environmental relationships within the organism. Primarily, this method has been applied in the analysis of qualitative transformations during development, i.e., to processes of differentiation proper, such as the appearance of the primary axial struc- tures, and later, the appear- an ovary will show, as a rule, only one lobe completely emptied of its eggs. The ovarian lobes (numbering from four (Continued on page 130) ~ = ———E— —— ance of the various organ rudiments. Thus it has been determined that the ectoderm of the amphib- ian gastrula seems to lack the innate factors Growth Correlations in Amphibia, Dr. V. Twitty 121 Ovulation and Egg Transport in the Frog, Roberts Rugh 121 Woods Hole Scene from Painting by zs 122 Franklin L. Gifford TABLE OF CONTENTS EditorialiePace! io... case eee 128 Book Review, Dr. Leonor Michaelis 129 Itemsvot, interesti«.2.<.0 ee ee 129 Comments on the Seminar Report by Dr. W. R. Duryee, Dr. M. E. Krahl SHR, eek THE ROCKS ON THE “PUBLIC BEACH” IN 19382 THE SAND ON THE “PRIVATE BEACH” IN 1932 Aucust 4, 1934 ] THE COLLECTING NET 123 ile: BIOLOGICAL EABORATORY COLD SPRING HARBOR GROWTH CORRELATIONS IN AMPHIBIA STUDIED BY THE METHOD OF TRANSPLANTATION (Continued from Page 121) necessary for the differentiation of neural tube, until these have been imposed upon the ectoderm by an extrinsic influence, in this case by the or- ganizer. The method is equally applicable, however, in the analysis of problems of growth, 1.e., to quan- titative aspects of development. The foremost of these problems pertains to the manner in which the growth of the component organs of the ani- mal is integrated within the growth of the whole. Although one may describe the growth of the or- ganism as the sum of the growth of its parts, it is scarcely possible to conceive that the latter grow independently of each other. It is thus conveni- ent, in any attempt to formulate the problem of orderly growth of parts, to distinguish between growth tendencies inherent in the parts them- selves, and regulatory mechanisms which exercise a coordinating influence. This distinction between intrinsic and extrinsic factors in the process of growth was recognized by Harrison (’24), who applied the terms “growth factor” and “regulatory factor”, respectively. The influence of intrinsic factors on the devel- opment of size is amply demonstrated by the ex- periments of Harrison and his students‘), who have shown that the growth of organs transplant- ed between slowly and rapidly growing species of salamanders is determined primarily by their ori- gin rather than by the size or developmental rate of the host organism. Perhaps the most striking case of this specificity in growth is that described by Twitty and Schwind (’31), for eyes and limbs exchanged between embryos of the large Ambly- stoma tigrinum and the smaller A. punctatum. The grafts parallel the normal donor organs in size with remarkable precision. In spite of the general adherence to specific growth rate which characterizes the results of the various experiments on heteroplastic transplanta- tion there are nevertheless discrepancies in several cases which reflect the operation of extrinsic in- * This report is based in part on material pre- sented by Twitty and Elliott in a current number of Jour. Exper. Zool., vol. 68. Readers of that article are referred to the present paper for a fuller discus- sion of the data therein presented. A comprehen- Sive account of the transplantation experiments on growth in amphibians will be given in a review ar- ticle to appear later. fluences associated with the organic environment of the host. In the discussion to follow, we shall review some of these factors, and others of prob- ably even greater importance, which have been demonstrated by transplantation experiments to affect the proportionate growth of parts. The importance of function on the size of transplanted organs is strongly evidenced in the results of Copenhaver (30), who found that hearts of 4. tigrinum attempt for a time to follow their specific growth rate when grafted to A. punctatum but later tend to regulate their propor- tions in accordance with the functional require- ments of their smaller hosts. Factors within this same category probably played a similar role in the experiments of Detwiler (’32). Portions of cord grafted from A. tigrinum to A. punctatum at first became larger than the adjoining segments but subsequently underwent regulation to dimen- sions proper to the host. In both of these cases, the grafts are placed into very intimate organic union with the rest of the animal and it is per- haps not surprising that they are unable to realize the independence of growth which characterizes such structurely discrete organs as the eye and limb. 4 Purely mechanical factors, if we are permitted to refer to them as such, may also alter the intrin- sic growth rate. This seems to be clearly the case for heteroplastically transplanted ears (Richard- son, '32). The tigrinum organ in the small punc- tatum, confined as it is between more rigid struc- tures within the interior of the head, does not be- come as large as under normal conditions ; where- as the punctatum ear on the large tigrinum is able to expand beyond its usual proportions. The in- ner ear in amphibian larvae is a thin-walled, weakly capsulated organ, whose exact dimensions are probably much more subject to the mechani- cal circumstances surrounding its development than is the case for more highly individuated or- gans. A third and very obvious environmental factor which might modify the inherent growth tenden- cies of transplanted organs, is the nutritive con- ditions within the host organism. Unfortunately, however, it is one of the most difficult to identify (1)See also Rotmann, ’31, ’33. 124 THE COLLECTING NET [ Vor. IX. No. 77 and evaluate. Granted that both donor and host species are maximally fed, there is at present no way of knowing that the concentration of food- stuffs in the blood stream is maintained at the same level in each. A possible interpretation of the results of Twitty and Schwind on 4. tigrinum and Ad. punctatum, in which the eyes and limbs grew equally well in their normal environment and when transplanted to the other species, is that the nutrient level in these two forms is practically identical. However, when we attempt to apply this interpretation to certain other results reported for these species, the task is less simple. Detwiler (’32), in his transplantations of por- tions of the cord from tigrinum to punctatum, found that in the earlier larval stages the graft exceeded not only the size of adjacent host seg- ments but also the size of corresponding segments in normal control larvae of the donor species. In the case of the cord at least, then, we must con- cede some differences in the organic environment provided by the two species. The question, as yet unanswered, is whether to identify this difference with nutritive factors or with some less tangible influence of the host. The interpretation is par- ticularly difficult in the case of the cord because of its complicated structural and functional rela- tionships. Detwiler (’30) has also described another in- stance where the graft was accelerated beyond its normal growth rate. This condition was noted in experiments with tigrinum and punctatum, in which he used material of the former species ob- tained from Tennessee, and evidently of different racial stock from the Illinois material previously used. Limbs transplanted from these embryos to punctatum exceeded the growth of the control tigrinum organs. Although the normal donor larvae grow less rapidly than the Illinois stock, their organs when grafted to punctatum grow more rapidly than grafts from the latter. This situation is particularly puzzling, although trans- plantations between the two stocks of tigrinum might prove enlightening. We are evidently deal- ing with a situation similar to that originally de- scribed by Harrison C24), who postulated the existence of “regulatory” factors in the organic medium provided by the host to account for his observations. Although the nature of these fac- tors is still obscure, they may yet prove to be es- sentially nutritive in character. Results contributed by the writer probably pro- vide a clearer example of the operation of nutri- tive factors in the control of size. Eyes of 4. tigrinum grafted embryonically to the smaller Triturus fail to realize their characteristic growth rate (Twitty & Elliott, 34). Their handicap is particularly great in the earlier stages of the ex- periment when the Triturus hosts are still utiliz- ing stored yolk for their growth. Later, when in- dependent feeding begins, the grafts show a par- tial recovery and threaten to overtake the normal eyes of the donor. It is believed that this initial retardation of the grafts is a direct result of in- ferior nutritive conditions provided by the hosts. The Triturus embryo is smaller than tigrinum and develops much less rapidly. By the time yolk absorption is complete in the former, the larvae of A. tigrinum have already doubled their length since the beginning of independent feeding. The fact that Triturus requires a longer period to con- vert a much smaller amount of yolk is strong evi- dence that the nutritive conditions are decidedly less favorable for the grafts than in their normal environment, and is quite sufficient to account for their slower growth. After the Triturus hosts have begun to utilize external sources of food, there is evidence that the nutrient level becomes somewhat higher. This is reflected not only in an increase in the growth rate of the Triturus larvae themselves, but also by a greater growth of the grafts relative to the latter. There is a period during which the transplants actually grow more rapidly than the normal tigrinum organs them- selves, but complete recovery from the initial handicap is not achieved. In summarizing the contribution of the various experiments reviewed above, we may state that, although genetic factors are of primary impor- tance in determining the size of heteroplastically grafted organs, the specific growth rate is never- theless susceptible to modification by functional, mechanical, and nutritive conditions within the organic environment of the host. It will be evi- dent, however, that these discoveries represent merely the first step in the analysis of growth regulation. The level of nutrient materials in the blood, functional relationships, etc., although sometimes capable of modifying the expression of intrinsic growth rate of individual organs, are scarcely adequate as the sole devices for integrat- ing their growth within the whole. Much more basic mechanisms must be in operation to account fully for the precise coordination by which con- stancy of proportions within the individuals of a type is preserved. We may assume that these factors are qualitatively similar in all species, and hence not always to be revealed by quantitative comparisons between different forms in the man- ner of the experiments summarized above. In this new category we are able to recognize mechanisms of at least two kinds, which appear to be fundamentally different. The first regulates the proportionate growth between structures inti- mately related to each other in their development. Its operation is clearly evidenced in the experi- ments of Harrison (’29) on the heteroplastic transplantation of the embryonic constituents of the eye. In an eye compounded of a punctatum cup and a lens of the larger tigrinum, the former is accelerated and the latter retarded in growth, to produce an eye which approaches harmony in —— Auvcusr 4, 1934 ] THE COLLECTING NET 125 the relative size of its parts. This case serves to illustrate the difficulty often encountered in dis- tinguishing sharply between intrinsic and extrin- sic factors in the control of growth. The “‘intrin- sic’ growth of the lens becomes an “‘extrinsic’’ influence when it modifies the growth of the cup. Therefore, the growth of the eye as a whole can not be regarded as the unmodified expression of intrinsic factors, but rather the resultant of an in- terplay between its component tissue characteris- ics. This statement for the eye undoubtedly ap- plies in varying degree to the majority of organs, and it is probably exceptional when the constitu- ent parts of a highly individuated structure are allowed a free and independent realization of their inherent possibilities for growth. Relative freedom in this respect is enjoyed primarily only by the organ as a whole. Its independence of en- vironmental relationships has been frequently demonstrated by heterotopic transplantations of total organ rudiments, which often develop as in- tactly and as rapidly as when in their normal posi- tion in the animal. Not only do the parts of an organ modify the growth of each other but the organ as a whole may alter the dimensions of structures with which it is intimately associated. The size of the eye determines, within limits, the size attained by the extrinsic ocular muscles (Twitty, 32) as well as the number of cells within the optic brain centers (Harrison, 29; Twitty, “32; Larsell, 29; et al). The familiar experiments of Detwiler (°33), Burr (°30), and others on the development of the nerv- ous system provide numerous instances of rela- tionships of this nature. The correlated growth in these cases is evident- ly due to dynamic interactions between the parts affected and appears to be dependent in most cases upon physical contiguity during develop- ment. Regulatory devices of this nature, which operate at only a restricted range within the ani- mal, do not account, obviously, for “integration at a distance’, between parts which are unrelated in their development. The latter phenomenon must be governed by a mechanism essentially dif- ferent in type. The existence of this second type of mechan- ism is clearly demonstrated by experiments de- signed specifically to create disharmonic size-rela- tionships between an organ and the rest of the animal, and by a study of their subsequent regula- tion. By “disharmonic” in this sense we do not refer to the unusual proportions produced hy em- bryonic transplantations between species of differ- ent growth rates, as a result of which the graft is smaller or larger than the normal paired organ of the host. In the latter case the difference in size is simply an expression of the difference in the inherent growth rates of the two organs. Both have developed synchronously, and in spite of their difference in size, are in the same stage of growth. Actual disharmony in growth stage is created only when one organ is given an initial growth handicap, or advantage, over its mate; when, in other words, we transplant organs to hosts older or younger than the donors. The results demon- strate very clearly the operation of forces which integrate the growth of the part with that of the whole. Grafts younger in growth stage than their hosts grow faster than under normal conditions ; those more advanced in growth stage than the ani- mal itself are retarded; so that in both cases ap- proximately normal relationships are ultimately reestablished. The absolute growth rate of the eye is thus partly a function of the environment in which its development takes place. This is il- lustrated lucidly in the experiment (Twitty & El- liott, 34) where the two eyes of a single animal are grafted, one to an older larva, the other to a younger. Although both grafts were identical, the former was accelerated and the latter retarded in its growth. The difference in their behavior can be attributed only to some condition, within the animal as a whole, which changes with the stage of development. The most satisfactory explanation which has heen offered for the nature of these changes is based upon Robb’s (29) application to the grow- ing animal of the principle of “partition coeffi- cients”. “As this expression is used in chemical terminology, it refers to the tendency of a sub- stance to distribute itself between two accessible solvents in proportion to its relative solubility in the two. In the case of the growing animal, par- tition phenomena between adjacent cells would result in an unequal but regulated distribution of available raw materials between the various tis- sues. The circulating medium which bathes the latter carries a limited amount of nutritive ma- terials, and the relative affinities for this food of the cells of different organs, or of different parts of one organ, will determine their relative growth and hence the characteristic body proportions of the animal” (restatement of Robb’s theory by Twitty and Schwind, 31). Huxley (32) arrived at a similar conception independently and it has been applied by Moment (°33) in his analvsis of growth in the rat. ; _ To adapt this idea to the explanation of regula- tive growth, it is merely necessary to postulate a gradual fall in the assimilative capacity of the body as a whole during larval development. This assumption is based on the generally accepted fact that true or percentage growth of the animal body decreases with age, which implies that the cells assimilate food less actively as their numbers in- crease. If this is true, then an eye grafted to an older animal would have an advantage over the rest of the animal in the competition for nutritive material, whereas an organ older than its host 126 THE COLLECTING NET [ Vor. IX. No. 77 would be handicapped in the struggle for avail- able food. In the former case the graft would be accelerated in its relative growth, in the latter case, retarded. These were the results noted in the original experiments of Twitty (30), and recently confirmed and extended. This interpretation enables one to form a fairly clear picture of the nature of growth regulation. According to it, the proportions within the body at any given moment are the product of an equi- librium between the competitive strengths (‘par- tition coefficients’) of the component parts which share the aie of growth. We know, how- ever, that the body-proportions often change as growth progresses. The vertebrate eye, for ex- ample, characteristically diminishes in_ relative size. The large chela of the crab increases with reference to the animal as a whole (Huxley). It is thus necessary to postulate that a general de- cline with age in the level of assimilation through- out the body does not affect all parts equally. Where the value for a given organ falls less rapidly than the average “for the entire system, “positively heterogonic”’ growth results 5 when the decline is more rapid in the part, “negatively heterogonic”’ growth characterizes the latter. Ac- cording to this conception, heterogonic growth (Huxley ) is at least partly the result of a differ- ential in the capacity of part and whole to assim- ilate food from a common medium. When this differential is increased or decreased by transplan- tation between individuals of different age, or dif- ferent specific grow th rates, the intensity of heter- ognic growth is altered accordingly. ‘Tt is even possible to reverse the normal relationship, chang- ing the heterogony of part to whole between posi- tive and negative values. With the “adoption of this theory, the differen- tiation between “intrinsic” and “extrinsic” factors in growth regulation largely disappears as a fundamental distinction. The various organs, in attempting to express their intrinsic growth rates independently, operate extrinsically on each other, limiting growth mutually through competition. The circulating medium plays a neutral role, since its food materials are equally available to all. This, of course, does not take into account the action of hormones, which may and often do af- fect the relative size of parts. Their influence is clearly subsidiary, however, to the more fund- amental mechanism already outlined. A larva de- prived of pituitary and thyroid, for example, al- though modified somewhat in its proportions, nevertheless attains them uniformly, in accord- ance with the orderly rules of growth. The accelerated growth of a young transplant- ed eye on an older animal has been attributed to its advantage over the host in securing nutrient materials. The question may be raised whether the relative distribution of food between the two would be affected by the level of feeding. We have tested this possibility under four different conditions. (1) Complete starvation, in which neither graft nor host increased in size (Twitty 30). This shows that the graft, in spite of its advantage, is unable to “‘parasitize” the host. (2) “Balanced feeding”. We apply this term to pe- riods of light feeding in which the amount of food was accidentally adjusted at the precise level necessary to maintain the body at a constant size. The graft, however, was able to secure enough nourishment to continue growth. This im- dependent growth of the part represents the max- imal degree of “‘positive heterogony” obtainable. (3) Light and (4) heavy feeding. Under the former condition, the amount of food adminis- tered, although small, was sufficient to support very slow growth of the larvae. Heavy feeding naturally caused them to grow rapidly. These two classes are grouped together because they give identical results. The growth of the grafts, rel- ative to the normal eyes of the hosts, is the same whether the latter are growing slowly or rapidly (Twitty and Elliott, ’34). These experiments shed further light on the manner in which food is partitioned in the grow- ing animal. The results with light and heavy feed- ing demonstrate conclusively that the percentage distribution of nutrient materials between part and whole is not affected by the rate of food in- take. Even under “balanced” feeding, when only the graft grows, the same apportionment is prob- ably observed; here, however, the quota allotted to the host is barely adequate to repair the losses of catabolism, whereas the transplant with its greater capacity for assimilation, is still able to register increase in size. These findings are in keeping with the analogy which we have adopted. If the partition of food is regulated by principles analogous to those which govern simple chemical systems, we should not expect the absolute amount present to affect its percentage distribu- tion between competing parts of the body. The latter point, however, raises a question to which our experiments give no satisfactory an- swer; namely, whether the level of nutrient mate- rials in the blood actually changes importantly with the intensity of feeding. We can readily con- ceive that growth operates to maintain them at a fixed concentration, its rate varying with the amount of food consumed. In seeking a simple analogy, we might compare growth, the forma- tion of new protoplasm, to the precipitation of formed substances from a saturated solution of reacting chemicals. The rate of precipitation is proportionate to the amount of fresh reagents added, whereas the concentration still in solution is unaffected. The results with “balanced” feed- ing are perhaps indicative that the concentration of food in the blood remains approximately at “saturation”? even when extremely small amounts are being eaten. Since the hosts themselves were Aueust 4, 1934 | fed merely enough to offset catabolic destruction, we might have anticipated that the quantity of available nutriment in the circulating medium would be practically depleted. This is evidently not the case, however, as is shown by the ability of the young transplants to continue their growth. In our efforts to interpret the causes underly- ing size-regulation in transplanted eyes, we have based our explanation upon differences between transplant and host associated with their ages. It is important to understand that by “age’’ we nat- urally do not refer merely to the absolute ages of donor and host. Handford (unpublished com- munication) has in fact shown that the results with homoplastically grafted eyes are essentially similar, whether the small and large individuals used for the exchange of eyes were of the same or different absolute ages. It is accordingly more accurate to apply the terms “‘growth stage” or “physiological age’, in designating the ditferences which we are attempting to express. Our assump- tion, restated, is that as true or physiological age increases the assimilative capacity of the cells de- clines and that this accounts for the accelerated growth of a younger organ on an older host. The validity of this assumption obviously de- pends in large measure upon the success with which it can be applied to other cases of regu- lative growth. An instance which immediately comes to mind is the regeneration of amputated appendages (see also Huxley). When the limb of a salamander larva, e.g., is cut off at its base, a blastema forms which later differentiates into a new limb. At first the regenerate is much smaller than the paired organ on the opposite side of the animal, but eventually it overtakes the latter in size. This is clearly comparable to the growth regulation of a young transplanted eye, and the same interpretation can be applied in both cases. The cells which are actively concerned in regen- eration are mostly primitive, undifferentiated, mesenchyme cells (Hellmich, ’29) which are “physiologically young” in the fullest sense of the term. Weiss (’27) and Milojevic (’24) have shown by transplantation experiments that the young blastema has many of the properties of an embryonic rudiment. We may thus think of the newly regenerated limb as a “young organ on an older animal”, just as properly as if it had been grafted from a much smaller larva. The application of this conception to the re- sults of Mangold (’31) raises more difficult ques- tions, and leads us to a closer scrutiny of the fundamental nature and significance of “physiol- ogical age”. Mangold, and also the present writer (Twitty & Elliott, 34), found that the initial size of the eye could be reduced by extirpating a portion of the optic anlage from the neural plate. This condition invites comparison with our ex- periments on transplantation, whereby the same THE COLLECTING NET 127 result is accomplished by replacing the eye from a smaller larva. There is, however, one important difference. The grafted eye is not only smaller, but younger, than the normal eye of the host, whereas in the case under discussion, the small eye has developed from material identical in age to that which gave rise to the normal eye. Its re- duced size is thus due, not to a younger age, but merely to its development from a smaller quanti- ty of embryonic material. Mangold found that by metamorphosis size reg- ulation was practically complete( ratio of test eye to normal eye about 9: 10). Our observations dis- closed the same tendency, but regulation proceed- ed less rapidly than in the case of small trans- planted eyes. This seems to suport our hypothesis that the tedency towards regulative growth de- pends, in part at least, upon differences in physiol- igical age between the part and the whole. The fact remains, however, that even in our exper- iments some size-regulation does occur and given sufficient time might have become practically com- plete. In order to harmonize this fact with our con- ception of the role of physiological age in the regulation of size-difference, it is proposed to make certain suggestions which, although specu- lative, introduce interesting possibilities. We should like to suggest, namely, that there may be a fundamental relationship between the size of an organ and its “physiological age’ —that an organ which is smaller than its mate of the same species is also “younger” physiologically even though it has developed from identical material. The means by which a given size might impose a correspond- ing physiological condition upon an organ is of course obscure, but possible mechanisms are not wanting. It is conceivable, for example, that as the organ grows, the arrangement and orientation of the cells undergo slight but definite changes resulting, among other possibilities, in a redistri- bution of the mechanical stresses upon the walls of each. Fischer has claimed that subtle relations of this nature are powerful enough to modify the physiological condition of cells in-vitro, to the ex- tent of rendering them capable or incapable of cell-division. Cells in the heart of a culture sel- dom divide, whereas those near the periphery, where one or more of their surfaces is exposed, multiply rapidly. “Die atisserste Zellzone einer Kultur ist in Wirklichkeit ihr “Stratum germin- ativum’”’, p. 99, ’30. He argues that this inhibi- tion in the central portions is not due to the ac- cumulation of metabolic products, or to inaccessi- bility to food. A statement by Fischer on p. 127 expresses clearly the important consequences of simple con- tactual relationships between adjacent cells. More- over, it is almost inevitable that these relationships (Continued on page 131) 128 THE COLLECTING NET [ Vor. IX. No. 77 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel Warner Parker, Margery Fuller Mitchell, and Annaleida Snyder Cattell. Printed by The Darwin Press, New Bedford BEACH ETHICS II Not long after the July 21st number of THE CoLLEcTING NET made its appearance we were roundly denounced (verbally, yet semi-publicly ) by a physiologist of some standing in the com- munity because of our comments on the Bay Shore Beach situation. In that number we wrote that “we may be criticised severely for bringing up the matter again...” We have been! Fur- ther, we will be! But as we also said we sincere- ly believe that it is desirable for everyone in Woods Hole to think about local bathing condi- tions. We were denounced for being unfair, for “‘self- ishly wanting to stir up a fight,” for “opening old wounds,” for attempting to ruin an “interesting magazine which had possibilities of doing some- thing really worthwhile.” We were warned that “such tactics’ would lose much of the support that we had earned from good biologists. The physiologist in question had a large and varied vocabulary and it would not be desirable to quote too precisely some of the things that he said. We categorically deny his statements! Further, we predict that as soon as enough time has elapsed to curtail adrenalin production that he will himself contribute to THE CoLLectinG NET. The individual, finally admitting we had a right to print our comments, in no uncertain terms called us “fools” for not getting the matter “off our chest’ by printing it in THE Woops Hote Loc. We rather agree; it should be printed there —too! Perhaps its editor can be prevailed upon to take up the matter in her paper. We wish to make our position clear! We sin- cerely believe that the beach situation should be of intimate concern to every scientific worker in Woods Hole; the discussion of it, therefore, falls within the scope of THE CottectinG Net. To the best of our knowledge and belief most people in Woods Hole object to the fence; to the best of our knowledge and belief, part of it is there, con- trary to law. If the section of the fence in ques- tion is not voluntarily removed within a reason- able length of time we shall address a letter to Chief Harold Baker of the Falmouth Police De- partment requesting its removal. If he does not consider it to be within the province of the town we shall appeal to the Commonwealth of Massa- chusetts. ELECTROKINETIC PHENOMENA AND THEIR APPLICATION TO BIOLOGY AND MEDICINE ABRAMSON, HArotp A., American Chemical So- ciety Monograph Series, 337 pp. Chemical Ca- talog Company, New York, 1934. The author undertakes to deal with all phe- nomena comprised under the term of electrokine- tic phenomena. This is: electric endomosis and kataphoresis, streaming potentials, and sedimen- tation potentials. The author is certainly entitled to this task on the ground of his many experi- mental contributions in this field, both to the phys- ico-chemical aspect of the matter and its biologi- cal applications. It is a very appropriate time to have a competent review of this field on account of the ever-increasing material. On the other hand, no other field of biology will offer such a difficult task as regards the fundamental science. The fundament is especially hydrodynamics, and the theory of the electric double layer, both in terms of the older theories as established by Helmholtz and Lamb, as in terms of the modern theory of the diffuse double layer as established by Gouy, and by Debye and Htickel. There is scarcely any other field in physi-cochemistry that would present so many difficulties to anybody not a professional physicist but a biologist, aS just those underlying the problems presented in this book. A certain compromise is necessary as to how far the author should resort to the funda- mentals, especially to its mathematical aspects. From the reviewer’s point of view the author has solved this compromise as satisfactorily as can be imagined. For the biologist, there will be of special inter- est the treatment of protein surfaces, of cells, especially red blood cells, and bacteria. Many tables and diagrams illustrate the text. A very attractive history of the discovery of the phenomena concerned is the beginning of the book. Then it deals with the theory and the methods, and finally surveys the application in all branches of biology in the widest sense, including especially immunology. Leonor MIcHAELIS Dr. and Mrs. Putneas W. WHITING have just returned to the Marine Biological Laboratory from a ten days’ trip in the White Mountains of New Hampshire. During six days of hiking they climbed eleven mountains over 4000 feet high, both in the Presidential Range and at Fran- conia. The nights were spent at small huts main- tained by the Appalachian Mountain Club. Dr. Whiting is a visiting investigator in genetics at the Carnegie Institution of Washington at Cold Spring Harbor. Avcust 4, 1934 ] THE COLLECTING NET 129 ITEMS OF Dr. H. S. Jennrncs, professor of zoology at Johns Hopkins University is working this sum- mer at the Hopkins Marine Station at Pacific Grove, California. Mrs. Jennings is at present in a hospital near Pacific Grove, recovering from a severe illness. The invertebrate zoology course has its usual high enrollment this year, opening on Tuesday with fifty-four in the class. Most of the students are coming from the mid-west and the middle At- lantic states, although there are three representa- tives from Canadian universities. Dr. and Mrs. Hupert B. Goopricu with their two children have left Woods Hole for their sum- mer home in the White Mountains, New Hamp- shire, where they will spend the rest of the sea- son. Juttan P. Scort, veteran photographer of out- standing scientific men is spending several days at Woods Hole. An exhibition of his photographs is being held in the Old Lecture Hall. Dr. M. v’Arcy MAGEE, a retired nemological surgeon is a guest at the Bureau of Fisheries at Woods Hole. Dr. Magee is interested in the con- servation of woods, wild life and streams, and is well known as the National Vice President of the Isaak Walton League of America. INTEREST On Saturday evening, August 4th, the second » “Mixer” of the season will be held in the Club- house at 8:30 P. M. A cordial invitation is ex- tended by the Club to all biologists, their family and friends. The informal reception for the foreign visitors at the Laboratory which was to have been given after the Friday evening lecture by Dr. Gary N. CALKINS was postponed because of the lecturer’s illness. The reception will be held on August 10, following Dr. CRAMPTON’s lecture. PROFESSOR A. S. PEARSE of the zoology de- partment at Duke University, visited the Labora- tory last Friday on his way to the S. Weir Mit- chell biological station at Bar Harbor, Maine. Dr. Pearse was formerly on the staff of instruction at the Marine Biological Laboratory and a special investigator at the Bureau of Fisheries from 1912-1924. Maryjyorre Hitt ALLEE, who is known in Woods Hole as the author of “Jane’s Island” ex- pects to have her newest book, “House of Her Own” published in the early autumn by Hough- ton Mifflin and Company. On Wednesday evening, about 8 o’clock, a large fish was noticed swimming close to shore at the Nobska Beach. The witnesses believed it to be a shark. THE MT. DESERT ISLAND BIOLOGICAL LABORATORY Recent arrivals at the Laboratory are as follows: Miss Jean Hibbard who is assisting her sister, Dr. Hope Hibbard; Mr. F. J. Knocke, Princeton University, who is to assist Dr. Dahlgren and Mr. Kaylor in their invertebrate survey; Dr. G. F. Sykes, Tufts Medical School who is spending only a few days’ visit at the Laboratory. Perhaps due to the unusually warm season this year the plankton of Frenchman’s Bay appears to be richer than in previous years. Especially large numbers of algae and diniflagellates are being re- corded daily. The laboratory’s beach wagon which was al- most completely wrecked last week has been re- paired and appears to be none the worse for its thrilling experience. Bill Crabtree, its driver, as well as the driver of the other car escaped injury except for a few minor skin bruises. Beginning on Wednesday, July 25, the labor- atory has arranged a weekly Visitors Day from 3 to 5 in the afternoon. Visitors will then be welcome to inspect the facilities for research, to examine the aquarium and other displays and to learn more about the research being conducted at the laboratory. It is hoped that all who are inter- ested in the activities of the island and in biology will visit the laboratory. The dog fish which were so plentiful early in the season are fast disanpearing. Not only the laboratory collector but the local fishermen re- port that during the past week the number of dogfish has decreased about 90%. Lobster picnics are now in full swing, and larg- er quantities than usual are being enjoyed because of the low price, from 20 to 30 cents a pound. 130 THE COLLECTING NET [ Vor. IX. No. 77 COMMENTS ON THE SEMINAR REPORT BY DR. W. R. DURYEE Dr. M. E. KRAHL Lilly Research Laboratories The rate at which oxygen is taken up by a given organism is conditioned by numerous fac- tors which one may group roughly as follows: (1) The nature, concentration, and distribu- tion of substrate in a condition to react with oxy- gen; (2) The amount of oxygen supplied, per unit of time, to this substrate; and (3) The speed with which the waste end- products leave the cells or accumulate in them. The water contained in protoplasm acts as a part of the solvent which determines the concen- tration of these materials. It also is an end-prod- uct of many biological oxidations. Hence it is im- portant to know the upper and lower limits be- tween which the water content of simple organ- isms may be varied without disrupting metabolic processes, and to know the direction in which oxygen consumption varies with increasing or de- creasing cellular water content. Dr. Duryee has attacked the problem by study- ing oxygen consumption following treatment with hypotonic and hypertonic salt solutions. His measurements have been made very accurately by an ingenious new method which has many possi- bilities of use in other reproduceable biological systems. It should, perhaps, be pointed out that the use of hypertonic and hypotonic salt solutions complicates the interpretation of his results for at least two reasons: First, in changing the salt con- centration a balance of ions is not maintained when the molar concentrations of the several salts are increased equally. The permeability of the cell to substances other than water can thus be altered. Second, hypotonic and hypertonic solu- tions have been shown by others to change the end-products of cellular metabolism. The results of Dr. Duryee’s current experi- ments with manitol and other non-electrolytes as hypertonic agents should bring out more clearly the role of the water. Actual measurement of the magnitude of the change in cellular water content should also be useful in correlating the data. OVULATION AND EGG TRANSPORT IN THE FROG (Continued from page 121) to six or seven) are quite distinct from each other. If, on the other hand, three male pi- tuitaries are injected it will subsequently be noted that the ovary is almost entirely emp- tied and the few eggs that remain are confined to a single lobe. These observations, supported by observations on the actively ovulating ovary, suggest that the ovarian lobes which receive the best blood supply are the first to be emptied of their eggs. It has been found that the male anterior pitui- tary is about half as potent as the female in re- spect to sex stimulation. About eighteen hours after pituitary injection (at laboratory tempera- tures) the eggs begin to leave the ovary. Such an ovary may be removed and placed in Holtfreter’s modification of amphibian ringers and the process of follicular rupture and egg emergence may be watched for a period of 10-12 hours. The initial rupture of the follicle is not a cata- clysmic process, and the time between initial rup- ture and ultimate dropping away of the egg may be from 4-10 minutes at laboratory temperatures. If an ovary is removed from a female about 20 hours after injection, and unruptured follicles are excised (isolated) from the ovary so that each egg is contained within its closed follicle sac, the process of follicular rupture and ege emergence can be observed in single cases. Since ovulation will continue in an excised ovary for such a long period (10-12 hours) the nervous and blood sys- tems cannot be essential for the latter stages of the process. The area of follicular rupture is pre-determined for the egg before maturation. Eggs which have received no yolk or pigment, but would be devel- oped for the following year, will show the region of ultimate rupture. ‘he position of this ultimate area of rupture explains why frog’s eggs are never liberated within the hollow lobes of the ovary. This region has no uniform relation to the animal or vegetal poles of the frog’s egg. The rupture area itself rarely exceeds half the diame- ter of the egg. This means, as observation also leads one to believe, that the smooth muscle cells of the cyst wall actively force the egg through the small aperture. If an ovary of an untreated, sexually inactive frog is excised and placed in ringers containing pepsin-HCI in the same concentration as in the human stomach, the ovarian follicles will rupture as they do during ovulation. This merely suggests that the first phase of ovulation may be in the na- ture of a digestive reaction. After the egg is liberated from its follicle, it is carried by coelomic cilia to the region of the os- tium. These cilia are found on the epithelium covering the liver, the pericardium, the lateral mesovarium, and the lateral and ventral perito- neum. They are absent on the digestive tract and mesenteries, the ovaries and oviducts, and the kid- neys. ; Aucusr 4, 1934 ] THE COLLECTING NET 131 The beat of these cilia is such that eggs are di- rected toward one or the other ostium. They are constantly beating, out of breeding season as well as during active ovulation. They are entirely ab- sent in the body cavity of males, and young fe- males in which the gonads have not yet matured enough to acquire eggs. These cilia may there- fore be regarded as a secondary sexual character, found only in sexually mature females. The fact that these cilia are constantly beating during the adult life of the female can be demon- strated by tying off the oviducts and examining after 12 hours for collected materials injected into the body cavity. If an egg is transplanted from the body cavity of an actively ovulating female to the body cavity of a sexually inactive female, this egg will be propelled to the ostium and through the oviduct just as it would during the breeding season. Such an egg can be fertilized, although the outermost layer of jelly may not be exactly normal. Usually the oviduct of the sexually in- active female is not as watery as that of one ac- tively ovulating. This fact may account for the slight abnormality in the outermost layer of jelly. If small buckshot, somewhat heavier than the frog’s egg, are introduced into the body cavity of a female they will ultimately be carried to the os- tium and through the oviduct. During their course through the oviduct they will acquire some of the jelly normally deposited upon the egg. The sex-stimulating factor of the anterior pitui- tary therefore seems to be responsible for the liberation of the egg from its follicle but is not necessary in respect to the transport of the egg and the deposition of oviducal jelly. GROWTH CORRELATIONS IN AMPHIBIA STUDIED BY TRANSPLANTATION (Continued from page 127) undergo gradual and systematic changes, however slight, during the development of complex organs. In an eye whose size has been reduced exper- imentally, by elimination of a portion of its rud- iment, our suggestion is that these relationships possibly tend to approximate those normally char- acterizing a younger: organ. Although it is futile at present to develop this conception further, it is perhaps illustrative of the type of approach. which it may become necessary to employ in the further study of problems of proportionate growth. In this discussion | have attempted to present a few conceptions which may be useful in the in- terpretation of problems of growth regulation. The hypotheses offered are based entirely on re- sults with amphibians, and moreover, for the sake of emphasis, no effort has been made to consider special cases of proportionate growth which might, and undoubtedly do, constitute exceptions to the principles outlined. BIBLIOGRAPHY Burr, H. S., 1930, Hyperplasia in the brain of Am- blystoma; J. Exp. Zool., vol. 55, pp. 171-191. Copenhaver, W. M., 1930, Results of heteroplastic transplantation of anterior and posterior por- tions of the heart rudiment in Amblystoma embryos; J. Exp. Zo6l., vol. 55, pp. 293-318. Detwiler, S. R., 1930, Some observations upon the growth, innervation and function of heteroplas- tic limbs; J. Exp. Zo6l., vol. 57, pp. 183-203. 1932, Growth acceleration and reg- ulation in heteroplastic spinal-cord grafts; J. Exp. Zool., vol. 61, pp. 245-277. 1933, Experimental studies on the development of the amphibian nervous system; Biological Reviews, vol. 8, pp. 269-310. Harrison, Ross G., 1924, Some unexpected results of the heteroplastic transplantation of limbs; Proc. Nat. Acad. Sci., vol. 10, pp. 69-74. 1929, Correlation in the devel- opment and growth of the eye studied by means of heteroplastic transplantation; Arch. f, Entw.-Mech., Bd. 120. S. 1-55. Hellmich, W., 1930, Untersuchungen iiber Herkunft und Determination des regenerativen Materials bei Amphibien; Arch, f. Entw.-Mech, Bd. 121. Huxley, J. S., 1932, Problems of relative growth; Dial Press, New York. Larsell, O., 1929, The effect of experimental ex- cision of one eye on the development of the optic lobe and optious layer in larvae of the tree-frog (Hyla regilla); J. Comp. Neur., vol. 48, p. 331. Mangold, O., 1931, Das Determinationsproblem. Dritter Teil. Das Wirbeltierauge in der Ent- wicklung und Regeneration. Ergebnisse der Biologie, Bd. 7.S 195-403. Milojevic, B. D., 1924, iiber Transplantationen, Beit- rage iiber die Determination der Regenerate. Arch. Wiss. Anat. u. Entw-Mech. Bd. 103. Moment, G. B., 1933, The effect of rate of growth on the postnatal development of the white rat; J. Exp. Zo6l., vol. 65, pp. 359-393. Richardson, Dorothy, 1932, Some effects of hetero- plastic transplantation of the ear vesicle in Amblystoma; J. Exp. Zo6l., vol 63. pp. 413-445. Robb, R. C., 1929, On the nature of hereditary size limitation. II. The growth of parts in relation to the whole; Brit. J. Exp. Biol., vol. 6, pp. 311-324. Rotmann, E., 1931, Die Rolle des Ektoderms und Mesoderms bei der Formbildung der Kiemen und Extremitaéten von Triton. Arch. f. Entw.- Mech., Bd. 124. S. 747-794; and 1933, Die Rolle des Ektoderms und Mesoderms bei der Form- bildung der Extremitaten von Triton. II. Opera- tionen im Gastrula—und Schwanzknospen-stad- ium. Arch. f. Entw.-Mech., Bd. 129, S. 85-119. Twitty, V. C., 1930, Regulation in the growth of en ears eyes; J. Exp. Zo6l., vol. 55, pp. 43-52. 1932, Influence of the eye on the growth of its associated structures, studied by means of heteroplastic transplantation; J. Exp. Zool., vol. 61, pp. 333-374. Twitty, V. C. and J. L. Schwind, 1931, The growth of eyes and limbs transplanted heteroplastically between two species of Amblystoma; J. Exp. Zool., vol. 59, pp. 61-86. Twitty, V. C. and H. A. Elliott, 1934, Relative growth of the amphibian eye, studied by means of transplantation. Vol. 68. Weiss, P., 1927, Potenzpriifung am Regeneration- splastom. I. Extremitaétenbildung aus Schwanz- blastem im Extremititenfeld bei Triton. Arch. f. Entw.-Mech., Bd. 111. 132 THE COLLECTING NET [ Vor. IX. No. 77 Low Cost -- Accessories for PROTECTION , SAVE TIME SAFELY Leitz The centrifugal force of even a small centrifuge of 2,400 R.P.M. and reasonable tube capacity calls for protection against accident or glass breakage. You can easily, and at remarkably low cost, protect r against serious accident. The International “Clinical Model” Centrifuge, with its built-in protective guard bowl, operates either a two or four tube head at a full 2,400 R.P.M. e with perfect safety. It has a maximum capacity oo umunator The Ultropak [luminator has ex- tended Microscopy into a_ field where any other method of obser- vation failed. It has become indis- pensible for observation of opaque objects of low reflecting power. A line of accessories for observations in polarized light, for Fluorescence Microscopy, for observations of specimens submerged in liquids and many special stands to suit every possible requirement will be dis- played at our exhibit during the month of August at R. G. Thomp- son's, Main Street. INTERNATIONAL CLINICAL CENTRIFUGE The “Clinical Model” was designed for the progres- sive General Practitioner and as an auxiliary cen- trifuge in the hospital and research laboratory I te where time and safety are important factors. It is ¢ 4 ¢ the smallest of the International line: but, in design Pp and workmanship, it is equal to the largest. Every International model can be operated at its DE T. 510 full capacity with adequate protection against mis- hap. Let us help you to select the model that will SOE ES Ed best meet your requirements. NEW YORK CITY Descriptive Bulletin upon Request BRANCHES: Washington, D. C., Chicago, IIl., San Francisco, Calif. Los Angeles, Calif. INTERNATIONAL EQUIPMENT CO. 852 Western Avenue Boston, Mass. Makers of Fine Centrifuges WE DO OUR PART — Aucust 4, 1934 ] THE COLLECTING NET 133 Dr. G. Gruebler & Co. (Prop. J. Schmid, Apoth. & A. Schmid, Chemist) Founded 1880 Microscopical Stains---Staining Solutions Physiological Preparations Highest Quality-Accurate Results - IMPORTED BY OS, INC,, NEW YORK Sole Distributors AKATOS, Inc. 55 VAN DAM STREET NEW YORK CITY Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, etc. Catalogs will gladly be sent on request. Please mention name of school and Spalteholz subjects taught, to enable us to : 5 a Transparent send the appropriate catalog. tee pai tory Preparations os a. Crav-Anams Company and LAN DAMS JOM INI Zoological 25 EAST 26th STREET NEW YORK Model of Human Heart Visit our display rooms and museum. 134 __THE COLLECTING NET | [ Vor. IX. No. 77 THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and MICRO SLIDES ay Se ak Jamun LS COVER GLASSES seripis and drawings cor the iceman and economical method of publishing biologi- DO NOT FOG 5 cal research, has been prepared by the Staff Ask your dealer —or write of The Wistar Institute Pres# and the cooper- ative efforts of more than fifty editors con- cerned in the editing of journals published by " 1 The Wistar Institute, and presents the con- N Ciay- ADAMS ComPANY sensus of opinion on many points relating to iN ; : ew YORK the mechanical preparation of manuscripts LINN 25 East 26th Street . and drawings for the printer and engraver. a ————— Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and (giving dealers name) to Reducing Valve Si oa rea ae eee submitted to editors, in order to offer as much combustions; in pH _ work; information and illustrative material on the with your oxygen burner, subject as is possible within reasonable limits. She ine models, for various _ It will save authors much time and expense purposes; with different in- in preparing papers for publication and tend iets for different gases, in- to expedite the publication of research. cluding HeS and NHs. Seay iD) Ask for folder NC Address Price $2.00 The Wistar Institute of Anatomy and Biology Hoke Inc. Thirty-sixth Street and Woodland Avenue 122 Fifth Ave. New York PHILADELPHIA, PA. COLD SPRING HARBOR SYMPOSIA 1 ON QUANTITATIVE BIOEOGY Volume I (resulting from conference-symposia of 1933 and dealing largely with surface phenomena) contains papers by Harold A, Abramson, D. R. Briggs, Robert Chambers, Barnett Cohen, Kenneth S. Cole, Hugo Fricke, Herbert S. Gasser, A. V. Hill, Duncan MaclInnes, L. Michaelis, Stuart Mudd, Hans Mueller, W. J. V. Osterhout, Eric Ponder, Theodor Svedberg, D. D. Van Slyke. From a book review: “Jf this initial high standard (Volume I) is maintained, it is diffi- cult to see how a worker in this field can do without these volumes...” Volume II will appear in the autumn of this year. It will contain papers and edited dis- cussions resulting from the conference-symposia of 1934 concerning some aspects of growth. Authors include: W. T. Astbury, Felix Bernstein, H. W. Chalkley, George L. Clark, Charles B. Davenport, M. Demerec, Hugo Fricke, J. W. Gowen, F. Gudernatsch, F. S. Hammett, Theo. L. Jahn, L. G. Longsworth, Hans Mueller, Charles Packard, Otto Rahn, Nicolas Rashevsky, Oscar W. Richards, Charles R. Stockard, Victor C. Twitty, Harold C. Urey, C. Voegtlin, C. P. Winsor, Sewall Wright, Ralph W. G. Wyckoff. The prepublication price of Volume IT, bound in cloth, is $2.90, cash with order. After publication, the price will be $3.35. The price of Volume I is $3.35, Persons purchasing Vol- ume II may obtain Volume I for $3.00. Address the Biological Laboratory, Cold Spring Har- boreal. NEY: J ————————————————— eee Avoust 4, 1934 ] THE COLLECTING NET SOOT TING (PE MOde BLE G& Achieve ‘Craditional Spencer Quality A tiny point of light at one end of a long, dark tunnel— a high power microscope on a rigid support at the other end—a lens under test in an especially constructed holder —and an expert to interpret the highly magnified image of the point of light, tell the story of — 1. Defective 2. Imperfect 3. Perfect Manufacture Design Lens With this apparatus, manufacturing errors are found and eliminated ... design faults are found, analysed and cor- rected ... that you may receive and continue to receive the full measure of Spencer quality. Products: Branches: Microscopes, Microtomes, Delineascopes, New York, Chicago, Boston, Visual Aids, Optical Measuring Instruments. San Francisco, Washington. UY NEW YORK 136 THE COLLECTING NET (Vou. 1X. Nowa Double the Value of the Instruction with a KOSB Balopticon In the first place, the KOSB Balopticon is an instru- ment of Visual Instruction—admittedly the most efficient teaching method. Secondly, it can be used in a room light enough so that students can take notes. They need not depend on memory. WE MAKE OUR OWN GLASS TO INSURE STANDARDIZED This combination of advantages marks this Balopti- > con as the last word in thorough and _ efficient teaching. PRODUCTION FOREVOUR Sry, apo This B & L Balopticon projects either lantern slides eB OR TAY or opaque objects on a transluscent screen. The screen is placed between the Balopticon and the audi- ence. The teacher always stands before the class and Complete details on the can teach easily and rapidly from the projected image. KOSB and other B & L Bal- opticons will be sent gladly on request. Write to the Bausch & Lomb Optical Co., 671 St. Paul Street, Roches- Ba ane usch & Lom OPTICAL INSTRUMENTS FOR THE SCIENCES AND B & L FRAMES Vol. IX. No. 7 SATURDAY, AUGUST 11, 1934 Annual Subscription, $2.00 Single Copies, 25 Cents. THE HYALINE PLASMA MEMBRANE OF THE SEA-URCHIN EGG Dr. RoBpertT CHAMBERS Professor of Biology, New York University Microdissection studies made by Dr. de Renyi and myself on epithelial tissues have indicated that the cells of simple epithelia, e. g., intestinal mucosa, ciliated epithelium and alveoli of the pancreas, are held together mainly by a thin cuticle-like membrane which lines one or the other side of the epitheli- um and to which the cells are more or less firmly attached. In contrast, the cells appear to be only slightly adherent to each other. This cuticle is stiffened by the presence of calcium and tends to be dis- solved by sodium or potassi- um salts. In a similar manner, it has been demonstrated that the pluteus of the sea urchin, also the gastrula and blastula of various echinoderms are coy- ered with a closely fitting cut- icle. When the cuticle is torn the cells in the vicinity of the tear fall away. When care is taken to injure only one cell of a blastula, by puncturing it, the injured cell begins to cytolyze (Continued on page 146) and loses its adhe- Rat, Dr. John W. Everett Dr. James F. Danielli Burr Steinbach INITIATION OF DEVELOPMENT AND METABOLISM IN SEA-URCHIN EGG Dr. Joun RUNNSTROM Professor University of Stockholin of Experimental Zoology, The sea-urchin egg is fertilized after the ma- turation divisions have been completed. The first change to be seen after penetration of the sperm is the elevation of the fertili- UM. H.W. Calendar TUESDAY, August 14, 8:00 P. M. Dr. Adrian Buyse: The differentia- tion of rat gonad primordia in normal adult and gonadectom- ized rat hosts. Dr. Caswell Grave: The accelera- tion of metamorphosis of Asci- dian larvae. Dr. E. R. and Mrs. E. L. Clarke: Observations on the formation of arterio-venous anastomoses. Dr. C. C. Speidel: Experimental study of striated muscle in vivo. Motion pictures to be shown 11:30, Aug. 15. FRIDAY, August 15, 8:00 P. M. Lecture: Professor Lee Foshay: “Studies on Alteration of Viru- lence in Bacterium, Tularense.” zation membrane. The fer- tilization is accompanied fur- ther by a series of changes such as an increase in the per- meability of its surface and an increase 1n_ viscosity, changes studied chiefly by American workers. All this probably means changes in the finer structure of the egg. It is to be expected that the changes in structure are ac- companied by changes in me- tabolism. Indeed, Warburg found by his work started in 1908 that the respiration of the sea-urchin egg increases about sevenfold following fer- tilization or treatment by a hypertonic solution. These re- Initiation of Development and Metabolism in the Sea-urchin Egg, Dr. John Runnstr5sm The Hyaline Plasma Membrane of the Sea- Uurchin Egg, Dr. Robert Chambers...... 137 Unusual Cytoplasmic Elements in tke Sur- viving Yolk-Sac Epithelium of the White The Tension at the Surface of Mackerel Oil, Injury Potentials in Scallop Muscles, Dr. H. ed by J. Loeb. Meanwhile our confirmed repeatedly sults had already been predict- Warburg's results have later been for the sea-urchin egg. knowledge of the mechanism of TABLE OF CONTENTS ISEAWeOS! Goon gnoanboudoDHoogouNCUEOdGAsbN 148 PGES Ohe eNECTCS CM. -ncletuis cyto- chromes — substrates-dehydrases + coenzymes The scheme indicates that the yellow enzyme also can be autoxidized. In cells with a high respiration it will gradually be oxidized by the phaeohemin enzyme. We find that the introduction of a diamine in- creases the respiration of the sea-urchin egg. Our scheme given above expresses the obvious idea that the diamine acts as a carrier in the cell. The strong increase of respiration by the diamine has perhaps something to do with the high po- tentials of these compounds as demonstrated by Michaelis. The results of the diamine experi- ments could suggest the hypothesis that the in- crease of respiration following fertilization is brought about by the formation of a reversible oxidation reduction system, a “carrier” absent in the unfertilized egg. This carrier would com- plete the respiratory system which in all other respects is ready for action in the unfertilized state of the egg. It is not possible today to de- cide between this more chemical hypothesis and the more physical one presented above. One pos- sibility would be that the diamine has the faculty of reacting through boundaries in the cell which separate the normal carrier from the phaeohemin enzymes in the unfertilized egg. I want to emphasize that the diamine experi- ments above all confirm our former conclusion 142 THE COLLECTING NET [ Vor. TX. No. 78 that the phaeohemin enzyme as well as the de- hydrase systems do not change following fertili- zation. It can be concluded that the block in the unfertilized egg lies somewhere in the chain of carriers present in the cell. About the nature of the block to be removed following fertilization, we could only give two alternative hypotheses. There is another fact I wish to point out. In the diamine experiments an increase equal to that following fertilization or by far surmounting this is brought about without any activation of the egg. We can conclude from this that a rise of respiration is perhaps not primarily connected with the activation at all. It is necessary to look around a little concerning this point on the work done with other material. We find then that in some forms like the starfish Asterias there is no change of respiration following the fertilization, as was found by Loeb and Wasteneys. Whi- taker recently even reported a decrease of the rate of oxygen consumption following the ferti- lization of the eggs of Chaetopterus and Cum- ingia, while in other material, such as Nereis, there is a slight, in Fucus, a considerable increase following fertilization. Though the data are in- complete they suggest, as Whitaker points out in a recent review of the field, that a rise of res- piration is not indispensable for activation. It seems possible that the rise of respiration is more an effect than a cause of activation. A series of facts point rather conclusively in this direction. It has just been reported by several observers, e. g., R. Lillie, Just, Batallion, than an artificial activation or even fertilization by sperm follow- ed by membrane formation is possible in sea water containing a fairly high percentage of CN. Only the further development does not proceed in the CN. As CN generally inhibits the respira- tion it is likely that such a result means that the fertilization of the sea-urchin egg may be pos- sible without increase of the respiration. In a number of experiments we fertilized the eggs of Paracentrotus in sea water containing CN and measured the respiration before and after the fertilization. A perfect membrane is formed but the respiration is about the same before and after fertilization. I then proceeded to fertilize the eggs of Paracentrotus in an atmosphere of puri- fied nitrogen. Before my first report appeared," a paper by E. B. Harvey was published which gave the proof that the eggs of Arbacia and other sea-urchins can be fertilized under anaero- bic conditions. This experiment is somewhat difficult because the sperm is very soon lamed by the lack of oxy- gen. Barron found later in Nereis a material which seems to be very suitable for such studies. My own experiments on anaerobic fertilization 1 Arkiv fiir Zoologi (Stockholm) B 20 No. 8, 1930. were carried out in the vessels of a Barcroft- Warburg micro-respiration apparatus. In our later experiments we used a special type of ves- sel with two sidearms. We bring a sperm-suspen- sion into one of these, while the eggs are in the main compartment of the vessels. The apparatus is brought as usual into a thermostat with a tem- perature of about 20° Centigrade. Then the air is replaced by purified nitrogen, a reading is made, the apparatus removed for a moment from the thermostat, the sperm-suspension in the side- arm is tipped into the egg-suspension. After 5-10 minutes readings are made again and one finds now that a positive pressure is developing, which indicates the formation of a gas. By a series of controls it was proved that the positive pressure is not caused by handling the apparatus or mixing the fluids. In further experiments it was proved that the gas'is COz and that it is formed by the breakdown of carbonates and_bi- carbonates present in the egg suspension. This was proved by determining the carbonic acid combined as salt at the beginning and the end of the experiment. We_ bring 2N H.SOx4 into one of the sidearms and tip this into the suspen- sion in one apparatus before fertilization; in an- other apparatus we do the same 10-15 minutes after the fertilization. The breakdown in the suspensions of living cells of the carbonic acid combined as salt to free CO» must be caused by the formation of an acid stronger than H»COs3. The COs» enters the gas-space and is measured by the increase of the pressure. The increased COs in solution is calculated. The acid-formation fol- lowing anaerobic fertilization is about 180 cm. or very nearly 0.01 milli-equivalents pro 1 cc. eggs. The volume of the eggs was determined by gentle centrifuging of the eggs in graduated thin tubes. A correction was made for the water in- terspace. Soon we found that the aerobic ferti- lization is also connected with an acid formation. A very great number of such experiments were made.! On the average 150 cm. acid is formed pro 1 cc. of eggs. Before the fertilization the carbonic acid combined as salt is determined in three parallel manometers. The same is done 10 minu- tes after fertilization. We find now a decrease in the content of combined carbonic acid’. Twenty to thirty minutes after fertilization we find an increase of combined COs again, probab- ly indicating an alkalinsation of the cell after the outburst of acid formation. We have formed the quotient between the free CO» formed and the oxygen consumed during different periods after the fertilization of the Paracentrotus egg. 1The tables giving typical experiments shown on slides during the lecture are found in J. Runn- strém, Biochem. Zeitschr. 258:257. 1933. Avéusr 11, 1934 } THE COLLECTING NET 143 X COs minutes — after fertilization Os 25.8 0 — 15 SS lS 12 13.1 15 — 30 ——— = (69) 18.9 7.6 30 — 45 (5 14.3 9.7 45 — 60 = 10:7 13.9 10.6 60 — 75 === = (HS 13),3) This quotient is not equal to the R. Q. The high value during the first quarter of an hour is due to the acid formation. The low value during this period is due to the formation of carbonic acid combined as salt. The true respiratory quotient is throughout equal to 0.95. Seventy- five minutes represents the period after which the first segmentation of the egg ensues. Up to the dissolution of the nuclear membrane 45 - 50 minutes after the fertilization the division 1s stopped in the nitrogen atmosphere. After the dissolution of the nuclear membrane, the division can proceed also under anaerobic conditions. | was eager to know if this period also is con- nected with an acid formation but did not find any indication of an acid formation comparable to that following fertilization. Following a principle advanced by Krebs, we determined in specially modified vessels both the free CO» and the carbonic acid combined as salt before and after fertilization. From these data the dissociation-nest can be calculated and from this (according to Michaelis) the pH _ value. From the change of pH the decrease of carbo- nate and bicarbonate, and thus the acid forma- tion, can be calculated. We studied also the morphology of the eggs fertilized under anaero- bic conditions both on the living and the fixed material. A fusion between the male and the female nucleus does not ensue. The whole structure of the protoplasma gets more anhomo- geneous. When the eggs kept in nitrogen are observed under a microscope with dark-field il- lumination, the protoplasma seems much brighter — the dispersion of lightis much stronger — than under normal conditions. Also the sections indicate profound changes. Apparently a strong gelification of the peripheral part of the cell is brought about. One gets the impression that the increase of respiration following fertilization is necessary for the maintenance of the normal structure of the protoplasma. The facts recorded for Paracentrotus eggs were confirmed with Psammechinus miliaris and Arbacia punctulata. We also found an acid for- mation in the eggs under the influence of hyper- tonic solutions under aerobic and anaerobic con- ditions. An increase of the concentration of the hypertonic solutions increases also the acid for- mation, as was shown for the Paracentrotus egg. Furthermore, the acid formation in the hyper- tonic solution is higher under anaerobic than under aerobic conditions?. The activation by an hypertonic solution of the Paracentrotus egg is rather poor and it has been a great pleasure for me to work on the Arbacia egg in Woods Hole. This egg can, as is well known, be easily activated by an hyper- tonic solution, particularly if one follows the directions given by E. E. Just. Under these con- ditions we could record an acid formation equal to or somewhat higher than that following ferti- lization. The period under which the acid is formed corresponds to about 15 minutes from the beginning of the treatment by the hypertonic solution. During this same period also the res- piration increases and arrives after 5 - 10 minutes at a level equal to that in fertilized eggs. It could be shown for the Arbacia egg also that the acid formed in the first period of the treatment gradually disappears. The conditions are very like them in the fertilized eggs, much more so than in the Paracentrotus egg, where it is more difficult to bring about an activation by means of a hypertonic solution, Loeb has stated that the hypertonic solution is ineffective unless it contains oxygen. It is possible that the explanation of this fact is simply that the acid formation is too strong during anaerobic conditions. The activa- tion seems to occur only if the acid formation corresponds to a certain optimum. If it is higher or lower the activation is reversed; cf. here the interesting studies of Tyler who reversed the fer- tilization in the Urechis egg by decreasing the pH of the sea water. We found further an acid formation on heat- ing the eggs to 38-41° for one to two minutes. The break-up of the eggs by means of distilled sea water causes an acid formation many times greater than that following the fertilization. These studies have been extended this summer to the Arbacia eggs, and new methods suggested by Mirsky have been used to break up the egg without changing the salt content of the medium. Also under these conditions a very strong acid formation takes place. Perhaps this acid formation corresponds to what has been called “acid of injury.” The facts suggest that 1¥For further details reported during the lecture cf. 1c. Biochem. Zeitschr, 258, 144 THE COLLECTING NET [ Vor. IX. No. 78 a mixture of substances kept apart in the unfer- tilized egg, an “opening of deors”’, can be made responsible for the acid formation following acti- vation of the egg. From different premises in- deed Dr. Mirsky arrives at similar ideas on the basis of quantitative experiments. These ideas are not new. They have above all been discussed by Ralph Lillie. The question is of interest if an acid forma- tion is a general feature of fertilization or if it is limited to the sea-urchin egg. Ralph Lillie, in- deed, on the basis of experiments on parthogen- esis, has arrived at the conclusion that acid for- mation is involved in the activation of the star- fish egg. I have had an opportunity to make only some few measurements on the starfish egg this year. The few measurements I made, however, seemed to confirm Lillie’s idea. A detailed com- parison between our results and those of Lillie’s would be very interesting. One finds parallels step by step. Experiments published in 1933 showed that iodoacetate does not impair the acid formation in an hypertonic solution. One must be careful, however, in drawing conclusions from negative results with iodoacetate, as the intact cells pos- sibly are not permeable for the iodoacetate at the pH 7-8 prevailing in these experiments. I have extended the experiments on the action of iodoa- cetate to the Arbacia egg. The concentration has been increased to 0.03 m, a concentration much higher than that which inhibits the lactic acid formation in muscle. In this concentration the iodoacetate penetrates very soon into the fertil- ized egg and stops the development. The un- fertilized eggs have been treated with the same concentration of iodoacetate in sea water. The eggs are shaken during the treatment. The eggs finally are broken up after 4-5 hours. The con- trol is intact after the same time of moderate shaking. The shaking is applied to facilitate permeation, Even after a treatment of 3-4 hours the eggs can be fertilized in the iodoace- tate. A membrane is formed and the develop- ment starts. The segmentation of the egg soon stops, however, generally during the first division in the stage just before dissolution of the nuclear membrane. The respiration goes up to the nor- mal value right after fertilization in iodoacetate, but gradually it drops to a value corresponding to about 60 - 75% of the normal respiration. This can be restored by addition of pyruvate, but the improvement in the development of the eggs at the same time is only very slight.. If the unfertilized eggs in iodoacetate are washed and transferred to normal sea water, a development ensues on fertilization. The larvae formed present a well marked inhibition of the animal part; the stomodeum is small, the oral arms are deficient. This fact proves that the iodoacetate must penetrate into the unfertilized egg. The unfertilized eggs were also treated with 0.03 m iodoacetate at pH about 6.0. The cytoly- sis ensues now much earlier than at pH 7.5 - 8.0. This is probably due to a faster penetration of the iodoacetate in the acidified sea water. Even eggs with a fairly anhomogeneous-looking cyto- plasma can be fertilized after a treatment with iodoacetate in acidified sea water, but the develop- ment stops soon after fertilization. Finally we found that the acid formation which follows on the break-up of the eggs is not impared by iodoacetate. Also fluoride is without any effect on the acid formation under these conditions. The next part of the lecture dealt with inhibition experiments with copper which, as shown by Frank Lillie, in the concentration 1:500,000 inhibits normal fertilization. The metabolic changes in- duced by hypertonic solution are not inhibited by copper in this concentration. The acid formation in broken-up eggs is unimpaired. We do not know the chemical nature of the acid formed under the conditions we have des- cribed. The experiments with iodoacetate and fluoride make it extremely improbable that the cluded also by numerous analyses of lactic acid cluded also by numerous analysis of lactic acid before and shortly (3,5, 10,15 minutes) after fertilization. The experiments with iodoacetate and fluoride exclude further a phosphorylation or dephosphorylation of hexoses. Direct experi- ments showed also that the content of inorganic phosphate does not change in any significant way following fertilization. Also the hydrolysis of the phosphate esters did not reveal any change in these compounds. It may be admitted, however, that important changes may remain unrevealed by this method. The content of inorganic phos- phate does increase indeed very definitely if the eggs are treated with an hypertonic solution under anaerobic conditions. Even in anaerobic fertilization an increase of inorganic phosphate was found but these changes seem to be due more to the anaerobic conditions than to the activation of the eggs. The conclusions from the actions of iodoacetate are valid only if the carbohydrate breakdown in the egg is of the same nature as in the muscle cell. This seems indeed to be the case; a decrease of respiration can as already meitioned be brought about by iodoacetate which can be compensated by the addition of pyruvate. The iodoacetate blocks the formation of certain carbohydrate break-down products, which can be substituted by pyruvate. The iodoacetate experiments exclude every enzyme action in which sulphydril groups are involved. This is also, as is well known, the case for certain intracellular proteolytic enzymes, the kathopsins. ; I finally shall try to summarize briefly the re- sults. Following fertilization an acid is pro- Avcust 11, 1934 ] THE COLLECTING NET 145 duced in the sea-urchin egg. This acid forma- tion seems to be very closely connected with the structural changes following the fertilization of the egg. The increase of respiration in the activated sea-urchin egg is not due to a change in the substrate or to a change in the phaeohemin or the dehydrase enzyme systems. It is due to the removal of a block in the chain of carriers intercalated between the enzyme systems men- tioned. The block is due to certain features of the structure of the ripe unfertilized egg or to the lack of a “carrier” which is produced in the activation process. I have spoken several times during this lecture about something seemingly as obscure as the structure of the cell. I wish to point out that the aim of the studies reviewed here has been an attempt to connect structural changes and metabolic activities. Still this is not much more than a program for work. But I feel very confident that our knowledge about the relation between structure and metabolism can be extended. I am glad to be able to mention finally that it will be possible through the work carried out this summer by Dr. Mirsky to meas- ure exactly certain structural changes following the activation of the egg. A similar point of view—the close connection between metabolism and structure—must be applied not only to the activation but also the the differentiation of the egg.t 1 cf. Se paper in THE COLLECTING NET vol. 1X. No. 4. CERTAIN UNUSUAL CYTOPLASMIC ELEMENTS IN THE SURVIVING YOLK-SAC EPITHELIUM OF THE WHITE RAT Dr. Joun W. Everett Instructor of Anatomy, Duke U niversity School of Medicine In the examination of the surviving cells of the rat yolk-sac epithelium, two heretofore unde- scribed cytoplasmic structures have been found. The one is a peculiar type of secretory granule; the other, an element which possibly is a Golgi apparatus in the living state. These (columnar) cells are found in the meso- metrial hemisphere of the visceral omphalopleure where it becomes extensively villated and vascu- larized about the twelfth day. The function of this epitheliunt is almost unknown, although it has long been supposed that it digests and absorbs the adjacent decidua capsularis. The presence of fat droplets in the cells between the tenth and fifteenth days would indicate fat absorption dur- ing that period. It is impossible at present to judge what functional significance may be as- signed to the structures under consideration. Immediately beneath the cuticular border of nearly every cell is a layer of small, refractile granules of varying size. Continuous observa- tion of a single set of them over a period of two hours has shown that each expands and contracts at a slow rhythm. They are not chondriosomes for they are not stained by Janus green. Typi- cally, they can be stained by only one group of the dyes used, the metachromatic thiazins. Stained with these dyes, the granules assume a pink color. Their secretory nature was determined from pre- served material, stained with iron haematoxylin. Frequently they were here seen to be connected by slender, blackened threads with the stalks of pear-shaped blebs projecting into the yolk-sac cavity. At a level of focus beneath the granules and above the nucleus, the typical cell of the 10 to 13- day epithelium shows an irregularly outlined, re- fractile vesicle, from which diverticula sometimes project into the paranuclear cytoplasm. This ele- ment slowly, but continuously changes its shape as evidenced by the three drawings presented, made from a single cell at intervals of five min- utes. The rate of movement is so slow that it is imperceptible unless periodic sketches are made. When the epithelium is stained with a metachro- matic thiazin dye or brilliant cresyl blue, the vesi- cle assumes a diffuse purple color. Suspended in this purple medium are scattered, dense blue rods in rapid brownian movement. That these rods are not artifacts is indicated by the brownian movement visible in unstained vesicles. A few of the rods are typically attached to the vesicle wall. Similar granules are stained red by neutral red, but with this dye coloration of the interven- ing fluid is not apparent. Nile blue sulphate colors the fluid metachromatically, but does not demon- strate the rod-shaped granules. Janus green does not enter the vesicle, but stains only the mito- chondria and (on occasion) the vesicle wall. The metachromatic color-shift in the vesicle does not appear to be the result of a high alkalin- ity. An alkalinity sufficient to give thionin and toluidin blue a purple color (i.e. pH>12.0) would completely shift brilliant cresyl blue and Nile blue sulphate to red. That does not occur. Perhaps the metachromasia is the result of solution of the dyes in a non-aqueous phase of the vesicle inter- ior. The position and configuration of the vesicle would suggest that it is a Golgi apparatus. Fur- ther evidence to that effect was found in material which had been preserved in Champy’s fluid, post- osmicated and stained with Altmann’s acid fuch- sin (Severinghaus’ method). This frequently produced a blackening of the walls of the vesicles and small granules attached to their inner sur- faces. These granules appear to be the rods seen in the vitally-stained cells. There was no other evidence of the apparatus in the osmicated epithe- lium. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 24.) 146 THE COLLECTING NET [ Vor. IX. No. 78 THE TENSION AT THE SURFACE OF MACKEREL OIL Dr. JAMES FREDERICK DANIELLI Commonwealth Fellow in Biology, Princeton University Harvey and Shapiro found that tension at the surface of the oil drop in mackerel eggs is ap- proximately 0.6 dynes. Mackerel body oil in bulk at the same pH as the egg has a tension of about 9 dynes. Consequently the low value found for the oil drop in the egg must be due either to a difference between the oil in the egg and the body oil, or to some surface-active material in the egg. A decision between these two latter alternatives is the object of this work. Large quantities of eggs were broken up in various ways, by crush- ing, freezing, etc., and the oil separated from the egg material by centrifuging. The surface ten- sion of this oil against the egg material was meas- ured with a Du Nouy tensimeter, the value ob- tained being 0.7 dynes. It was shown that this low value was due to an adsorbed layer of pro- tein-like material. The denaturation of this adsorbed material at a hydrocarbon-water interface was investigated and it was shown that small quantities of mack- erel oil dissolved in the hydrocarbon markedly retard the rate of denaturation; and that denatur- ation is prevented entirely when the interface is saturated with mackerel oil. It was also shown that other surface-active substances, such as leci- thin, palmitic acid, and cholesterol when present in small quantities in the oil phase decrease the rate of denaturation; it thus seems probable that the denaturation of protein at the oil-water inter- face is prevented when the interface is saturated by surface-active compounds which have hydrated polar groups, so that protein is no longer directly in contact with the hydrocarbon surface, but with a surface which in many ways compares with that of other protein molecules. It was also shown that in fundulus eggs and in starfish eggs there are protein-like substances which have a high surface-activity. Previously the low surface tensions obtained for cells had seemed incompatible with the view that the cell membrane is lipoidal in character. In the presence of such extremely surface-active material as has been found in these eggs the low values obtained for surface tension of cells can be understood, since any lipoid would necessarily have a protein layer adsorbed upon it. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 31.) THE HYALINE PLASMA MEMBRANE OF THE SEA URCHIN EGG (Continued from page 137) siveness to the overlying cuticle upon which it falls into the blastocoele and its place is soon occupied by the bulging of the neighboring cells into the vacated space. The cuticle has been traced to the so-called hyaline plasma layer or membrane which is pres- ent in the fertilized egg and which has its origin apparently by an exudation of material from the ege after fertilization. In the presence of cal- cium of the sea water, the external border of the exuding material is precipitated to form a thin granular outer border with a hyaline, very soft, jelly-like material beneath. This material first appears at the site of sperm entry in the form of an excrescence, the so-called exudation cone. The base of the exudation cone spreads slowly to form the hyaline plasma layer which encircles the egg within 8 to 10 minutes after fertilization. The functions of the hyaline plasma layer are suggested to be as follows: 1. It forms in the cleavage furrow and pre- vents the freshly formed plasma membrane of the opposite walls of the furrow from running together if the walls should come into contact. 2. The outer border of the hyaline plasma layer is precipitated by the calcium of the sea water into a relatively stiff and coherent mem- brane to which the outer surfaces of the blasto- meres adhere. Because of this fact it is suggest- ed that the appearance of the blastocoele occurs by a rounding up of the inner borders of the very fluid blastomeres, while the blastomeres become wedged together as they remain attached to a cuticle surrounding the entire blastula. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 31.) ns a ite Aueust 11, 1934 | THE COLLECTING NET 147 INJURY POTENTIALS IN SCALLOP MUSCLES Dr. H. Burr STEINBACH National Research Fellow, University of Chicago - When a cut is made across the striated portion of the Pecten adductor muscle, an injury poten- tial of considerable magnitude (30-40 mv.) may be led off from the uninjured to the injured sur- face. If we assume that the magnitude of the injury potential may be used as an index of the degree of injury in any particular instance, then when the injury potential decreases due to some change taking place at the cut surface, we may speak of recovery at that surface. Such a recoy- ery, or decrease of injury potential has been shown to take place at the injured surface!. This “recovery reaction” requires the presence of cal- cium either in the tissue or in the applied solu- tion. There is good reason to believe that this recovery process is closely related to the film for- mation observed in various eggs, plant cells and other forms of living material. The calcium ef- fect on the injury potential appears to be actively antagonised by potassium ion. That is to say, the injury potential is maintained at its original magnitude when the cut surface is bathed with a solution containing excess potassium, or the in- jury potential may be restored in a “recovered” muscle when the recovered surface is treated with potassium. It was found, moreover, that when the cut surface was treated with solutions containing varying ratios of potassium and calcium, the measured potential difference would increase reg- ularly with increase in relative potassium concen- tration. When the p.d. values are plotted against the logarithm of the mol fraction of potassium, a straight line results. Since the cut surface often becomes approximately 30 mv. more negative to the uncut surface for a hundred-fold concentra- tion of potassium, it was thought that the normal injury potential might arise because the injured surface could act, under some conditions, as might a potassium ion electrode. Concentration potentials were estimated, as oc- curring across the cut surface, by measuring the potential difference between the tissue and vari- ous concentrations of electrolyte solution bathing the cut surface. The potential differences meas- ured were converted to relative values in the usual manner. Potential differences estimated in this manner have no exact physico-chemical significance as concentration potentials except in certain special cases. However, for the sake of simplicity, I have treated them as though they were such. The results showed that dilute solu- 1 Steinbach, H. B., 1933. Injury potentials in scal- lop muscles. J. Cell, & Comp. Physiol. v. 3, p. 203. tions of potassium chloride are electropositive to concentrated solutions (9 my. for 1/100 concen- tration ratio) when the concentration potentials are estimated in the manner described. Dilute solutions of sodium, magnesium and calcium chlorides are negative to more concentrated. Thus although the cut surface behaves as might an imperfect potassium ion electrode, it does not behave as an interface selectively permeable to cations in general. Consideration of these and other results indi- cated that the injury potential might be qualita- tively described as a potassium concentration po- tential, but that quantitatively the concentrations potential values were much too low. However, the marked effect of calcium on the normal injury potential suggested that some calcium might be essential in the medium in order that the surface might function properly as a potassium electrode. Experiment showed that this is indeed the case. If calcium is present in a certain minimal con- centration or over (ca. 0.003 m. or over) the potassium concentration potential is greatly in- creased. The magnitude of the increase varies somewhat with the time of exposure but the con- centration potential is always more than doubled by the presence of calcium. In order for the cut surface to develop a po- tassium concentration potential of a magnitude comparable to the normal injury potential, a cer- tain amount of calcium must be present in the applied solutions. Given this amount of calcium, the potassium concentration potential is indepen- dent of the concentration of any ion other than potassium over a wide range of concentrations. Sodium, magnesium, chloride or excess calcium ions have no appreciable effects. The concentra- tion potentials estimated in this manner are inde- pendent of the osmotic pressures of the solutions used within the concentration ranges used. The calcium effect on the potassium concentration po- tential cannot be demonstrated on the uninjured surface of the muscle, even though the concen- tration potential of pure KCI solutions is the same at either the cut or uncut surface. These experiments lend further support to the conclusion that, in Pecten muscle, the injury po- tential arises at the injured surface because of some reaction occuring there and that the injury potential may be similar to a potassium concen- tration potential. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 31.) 148 THE COLLECTING NET [ Vor. IX. No. 78 THE RELATIVE EFFECTS OF INCREASED CARBON DIOXIDE AND DIMINISHED OXYGEN UPON THE HEART RATE OF YOUNG TROUT Dr. CHARLOTTE HAywoop, THELMA O. STEVENS, Department of Physiology, _ This study has made use of the fry of the brown trout, the Loch Leven trout, and the brook trout within two months after hatching, at which time the heart may be readily observed through the body wall. The method consisted in observ- ing with the binocular microscope two or three of the young fry, placed in a little water in an Engelmann chamber which bore a cover that could be sealed on tightly. The chamber was kept at a constant temperature of about 6° C., as were also the gases that were passed through it. These gases were carbon dioxide, nitrogen, and a mix- ture of either of these gases with air. The frequency of the ventricular beat was de- termined from time to time before and during exposure to the gas to be studied. It was found that both increased carbon dioxide tensions and diminished oxygen tensions slowed the heart rate. With carbon dioxide concentrations of as low as 5 or 10% the heart rate began to slow at once and was diminished almost to one-half at the end of an hour. On the other hand there was usually a lag before lowered oxygen tensions began to re- tard the heart rate, and an extensive reduction in the oxygen tension was required to produce a re- tardation similar to that obtained with moderate amounts of carbon dioxide. For example, an HeLen M. TEWINKEL, AND MARGARET SCHOTT Mount Holyoke College oxygen tension of 15 mm. Hg (about one-tenth of the normal oxygen tension) lowered the heart rate to one-half in 1 to 2 hours, which could be done by a carbon dioxide tension of 75 mm. Hg. 80 mm. oxygen (about one-half the normal ten- sion) usually failed to reduce the rate to half its original value during an exposure of 8 hours or longer. Recovery of the heart frequency was studied following equal decrements in 20% carbon diox- ide and in purified nitrogen. After carbon diox- ide, the frequency increased gradually, but stead- ily, until the original rate was restored in 50 to 100 minutes. The recovery following oxygen deficiency was less uniform: a few of the fish died; the surviving fish began recovery with a quick rebound at first, but this was usually not well sustained and ultimate recovery did not occur until much later. These differences in recovery suggest that the mode of action of increased car- bon dioxide and of diminished oxygen upon the heart frequency of the young trout is not the same. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 31.) NOTES ON THE BEHAVIOR OF ASTERS Dr. Henry J. Fry Visiting Investigator, Department of Anatomy, Cornell University Medical College A discussion of the significance of asters must deal with the puzzling fact that in cells of the same type—spermatocytes, for example—asters may be present in those of one species, but absent in those of another. This holds true in a number of cell types, and raises questions con- cerning the function of asters, since cells carry on similar activities whether they occur or not. This preliminary report, which attempts to analyze this situation, deals only with normal mi- totic figures. Sperm-asters, cytasters and nuclear monasters of artificially activated eggs, uni- and multi-polar figures, and other unusual conditions, are not discussed, not because they are not signifi- cant, but because any reconsideration of the sub- ject is simplified if attention is at first confined to the usual bipolar mitotic figures. An examination of the cytological literature de- scribing normal mitosis and cell division shows that, with rare exceptions, asters occur only if the two following conditions are satisfied: first, dur- ing the time when the spindle becomes organized its ends must be more or less sharply focalized; second, these focalized spindle-ends must lie in cytoplasmic areas of considerable size (Group I, Figs. 1 and 2). If the spindle-ends arise as blunt structures, asters are absent even when they lie in large cytoplasmic areas (Group II, Figs. 3 and 4). If the spindle-ends are sharply focalized, but cytoplasmic areas are not available (Group II1), as when the spindle-tip is near the cell wall (Fig. 5) or within the nuclear membrane (Fig. 6), as- ters are again absent. If both conditions are un- fulfilled there are of course no asters (Group IV, Figs. 7 and 8). The chart lists the chief classes of cells in which each of these major mi- totic configurations occurs. Considering typical cases where the spindle-end is focalized, the presence or absence of a cyto- plasmic arez may account for the fact that the large epithelio-muscle cells of Hydra have asters (Continued on page 154) Aveusr 11, 1934 } THE COLLECTING NET 149 THE WOODS HOLE OCEANOGRAPHIC INSTITUTION The Atlantis sailed early Monday morning for a cruise of a week or more to the region off Georges Banks. © The scientists on this trip in- clude geologists who will study the formation of the Atlantic coastal shelf. Core samples from the bottom are to be taken for study. Dr. Hen- ry C. Stetson and four assistants comprise the group of geologists. The preceding Atlantis cruise, a short one, was chiefly for the purpose of continuing the studies of the bacteriology of the sea under the direction of Dr. Selman A. Waksman. Important contributions to the diffi- cult study of the aquatic nitrogen cycle are con- tinuing to come out of this series of investiga- tions. “As the oceanographic world is well aware, the work is being so correlated with planktonic studies and with physical and chemical ocean- ography as to promise a solution in the near fu- ture of some of the fundamental problems of oceanography. Such an outcome would hardly be possible but for the varied and comprehensive research programs made feasible by the equip- ment of the Oceanographic Institution and the versatility of its staff. Dr. H. B. Bigelow, director of the Oceano- graphic Institution stated in a recent report: “Experience on long cruises and in heavy weath- er has proved that the Atlantis is an excellent sea boat; that no mistake was made in the selection of an auxiliary sailing ship for our particular work; and that her gear is well designed for modern deep-sea exploration.” The weekly staff meetings at the Oceanogra- phic Institution have upheld the precedent of previous summers—a precedent crystallizing into a laboratory tradition as well characterized as is that of the Marine Biological Laboratory. The staff meetings are well attended. Sometimes as many as fifty are present. Held in the commo- dious library, they are pervaded by a congenial atmosphere conducive to free and intimate dis- cussions. They are doing much to further the team work so necessary for successful pursuit of an oceanographic program where the efforts of many specialists must be combined for attack upon the complex “problem of the sea.” At the meeting this week, Dr. Selman A. Waksman spoke on “The Role of Bacteria in the Sea.” This paper discussed in detail the me- thods and results of his recent experimental work. A shorter resumé of this same work was given at the Marine Biological Laboratory Tues- day evening, August 7, under the title, ‘Origin and Nature of Organic Matter in the Sea Water and Sea Bottom.”” A summary of the paper will appear in THE CoLLectinG Net. Marcery MitcHELL THE BOARD OF REVIEW OF THE MARINE BIOLOGICAL LABORATORY The Board of Review of the Marine Biologi- cal Laboratory which was provided for in 1924 functioned for the first time this summer on July 21. This Board is to meet every ten years to make a study of the work at the Laboratory to insure its national character and maintainance on a high plane. If the Board feels at any meeting after investigations that the Marine Biological Laboratory is not performing valuable services in biological research, they are obliged to file a written memorandum with the Trustees in charge of the endowment fund giving reasons for such a decision. After five years if two-thirds of the Committee still maintain that their decision is correct, the endowment funds may be used for purposes within the field of biological research as indicated by that Committee of Review. Those present at the meeting were Dr. Gary N. Calkins representing Columbia University, Dr. Wesley R. Coe representing Princeton Uni- versity, Dr. Frank R. Lillie representing the University of Chicago, Dr. Alfred C. Redfield representing Harvard University, Dr. Fernandus Payne representing the National Research Coun- cil, Dr. T. H. Morgan representing the Ameri- can Association for the Advancement of Science, and Dr. Charles R. Stockard representing the National Academy of Sciences. Dr. Conklin and Dr. Redfield were elected Chairman and Secre- tary respectively. The endowment funds of the Marine Biological Laboratory were set up as a trust on the advice of the administration in order to avoid reorgani- zation of the Board of Trustees of the Labora- tory such as would have been necessary if they were to assume charge of the investment of the endowment funds. The Board of Review was established at the same time to insure permanent protection of the purposes of the trust. —B. A. The supply department reports an unusually large demand this summer for arbacia. During the week of July 30 to August 4, 7030 were used by workers at the Marine Biological Laboratory, while previous weeks in the middle of July reached an even higher mark. On August 2 alone 1608 sea-urchins were ordered. JuLtan P. Scott is spending the remainder of the summer at Woods Hole exhibiting his photo- graphs of scientists in the Old Lecture Hall. His collection consists of twenty-three hundred pic- tures, but only one hundred and fifty are exhibit- ed. Mr. Scott came to Woods Hole in 1919 as laboratory assistant to Dr. Leo Loeb, and started the hobby which is now his chief interest. The first photograph in his collection was that of Dr. Frank R. Litrie. Mr. Scott travelled around the world once and has made three trips to the Orient, the last one terminating a year ago. 150 THE COLLECTING NET [ Vor. TX. No. 78 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel Warner Parker, Margery Fuller Mitchell, Barbara Allee, and Anna- leida Snyder Cattell. Printed by The Darwin Press, New Bedford THE BEACH QUESTION IN THE LIGHT OF A CHANGING ENVIRONMENT (An Editorial Contributed by Dr. Henry McE. Knower) Community problems of the adjustment of needs, claims, “rights” of the individual are of course inevitable subjects of discussion for the Woods Hole Forum. They are again brought to the front by the reopening of the “Beach Pro- blem” in THe Cotitectine NET. I agree with the editor that the beach problem is a serious one and would like to see something done to improve conditions there. It is indeed an important need! But I cannot join in attack on the beach lot owners nor approve of the more extreme method advocated by some of calling in the Town to take over the beach as public pro- perty. It seems that history will give us a good back- ground for discussing the best attitude to take so I will first give a brief summary of the history of some of the factors involved in the situation, and later take up the “beach question’’ itself. We will divide the history into a consideration first of the development of the /nternal Environ- ment or Laboratory proper, and second to the general adjustments of the community under the head of External Environment. In the absence of a chairman, I suppose any- one venturing to address the Forum must intro- duce himself and offer some excuse for speak- ing. So I offer an exceptionally long experience in the community from earliest times, and parti- cipation in many cooperative efforts of its history. Looking back over forty years, (1 came first in 1892) there is much change to record; though, I hasten to add, something has kept the essential fundamental spirit and working of the place re- markably the same through it all. Changes have affected us mostly through modifications in the External Environment; while the vital, deeper, Internal Environment has been touched only in- directly. Let us start with the Internal Environment. It has been pointed out many times before that adherence to a strict independence of action with an exceptionally fine cooperative spirit among the strongly individualistic Woods Hole commun- ity is the principle that has preserved the best features of the Marine Biological Laboratory. In the early period following the establish- ment of the Laboratory in 1888, as well pictured in records and photographs preserved in the Marine Biological Laboratory library the situa- tion in Woods Hole was the presence of a small group of biologists in a community of New Eng- land fishermen. There was no Town and Gown; only the Town. The Woods Hole residents were self- sufficient, self-contained. They were kind and quite patient with these summer visitors to their home who overran the place, at times a bit in- considerate of local prejudices, but proving themselves in time more and more helpful rather than harmful as some of the fishermen’s com- munity had feared. There gradually grew up a mutual respect and cooperation which enabled the visiting biologists to devote themselves to their special professional problems in the laboratory and postpone social and community involvements and adjustments which were later to intrude more and more. For some time a simple building and sparse equipment was enough for the small group of biologists. They had to fend for themselves and their work did not seem to require more ade- quate facilities. The problem of supplying microscopes, books, journals, apparatus, special equipment, was met by loans or personal ar- rangements for the general need. A collecting department was soon organized. A Mess was established, and managed by various amateur experimenters, often “not so bad”. New wooden buildings came into being with running water, gas, and later electricity from contribu- (Continued on page 157) CURRENTS IN THE HOLE At the following hours (Vaylight Saving Time) the current in the Hole turns to run from Buzzards Bay to Vineyard Sound: Date P. M. August : Braz August : 6:31 August Di: Ze August : 7:54 August oie ee CHa eoroe PN E NOY a teen 227 Aupust 17 eee O42 LOEAM PAUSTISE MG ieeeeen: + L035) 120 In each case the current changes approxi- mately six hours later and runs from the Sound to the Bay. It must be remembered that the schedule printed above is altered somewhat by wind conditions. Prolonged winds sometimes cause the turning of the cur- rent to occur a half an hour earlier or later than the times given above. The average maxi- mum velocity of the current is five knots. Aveusr 11, 1934 } THE COLLECTING NET 151 LEEMS (OF Dr. IrvinGc LANGMuIR, Nobel Laureate, who is associate director of the research laboratories of the General Electric Company in Schenectady arrived at Woods Hole by plane on August 7th, as guest of Dr. G. H. A. Clowes. On Wednes- day evening he delivered a lecture on “Mono- molecular films at the interface between oil and water” in the auditorium of the Marine Biologi- cal Laboratory. Dr. Langmuir won the Nobel prize for chemis- try in 1932 and has made outstanding contribu- tions to science in the field of atomic structure and on the theory of chemical valence. He has worked on vapor pressure of metals, surface ten- sion, production of high vacua, acoustic devices for submarine detection, electrical discharges in gases, and upon many phases of physics and chemistry. Friends of Dr. Gruman A. Drew will be sor- ry to hear of the continued serious illness of the former Assistant Director of the Marine Biologi- cal Laboratory, who is at his home in Eagle Lake, Florida. He came from the University of Maine in 1911 to fulfill the position as assistant director, and remained until 1926. Earlier he had been an instructor in the invertebrate zoology course from 1901-1909. Dr. Drew is remembered by biolo- gists for his work on the habits and embryology of mollusca and for his careful planning of the newer buildings of the Laboratory. Dr. Paut S. Gattsorr, director of the U. S. Bureau of Fisheries, left Woods Hole on August 2nd to visit the Chesapeake Biological Station at Solomon's Island, Md., where he delivered a lec- ture on “The Pearl Oyster Reefs in the Hawaii- an Islands.” Dr. Galtsoff spent some time in Hawaii in 1930 when he was sent by the Bureau of Fisheries to study the depletion of the pearl oysters there. After delivering his lecture he attended the Convention of Pearl Oyster Grow- ers in Baltimore, and returned to Woods Hole on August 9th. Mr. Martin G. LARRABEE, a scholar on the Johnson Foundation at the University of Penn- sylvania during the past year, has been awarded the George Leeb Harrison Fellowship for the coming year at the University. Mr. Larrabee who has been working at the Marine Biological Laboratory left Woods Hole on August 5. We wish to make apologies for having printed upside down Figure 6 of Professor John Runn- strom’s paper on “The Physiology of Determina- tion in the Sea~Uurchin Development.” INTEREST Dr. Lyte V. Beck has been appointed to a position on the staff of the Cancer Research Laboratories at the University of Pennsylvania School of Medicine where he will carry on cyto- logical work under the direction of Dr. Ellice Mc- Donald. Dr. Beck was formerly an assistant in the Eh Lilly Research Laboratories. Dr. Asa O. WeEESE of the department of zool- ogy at Oklahoma University, is a member of the faculty at the Rocky Mountain Biological Sta- tion, and is continuing his work in ecology there. Mr. Lupvic BrowMan, curator of the depart- ment of zoology at the University of Chicago, was obliged to stop in Providence while on his way to Woods Hole in order that Mrs. Brow- MAN might be operated on for a sudden attack of appendicitis. Mr. Browman is a student in the invertebrate zoology course. Dr. Duncan S. Jounson, professor of bot- any, and director of the laboratory and botanical gardens at Johns Hopkins University, and Dr. Urric DAHLGREN, professor of biology at Princeton University who formerly spent many summers at Woods Hole, are working at the Mt. Desert Island Biological Laboratory. Epwarp DeLAMATER who was a member of the empryol- ogy course at the Marine Biological Laboratory this summer, has been appointed assistant to Dr. JOHNSON during the coming winter. Mr. Manley L. Natland of the Shell Oil Com- pany of Long Beach visited the Scripps Institu- tion in connection with his foraminiferal research ine in the part of the Gulf of Catalina near La olla. MEETING OF THE GENETICS SOCIETY The Genetics Society of America will hold a meeting at the Marine Biological Laboratory on Wednesday and Thursday, August 22 and 23. The program which includes a number of papers by visiting geneticists as well as by workers in residence here, will be printed in the next issue of THE CoLLectine NET. The Genetics Society was formed in 1932 by the re-organization of the Joint Genetics Sec- tions of The American Society of Zoologists and The Botanical Society of America. All biologists, whether members of the Socie- ty or not, are cordially invited to attend the ses- sions, including the Clam Bake for which tickets may be purchased from the secretary, Dr. P. W. Whiting, before Wednesday, August 22, at $1.25 each. 152 THE COLLECTING NET [ Vor. 1X. No. 78 [TEMS Dr. and Mrs. Paut RezNrKorr left Woods Hole on July 30 for Northfield, Mass., where Mrs. Reznikoff and their daughter, Camilla, are to spend a fortnight. Dr. Reznikoff left after a day or two to resume his duties at the New York Hospital of the Cornell University Medical Col- lege, where he is assistant professor of medicine. Dr. Borts Epurussi, who has been in Ameri- ca since January on a_ Rockefeller Foundation Fellowship and who spent the earlier part of the summer at the Marine Biological Laboratory, left Woods Hole on Friday. He is returning to France and continue his work in genetics at the Institut de Biologie in Paris. Dr. GEorGE F. PApENFuss of South Africa who has been working in the department of bot- any at Johns Hopkins University for several years, has received a James Buchanan Johnston scholarship, and will be at working on algae at Gotenborg, Sweden during the coming year. He is now visiting the small island, Heligoland, off the coast of Denmark. ater 6 0am 0-0 a 0D aD a OD ED At the Criterion Theater in Bar Harbor, last Sunday, July 22, the new film, “Mt. Desert Is- land and Its Marine Life,” taken by the labora- tory last year was shown toa packed house of over 1100 people. The pictures were taken by Norman McClintock, well known photonaturalist, and were made possible by support from the Biological Laboratory, the Marine Development Commis- sion, the Bar Harbor Chamber of Commerce, Rutgers University and Mr. Dorr, Superintendent of Acadia National Park. Dr. Walter Damrosch and Mr. Chester Westcott welcomed the audience on behalf of the summer residents and business interests of the island, and Dr. Cole spoke on be- half of the laboratory, explaining why and how the film was made. The audience applauded sev- eral scenes, including those from Cadillac Moun- tain, the rhythmic movements of the barnacle, be- havior of the tube feet of starfish, locomotion of sea slugs, nesting habits of the herring gull, etc. The Criterion management kindly presented a news reel and a Silly Symphony to conclude a most interesting program. Many favorable com- INDE RE Sd Dr. Marcaret SUMWALT who has just com- pleted the past year’s work on a National Re- search Council Fellowship at the University of Pennsylvania, arrived last week at Woods Hole and will spend the rest of the summer here. Witt1Am Doyte, who worked last year at the Marine Biological Laboratory received his Ph.D. degree from Johns Hopkins University in June and was also awarded the Adam T. Bruce Fel- lowship for research next winter at the Univer- sity. Dr. Doyle is at present at the Tortugas laboratory and will come to Woods Hole later in the season. Dr. AuTe Ricnarps .of the department of zoology at Oklahoma University, and his wife, Mrs. Mitprep H. RicHarps, are spending the summer at the Rocky Mountain Biological Sta- tion at Gothic, Colorado. Dr. Richards is a di- rector of the Station and also on the faculty for instruction. He and Mrs. Richards have done extensive research in cytology and genetics, and formerly spent several summers at the Marine Biological r. aboratory. | The Mt. Desert Island Biological Labora | oy ments have been received, and it is hoped that more such films can be prepared, especially for use in schools and colleges. The laboratory would be glad to receive inquiries from those who might be able to use such films at very moderate ex- pense. Recent visitors at the laboratory have included Dr. Emil Witschi, of the University of Iowa, and his son, who were on their way to Woods Hole, Mass., after a motor trip through Canada; Dr. and Mrs. Boris Ephrussi, of the Institute for Physico-chemical Biology in Paris, who were on their way back to France after spendirig eight months at the California Institute of Technology and at Woods Hole; Dr. and Mrs. Gordon Gates, of Rangoon, Burma, who are vacationing in Maine previous to their return to Burma in the fall; Dr. Walter Gilbert, Secretary of the Car- negie Institution of Washington, and Mrs. Gil- bert, who were particularly interested in the tissue culture work of Dr. Lewis; and Dr. Wayne E. Manning, of Smith College, who hopes to do some botanical collecting in this region. —---. Aucust 11, 1934 ] THE COLLECTING NET 153 On July 25th the lowest summer tide of two years (—3.1 feet) attracted every one at the laboratory to the beaches at 7 a. m. Unusual opportunities for collecting seldom seen animals and plants were afforded by the drop of 11.4 feet from high water. Several unusual forms were collected including the rare nemertean, Mal- acobdella, found only in the clam, Pholus; the polychaete, Lapraea never before recorded from this region, and the rare holothurian, Psolas. Giant starfish, Asterias, Crossaster and Solaster, A. S. Johnson, comptroller of Rutgers Uni- versity, arrived on August 4th to spend the remainder of the month with his family. For visitor’s day on August Ist some 50 dif- ferent species of animals were displayed in the aquaria which had been especially illuminated for the purpose. Forty-one visitors went througla the laboratory, inspecting the aquaria and the individual demonstrations arranged by the vari- our workers. Of special interest were the tissue Nereis 33 inches long, many doodles (Echiurus) culture preparations of Mrs. Lewis and Dr. were some of the other specialties. Hibbard. News from the Scripps Institution of Oceanography fe > > OP 1 Mr. Roger Revelle left for Bremerton, Wash- ington, to join the U. S. S. Bushnell which was scheduled to leave for the Aleutian Is- lands. The plan is that the Bushnell will occupy a series of oceanographic stations along a line from the Aleutian Islands to Pearl Harbor, Hawaii. At each station collections of water samples at dif- ferent depths to a depth of about 2,000 fathoms will be made and records will be made of the sub-surface temperatures. One of the purposes of the work is to get information on those factors in sea water which affect the velocity of sound in sea water. The information is needed so as to de- termine the precise depths of the sea bottom be- low the surface along the lines run across the North Pacific by the U. S. S. Ramapo. Mr. Re- velle will assist in collecting water samples and in recording the temperatures. He will also collect samples for determining the oxygen in the waters at various depths and he will ship back to the In- stitution water samples which will be chemically studied after their receipt at the Institution. It is also his intention to make some biological collec- tions, especially the plankton organisms and, if possible, he will collect some bottom samples. During the past few years the U. S. Navy has been very active in the study of a number of the oceanographic features of the Pacific. The U. S. S. Ramapo has run many lines of echo-soundings across the North Pacific. The Navy has taken the lead in an intensive oceanographic survey of the Gulf of Panama and in the waters along and off the coast of Costa Rica, and it has done much work in the waters adjacent to the Aleutian chain of Alaska. The line of stations which the U. S. S. Bushnell intends to occupy between the Aleutian Islands and Pearl Harbor is in general accord with the plan for more ex- tended oceanographic work, and the results should make an important contribution to knowledge of a part of the North Pacific on which we at present have little information. TO Dr. Parker D. Trask of the U. S. Geological Survey visited the Scripps Institution. He came especially to have conferences with Professor E. G. Moberg and Mr. Roger Revelle about some of the chemical features of sea water with reference to the formation of limestone. Mr. Horace R. Byers, instructor in meteoro- logy for the Transcontinental Air Company at Glendale, was a visitor at the Insitution, coming to see Professor G. F. McEwen regarding a paper which he is about to submit for publication on the influence of climatic factors on human ac- tivities. His paper shows how the modern quanti- tative meteorological ideas can be used to carry out more precise quantitative studies of human geography. Prof. J. Edward Hoffmeister, University of Rochester, who is on his way back from about six months’ field work in the Fiji Islands, study- ing the corals, coral reefs, and geology, recently came to the Institution particularly for con- ference with Director T. W. Vaughan regarding the scientific results and preparation of the mate- rial for publication. Prof. Gordon H. Ball and Mr. Boris Kriches- ky of the University of California at Los Angeles visited Scripps Institution, consulting with the heads of the different laboratories in regard to work Mr. Krichesky hopes to do in connection with his studies. Prof. and Mrs. Maurice James of the Univer- sity of Colorado left La Jolla on their return trip last Saturday after spending six weeks in study and research at the Scripps Institution on the taxonomy, distribution, and ecology of the in- vertebrates of the intertidal zone in the La Jolla region. 154 THE COLLECTING NET [ Vor. IX. No. 78 NOTES ON THE BEHAVIOR OF ASTERS (Continued from page 148) while the small gland cells do not; the large pan- creatic acini cells of the dogfish have asters but the small brain cells do not; and the large blas- tomeres of Cumingia embryos have asters, where- as the small ones of the same embryo do not. Similarly, considering representative cases where cytoplasmic areas are present, the shape of the spindle-tip may account for the fact that develop- ing Drosophila eggs have asters, while those of another fly, Muiaster, do not; in the gregarine, Monocystis, the first division figure which arises after two individuals have encysted, has asters, while subsequent figures do not; and the polar- body figures of most oocytes have asters, whereas the others do not. Several hundred studies have been examined, and no case was found among them—though cer- ine ees, Spinpte-Enpd Focatizend CYToPLasmic AREA PRESENT aT Spinpie -Enp Large blastomeres Few spermatocytes Few somatic animal cells Some protista CyTopLasmic Area Not Present AT Spinpte -Enp Small blastomeres Most spermat acytes Most somatic animal cells CONDITIONS UNDER. WHICH tainly such may exist—where a sharply focalized spindle-tip, arising in a cytoplasmic area of con- siderable size, is without an aster; nor, on the other hand, where a blunt spindle-end arising in such an area has an aster. There are, however, some half-dozen cases of a doubtful character, such as a cell in which it is difficult to decide whether the spindle-end is blunt or slightly focal- ized. It is to be remembered that sharply focal- ized spindle-ends frequently become blunt when the chromosomes move toward them; hence as- ters which arose about focalized spindle-tips are associated with blunt ones during the late history of the figure; this, however, does not affect the conditions under which asters arise. It must also be kept in mind that in the case of intra-nuclear mitotic figures, the nuclear wall may inhibit the Most odcyte =) Few protista development of asters in the same way as the cell wall does in cases where the mitotic figure lies in the cytoplasm (cf. Figs. 6 and 5, and also 8 and 7). kinally, the evidence from sperm-asters, cyt- asters, and other unusual configurations, shows that asters can arise under conditions other than those discussed here. Another fact about asters difficult to explain is that they supposedly have two modes of origin, in one case arising de novo without reference to pre-existing asters, in the other arising by the di- vision of pre-existing ones. If an aster divides, it acts like a self-perpetuating cell component, sug- gesting that it plays some controlling role in the cell. However, a re-examination of a number of supposed cases of astral division shows that while SPINDLE-END BLUNT 3 4 Few sperm atocytes Few odcytes Few protista Somatic plant cells Ma ny protista ASTERS ARE PRESENT IN CELLS asters may sharply elongate, when the spindle- end widens, they do not divide. Old asters disin- tegrate from the center outwards; in some in- stances the new asters do not arise until all ves- tiges of the old aster have disappeared ; in others the new ones arise in the disintegrating central area of the old aster, while the peripheral portion of the latter still persists. The most generally accepted function of asters is that they play some kind of role in cell division. The most significant fact to be considered in this connection is that the majority of animal cells divide without asters. If one considers all types of animal cells—protozoa, oocytes, spermatocytes, blastomeres, and the innumerable classes of som- atic cells of invertebrates and vertebrates—asters eee Aucust 11, 1934 } THE COLLECTING NET 155 are found to be present in only a few cell types, and these are the large ones. Asters exhibit max- imum size and distinctness in eggs and early blas- tomeres, but they are absent or vague in most spermatocytes, and absent in the majority of somatic cells. Our thinking about animal cells has been shaped by the fact that most of our studies of asters have been concerned with eggs and large blastomeres, which are in reality atypical, owing to their large size. Actually, the majority of ani- mal cells carry on their activities, including mi- tosis and division, in the absence of asters. In cases where asters are present, the follow- ing facts suggest that they may not, in many cases, be concerned with cell division. (1) In Pennaria eggs the volume of the first cleavage figure is only about 0.15% of the volume of the egg, and the asters are therefore probably not concerned with cleavage; their volume seems to be related to the spindle volume rather than to cell volume. (2) In Cuwmingia eggs, during both polar-body cycles and the first-cleavage cycle, astral size is clearly related to spindle size, and it is independent of whether minute polar bodies are being pinched off or large blastomeres are being formed. If asters play some part in cleay- age here, it is hard to explain why the mitotic figure associated with that process has exactly the same size as the one associated with pinching off the first polar body. (3) If asters play some role in the process of cell division it would be ex- pected that, from one species to another, division would occur at approximately the same structural phase. But in Cumingia eggs cleavage occurs when asters are in late anaphase; in Arbacia eggs they are in telophase; and in Pennaria eggs they are in prophase of the second cleavage cycle. (4) The relation between astral structure and the time of cleavage may vary from one division to the next within the same egg, thus suggesting that the two are unrelated. In Arbacia eggs 50% first cleavage occurs when 80% of the eggs are in telophase, but 50% second cleavage occurs when only 40% are in telophase. (5) In some cells, such as those of the dogfish pancreas and Rom- alea spermatocytes, asters are present during the early mitotic phases, but completely disappear be- fore the cell divides. (6) In certain eggs, such as those of Echinarachnius, where asters are at max- imum size during first cleavage, low temperatures can practically suppress them without stopping cleavage. It has also been suggested that asters may play some role in the localization of organ-forming materials ; this may hold true in certain cases, but astral behavior in other instances is hard to ex- plain on this assumption. It is difficult to see, for example, why developing Drosophila eggs require asters for the localization of materials, while those of another fly, Miaster, do not; why epi- thelio-muscle cells of Hydra require them but its gland cells do not. Again, if asters of the polar- body figures in most animal eggs are involved in the localization of materials, it is hard to explain why the most highly differentiated phylum of all, the vertebrates, has anastral polar-body figures in many cases. To summarize briefly: First, asters of normal bipolar mitotic figures usually arise only if the spindle-end is sharply focalized and lies in a cyto- plasmic area of considerable size—which probably accounts for the sporadic manner in which they may be present or absent in cells of the same type which are carrying on the same activities. Second, in a number of cases studied, asters do not divide but disintegrate at the same point—the spindle- end—where they were organized. And third, they are absent in most cells; but in those in which they do occur, it appears that in many instances they do not play a role in cellular activities such as cell division or the localization of organ-form- ing materials. It is therefore tentatively suggested, as a work- ing hypothesis, that asters, when present, usually represent a secondary effect of the forces in- volved in the formation of the spindle, and that they probably do not play an essential rdle in cellular activities. (This article is based upon a seminar report pre- sented at the Marine Biological Laboratory on July 24.) A class for the study of Russian has been or- ganized by a group of students and workers at the Laboratory, with Samurt A. Corson of the University of Pennsylvania as instructor. The class will hold its meetings on Tuesday and Thursday evenings at 7 P. M. in the Old Lecture Hall, the first one being on July 31st. So far the group is studying chiefly the elements of the language with a view to enabling them to read Scientific Russian and to enter into simple conver- sation. A number of the students expect to at- tend the meetings of the International Physiologi- cal Congress to be held next summer at Moscow. Dr. AtFreD Otto Gross of Bowdoin College is spending the summer with DonaLp Macmit- LAN on a trip of exploration in the Arctic; the cruise is made in Macmillan’s own small ship, Bowdoin. There are five or six in the party, in- cluding students from Bowdoin College of which Macmillan is an alumnus. They left in June and are exploring along the coast of Labrador, while Dr. Gross is collecting biological material. Dr. Gross has been professor of biology at Bowdoin College since 1922. He has twice contributed to Tue Cottectrnc Net material concerning the last heath hen, 158 THE COLLECTING NET [ Vor. 1X. No. 78 LIST OF BIOLOGICAL BOOKS SUBMITTED BY PUBLISHERS AS BEING PUBLISHED IN AMERICA SINCE SEPT. 1, 1933 Abramson, H. A. Electrokinetic Phenomena. 1934. The Chemical Catalog Co., Inc. : Appleton, J. L. T. Bacterial Infection. Febiger. 7 Bache, Louise F. Health Education in an Ameri- ean City: an Account of a Five-year Program in Syracuse. Feb. 1934. Doubleday Doran. Bailey and Greene. Laboratory Manual in Biology. 1934. Allyn and Bacon. Baker, J. R. and J. B. S. Haldane. Biology in Everyday Life. 1933. Geo. Allen and Unwin. London. Barbour, Thomas. Reptiles and Amphibians. Apr. 1934. Chicago Field Museum of Natural His- tory. Barcrnes Joseph. Features in the Architecture of Function. Cambridge Univ. Press. 1933. Barlow, Nora. A Diary of the Voyage of H. M. S. Beagle by Charles Darwin. Macmillan. Barton-Wright. Kecent Advances in Botany. P. Blakiston’s Son and Co., Inc. Barton-Wright. Recent Advances in Plant Physi- ology. znd Ed. P. Blakiston’s Son and Co., Inc. Becker, E. R. Coccidia and Coccidiosis of Domes- ticated Game and Laboratory Animals and of Man. Mar. 1934. Collegiate Press, Inc. Ben Meyr. Your Germs and Mine: the Story of the Good and Bad Microbes. May 1934. Double- day Doran. Bergey. Manual of Determinative Bacteriology. Mar. 1934. Williams and Wilkins Co. Bigelow, Henry B. Studies of the Waters on the Continental Shelf—Cape Cod to Chesapeake. I: The Cycle of Temperature. Woods Hole Ocean- ographic Institution. Brown, Harcourt. Scientific Organizations in 17th Century France. Williams and Wilkins Co. Brown, Dr. Porter. The Art of Becoming a Moth- er. Dec. 1933. Eugenics Publishing Co. Burke, Edmund J. Lectures in Biology, Zoology. Fordham Univ. Press. Burton, Maurice. Sponges. British Museum. Calkins, Gary N. Biology of the Protozoa. Lea and Febiger. Carnegie Institution of Washington. Contributions to Embryology. W. F. Roberts and Co. Carr, W. H. Trailside Transformation 1934. Am- erican Museum of Natural History. Chapman, F. M. Autobiography of a Bird-Lover. Appleton-Century. Coker and Totten. Trees of the Southeastern States. June 1934. The Univ. of N. C. Press. Cold Spring Harbor. Symposia on Quantitative Biology, Vol. 1. 1933. Cole, E. C. Introduction to Biology, 1933. Wiley. Cole, Wm. E. The Teaching of Biology. Appleton Century. Creaser and Ortenburger. Decapod Crustaceans of Oklahoma. Univ. of Oklahoma Press. Crowther, J. G. Biology in Education. 1933. Heine- mann. Curran, C. H. The Families and Genera of North American Diptera. John D. Sherman, Jr. 1934. Curtiss and Guthrie. Textbook of General Zoology. Wiley. Cushnym, Edmunds and Gunn. A Textbook of Pharmacology and Therapeutics. 1934. Lea and Febiger. Curtis, Caldwell and Sherman. Apr. 1934. Ginn and Co. Curtis, F. D. and others. Biology for Today. 1934. Ginn and Co. Lea and Biology for Today. Darlington. Recent Advances in Cytology. P. Blakiston’s and Son, Inc. Dice, L. R. Preparation of Scientific Specimens of Mammals in the Field. Univ. of Michigan. Ditmars, Raymond. Reptiles of the World. Mac- millan. Dorf, Erling. Pliocene Floras of California. Car- negie Institution of Washington. Dougan, Lewis M. Stories of Outdoor Science. Lyons and Carnahan. Fiedler and De Beer. German Reader for Biology Students. Oxford Univ. Press. Fischer, Martin H. and Marian O. Hooker. Lyophillic Colloids. Charles C. Thomas. Fitcher, Palmer Howard. Giants and Dwarfs: a Study of the Anterior Lobe of the Hypophysis. Harvard Univ. Press. 1933. Freeman, G. L. Introduction to Physiological Psychology. 1934. Ronald Press Co. Furbay, John H. Nature Chats. A Year Out of Doors. Science Press. Gadow, H. W. The Evolution of the Vertebral Column. Cambridge Univ. Press. Gould, Pocket Medical Dictionary. P. Blakiston’s Son and Co., Inc. Gregory, Wm. K. Man’s Place Among the Arthro- pods. Oxford Univ. Press. Gudger, Eugene W. Archaic Fishes. Article V. The Natural History of the Frilled Shark. Ameri- can Museum of Natural History. Haggard, Howard W. Mystery, Magic and Medi- cine; the Rise of Medicine from Superstition to Science. Nov. 1933. Doubleday Doran. Harris, J. A. Physico-Chemical Properties of Plant Saps. 1934. Univ. of Minnesota Press. Harrow and Sherwin. Chemistry of Hormones. Apr. 1934. Williams and Wilkins Co. Hartman and Strauss. Anatomy of the Rhesus Monkey. Nov. 1933. Williams and Wilkins Co. Harvey, E. Newton and Arthur K. Parpart. Lab- oratory Directions in General Physiology. Holt. Henrici, Arthur T, The Biology of Bacteria. Heath. Hiss, John Martin. New Feet for Old. Sept. 1933. Doubleday Doran. Hogben, Lancelot. Nature and Nurture. Norton. Homes Harry N. Introductory Colloid Chemistry. iley. Hunter, G. W. Science Teaching at Junior and Senior High School Levels. May 1934. Ameri- can Book Co. Huxley, Julian S. and G. R. De Beer. The Ele- ee of Experimental Embryology. Macmil- an. Jackson and Warfell. Notes on the Occurrence of Mammals in the Regions Adjacent to the Salt The Plains of Northwestern Oklahoma. Univ. of Oklahoma Press. Jean, Harrah, Herman and Powers. Introductory Course in Science for Colleges. Vol. 1—Man ae the Nature of His Physical Universe. Mar. Vol. 2—Man and the Nature of His Biological World. May 1934. Ginn and Co. Jennings, H. S. The Universe and Life. Yale Univ. Press. Koch. Practical Methods in Biochemistry. Mar, 1934. Williams and Wilkins Co. Kofoid, Charles A. Termites and Termite Control. Univ. of California Press. Lake, Harley and Walton. Exploring This World of Science. 1934, Silver, Burdett and Co. Lenard, Philip. Great Men of Science. Macmillan. Aveust 11, 1934 ] THE COLLECTING NET 159 Levine, Max. Laboratory Technique in Bacteriol- ogy. Macmillan. : Lobel, Josef. Medicine—A Voyage of Discovery. Farrar and Rhinehart. Loeb, Leonard B. and A. S. Adams. The Develop- ment of Physical Thought. 1933. Wiley. Lubbock, Constance. The Herschel Chronicle. Macmillan. Lucas, F. A. The Hall of Dinosaurs. Nov. 1933. : American Museum of Natural History Reprint. Lutz, F. E. Invisible Colors of Flowers and But- terflies. 1933. American Museum of Natural History. MacDougal, D. T. and Earl B. Working. The Pneumatic System of Plants—Especially Trees. Carnegie Institution of Washington. Macgintie, Harry D. The Trout Creek Flora of Southeastern Oregon. Carnegie Institution of Washington. Mayers, L. H. What We Are and Why. 1933. Dodd, Mead and Co. Mills, H. B. A Monograph of the Collembola of Iowa. May 1934. The Collegiate Press, Inc. Miner, R. W. Diving in Coral Gardens. Nov. 1933. American Museum of Natural History. Morgan, T. H. Embryology and Genetics. 1934. Oxford University Press. Needham, J. G. and J. T. Lloyd. Life of Inland Waters. Comstock Publishing Co. Needham, J. G. and P. R. Needham. A Guide to the Study of Fresh Water Biology. Comstock Publishing Co. Newman, B. M. Advanced Biology. 1933. College Entrance Book Co. Newsholme and Kingsbury. Red Medicine: Social- ized Health in Soviet Russia. Dec. 1933. Double- day Doran. Nicholson, Daniel. and Febiger. Ortenburger and Bird. The Ecology of the West- ern Oklahoma Salt Plains. Univ. of Oklahoma Press. Ortenburger ard Ortenburger. Howard Atwood Kelly. Univ. of Oklahoma Press. Packard, Earl L., Remington Kellogg and Ernst Huber. Contributions to Paleontology: Marine Animals. Carnegie Institution of Washington. Park and Williams. Pathogenic Microorganisms. Lea and Febiger. Parsons, L. M. Introduction to Biology. 1933. Mac- millan. Pearl, Raymond. Constitution and Health. Kegan, London. Pearl, Raymond and R. D. Pearl. The Ancestry of the Longlived. Johns Hopkins Press. 1934. Peterson, R. T. A Field Guide to the Birds. Apr. 1934. Chicago Field Museum of Natural His- tory. Plath, Otto E. Bumblebees and Their Ways. Mac- millan. Plimmer, R. H. Organic and Bio-Chemistry. 5th Ed. Longmans. Rahn. Physiology of Bacteria. P. Blakiston’s Son and Co., Inc. Ramsey, G. F. Project Making in Elementary Science. 1934. American Museum of Natural History. Reid, Mary E. and Marjorie E. J. Chandler. The London Clay Flora. British Museum. Robbins and Ramalay. Plants Useful to Man. P. Blakiston’s Son and Co., Inc. Robbins and Rickett. Botany. June 1934. D. Van Nostrand Co., Inc. , Roberts, T. S. Bird portraits in Color: 295 North aan Species. Aug. 1934. Univ. of Minn. ress. May Laboratory Medicine. 1934. Lea Paul Robinson, Dr. W. J. Our Mysterious Life Glands. Feb. 1934. Eugenics Publishing Co. Robinson, W. W. Ancient Animals. Macmillan. Romer, Alfred S. Man and the Vertebrates. Univ. of Chicago Press. Rose, Mary S. ‘The Foundations of Nutrition. Mac- millan. Roule, Louis. Fishes: Their Journeys and Migra- tions. Norton. Sansome and Philip. Recent Advances in Plant Genetics. P. Blakiston’s Son and Co., Inc. Saunders, Charles F. Useful Wild Plants of the U. S. and Canada. McBride. Schwesinger, G. C. Heredity Macmillan. Scott, Wilfred W. Editor. Standard Methods of Chemical Analysis. 2 vols. 4th Ed. Van Nos- trand Co. Sharp, Lester W. Introduction to Cytology. 3rd Edition. McGraw-Hill Book Co. Sherman, Henry C. Food Products. Macmillan. Sherringion, Charles. The Brain and Its Mechan- ism. Macmillan Co. 1933. Sherwood, G. H. The Museum in Education. 1934. American Museum of Natural History. Shull, Larue and Ruthven. Principles or Animal Biology., 4th Edition. McGraw-Hill Book Co. Silberstein, Ludwik. Causality: a Law of Nature of a Maxim of the Naturalist. Macmillan. 1933. Skilling, W. W. Tours Throughout the World of Science. McGraw-Hill Book Co. Snedecor, G. W. Calculation and Interpretation of and Environment. Analysis of Variance and Covariance. Jan. 1934. The Collegiate Press, Inc. Smallwood, Reveley and Bailey. New Biology. 1934. Allyn and Bacon. Smith. Recent Advances in the Study of Plant Viruses. P. Blakiston’s Son and Co., Inc. Sternberg, Charles H. Life of a Fossil Hunter. Jensen Printing Co. Sullivan, J. W. N. The Limitations of Science. Oct. 1933. The Viking Press, Inc. Talmey, Dr. Bernard S. Love: The Science of Sex Attraction. Eugenics Publishing Co. Tilden, J. E. The Algae and Their Life Relations. Univ. of Minnesota Press. Viskery, Hubert B. et al. Chemical Investigations of the Tobacco Plant. Carnegie Institution of Washington. Warden, Jenkins and Warner. Introduction to Comparative Psychology. 1934. Ronald Press Co. White, E. G. Textbook Mosby. Whitlock, H. P. Jade and the Antique Use of Gems. Dec. 1933. American Museum of Natural History. Winslow, C. HE. A. A City Set on a Hill: the Sig- nificance of the Health Demonstration at Syra- cuse. Jan. 1934. Doubleday Doran. Wistar Institute Style Brief. 1934. Wood and Tinker. Fifty Years of Bird Migration in the Ann Arbor Region of Michigan. Univ. of Michigan. Wright, A. H. and A. A. Wright. The Handbook of Frogs and Toads. Comstock Publishing Co. Wyeth, F. J. Elementary General Biology. Bell. G. Yonge, C.M. A.B. C. of Biology. 1934. Routledge. Catalogue of the Books, Manuscripts, Maps and Drawings in the British Museum. British Mu- seum. Papers in Physical Oceanography and Meteorology. Vol. Il. Woods Hole Oceanographic Institution. Papers of Tortugas Laboratory. Vol. XXVIII. Car- negie Institution of Washington. of General Biology. 1933. 156 THE COLLECTING NET [ Vot. IX. No. 78 THE COURSE IN INVERTEBRATE ZOOLOGY Dr. Expert C. CoLe Director of the Course, Professor of Biology, Williams College The course in invertebrate zoology, beginning this year on July 31, is designed for students hav- ing a substantial training in biology and who, pre- sumably, have a professional interest in zoology. An attempt is made to secure as wide an institu- tional representation as possible, so far as this is consistent with the acceptance of thoroughly qual- ified students. Forty-four colleges and universi- ties are represented in the present class. Twenty- nine women and twenty-five men are enrolled. The personnel! of the staff of instructors remains the same as that of last year, with one exception. Mr. Seymour M. Farber, a member of the class in 1930 and winner of a CoLLectinG Net Schol- arship in that year, is serving as laboratory assist- ant in place of Mr. Stewart Rankin who held that position last year. As usual, representatives of important inverte- brate phyla are studied in the laboratory. The regular lectures of the course include not only detailed information which will be of use to the student in the pursuance of his laboratory work but also those general considerations which aid him in securing a broad viewpoint of the subject as a whole. In lectures and laboratory work both the physiological and the morphological aspects of the field are kept in mind. From time to time spe- cial lecturers will be invited to address the class on subjects having a close relation to the work of the course. The work of this year includes eight field trips, covering such habitats as mud and sand flats, rocky shores, brackish pools, marshes, wharf piles, and open water where the dredge and net may be used. Although the securing of a large number of forms is desirable, this aim is subor- dinated to a more important one—the study of the animal in relation to its environment. So far as is possible, invertebrate forms are identified in the field, but further work of this sort is car- ried on in the laboratory after each trip. Since the check list of forms carries the names of 327 species, it is obvious that in a course of this sort more attention must be paid to this list of ani- mals than to those species which are of interest chiefly to the specialist. It is planned to prepare a public exhibit of forms secured on a represen- tative field trip. This will take place about August 13th, and the exhibit will probably be located in the vestibule of the Main Building, as in previous years. Although the work of this course is intensive, it does not require more time and effort than a well-prepared and industrious student may rea- sonably be expected to devote to it. At the close of the course on Sept. 6th, it is believed that each student will have acquired some measure of ac- quaintance with a wide variety of invertebrate animals, and the environments in which they dwell. SPINDLE FIBER ATTACHMENTS: DO THEY CHANGE WITHOUT CHROMOSOMAL INVERSIONS OR TRANSLOCATIONS? Dr. E, ELEANOR CAROTHERS Department of Biology, University of Pennsylvania The report is based on the behavior of the sex- chromosomes in the spermatogenesis of a single male of Trimerotropis coquilletti. All are, there- fore, direct descendants of one x-chromosome which the individual received from his mother. In adjoining first spermatocyte metaphases, the x-chromosome may have terminal, median or sub- terminal spindle fibers. In some cases fibers at all three loci on a single chromosome function simultaneously, and if one fiber leads to the op- posite pole from the other two, the x-chromosome is flattened out and held near the center of the equatorial plate, with clear indication that a pull is being exerted at these loci. By inversion a v- shaped chromosome might be derived from a rod- shaped one or vice versa, (a definite relationship exists between the shape of a chromosome and the location of its spindle fibers as was pointed out by McClung in 1914). Conceivably the extra points of attachment might have been acquired by translocations from other members of the com- plex, but in that case a cell in which the x-chrom- osome has fibers arising at three points should show some of the other chromosomes of the com- plex behaving abnormally because of the loss. Such is not the case. The more logical conclusion seems to be that this chromosome gives rise to ex- tra spindle fibers due to some change within it- self. Evidence is offered to support the theory that the fibers which pass from the chromosomes to the poles and those which connect the daughter chromosomes at anaphase (interzonal fibers), are of chromosomal origin, hence differ both in origin and function from the astral radiations which pass from pole to pole and are of cytoplasmic origin. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on July 24). Se Aveusr 11, 1934 } THE COLLECTING NET 157 A CRUISE WITH THE COAST GUARD IN GREENLAND On Friday August 10th, Lieutenant Com- mander E. H. Smith, former captain of the Coast Guard base at Woods Hole, showed moy- ing pictures for the Children’s School of Science, entitled “A Cruise with the Coast Guard to Greenland.” The first film illustrated the work of the In- ternational Ice Patrol along the coast of New- foundland in the North Atlantic where the U. S. Coast Guard protects ships from the hazzard of icebergs. The bergs exhibit beautiful colors in the sunlight, are very dangerous, and carry eight- ninths of their bulk below the surface of the water. Commander Smith had excellent moving pictures of the icebergs showing striking con- trasts between dark shadows and glistening white ice. In 1928 Commander Smith was sent on a trip of oceanographic investigation to Greenland on the Coast Guard cutter, Marion. They stopped for a few days at Boston to gather together the necessary oceanographic equipment in order to study the salinity and chemistry of the water, and the changes in temperature and currents. Dur- ing eight weeks of continuous work, observations of the water were made every thirty or forty miles. Progressing through the Gulf of St. Lawrence the expedition reached Baffin Bay, the rendezvous of seals, walrus, and polar bears. One cub bear was caught after a great deal of trouble on the part of the crew. An 800-pound mother bear and her second cub had to be sacrificed in order that the remaining cub could be captured. It was named ‘‘Marian,’ weighed 400 pounds, and was wild and savage attempting to bite its captors. A cage was built for Marian, and when the expedition stopped at Martha’s Vineyard on the return voyage the cub was a popular attrac- tion for all the children of the Island. It is now in the zoo at Washington. Greenland is a colony of Denmark, and since the Esquimaux are no longer able to support themselves adequately by catching seals and whales, the officials from Denmark undertake to educate and govern them. In Greenland they found some of the glaciers travel 60-70 feet daily, and rose to a height of. 70 feet above the water. The Marion and her crew were in this region from January until May and were unable to navigate the waters by boat during the winter because of the unbroken stretches of ice. The expedition alsc stopped at Labrador and the men spent a day examining the vegetation which grows profusely in the summer. Com- mmander Smith showed picturesque views of the mountains, fjords, and icebergs of Labrador. THE BEACH QUESTION IN THE LIGHT OF A CHANGING ENVIRONMENT (Continued from page 150) tions secured by members of the Corporation or by creating a deficit on faith in the future. These conditions persisted while the number of investi- gators steadily increased, and many new methods were introduced calling for more space and more varied equipment. So the Marine Biological Laboratory was greatly enlarged in 1916 and again in 1925 in- -corporating a thoroughly modernized Jnternal Environment for the work. At the same time much more and better general facilities were sup- plied to serve the larger and more diverse crowd of workers. The collecting department became an extensive business; a large auditorium ac- -commodating 500 persons was added; and an exceptional library, where books and journals are “made easily available in ample space, and which is rapidly becoming one of the best in the world for biologists, was gradually built up from the very modest beginnings housed in one small “room in the ’90s. We have sketched, thus briefly, some of the “main developments in the laboratory concerned with the provision of facilities for the profes- sional work proper, chiefly to point out the con- siderable burden of special “problems” which had to be met here and the unique spirit of co- _ and unselfish devotion which brought about the splendid result we now enjoy. For the creation and development of this ex- tensive and adequate /nternal Environment, the Laboratory proper, which is proving favorable for so much effective work, we must thank a group of devoted, individualists who sacrificed much time and effort for the good of the cause. Some of that very group still continue to bear the burden of responsibility they have long car- ried; and gratitude is due them we think, not only for the unusual constructive result, but, especially, for the example of adherence to the fine principles of independent self-reliance and unselfish cooperativeness which have brought it all about. In reflecting on the possible application of their successful methods to the management of some of the difficulties which arise for us in the more external relations of the Laboratory we must advise adherence here also to the same self- contained independence from outside interference with continued cooperative effort among our- selves. This discussion will be continued in the forth- coming number of THE Cottectine Net. This second part will be devoted to the External En- vironment, of which the beach problem is a special case. LOO Re, Ai . TEE, Books Reduced 123: to 2-3, OEE Of fice Street Call at our on Water Die COLE CRIN GIN ET WOODS HOLE, MASS. oO & MICRO SLIDES COVER GLASSES DO NOT FOG Ask your dealer—or write (giving dealers name) to ‘Cray- ADAMS Hl 25 East 26th Street NEW YORK R oN oN 020) 0 ee 0 0 0) ODO Ces LEA & FEBIGER PUBLICATIONS NEW WORKS AND NEW EDITIONS On Exhibit August 20-25 Mr. W. D. Wilcox, Manager of the College Department, will be in personal charge. 2nd edition 2nd edition | Appleton on Bacterial Infection, | Bell’s Text-Book of Pathology, Boyd’s Text-Book of Pathology, 2nd edition Calkins’ Biology of the Protozoa, 2nd edition Cowdry’s Histology Cushny’s Pharmacology’and Therapeutics, 10th edition Fishberg on Hypertension and Nephritis, | 8rd edition Gershenfeld’s Bacteriology and Sanitary Science, 2nd edition Grafe on Metabolic Diseases and Their Treatment Kuntz on The Autonomic Nervous System, 2nd edition Nicholson's Laboratory Medicine, 2nd edition Park and Williams on Pathogenic Microorgan- isms, 10th edition Starling’s Physiology, 6th edition Weil's Text-Book of Neuropathology Wiggers’ Physiology in Health and Disease ! AND OTHER STANDARD TEXT-BOOKS LEA & FEBIGER 600 WASHINGTON SQ. PHILADELPHIA, PA. 3 0 OS OE Se OO OE 4% COLLECTING NET [ Vor. IX. No. 78 Turtox Microscope Slides Over two thousand different mic- roscope slides for Biology and related subjects are made in the Turtox slide laboratory. Specialized technicians use modern methods to produce - slides which any teacher may well be proud to show. Ask for your copy of the 194-page, illustrated Turtox Microscope Slide Catalog GENERAL BIOLOGICAL SUPPLY HOUSE Incorporated 761-763 EAST SIXTY-NINTH PLACE CHICAGO COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY Volume I (resulting from conference-sym- posia of 1933 and dealing largely with surface phenomena) contains papers by Harold A. Abramson, D. R. Briggs, Robert Chambers, Barnett Cohen, Kenneth S. Cole, Hugo Fricke, Herbert S. Gasser, A. V. Hill, Duncan Mac- Innes, L. Michaelis, Stuart Mudd, Hans Muel- ler, W. J. V. Osterhout, Eric Ponder, Theodor Svedberg, D. D. Van Slyke. From a book review: “If this initial high standard (Volume I) is maintained, it is diffi- cult to see how a worker in this field can do without these volumes. . .” The price of Volume I is $3.35. Volume IL will appear in the autumn of this year. The prepublication price of Volume II, bound in cloth, is $2.90, cash with order. After publi- cation, the price will be $3.35. Persons pur- coeee Volume II may obtain Volume I for $3. Address the Biological Laboratory, Sold. ‘Spring Harbor, L. I., N. Y. ee | _— Aucust 11, 1934 ] THE COLLECTING NET 161 Natural Convergence of Eyes maintained by SPENCER WIDE-FIELD MICROSCOPE Eliminates Cyestrain .. Increases Comfort ’ 1 fi Spencer Wide-field Binocular Microscope is the only one in which the prisms are arranged to permit the axes of the oculars to converge at an angle of 8° which is the normal angle of convergence of the eyes Folder M-55 gives complete description and prices of the Spencer series of Wide-field Binocular Microscopes. We will be pleased to send you a copy. BUFFALO at convenient reading distance. This exclusive patented Spencer feature is one of great importance to one whose work necessitates the constant use of a wide-field microscope. The ordinary construction requires a convergent an- gle of the eyes of 16° which imposes an eyestrain that is excessive. The inten- sive eyestrain from working at this ocular convergence angle of 16° is eliminated . . and it just naturally results in protection for the eyes and increased comfort for the observer. NEW YORK 162 THE COLLECTING NET [ Vor. IX. No. 78 BIOLOID Embedding Paraffine BIOLOID* Paraffin has been specially prepared for embedding and it will be found far superior to the quality ordi- narily offered. It is processed from the best domestic wax; it is pure white in color, free from excess oil, and practical- ly neutral in reaction. The melting points have been carefully checked in accord- ance with the methods of the American Society for Testing Materials. Each cake is individually wrapped in glassine paper and packaged in a substantial carton. It is available with the following melting points : ; au ) BO-52560125-125— ks) RES HAC) (CUZCO 195) 56-58°C (133-135° F.) Price NVA Ey pate GG leetstetetetec etece cats safer iezel Syezo TOADS CAKES W eratereieiaton: lela) sn10/0 2.40 25— 1) NP WiGHKES aia lelalelsieleisialcln« 5.50 1O0—— 0) ABS MCAe Siar ierete eielectsisie's- 20.00 * ‘Bioloid” is the registered tradename for Will Cor- poration staining, mounting, embedding clearing, and fixing media. WILL CORPORATION LABORATORY APPARATUS AND CHEMICALS ROCHESTER, NY. THE VOLUMETTE An instrument designed to deliver repeat- edly quantities of fluid in accurate prede- termined doses. ADV ANTAGES Accuracy — Durability — Ease in Cleaning Quietness — Speed Write for Bulletin No. 486 re Motor Oper- ated Volumette. For Hand Operated Vol- umette, advise requirements. EIMER & AMEND Est. 1851 Inc. 1897 Headquarters for Laboratory Apparatus and Chemical Reagents Third Ave., 18th to 19th St., New York, N. Y. THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and 37 plates, published January, 1934 This guide for authors, in preparing manu- scripts and drawings for the most effective and economical method of publishing biologi- cal research, has been prepared by the Staff of The Wistar Institute Press and the co6per- ative efforts of more than fifty editors con- cerned in the editing of journals published by The Wistar Institute, and presents the con- sensus of opinion on many points relating to the mechanical preparation of manuscripts and drawings for the printer and engraver. Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. Aveust 11,1934] == ===~=THE COLLECTING NET eae 163 Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, etc. Catalogs will gladly be sent on request. eri oncl Please mention name of school and : palteholz subjects taught, to enable us to . . 7 Life History Transparent send the appropriate catalog. of Chick Preparations Human Amy Ay INOW Cow A age y oa : JLAY--\ADAMS VOMPANY oological E 25 EAST 26th STREET NEW YORK Model of Human Heart aa = Visit our display rooms and museum. New Microscope Model H With low fine adjustment in case. Stand HCE Complete Abbe Illuminating Apparatus Revolving mechanical stage E movement 50 x 70 mm. $245.00 Quadruple nosepiece 11.00 Inclined binocular body 79.00 Without optics $335.00 f.o.b. N. Y. A copy of catalog Micro 488 will be supplied upon request. CARL YZ Ee les: 5).4 Nees Atlin AN em we 728 Se. Jelillll Street: 468 5 N E W YO _R_K cari7fisspb OS ANGELES a femme __ THE COLLECTING NET _ [ Vou, IX. No. 78 WE MAKE OUR OWN GLASS TO INSURE STANDARDIZED PRODUCTION FOR YOUR GLASSES, INSIST ON B aL ORTHOGON LENSES AND B a L FRAMES Write for complete in- formation and details of the accessories avail- able for different types of work. Bausch & Lomb Optical Com- pany, 671 St. Paul Street, Rochester, New York. OP ICAL INSTRUMENTS FOR’ WILL NEVER FORGET Only the Camera remembers exactly—Notes and sketches do not! Your microscope observations can be recorded by the camera—in most cases—as easily and as quickly as you can take snapshots of events you want to remember in every day life! B & L Photomicrographtc Equipment GBVP represents the finest photomicrographic equipment money can buy. Spring suspension, mounted in live rubber at all four corners elimi- nates vibration and gives the instrument its remarkable stability. Its permanently aligned structure whereby the illuminating unit is mounted on the same bed which carries the microscope plate makes for rapid observations under identical conditions. The elements are always in position, ready to use. You need not be an optical engineer to use this equipment. Bausch s Lomb HE SCIENCES Vol. IX. No. 8 SATURDAY, AUGUST 18, 1934 Annual Subscription, $2.00 Single Copies, 25 Cents. THE MARINE ZOOLOGICAL LABORA- TORY ON THE ISLES OF SHOALS Dr. Lowett E. NoLanp Associate Professor of Zoology, University of Wisconsin Ten miles out from Portsmouth, on one of the historic Isles of Shoals, the University of New Hampshire is conducting the seventh session of its marine zoological station. he laboratory is located on Appledore Island, once the home of the artist and poet- ess, Celia Thaxter. The build- ings were formerly a part of a group of dwellings about a large summer hotel, Apple- dore House, which was de- stroyed by fire about twenty years ago. Courses are offered in histology, embryology, com- parative anatomy, invertebrate zoology and elementary zool- ogy to a group of about forty students, mostly undergradu- ates. Dr. C. F. Jackson, dean of the College of Liberal Arts at the University of New Hampshire and chairman of the zoology department there founded the station seven years ago, and is now its director. are Mrs. Jackson; Dr. Norman Arnold, of Dart- Il. (Continued on page 174) mouth College; and THE EFFECTS OF X-RADIATION ON CELL STRUCTURE AND GROWTH GEORGE L. CLARK Department of Chemtstry, Massachusetts Institute of Technology MRM. B. EU. Calendar TUESDAY, August 21, 8:00 P. M. Dr. Morris H. Harnly: “The tem- perature-effective periods and ‘growth curves’ of the vestigial wings of Drosophila melano- gaster.” Dr. P. W. Whiting: ‘Selective fer- tilization in Habrobracon.” Dr. B. R. Speicher: ‘Maturation, fertilization and cleavage of Habrobracon eggs as_ revealed by the Feulgen reaction.” Dro oH. Hy Plough Vand) Mr Pal, Ives: ‘Mutations and modifica- tions in Drosophila induced by sub-lethal high temperature.” THURSDAY, August 23, 8 P. M. Lecture: Dr. F. Baltzer: “Experi- ments on Sex-development in Bonellia.” FRIDAY, August 24, 8:00 P. M. (Continued on page 176) On the staff IT. The undoubted value of X-rays as a research tool to the biologist in the field of genetics has been convincingly demonstrated in the past few years by the work of H. J. Muller of Texas and a very considerable number of other workers. Much less well known are the great possibil- ities of cell studies depending upon an increasingly quanti- tative evaluation of character- istic radiosensitiveness which may well become the most powerful method of patholog- ical diagnosis. | Uncertainty remains as to fundamental differences between normal and pathological tissues, and the mechanism involved in radiobiology and therapy of cancer cells: It has seemed desirable, therefore, to sum- marize briefly our present knowledge of the biological effects of X-radiation, under the following heads: Bactericidal and General Lethal Effects. Effects on the Hereditary Material. The Marine Zoological | Editorial Page Dr. W. C. Allee The Effects of X-radiation on Cell Structure and Growth, Dr. George L. Clark Laboratory of the Isles of Shoals, Dr. Lowell E. Noland..... 165 Trustees of the Marine Biological Laboratory 175 Cooperation Among Marine Laboratories, TABLE OF CONTENTS Items of Interest Permeability of Amoeba Proteus to Water, Dr. Coleen Fowler The Genetics Society of America—Program 181 Woods Hole Choral Club Concert Wolfinsohn-Wilson-Wagenaar Trio Concert. .182 The Beach Question in the Light of a Chang- 177 ing Environment II, Dr. H. McE. Knower. .183 DORMITORY FOR MEN, FOR GIRLS AND FOR THE FACULTY ON APPLEDORE ISLAND 1 . HL AN u JACKSON HALL ON APPLEDORE ISLAND Aucusr 18, 1934 ] _THE COLLECTING NET 167 Sie BIOLOGICAL LABORATORY COLD SPRING HARBOR THE EFFECTS OF X-RADIATION ON CELL STRUCTURE AND GROWTH A GENERAL SURVEY OF RADIOBIOLOGY (Continued from page 165) Ill. Effects on Normal Cells. IV. The Radiosensitiveness of Cells. V. X-Ray Effects on Human Tissues. VI. The Stimulating Effect of X-Rays. VII. Medical Implications of Cell Radio- sensitivity and Reactions to Radiation with Special Reference to Cancer. VIII. A New X-Ray Method of Pathological Diagnosis. I, Bactericidal and General Lethal Effects of X-Rays This topic is first considered since the first sug- gested possible biological application of the new rays, following announcement of their discovery by Roentgen in November, 1895, was directed to bactericidal properties. The rays were considered to be present in sunlight. Since the sun’s rays were known to have bactericidal properties and were beneficial in treatment of diseases, especially tuberculosis, it was natural to turn to these rays as possessing possibly even more powerful physiological effects. On January 29, 1896, T. Glover Lyon made definite suggestions in a letter to the London Lancet. But on February 17, 1896, in another letter he announced negative re- sults on tuberculosis and diphtheria bacteria. This was the beginning of experiments on every type of organism in laboratories throughout the world, an account of which would occupy an entire vol- ume. Some of the experiments were highly fav- orable, but the great majority were negative; positive bactericidal effects in one laboratory were entirely disproved in another. Thus began the great science of X-ray therapy under these highly unfavorable conditions. While it became nearly universally agreed that bacterial cultures were nearly inert except to relatively enormous doses of radiation, the tremendous effect of rays on liv- ing tissue also became apparent. In 1902 Pusey and Caldwell stated in their book the following: “The fact that organisms in living tissues can be destroyed by exposure to X-rays while the same organisms in inert cultures are uninfluenced by X-ray exposure, proves positively that it is not the influence of the X-rays per se that causes de- struction but that the tissues themselves doubtless under the conditions of activity excited by the X- rays play the important role in the germicidal process.” ; It would be profitless to enumerate the long series of experiments on bacterial cultures. The reputable results agree in showing logarithmic death rates for very large dosages, as exemplified by results by the writer on B. coli and Erythro- bacillus prodigiosus. The irradiation was effected with a tungsten target tube using a potential of sixty-five kilo volts and a current of three to four milliamperes. At certain time intervals test tubes were removed and dilutions plated out to determine the total counts. The analytical results are summarized in Table I. The rate of sterilization may be noted when the bacterial count is plotted logarithmically against time of irradiation. The following conclusions may be derived from these data : (1) X-rays act like sterilizing agents upon cultures of 5. coli and Erythrobacillus prodigio- sus, in that the curves are characteristic steriliza- tion or death-rate curves showing that the total counts decrease logarithmically with time. (2) In this experiment 5. coli did not show variation or mutation when it was treated with X-rays. (3) With increasing irradiation Erythrobacil- lus prodigiosus showed a tendency toward lack of ability to produce its characteristic red pigment. By allowing the organism to grow on the plate for a period of five days the greater portion of the colonies produced their pigment. If a trans- fer of a white colony is made to an agar slant the characteristic pigment is produced in twelve hours. In only one case was it necessary to make a second transfer in order to bring about the de- velopment of the pigment. The above is another example of the variation tendency of Erythroba- cillus prodigiosus. Cultures of simple organisms have been used frequently to discover the effects of various X- ray wave-lengths, generally indicating as in all other cases a biological effect independent of wave-length under comparable conditions of ener- gy absorption. Wyckoff! found that rays kill B. coli and B. aertryke in linearly exponential fash- ion. He considers that death results by the ab- 1 J, Exp. Med. 52, 485, 769 (1930), 168 THE COLLECTING NET [ Vor. TX, No. 79 TABI we SUMMARY OF ANALYTICAL RESULTS B. Coli Data s 1 Time of Total count aa € irradiation per cc. Remarks number in minutes (24 hrs.—37° C.) 1 0 70,000 2 5 70,000 3 15 17,000 Single colonies were picked from plate of sample number 4 30 9,000 6 and morphology and cultural characteristics were ex- 5 45 1,700 amined. All tests were characteristic of the original. 6 60 10 7 90 0 Erythrobacillus prodigiosus Data Observations of Glucose-Agar Plates Sample ee ie 2nd day 3rd day 5th day pnnen irradiation ; in minutes Tot Couey ; Pigment of ‘ s per cc. Pigment Polonios Pigment of colonies 20° C. al 0 800,000 red 4:\0 (PAN OA Sus 5O07C 2 5 600,000 red iit | oo000DuCO0dCOSTO: 0 3 15 480,000 red Fed’ §_ ieleiescateterctet ese Meeaene = 4 30 400,000 red 140 NB Goopscodondooon0]~ 5 45 300,000 red TEC 5 | Winierenstonetesokehely eles ane 6 60 45,000 80% white* 50% white 90% red 20% red 50% red 10% white 7 90 400 100% white* 80% white 60% red, 40% white 8 120 (Mn amaraurin con .cnommopdrOmne he Vodiadbocoonaoc seu” * All white colonies that were fished from the plates (12 in number) and streaked on agar slants produced luxuriant red pigmented growth at 20° C. in twenty-four hours with the exception of one colony fished from sample number 7 which failed to give red pigment until it was transferred to a second agar slant. sorption of a single X-ray quantum of energy. Since only 1 in 20 of the absorbed quanta kills, the sensitive cell constituents whose destruction leads to cell death must have a volume which is less than 0.06 of the bacterium itself. Closely re- lated are experiments with yeast. Wyckoff and Luyet? find that unlike bacteria, yeasts can be in- jured, without being killed outright. Injury is followed by an extraordinarily large number of two-celled colonies which on prolonged incubation ultimately die without further budding. Of course, the final absolutely general effect of X-rays on living organisms is a lethal one. The quantity of radiation required for death is so defi- nite for each particular type of cells that a lethal count serves as a quantitative dosimeter of the radiation, as demonstrated by Packard's paper in these symposia. Five years ago, Wood showed that for Drosophila eggs 50% are killed by 190 r and 90% by 500 r. As a further indication of the lethal effects, Table 2 shows the results of ex- periments in the writer’s laboratory on hen’s eggs. 2 Radiology 17, 1171 (1931). Il. Effect of X-Rays on Hereditary Material. The experiments with bacteria have shown that the only observable biological effect is death, while in yeast it is indicated that reproductive processes may be greatly affected by irradiation energies far smaller than required for death of the organ- ism. This brings us to a brief discussion of X- rays in the science of genetics, already recognized by all biologists. In 1903 Albers-Schonberg dis- covered the sterilizing effect of X-rays. From that day to this the controversy has raged as to what effect may have been suffered by those cells irradiated but not sterilized. The controversy has taken the principal direction of whether or not sterilization of human beings by X-rays is safe, or whether cells which have suffered changes in the hereditary material may not produce ab- normal young when reproductive powers are re- established. Muller, outstanding authority in the field, has the following to say about X-ray meth- ods in the hands of the geneticist. Speaking of the gene he says, “It is evident that one chief method of attack in this new type of physiologi- ; | Aveusr 18, 1934 } THE COLLECTING NET 169 TABLE 2. Eacs 180,000 P. K. V.5 MA. 4% Cu 1 Al 50 cm. Dist. Effective A.U. 0.22 No.of No. Dead No. of Time Rayed “R” Units Eggs Inf. Germs Chicks 2 min. 34 sec. 38.50 LO) 2 5 3 5 min. 9 sec. 17.25 9) 52 3 4 7 min. 43 sec. 115.75 10 al 5 4 16 min. 18 sec. 154.50 10710) 3 af 12 min. 52 sec. 193.00 10 O 7 3 15 min. 27 sec. 231.75 10 3 4 3 18 min. 1 sec. 270.25 10550 6 4 20 min. 36 sec. 309.00 10 O 8 2, 23 min. 10 sec 347.50 Gal 4 3 25 min. 45 sec 386.25 co 6 al 28 min. 19 sec 424.75 @ il 4 1 36 min. 3 sec 540.75 {hy al 4 1 43 min. 46 sec 656.50 i 5 al 51 min. 30 sec 772.50 tf tf 0 59 min. 13 sec 888.25 ff ft) ff 0 66 min. 57 sec 1004.25 ie pel! 6 0 77 min. 15 sec. 1158.75 Gj al tf 0 cal analysis, a method which must be of high eventual importance to pathology, as well as to physiology and embryology, would be the altera- tion or the excision of individual genes, one at a time, out of these thousands of genes, followed by intensive embryological, physiological and physicochemical study of the effects thereby pro- duced on the organism. In other words, if we had the ability to change individual genes, we should have, in effect, a scalpel or an injecting needle of ultra-microscopic nicety, wherewith to conduct the most refined kind of vivisection or biochemical experiments on our experimental ani- mals, not experiments in which gross parts are removed, injected or otherwise changed, but ex- periments in which the finest most fundamental elements of the body fabric are separately at- tacked. Changes in the genes which have arisen spontaneously and are already at hand, can of course be used in such a study, but many of the most instructive types of these have already been largely weeded out by a process of natural dying off, in other words, natural selection, before we can find the individuals containing them, while many of those still existing be scattered far apart and concealed. Moreover, in nature gene changes arise very rarely. Thus the advantage which we would have if we were able to manufacture our own supply of them is evident. But apart from the action of radiation no generally effective means of artificially producing such gene changes, or gene mutations, as they are called, has as yet been satisfactorily demonstrated, that is, demon- strated in such a way that most specialists in heredity are willing to agree that the case is proved. Hence, the question of the production of changes in the hereditary material by means of roentgen or radium rays becomes all the more urgent.” % Science of Radiology, C. C. Thomas, Springfield, Iil., 1933. C. R. Bardeen, in 1906, first found that the ova of a toad fertilized by spermatazoa exposed to X- rays developed abnormally, and he concluded that the action of the rays must be on those unknown substances in the nucleus, or the protoplasm most intimately associated with the nucleus which con- trol the morphogenetic activities of the cell. Thus Bardeen was lacking only the concept of the chromosome. Most of the work between 1905 and 1925 has limited value largely because a large preponderance of new gene mutations probably escaped attention since almost without exception these are recessive and can not be detected until the third generation. Visible alterations on chromosomes of irradiated cells starting with the work in 1905 of Koeneke on lilies, were recorded. Mohr in 1919, reported irradiation of locusts often caused a chromosome to enter the wrong cell (non-disjunction). It has remained for the work of Muller and associates especially on Dro- sophila to found a technique and an entire radio- genetics. Briefly outlined some of these facts are as follows: (1) There are two kinds of alterations in the structure of hereditary material: (a) intrachro- mosomal or change in linear arrangement of genes within the chromonema which forms the basis of the chromosome, by simple loss, simple translocation, mutual translocation, inversion, de- letion and complex combinations of these; (b) a change in the composition of the genes them- selves. Changes of both kinds can be produced in abundance in Drosophila, 150 or more times as abundant as in controls. Over half of sperm re- maining capable of taking part in fertilization and development are genetically affected. Mutated genes and changed chromosomes are inherited in accordance with usual Mendelian and chromoso- mal properties. (2) Visible mutations extend to bodily form and size, colorations, conformation of individual organs, fertility, reaction to light and gravity and heat tolerance. Morphologic changes are often obscure and secondary to physiological changes. At least 80% are lethal and most of the rest detri- mental. Changes are distinctly random hence a very few may happen to be beneficial. The chro- mosome changes usually are more detrimental than gene mutations. (3) The mutations and changes are produced by X-rays in germ cells of all types, spermatozoa whether irradiated in the testis or after reception by the female, spermatogonia, mature eggs, oocytes and oogonia. If an embryo is irradiated the adult may show a patch of mutated tissue, in the midst of normal tissue, derived from a single embryonic cell. A great variety of plants and animals show results confirming those on Droso- phila, including wasps and other insects, barley, tobacco, jimson weed, maize, wheat, cotton, prim- roses, snapdragons, petunias, mice, frogs, etc. 170 THE COLLECTING NET [ Vor. IX. No. 79 Snell, last year, demonstrated that a quarter of functional sperm of mice after irradiation with only 400 roentgens, carries chromosomal translo- cations detected through inheritable infertility in the first and later generations; and that over 507% of embryos die in utero when one parent comes from an affected line; and morphological changes such as misshaped spleens and dwarfism. The implications of such results in terms of human beings are greatly stressed by Muller. (4) Chemical and molecular — structural changes in reproductive cells still escape us and await the joint efforts of chemists, physicists and biologists. Ill. The Effects of X-Rays on Normal Cells. Fricke in a paper in these symposia has con- sidered the physical basis of radiobiology. The very first step in a long series of events attendant upon irradiation of cells is a purely physical one —the collision of radiation quanta with electrons in the atoms and molecules of the chemical sub- stance of a cell, and the release of these photo- electrons, leaving ionized atoms, or the raising of electrons to higher energy levels, leaving an acti- vated atom or molecule. The ionized or activated atoms have properties no longer to be associated with the original unirradiated atom. The proto- plasm, the protein molecules, are of necessity changed and usually broken down to other sub- stances ordinarily simpler. These degradation products or molecular fragments must be as for- eign to the cell in its normal structure and func- tioning as though we had introduced materials from the outside by some micro-inoculation. Un- der these circumstances the morphology and phys- iology of the cell must change in such a way that at least the gross features can be recognized by the histologist and the physiologist. Let us try to enumerate these identifiable steps in the series of changes, depending in part upon the clear sum- mary of Packard in the book “The Science of Radiology”. (1) All changes follow a latent period after irradiation which may be very short if our meth- ods of detection are sufficiently sensitive for min- ute structural, chemical and biological changes, but may be very long if we judge only by death or a gross phenomenon such as skin erythema. (2) Translucent protoplasm first becomes tur- bid and then granular indicating that the state of aggregation of protein molecules completely changes, because of disturbance in distribution of electric charges making for coagulation. The same effect may be produced in vitro, by chemical effects, and unfavorable media. Obviously since irradiated cells are known to return to normal unless subjected to unbearable dosages, the pro- cess can be reversible. One theory due to Des- sauer held by a number of radiologists was coagu- lation by minute points of radiation heat gener- ated in the tissue from the conversion of kinetic energy of the photoelectrons. (3) Many workers have demonstrated the shrinking and clumping of chromosomes, moving irregularly to daughter cells, some lagging behind, and some entering the wrong cell. In more ad- vanced stages the chromatin falls to pieces and the pieces swell up and fill the cell body. After very heavy X-ray dosage, the protoplasm is vac- uolized ‘and mitochondria fragmented; and of course death follows. (4) Often the marked swelling of irradiated nuclei and the ballooning of the cytoplasm are easily observed and must be interpreted as an in- creased capacity of these structures to absorb water through an altered cell membrane. While this system is too complex for complete under- standing and explanation it is logical to believe that intracellular changes produce new electro- lytes by decomposition of salts, proteins and fats and that water is drawn in by simple osmosis. (5) The viscosity of proplasm is increased following irradiation. (6) The hydrogen ion concentration is in- creased for the protoplasm. This effect may last for only a very short time in the case of the skin to hours and days, for example in lymphoid tis- sue. The importance of this in therapy will be- come readily apparent when it is considered that in cancer the blood becomes decidedly more al- kaline. (7) Cell respiration is decreased probably be- cause oxidation enzymes are injured. Aerobic and anaerobic glycolysis is increased. (8) A gross effect in tissues is the actual closing of blood vessels and a disturbance of the vascular supply. (9) Curtis, in work on such invertebrates as coelenterates, planarians and annelids, and Butler with the vertebrate amblystoma, have shown that the normal ability of these species to regenerate lost parts is completely destroyed when subjected to X-radiation. Hence X-rays afford a method of studying experimentally process in regeneration as compared with differ- entiation in normal embryonic development. (10) .The rate of cell division is changed. It is a universal finding that cells which are in mito- sis at the time of exposure to X-rays complete the division even with inordinately high dosages up nearly to 100,000 roentgens, while other cells are prevented from undergoing mitosis. They lose motility and then may literally explode. Af- ter awhile they may regain apparent normal be- havior, but occasionally after a few further divi- sions may die suddenly. (11) We have thus enumerated the best known morphological, chemical and physiological changes following irradiation. The question naturally arises as to whether these are interde- pendent and parallel, or merely different aspects the differentiation — et Aveust 18, 1934 } THE COLLECTING NET 171 equally important. In consideration of all facts, particularly from the chemical point of view, we would agree that there are primary and secondary biological effects. The primary effects are: a. Increase in hydrogen ion concentration of protoplasm which may account for coagulation. b. Increase in cell membrane permeability. c. Increase in viscosity, a consequence of hy- drogen ion concentration change. d. Change in respiratory rates. The actual changes in form observed for irradiated cells are a consequence of these effects. IV. The Radiosensitiveness of Cells. We come now to a subject which in the opinion of the writer is the most important and _ signifi- cant in the entire field of radio biology. It is the basis of one of the most important applications of radiation in biology, of identification of cells, of medical diagnosis and of X-ray therapy. Prom- inent among a large group of experimenters who have demonstrated characteristic radiosensitive- ness of cells is Dr. A. U. Desjardins of the Mayo Clinic. The following classification is based upon his work. In 1904 Bergonie and Tribondeau an- nounced the principle that young or immature cells are more radiosensitive than old or adult cells. This was the beginning of a vast number of observations ranging from effects of irradiat- ing a pregnant uterus, to single cells. To inter- pret the principle to the end that sensitiveness varies directly with the reproductive capacity of the cell and inversely with the degree of differen- tiation, is perhaps not so satisfactory except in a general sense, since there are notable exceptions as Packard has shown. Indubitable as is the relation of the age of cells to radiosensitiveness, analysis of the experiments made to test the susceptibility of different organs and tissues brings out the even more important fact that each variety of cell in the body has a specific sensitiveness, or rather a specific range of sensitiveness, to radiation. This is not intended to imply that all cells of one kind, such as lymph- ocytes or squamous epithelial cells, react in pre- cisely the same way to a given dose of rays. A certain measure of variation in reaction must necessarily occur, because different cells of the same kind are struck by the rays while in differ- ent stages of metabolism. Other still unknown factors also may play a part. However, if allow- ance be made for such variation, and if reaction time be taken as a criterion, the specific sensitive- ness of each kind of cell looms up as the domi- nant single fact of radiology and: deserves to be recognized as a law. And yet, if we may judge by present-day writings, the existence of such a law and of its medical and biological implications is not at all realized. For years much has been made of the dogma that pathologic cells are more radiosensitive than normal cells of the same kind, is wholly invulnerable to radiation ; but, as Lazarus-Barlow and others have shown, the foundation on which this dogma rests is tenu- ous and insecure. The physiologic condition of cells undoubtedly has some influence on their sen- sitiveness, but, as I shall bring out presently, such influence is small as compared with the specific natural susceptibility of each variety of cell. Al- though the factors responsible for such specificity have not yet been determined, the sensitiveness peculiar to each kind of cell appears to be related chiefly to the natural life cycle. Thus the lymph- ocytes, whose metabolic cycle among human cells is the shortest, are also the most radiosensitive, and the nerve cells, whose life cycle is the longest, are also the most resistant to irradiation. But to this question as to many others the final answer has not been given. When a living tissue or organ is exposed to roentgen rays or radium, a more or less important proportion of cells may subsequently exhibit tem- porary inhibition of metabolic activity or complete and permanent disintegration, or may not show any deleterious effect. Moreover, if the time in- tervening between irradiation and perceptible re- action is taken as a criterion, it will be found that certain species of cells react more rapidly than others to a given dose, or the degree of reaction to the same dose is greater for some kinds of cells than for others. According to our present knowl- edge cells may be classified, according to their radiosensitiveness, in the following order: Lymphoid cells (lymphocytes). Polymorphonuclear and eosinophile leukocytes. Epithelial cells : (1) basal epithelium of certain secretory glands, especially the salivary glands; (2) basal epithelium (spermatogonial cells) of the testis and follicular epithelium of the ovary; (3) basal epithelium of the skin, mucous membranes, and of certain organs, such as the stomach and small intestine; (4) alveolar epithelium of the lungs and epi- thelium of the bile ducts (liver), and (5) epithelium of tubules of the kidneys. Endothelial cells of blood vessells, peritoneum. Connective tissue cells. Muscle cells. Bone cells. Nerve cells. pleura and Although the difference in susceptibility be- tween the most sensitive and the least sensitive varieties of cells is considerable, none of the cells all cells, what- ever their variety, may be destroyed or injured if exposed to a sufficiently large dose of rays, especially if doses within the therapeutic range are disregarded. 172 THE COLLECTING NET { Vor. IX. No. 78 Closely connected with sensitiveness is the power of recovery, obviously a phenomenon of enormous importance in therapy. The protozoan paramoecium can recover after X-ray dosages as high as 60,000 roentgens. Numerous experiments have demonstrated that best recovery is associated with low reproductive and metabolic rate of the cells. Hence susceptibility and recuperative powers are inversely proportional. V. Observation of X-Ray, Effects on Human Tissues Action on the skin. According to the dose applied, it is customary to recognize four degrees of skin reaction. First degree: no visible inflammation but epila- tion followed by tanning; lasts from two to four weeks and is followed by complete recovery. Second degree: moderate erythema with defi- nite vascular dilation (hyperemia) and a sensa- tion of increased temperature in the treated area ; epilation and pigmentation; duration from six to twelve weeks with eventual recovery and disap- pearance of the tanning. Third degree: erythema of reddish-blue color with vesiculation; epilation; loss of papillae and of sweat and sebaceous glands; pain; duration from eight to fifteen weeks; heals with epilated thin scar and often develops telangiectasis in an atrophic scar; danger of late reaction years after the injury. Fourth degree: deep reddish-blue erythma with vesiculation and necrosis of cutis, developing into an ulcer; extremely painful; prognosis as to com- plete recovery doubtful. In most cases, wide ex- cision is the only remedy. It is obvious, of course, that between these four degrees, there are all types of variation possible. Numerous histological studies of the irradiated skin have been undertaken; they lead to the con- clusion that the acute roentgen reaction of the skin is essentially a degeneration of the epithelium (germinative layer), combined with inflammatory processes (interstitial edema and leukocyte infil- tration). There are also changes in the capilla- ries, namely, dilatation and later thickening of the walls. This picture is most characteristic in the base of a typical roentgen ulcer. Entirely differ- ent from this acute reaction are the findings in the skin of radiologists following the cumulative effect of numerous small doses of roentgen rays. Marked blood vessel changes are absent. There is hyperemia, changes in the blood distribution, and hypertrophy of the epithelium (hyperkerato- sis). Hair papillae, sweat and sebaceous glands have almost disappeared. Usually, there is ede- ma in the corium accompanied by atrophy: of the elastic elements. Microscopically, an acute ulcer can present the same changes and that, undoubt- edly, explains the tendency of both to terminate in malignant degeneration (roentgen carcinoma ) . MUCOUS MEMBRANES The mucous membranes, for instance, in the mouth and trachea, show essentially the same re- action to roentgen rays as the skin, with two ex- ceptions. The latent time is shorter and so is the regenerative period. Mucosa covered with cylin- dric epithelium is much less radiosensitive than that with pavement epithelium. This can be dem- onstrated by irradiating the oesophagus and tra- chea with an identical dose of roentgen rays. While no reaction can be observed in the trachea, the upper layer of the mucosa in the oesophagus shows marked signs of injury. THE BLOOD Great practical importance must be accorded to the changes of the blood and the blood forming organs following irradiation. Roentgen rays of short wave length in moderate and _ toleration doses cause a leukopenia which is often preceded by a brief period of leukocytosis. Depending greatly upon the resistance of the individual, it takes from two to six months before the blood has completely recovered. The polynuclear leuko- cytes seem to return to normal more slowly and a relative lymphocytosis is, therefore, common, As a rule, the erythrocytes remain intact; it ap- pears, however, that the red blood corpuscles of normal people are more resistant than those of cancer patients. BONE MARROW In the bone marrow, the histological changes are diagnostic within two to three hours after the exposure. There is pyknosis of the nuclei of all white blood cells beginning with the lymphocytes, while the polynuclear cells are most resistant. If the dose was high enough, only the mature red blood corpuscles remain intact within the capil- laries of the marrow. Regeneration is possible even after intensive exposures. The first changes in the blood of radiologists consists of a leuko- penia with relative lymphocytosis. SPLEEN AND LYMPH GLANDS The spleen and lymph glands display identical changes, namely, an enormous pyknosis of the nuclei in the germ centers, and soon afterwards, throughout the lymphatic tissue. Phagocytes take up the chromatin pieces and there are very few cells left in from twelve to fourteen hours after irradiation. Within twenty-four hours, regenera- tion usually begins and complete recovery takes place, provided no lethal dose has been applied. SEX GLANDS The sex glands, i. e., testicle and ovary, are very sensitive to radiation. A relatively small dose destroys the function of the ovary while about twice that dose produces an aspermatogene- sis. In the testicle, a slow degeneration of the germinative epithelium takes place while the in- ee ee ee ee Ce ee ———E———) ) a -eD A meeting of the Corporation and a meeting of the Board of Directors were held on July 21. Professor Harold C. Urey was elected a member of the Board. He is the first chemist to have a place on it. Dr. and Mrs. Stuart Mudd and their three chil- dren have been at the Laboratory since the first of the month. Their guests have included Dr. Robert Nugent, of the University of Arizona; Dr. and Mrs. Kenneth Appel, of Philadelphia; and Miss Anne Taylor, of Greenwich. Recent visitors further include Dr. Melvin Mooney, physicist in the laboratories of the U. S. Rubber Company; Dr. J. W. Gowen, of Rocke- feller Institute at Princeton; Dr. Sewell Wright, of the University of Chicago; Dr. Charles R. Stockard, Professor of Anatomy at Cornell Med- ical School; Dr. L. G. Longsworth, of Rocke- feller Institute; Dr. and Mrs. Ralph W. G. Wyckoff, of Rockefeller Institute; Dr. P. S. Henshaw, of Memorial Hospital, New York City; Dr. Frank Blair Hanson, Assistant Direc- tor of the Division of Natural Sciences, Rocke- feller Foundation, Mrs. Hanson and their fam- ilv; Professor John S. Nicholas, of Yale; and Mrs. Wanda K. Farr, of the U. S. Department of Agriculture and Boyce Thompson Institute. Professor and Mrs. Robert Gaunt and Mr. and Mrs. William Parkins visited Dr. and Mrs. Swingle at Lake Winnesquam, New Hampshire, . Aveust 18, 1934 ] THE COLLECTING NET 181 THE GENETICS SOCIETY OF AMERICA PRESIDENT, Sewall Wright, University of Chi- cago, Vicr-PRESIDENT, Donald F. Jones, Connecticut Agricultural Experiment Station. SECRETARY-TREASURER, P. W. Whiting, Carne- gie Institution of Washington, Cold Spring Harbor. PROGRAM OF THE SUMMER MEETING MARINE BIOLOGICAL LABORATORY, Woods Hole, Mass. AUGUST 21-23, 1934. Tuesday Evening, August 21, 8:00 P. M., Auditorium Regular weekly meeting of Marine Biological Laboratory for presentation of scientific papers. 1. The Temperature-effective Periods and “Growth Curves’’ of the Vestigial Wings of D. melanogaster. (12 min.): Morris H. Harnly, Washington Square College. 2. Selective Fertilization in Habrobracon. (12 min.) : P. W. Whiting, Carnegie Institution or Washington, Cold Spring Harbor. 3. Maturation, Fertilization and Cleavage of Habrobracon Eggs as Revealed by the Feulgen Reaction. (12 min.): B. R. Speicher, Carnegie Institution of Washington, Cold Spring Harbor. 4. Mutations’ and Modifications of Droso- phila Induced by Sub-lethal High Temperature. (12 min.) : Harold H. Plough, Amherst College. Wednesday Morning Session, August 22, 10:00 a. m.: Auditorium. 1. Impaternate Daughters of Females Heter- ozygous for a Sex-linked Gene in Habrobracon. (5 min.): P. W. Whiting, Carnegie Institution of Washington, Cold Spring Harbor. 2. Localization of the Sex Gene in the X- chromosome of Trimerotropis rebellis. (5 min.) : E. Eleanor Carothers, University of Pennsyl- vania. 3. Sex Expression in Certain Dioecious Plants. (15 min.) : Walter F.-Loehwing, Univer- sity of Iowa. 4. A Difference between Reciprocal Hybrids in Mice. (15 min.): A. Marshak, Harvard Uni- versity. Announcements and Short Business Meeting, 11:00 a. m. 5. Gametophytic Genes in a High-waxy Strain of Maize. (15 min.): Wm. H. Eyster, Bucknell University. 6. Location of Genes Responsible for Pollen Tube Abnormalities. (15 min.) : J. T. Buchholz and A. F. Blakeslee, Carnegie Institution of Washington, Cold Spring Harbor. 7. The Effect of the Gene Vestigial on Facet Number in Bar-eyed Flies of Drosophila. (15 min.) : O. S. Margolis, Washington Square Col- lege. Wednesday Afternoon Session, August 22, 2:00 p. m., Room 130, Brick Building. Demonstrations. 8. (2 p. m.) Mottled Eyes in D. melanogas- ter. Elizabeth H. Gay, Carnegie Institution of Washington, Johns Hopkins University. 9. (2:30 p. m.) Variation of Spindle Fibre Loci within the Individual. E. Eleanor Caro- thers, University of Pennsylvania. 10. (2:30 p. m.) Localization of the Sex Gene in the X-chromosome of Trimerotropis re- bellis. E. Eleanor Carothers, University of Penn- sylvania. g 11. (3:00 p. m.) Maturation and Early De- velopment of Fertilized and Unfertilized Eggs of a Grasshopper, Melanoplus differentialis. Elean- or H. Slifer, University of Iowa. 12. (3:30 p.m.) Maturation, Fertilization and Cleavage of Habrobracon Eggs as Revealed by the Feulgen Reaction. B. R. Speicher, Car- negie Institution of Washington, Cold Spring Harbor. 13. (4:00 p. m.) Octoploidy and Diploidy in Miastor. Alfred F. Heuttner, Washington Square College. Wednesday Evening, August 22, 6:00 p. m.: Clam Bake Thursday Morning Session, August 23, 9:00 a. m.: Auditorium 14. The Effect of X-rays on the Chromo- somes of Tradescantia gigantea. (15 min.) : Her- bert Parkes Riley, Harvard University. 15. Heat-induced Mutations in Drosophila and Their Bearing on Problems of Evolution. (15 min.) : Victor Jollos, University of Wiscon- sin. 16. Inheritance of a Number of Mutations Resulting from Aging Seed. (15 min.): J. L. Cartledge and A. F. Blakeslee, Carnegie Insti- tution of Washington, Cold Spring Harbor. 17. Male Crossing-over in Drosophila Fol- lowing Exposure to Heat. (5 min.): Harold H. Plough, Amherst College. 18. The effect of Temperature on the Com- plex of Genes for Vestigial Wings in D. melan- ogaster. (15 min.): Morris H. Harnly, Wash- ington Square College. 192 THE COLLECTING NET 19. Effects of Temperature on Isogenic sc 1, sec 5 and sc 1/sc 5 Stocks of D. melanogaster. (15 min.) : George P. Child, Newark, N. J. 20. Does Evidence from Phenogenetics Sup- port the Idea of a ‘Master Reaction’? (15 min.) : Louis Powsner, Washington Square College. 21. A New Aspect in Human Heredity. min.) : Felix Bernstein, Columbia University. 22. Coil-springs and Chromosomes. (15 min.) : Harry H. Laughlin, Eugenics Record Of- fice, Cold Spring Harbor. (20 23. The Biological Meaning of Race. min.) : Mare A. Graubard, Columbia Univers Thursday Afternoon Session, August 2:00 p. m.; Auditorium. Seminar: Genetics and Development. Speaker : Emil Witschi, University of Iowa. § ject: Sex Genes and Embryonic Sex Dif entiation. Discussion led by B. H. Willier, University Rochester. 7 WOODS HOLE CHORAL CLUB CONCERT The eighth annual concert of the Woods Hole Choral Society will be held at 8:30 on next Mon- day evening, August 20, in the auditorium of the Marine Biological Laboratory. The Woods Hole Choral Society was organ- ized in the summer of 1926, by a group of music lovers interested in choral singing, the first con- cert being given the following summer. The chorus ‘is composed of thirty to fifty voices and is open to all members of the community interest- ed in choral singing after conference with the di- rector. Meetings are held twice weekly in the M. B. L. Club throughout the summer season. A number of the original members are still singing with the chorus, although the membership neces- sarily changes somewhat from season to season owing to the fluctuating character of the summer population. During the present season there has been great interest shown—the membership is above the ayerage, the rehearsals have been well attended and much enthusiasm evidenced for the music selected. The choral society was most fortunate from the start in securing the services of Ivan Gorok- hoff as director, without whose skill as a musician and superlative qualities as a trainer and leader, the consistently high standard of the programs would have been impossible. The program for this season’s concert has been chosen with care and presents an_ interesting variety with a marked predominance of songs of a tuneful character. The numbers have been arranged in groups. The first group consists of four chorals of sacred music, the style of each of which is markedly different. It includes a simple and beautiful “Ave Maria” by Arcadelt, a mag- nificent chorale by César Franck, and two Rus- sian numbers remarkable for their rich har- monies. Following this there will be a group of three tuneful Schubert dances and two familiar folk songs by Brahms. Gounod’s “Chorus of Bacchantes” will be followed by two Russian folk songs, both characterized by care-free rhythms. The concluding numbers will be adaptations by Holst of English folk songs, one of which is the popular “Swansea Town” whose dramatic trayal of a storm at sea and its references t adventurous life of sailors make it a fay with Woods Hole singers and audiences. Through the generosity of friends who are terested in promoting choral singing in Hole, it has been possible for the first time t duce the price of tickets to permit a larger n ber of music lovers to attend the concert. price of tickets will be fifty cents for r seats and twenty-five cents general admiss The concert will be given for the benefit of 1 CotLectinG Net Scholarship Fund. Tickets be obtained in advance at the office of THE | LecTING Net and the Marine Biological Lal tory, and will be on sale at the door on evening. THE WOLFINSOHN-WILSON-WAGENAAR T CONCERT FOR THE COLLECTING NE’ SCHOLARSHIP FUND Scholarship Fund will be given by the sohn-Wilson-Wagenaar Trio of New Yor! cians on Wednesday evening, August 23, it Marine Biological Laboratory Auditorium. concert is presented as a special favor te Fund, for the Trio’s regular schedule of grams here has been completed. The program will begin at 8:30 with t zart B-flat Major trio in three movements, 4 gro moderato, Larghetto, and Allegretto, a arranged in the usual four parts, the first last consisting of formal trios, the second third of solos by Miss Nancy Wilson, ‘cellist Mr. Wolfe Wolfinsohn, violinist, of the Miss Wilson’s offering, a ‘cello sonata by at eighteenth century Italian composer, Sammai constitutes the second part. Mr. Wolfinsohi offer three shorter numbers for the third sect The first is a folk-dance, the work of a wo composer of the late eighteenth and early teenth centuries, Anna Maria Paradis, and is titled, “Sicilienne.’’ Following this Mr. We sohn will play Fritz Kreisler’s famous Viennoise” and the De Falla “Spanish D which he has already played at a Community Avevsr 18, 1934 J THE COLLECTING NET 183 Hall concert here. Bernard Wagenaar, pianist of the trio, will accompany both Miss Wilson and Mr. Wolfinsohn. The final number will be the five movements of the Dvorak “Dumky” trio. This number is patterned after the “Dumky,” a type of Russian poetic literature in which several poems, each al- ternately mournful and gay, follow each other in a sort of cycle. The movements of the trio re- flect the individual poems by their titles, Lento- allegro, Andante-vivace, Andante, Allegro, and Lento-vivace. This work too has been presented in Woods Hole already this summer, and was very well received. Mr. Wolfinsohn, violinist of the South African by birth, an Englishman by educa- tion. He studied under Hans Wessely at the Royal Academy of Music in London. At present he is leader of the Stradivarius Quartet of New York, all of whose members play on instruments made by the renowned Antonio Stradivari of Cremona. Mr. Wolfinsohn is not risking his Stradivarius in the damp climate of Woods Hole, but the instrument he will play is the handiwork of one of the Master’s pupils, Balestrae, and dates from 1765. aa, ss El Miss Wilson, ’cellist, is well known in Woods Hole as the daughter of Professor E. B. Wilson of Columbia, and has played here many times in the past few years. She is a pupil of Pablo Casals, and was active in many chamber music organizations before her marriage to Robert Nathan, one of the younger New York novelists. Since her marriage she has concentrated on solo work. Mr. Wagenaar is composer, violinist and pro- fessor as well as pianist. Toscanini led the New York Philharmonic orchestra in his symphony last season, and the trio of which he is a mem- ber played a ’cello sonatine of his here on August 12. Mr. Wagenaar holds a professorship of composition at the Juillard School of Music in New York. The hour has been set fairly late in the eve- ning especially to avoid conflict with the clam- bake for the visiting geneticists. The clambake will be over before the concert begins, and it will be possible to attend both affairs without diffi- culty, Tickets are priced at fifty cents, a dollar, and up, and will be on sale at the office of Tur Cor- LECTING Nev? and at Robinson’s Pharmacy in Falmouth. : THE BEACH QUESTION IN THE LIGHT OF A CHANGING ENVIRONMENT. II. H. McE. The Internal Environment or Laboratory proper which we sketched in the last issue as the special field of the biological workers is, of course, fairly definitely organized for a relatively limited scope of problems, the needs of the workers not being too far apart, however diverse their interests. On the other hand, improvements in the Labora- tory and in living conditions attracted a steadily increasing number of workers, and soon families and friends and other outsiders also began to come and spend more time. Thus were created many new and varied influences intimately affecting the work from without in what we have called the Ex- ternal Environment, in which the requirements, quite different from the simple surroundings of the first days, were of much more extreme range in ac- cord with the great diversity of individuals. We may roughly group the problems of this Ex- ternal Environment under three heads: first, condi- tions of living for the individual and for the family; second, various methods of relaxation, diversions, sport; and third, the social problems of contact, in- terchange of ideas, and enjoyments of general com- munity nature. In the first years little thought was given to liv- ing conditions, a room and a bed in the old stone building or in some native residence sufficing for the few hours spent outside of the Laboratory. When, however, wives and small families began to come, all sorts of adjustments were required to care for these and the limited facilities and endur- ance of the town were put to severe tests. It must be said of the residents that they have maintained a kindly, permissive attitude through- IKNOWER out, refraining from interference and even encour- aging a steady expansion of the numbers of sum- mer visitors, with many encroachments on what might have been considered their peculiar privi- leges. The writer has learned respect and admira- tion for some sterling characters who have grown strong and independent among the natives of Woods Hole, I might say in spite of the scientists and their ideas, or other deterrents to their type of individualism. It is true, however, that their early suspicions of the moral and social standards of the invading biologists were satisfactorily proved to be mistaken, and a lasting friendship grew up, with also, be it noted, a pleasing material profit always on the side of the home town. As with many tribes, home-making fell largely to the wives, with the increase of population in Woods Hole. The reactions of both the native wives and those of the biological immigrants to the crowded conditions gradually resulted in more local com- forts, necessities indeed for the new arrivals from the cities until bath tubs, good running water, good ventilation, and aH sorts of conveniences to ease the job of the house-keeper became common, The native men continued their fishing and daily rou- tine; it was generally their wives who met more freely the needs of the biological housewives whose husbands were too much absorbed at the Labora- tory to be of much use at home. Oh, there was ex- ceptions among the males, of course! But I want to take this occasion to pay a marked tribute of admiration and affection to the wives of the biolo- gists who have unselfishly done so much here to help make possible and easier their husbands’ work THE COLLECTING NET [ Vor. 1X. No, 79 184 and careers adopted by themselves. They impro- vised suites in old houses; turned the ‘Corner House” once on the site of the present apartment building into livable quarters, and later did the same for the Dexter House; they even transformed barns into dwellings. All this for husbands and children, and for these again, they brought about an adequate flow of supplies, eggs, milk, fresh veg- etables, and fruits, up to then extremely scarce for the summer people, yet so vital to healthy living. The whole town profits from this today. Finally, one after another took over the considerable cares and social duties of hostess in the new summer homes which were acquired from time to time by more settled members of the crowd. All of this inevitably led to their further partici- pation with the biologists themselves in the organi- zation and enjoyment of other factors of this Ex- ternal Environment which contributed so much to the well-being of the working biologists in provid- ing relaxation, social contacts, healthy rest, and physical repair and upbuilding. Many of these women were carried off into ma- trimony when already headed for a career in biolo- gy, and some indeed managed to keep up this work along with their obligations to the cominunity; but whether they managed a full return to the techni- cal working out of professional plans or not (as at least one nobly did after she had arrived at the Grandmother stage) all must be granted our last- ing gratitude for many vital and constructive things brought in to make our External Mnviron- ment better, and for constantly fighting against in- fluences which might disturb the quiet and effec- tiveness of the laboratory work. Tnis brings us to the first period of beach usage. In 1892 we swam mostly off the Fisheries wharf, though the Big Beach was regularly used and an occasional trip to Nobska was not unusual. Bath- ing was at that time an athletic exercise, swimming as contrasted with the modern beach sun-bath. It had a favored place with sailing, bicycling and tramping. Later on tennis came in and a golf course fol- lowed. At first there was only the Big Beach, but by 1910 a few houses were occupied by biologists, pioneers at the periphery of the town, upon Crow Hill which then seemed tar into the country. Swim- mers who used the Little Beach and the hospitable Lillie’s pier nearby were quite apart. With this segregation there were fewer people on the Big Beach and no crowding even when all the swimmers came there. Subscriptions provided a tennis court as well as rafts off the beaches. One would like to dwell upon this period of grad- ual growth and expansion, with healthy laboratory activity; pleasant, but not too responsible social contacts; informality; old clothes; plenty of space along the shore and in the woods, freely open to us by the Fays, hospitable owners. There are many interesting aspects to this situation, where every- one showing an interest in our aims and work was welcome. The very attractiveness and possibilities of the situation with other influences were, how- ever, bringing about great changes which culmin- ated in 1916 with the opening of the new brick Laboratory building, which we owe to the generos- ity of Mr. Charles R. Crane, just another debt to Mr. Crane to be added to previous years of sympa- thetic moral and material support. The donation, which made it possible for the first time to plan ahead deliberately for the Laboratory, was made in 1913 and at the same time opportunities were ar- ranged for a corresponding development of the Ex- ternal Environment, so that the Laboratory work- ers might be assured of suitable conditions for liv- ing. Mr. Crane presented a building for the M. B. L. Club and arrangements were made with the Fay Estate for the sale of beach lots bordering the Big Beach, not too far from the center of town. Then completing full provision for numbers expected, twenty acres of wood lots on Crow Hill were pre- sented to the Laboratory to be sold to biologists. All this assemblage of marvelous opportunities led quickly to active centrifugal movement to es- cape from the then so-called ‘‘crowded center.” Homes sprang up along the beach, and on the Hill. The segregation around the Little Beach, Lillie’s pier and elsewhere tended still more to leave the Big Beach freer. More buildings were opened for dormitories and Mr. Crane gave the Dexter House for apartments, which carried a group of swimmers to Nobska Beach. But this was war time, which meant a Naval re- - serve station near the Mess Center and a good many new additions to the Beach crowd. Automo- biles began to appear in numbers more and more during the period, bringing many outsiders (rank outsiders!) to the town and on to the beach. With the building of the row of bathhouses the beach problem was now set; though the biologists who had built along the shore long continued to wel- come bathers off their beaches. By 1925 the scientific value of the Laboratory had become so universally recognized as to bring further donations from the large financial founda- tions resulting in the addition of the western Ell of the brick building, with library expansion and con- siderable development of other features of the plant. In addition the new dormitories and apartment houses were opened and very considerable exten- sions of the Mess Hall and kitchens were installed, all of which turned the tide from the centrifugal migration which was then going on to a centripetal movement. By this time the arrival of more autamobiles bringing many casual visitors, and the follow-up of the naval reserve by the coast guard station, were further causes contributing to a real crowding on the Big Beach, another contributing factor being the shutting off of much shore line and other beaches by persons outside the biological commun- ity who bought up the property. The new sun- bathing habit on the shore, out of the water, in- creased the crowding on the beach. The owners of the northern beach lots, now sep- arated by the fence, continued to permit the in- creasing crowd to use their beach front, but it must be confessed that observers who realize the return made for this privilege can only sympathize with the owners and agree in their reaction, expressed by the fence. I regret that requirements of space will make it necessary to postpone a proper analysis and con- clusions of our sketch of Environmental history in its bearing on the “beach question.” If I have taken considerable trouble in this paper it is because I think the editor’s honest impulses in championing what he considers abuses of the pub- lic should be appreciated even though they are per- haps a bit too heated. I think also that he should be thanked rather than censored for a usually calm and fair forum in THE COLLECTING NET, However, we still believe that a thorough examina- tion of facts will result in sympathy and coopera- tion with the beach lot owners rather than blame for them. We shall hope to find indications for future adjustments too. Avucust 18, 1934 ] THE COLLECTING NET 185 Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, etc. Catalogs will gladly be sent on request. Please mention name of school and | ae Spalteholz subjects taught, to enable us t é r S Transparent eral the apRonEAte catalog. me ety Preparations | x Human =e ea ¢ arena Sa? end Crav-Apams Company ooisescal 25 EAST 26th STREET NEW YORK poe ee een eae Visit our display rooms and museum. Dr. G. Gruebler & Co. (Prop. J. Schmid, Apoth. & A. 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Also | Bell’s Text-Book of Pathology, 2nd edition other models for other fuel i Boyd’s Text-Book of Pathology, 2nd edition gases. 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Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. 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Mun Hedical Nicroscope EATURING the exclusive Spencer constructional advantages: (1) Balanced Optical System, (2) Metal Mounted Achromatic Oil Immersion Objective Lens, (3) Dual-Cone Nosepiece, (4) Fork-Type Substage, (5) New Rhodium Plated Parts. Write for Folder M-61-S for complete description and prices. S | 'S j ans Company R : BUFFALO PNEW YORK | bd > ——4 = ee Vol. IX. No. 9 SATURDAY, AUGUST 25, 1934 Annual Subscription, $2.00 Single Copies, 25 Cents. INTERLABORATORY COOPERATION AND SCIENTIFIC EQUIPMENT Dr. SAMUEL E. Ponp Technical Manager of the Marine Biological Laboratory Exchange of information to avoid unproduc- tive duplication of effort is one thing; a practical medium for conducting such an exchange is an- OBSERVATIONS CONCERNING THE CREMISTRY OF CELL GROWTH AND DIVISION CarL VOEGTLIN The investigation of the chemistry of the growth and division of cells must take account of at least three attributes of living cells: (1) their complex physical organization, (2) their complex other. The articles by Dr. R. Gueblannse ands Drei VWeeel. Cole which appeared in the June 30 and July 28 issues of Tue CoLLecTING NET sug- gest several ways of improv- ing the prosecution of re- search in marine biology through conferences, sympos- ia and coordination. While it is true that knowledge may be advanced by group discussions of problems, there are several aspects of any problem which require a special treatment. The exchange of ideas which may be brought about between a few pairs of interested workers as against the ex- MM. B. EU. Calendar TUESDAY, August 28, 8:00 P. M. Lecture: Dr. C. G. Hartman: “The time of ovulation in the men- strual cycle.” The usual all-day _ scientific meeting for the presentation and discussion of results obtained in Woods Hole during the current season will be held this year on Friday, August 31. Those having papers to present are requested to hand their titles to Mrs. Sepulveda in the Administration Office not later than Monday. Persons desir- ing to have abstracts published in the Biological Bulletin should submit these in a form suitable for publication, with their titles. Except in unusual cases, the length chemical organization, and (3) the time relations between the different morphological and physiological events as- sociated with growth and divi- sion. This frank admission of the difficulties inherent in the investigation of this problem, therefore, will not raise un- reasonable hopes of its early solution. However, I believe that a systematic attack from the chemical standpoint will produce knowledge of great value to biology and to medi- cine in particular. I must confine my remarks largely to some recent obser- vations made at the National change of ideas through a larger group, is not touched words. of abstracts should not exceed 300 Institute of Health with ref- erence to certain oxygen con- upon by the two authors re- ferred to. It may be of interest, therefore, to consider the spread of ideas and in particular an aspect of technical (Continued on page 204) suming animal cells, leaving out of consideration plant cells and anaerobic organisms. For the purpose of this analyti- cal discussion I shall consider cell growth and TABLE OF Observations Concerning the Chemistry of Cell Growth and Division, Dr. Carl Voegtlin 189 Interlaboratory Cooperation and Scientific Equipment, Dr. Samuel E. Pond.......... 189 Origin and Chemical Nature of the Organic Matter in Sea Water and Sea Bottom, Dr. S. A. Waksman and Dr. Cornelia L. Carey 198 Studies on Evolution in the Islands of the South Seas, Dr. Henry E. Crampton (Summary) CONTENTS Techniques for Experimental Work at the Marine Biological Laboratory, Dr. Henry J. Fry Editorial Page J UREVTAISI CG) by od Bal) col) Merona hee ne teen Gece oc0, EAC 203 Report of Meeting of the Genetics Society 205 The Effects of X-Radiation on Cell Struc- ture and Growth, Dr. George L. Clark (continued ) THE FOREPEAK OF THE “ATLANTIS” IN MARCH A photograph taken at sunrise durirg the first week of Spring by Charles E. Renn, microbiologist at the Woods Hole Oceanographic Institution. At the time the research ketch was off Portland making one of its chemical and_ biological surveys of the Gulf of Maine. The ice was over a foot thick in places and Mr. Renn estimates that the section of the vessel shown in the photograph is encased in over thirteen tons of ice. aT Aucust 25, 1934 } £ THE COLLECTING NET 191 NY. State Fish Hatcheries - SSS Gh Cold Carnegie Institution - Blackford Hall dormitory ICAL Spring Laboratory buildings * Main Building » Residences tt aan a! ta Harbor THE CHEMISTRY OF CELL GROWTH AND DIVISION (Continued from page 189) cell division more or less separately, and apart from cell differentiation and organization of cells into tissues. Cell growth and_ division are of course interdependent biological func- tions in the sense that both are needed for proliferation. To avoid confusion in the use of these terms I shall base the definitions on the us- ual sequence of physiological events. Cell growth involves assimilation of food, synthesis of proto- plasm and consequently increase in size. Cell divi- sion is defined as the actual process of fission of the nucleus and cytoplasm. From the technical standpoint it is essential for the study of these problems to select the most suitable biological material and to control the physical, chemical and biological conditions as far as possible. Ideal conditions for chemical studies could be furnished by free living cells, growing in a medium of exactly known composition, where each cell could be kept under continual observa- tion as regards increase in size (volume) and the time and conditions under which cell division oc- curs. An approach to such an ideal experiment has been made recently by Mast and Pace (1933), who obtained rich cultures of Chilomonas paramecium under apparently sterile conditions on a medium composed of inorganic salts. In the case of other protozoa it is not yet feasible to study growth on a medium of exact composition, nor is this possible with cultures of metazoan cells. Working with tissue cultures Dr. Earle at the National Institute of Health has at least succeeded in defining cultural conditions more rigidly by using normal blood serum, which has a more or less constant compo- sition, and adjusting the gas phase to definite ten- sions of Oy and COs and thereby the pH of the liquid phase. It is thus possible to study the action of added chemicals on cell proliferation, using this term as including both growth and division. As regards the chemical requirements for the growth of the organism as a whole we are better informed in the case of the higher animals than in the case of the lower organisms. This is prin- cipally due to a lack of suitable microanalytical methods. Protein synthesis: Now as far as cell growth is concerned, I shall first deal with the synthesis of intracellular protein with reference to the function of proteinase or cathepsin. This intra- cellular enzyme occurs in the tissues of the higher animals and causes the partial hydrolysis of gen- uine proteins. For a long time it was known that the hydrolytic action of the corresponding enzyme in plants (papain type) is favored by the pres- ence of such substances as H2S or cyanide. Fol- lowing the discovery of glutathione it was found (Grassmann and Waldschmidt-Leitz and their co- workers) that reduced glutathione also “‘ac- tivates” cathepsin and papain whereas the discelphide form of glutathione is inactive. Soon afterwards Voegtlin, Maver and Johnson showed that SH groups attached to protein had the same “activating” effect. They also found that preliminary oxidation of the SH groups in tissue extracts by H2Os, traces of copper salts or aera- tion, completely inhibits the proteolytic action. Since Hopkins and Elliott (1931), experimenting with mammalian liver tissue, had shown that a reversible oxidation of glutathione results from changes in Og tension, according to the equation 2R.SH+0=]R.S—S.R+ H:20, it occurred to us that the action of cathepsin upon tissue pro- teins may be a function of the Oz tension. Ex- periments with extracts of normal and malignant tissues, buffered to a pH range slightly on the acid side of neutrality, showed that under anaero- bic conditions (air replaced by purified Noe) proteolysis was accelerated as compared with the same extracts exposed to atmospheric air. Sim- ultaneous estimations of organic SH compounds indicated a greater concentration in the anaerobic as compared with the aerobic digests. The next step was to ascertain whether oxy- genation of an anaerobic tissue digest would fur- nish evidence of resynthesis of protein. For this purpose various tissue extracts, again adjusted to a pH near neutrality, were digested under anaero- bic conditions for a few hours and the degree of proteolysis measured. These digests were then submitted to oxygenation by purified air or oxy- gen for a few hours, samples being taken for 192 THE COLLECTING NET [ Vor. IX. No. 80 analysis at regular intervals. The results obtained clearly indicated resynthesis of protein during the oxygenation period, simultaneously with a decrease in the concentration of SH compounds. Moreover, it was shown that the process could be reversed once more by subjecting the oxygenated digests to a second anaerobic period of digestion. Similar results were obtained with digests of heat-coagulated eggwhite and papain. Shortly afterwards Rondoni (1933) reported similar re- sults. I have called attention to the fact that all these experiments were carried out in the presence of buffers, which held the pH constant near neutral- ity, in other words, near or not far removed from the pH of tissues and blood. This was done in or- der to simulate physiological conditions as much as possible and proved to be one of the principal reasons for our success in demonstrating the re- versibility of the action of proteinase, as will be shown presently. A review of the many papers by other workers dealing with the action of cathepsin will reveal the fact that practically all such studies were made at a pH between 4 and 5, simply for the reason that the lytic activity of cathepsin in this strongly acid range is most pro- nounced. Therefore, it was of importance to study the influence of variation in pH on the synthetic process. The results justified the conclusion that, all other conditions being the same, a decrease in pH from neutrality to about 6.5 to 6, or an in- crease into the definitely alkaline range prevents resynthesis of protein by oxygenation. Inhibition of protein synthesis is also observed if small amounts of copper salts are added to the digests before oxygenation is begun. In all these experiments the methods used for the demonstration of the synthesis of protein were those commonly used for studying the lytic action of proteinase. Nevertheless, it is necessary to furnish further proof of actual synthesis than that already offered. For instance, it might be ,ob- jected that the so-called resynthesized protein had its origin in a non-enzymatic oxidation of SH containing protein cleavage products, which re- acting with oxygen would be oxidized to disul- phides of correspondingly larger molecular size. No evidence to this effect was obtained in suit- able experiments carried out in the absence of proteinase. An effort was also made to secure ev- idence as to whether or not the resynthesized pro- tein has the same chemical composition as the original undigested protein. The quantitative esti- mation of the various amino acids in the acid hy- drolysates of the original and resynthesized pro- tein should serve this purpose. However, cystine is the only amino acid that can be estimated ac- curately by means of the highly specific Sullivan method, whereas the methods for the other amino acids are lacking in specificity or accuracy. The Sullivan method was therefore chosen for the estimation of cystine in (1) the original protein, (2) the protein residue remaining after anaerobic digestion, and (3) the protein obtained following oxygenation, Our results indicate that the cystine content of the resynthesized protein is of about the same order as that of the undigested protein. Of course I am fully aware that all this ev- idence is not strict chemical proof of protein syn- thesis. However, it is well to point out that there is some physiological evidence which is in har- mony with the view that the oxygen tension is a physiological factor regulating the cleavage and synthesis of tissue proteins. Thus it is known that prolonged exposure of animals to a considerably decreased Oz tension inhibits the growth rate of normal and cancerous tissues. Also, Mothes inde- pendently has obtained very interesting results on plant tissues, showing a similar controlling influ- ence of the oxygen tension on protein metabol- ism. Before leaving this subject I should like to make a few remarks concerning the biochemical mechanism. The experimental facts suggest that. the action of cathepsin depends upon the presence in the enzyme of a reversibly oxidizable group. This hypothesis would explain the controlling in- fluence of the oxygen tension in the presence of suitable oxidation catalysts. For instance, the unique role of traces of copper salts in the oxida- tion by molecular oxygen of reduced glutathione, cysteine, and the SH group in coagulated crystal- line egg albumin has been carefully studied in our laboratory. Reduced glutathione is oxidized to the corresponding disulfide; cysteine under the same conditions suffers a molecular breakdown with the formation of some sulfuric acid, COs. and ammonia ; protein sulphydryl is oxidized with com- plete disappearance of the nitroprusside test. It is also peculiar that the intracellular proteinases are the only proteolytic enzymes whose action has been shown to be influenced by oxidative changes. They are also easily inactivated by very low concentration of monoiodoacetic acid, a com- pound which is known to react with various SH compounds. Pepsin, on the contrary, is not in- fluenced by changes in Os tension, SH com- pounds or iodoacetic acid. This evidence suggests that variations in Oz tension influence the hypo- thetical reversably oxdizable enzyme group either _ by direct interaction or indirectly by shifting the apparent oxidation-reduction system of tissues in which the organic sulphur compounds play a prominent role. The inhibiting action of decreasing the pH upon the synthetic action of cathepsin calls for some comment. Since the work of Pasteur it is well known that diminution in Os tension in- creases cellular glycolysis with the formation of acidic products and hence a tendency to decrease in the pH of the biological system. The recent work of Bumm and Appel (1932) indicates that > ~aeieaicadied eae Avcusr 25, 1934 } THE COLLECTING NET 193 reduced glutathione, but not the corresponding disulphide, increases the aerobic glycolysis of mammalian tissues 7 vitro, whereas it has no ef- fect on the rate of Oz consumption, as had been previously shown by Voegtlin, Rosenthal and Johnson. Since it can be assumed that the chem- ical activity of Oz in tissues is proportional to its tension, it would follow that lowering the Oy» ten- sion would tend to keep the tissue SH system in the reduced state and thus increase glycolysis. And if the rate of glycolysis is sufficiently in- creased the tissue buffer capacity would no longer prevent a shift to a lower pH. Consequently there would be a tendency to proteolysis. This comment serves the purpose of calling attention to the pos- sibility of the coupling of various biochemical re- actions, in the case under consideration: (1) ox1- dation-reduction, (2) proteolysis-protein synthe- sis and (3) glycolysis. So far I have dealt with one requirement for cell growth, namely the synthesis of cellular pro- tein. The equally important synthesis of the spec- ific nucleoproteins and of nucleic acid has not been studied. However, the enzymatic cleavage of nucleic acid into its components has received some attention in recent years. In this connection I also refer to a paper by Hammersten, who carried out an important physico-chemical study of nucleic acid and its reactions with histones and amino acids. Studies on Amoeba: We may now pass to some observations made on a living cell, Amoeba pro- teus. Dr. Chalkley will discuss its physiology and the advantages in selecting this organism for chemical studies on growth and division. | shall confine myself to the discussion of the results of eur joint work from the biochemical aspect, first the action of glutathione. We had shown long ago that glutathione is concerned in the pharmacolog- ical action of certain organic arsenicals, the toxic action of such heavy metals as gold and copper, and the toxic action of cyanide. All these sub stances react 1m vitro with glutathione and similar sulphur compounds either by forming condensa- tion products, as for instance: R.As:O+2R.SH= R.As(SR)2+H20O, or by affecting the state of oxidation of glutathione: 2RSH=k.S—S.R. We have found that the toxic action of these sub- stances can be prevented if the ofganism is sup- plied with an extra amount of glutathione. Therefore, we assumed that these poisons interact chemically with the organic sulphydryl system of the organism. This pharmacological evidence indi- cates furthermore that the organic sulphur sys- tem of the organism is essential for the mainten- ance of life. We had also found that the reduced glutathione content of rats decreases during em- bryonic life and after birth. Similar observations were made simultaneously on chick embryos by Murray. We therefore suggested a possible rela- tion between growth rate and glutathione content, realizing, of course, that direct proof was neces- sary to establish such a relationship. The observa- tions of Baker, working with tissue cultures, and particularly the studies of Hammett on plant roots and Paramecium then showed that gluta- thione and certain other SH compounds apparent- ly increases cell proliferation. At that time Dr. Hammett regarded the SH group as the natural chemical stimulus for mitosis. I emphasize that our observations on amoebae were made on isolated cells, the response of each cell to the chemical under consideration being as- certained. This procedure we regard as essential for exact studies. In order to eliminate individual variation it is necessary to work on a sufficient number of cells to make results statistically sig- nificant. It is obvious that growth and division studies must be preceded by a careful toxicolog- ical study in order to select subtoxic concentra- tions of chemicals. From preliminary experiments it seemed likely that the action of glutathione upon cells of dif- ferent ages might differ. In other words, small cells which have just gone through division, and larger cells which have gone through division earlier and have grown to larger size, might show a different response to glutathione. This assump- tion was verified by numerous experiments, which clearly showed that the per cent of nuclear and cytoplasmic division in glutathione solution as compared with the saline control was greater with larger than with smaller cells. Since we have found that the per cent of division of the control cells is also a function of cell size, it follows that cell division in d4moeba depends on cell size, irre- spective of whether the cells are immersed in con- trol or glutathione solution. Therefore, “the much greater chance of a larger cell to exhibit signs of cell division, as compared with a small cell, must indicate that the chemical state (to use a word with broad meaning) of the small cell and partic- ularly of its nucleus differs from that of a larger celi.” I shall return later to this fact that a certain cell size is required to demonstrate the stimulat- ing action of glutathione on cell division. Further evidence showed that nuclear division may occur without subsequent cytoplasmic division, indicat- ing the relative independence of the two pro- cesses. The apparent stimulating action upon cell division is exerted by glutathione in the SH or disulphide form. It can readily be shown that un- der aerobic conditions SH glutathione is rapidly oxidized to the disulphide in the suspension me- dium. Therefore, it would seem that the observed physiological action of glutathione depends not so much, if at all, on its modifying action of the cell environment, but rather upon the interaction of absorbed glutathione with protoplasm. Since these observations seemed to suggest that the observed stimulating action of glutathione is related in some way to an influence, either direct 194 THE COLLECTING NET [ Vor. IX. No. 80 or indirect, upon the cell nucleus, a more detailed analysis was undertaken of the glutathione action upon Amoeba under survival conditions. In these experiments the total cell volume and nuclear vol- ume of each cell were measured daily for a pe- riod of three days. In view of the fact, previously mentioned, that copper salts are extremely effec- tive catalysts for the oxidation of glutathione, and the well-known oligodynamic (growth inhib- iting) action of copper, similar experiments were also made with highly diluted solutions of copper salts. The results may be summed up as follows: In the control cells withdrawal of external food supply results first in inhibition of cell growth, then in nuclear growth. Addition of glutathione to the suspension medium enhances the growth of the nucleus apparently at the expense of the cytoplasm. Treatment with CuCl, inhibits all growth and particularly that of the nucleus, in- hibits nuclear division and probably decreases the rate at which food reserves from food vacu- oles are assimilated. Glutathione has a contrary effect on growth and food assimilation and in- creases the percentage of division in a given group of cells, particularly in those of medium size. All these effects are also functions of orig- inal cell size. Chalkley has found that under ordinary cul- tural conditions nuclear division normally occurs when a calculated nuclear volume of about .00002 cubic millimeters is reached. Now, since gluta- thione stimulates and copper inhibits nuclear growth, it might be inferred that the apparent stimulation and inhibition of nuclear division by glutathione and copper respectively are primarily due to the influences these chemicals exert upon nuclear growth. As the nuclear volumes of divid- ed cells at the exact time of division were not ob- served, these volumes were approximated by tak- ing for each cell the mean between the nuclear volume at the observation preceding division and the sum of the nuclear volumes of the daughter cells at the first observation after division. The averages of these means were for 27 cells that divided in control solution .0000205; for 25 in glutathione .000018; and for two cells in copper solution .000018. We conclude, therefore, that two intrinsic characteristics of Amoeba control nuclear division, namely, (1) the rate of nuclear growth and (2) the apparently definite average optimum nuclear size at which nuclear division is most probable. These factors seem to control the action of both glutathione and copper and we may say that glutathione stimulates nuclear division by accelerating nuclear growth, apparently by facil- itating transfer of cytoplasmic material to the nucleus. So far there is no clear evidence that glutathione can cause nuclear division in Amoeba of a smaller nuclear size than the optimum nu- clear size at which controls divide. In other words, our experimental evidence does not indi- cate that glutathione can specifically elicit mitosis, i.€., it is not a mitotic stimulus in the strict sense. I am rather inclined to attribute to glutathione and copper regulatory functions of metabolic re- actions which are primarily concerned with cell growth. In this connection | again refer to the evidence relative to the action of proteinase. Probably enzymes concerned in the metabolism of the nucleus are also involved. Work along this line is under way. The last topic I wish to discuss is the action of chemicals on the process of cell division per se and therefore separate from growth. This work was made possible through a careful morpholog- ical and physiological study of Amoeba proteus by Drs. Chalkley and Daniel, the results of which Dr. Chalkley will report himself. Suffice it to say, that it is possible to select with ease from a stock culture cells which will go through the division process within the next half hour. Using such dividing cells Daniel and Chalkley established the fact, already known for some other cells, that cell division is a function of temperature. The tem- perature coefficient suggests an underlying chem- ical process, though | am reluctant to consider such evidence as conclusive proof. Certain observations on glycolysis in malignant tissues at our Institute made it desirable to study the effect of acids, particularly lactic and pyruvic acids upon dividing amoebae. Chalkley and Dan- iel recently completed this work. They find that a considerable increase in the hydrogen ion concen- tration of the suspension medium produced by HCl does not influence the division process; whereas lactic acid and pyruvic acid solutions of the same pH, while unable to prevent nuclear division, prevent cytoplasmic division. The latter effect can be modified or abolished by the addition of CaCl, to the medium in the case of lactic acid, and by the addition of CaCl. or NaCl in the case of pyruvic acid. The whole evidence is brought into relation with the observations by Heilbrunn and Daugherty (1932) on viscosity changes produced by physiological cations and the well-known observations of Jacobs on the permeability of cells to organic acids. Furthermore, these experiments indicate that it is comparatively easy to hibit cytoplasmic division in Amoeba by means which probably af- fect the physico-chemical condition of the cell membrane and cytoplasm. Recently Dr. Chalkley and I have studied the action of certain chemicals on nuclear division in dividing Amoeba. This is obviously a more diffi- cult subject. First of all, inhibition or accelera- tion of nuclear division could only be expect- ed from exposure of cells to chemicals which from pharmacological experience penetrate with great rapidity and yet which do not manifest a tox- ic action in concentrations sufficiently high to in- fluence the division of the nucleus. In view of the 4 4 e Aveusrt 25, 1934 i] THE COLLECTING NET 195 physiological importance of cell respiration we first determined whether exposure of dividing amoebae to subtoxic concentrations of the class- ical respiratory poisons had any influence upon nuclear division. The action of H.S or HCN, CO and trivalent arsenic was studied. The first three of these substances are known to exert their biological action very rapidly. Briefly it was found that HeS and HCN when added in certain concentrations to the suspension medium greatly delayed nuclear division of cells in early prophase. The inhibition was less marked or ab- sent in the later stages of mitosis. Cells whose nuclei had failed to divide during exposure to H2N or HCN, completed mitosis when trans- ferred to saline solution, indicating that the in- hibition is reversible and therefore is not com- plicated by an irreversible toxic action upon the cells. Particular attention is called to the fact that HeS, a sulphydryl compound, is shown to inhibit mitosis. So far no evi- dence has been secured which would indicate that this chemical can exert a stimulating effect in any concentration. The inhibiting action of H2S and HCN is in striking contrast with the inactiv- ity of CO (absence of light) and trivalent arsen- icals. There is no reason to assume that CO does not rapidly penetrate cells. However, lack of rapid penetration perhaps may account for the negative results with the two forms of otherwise pharmacologically potent arsenicals. Ethyl ether delays the mitotic process and has a tendency to produce vacuolization of the nucleus. Methylene blue, H2Oxz, iodoacetamide and cupric salts ex- hibit no influence on mitosis in subtoxic concen- trations. These results are of interest for several rea- sons. First, because they demonstrate the feasibil- ity of studying under controlled conditions the action of chemicals upon the mitotic process as a whole and, if desired, on its several phases. Sec- ond, because the action of certain substances on mitosis may throw light upon the nature of the underlying chemical processes. Take for instance the failure of high concentrations of CO to in- fluence mitosis. According to the well-known work of Warburg and his collaborators, the ac- tion of CO upon cells is due to its chemical affin- ity for the hemin-containing respiratory enzyme, 1.e., the oxygenase, which apparently activates molecular oxygen preparatory to its metabolic utilization. This enzyme has been found in all or- ganisms examined and presumably also occurs in Amoeba. The negative results with CO, there- fore, suggest that mitosis in this organism is not concerned with an oxidative reaction controlled by Warburg’s respiratory enzyme and, further- more, that mitosis can proceed normally in the absence of oxygen, unless the small residual traces suffices. The same reasoning applies to cy- tochrome which, according to Keilin, is also inac- tivated by CO. Incidentally it is of interest, that Bodine and Boell quite recently have shown a curious difference in the action of CO on embry- onic cells of Melanoplus differentialis. They find that the oxygen consumption of growing and dividing cells is diminished by CO. On the other hand, cells in the physiologically resting stage (blocked), and showing no growth or division and a very low Oz consumption, respond tu CO with increased Oy consumption. This curious dif- ference in response to CO indicates that the met- abolic state of the two types of cells must be dif- ferent. The inability of H2O., methylene blue, copper salts and iodoacetamide to influence mitosis is in- teresting, because one might expect that these substances, by acting as oxidizing agents, would modify certain phases of cellular oxidation-reduc- tion. Ether, as other anaesthetics, decreases Os consumption and inhibits mitosis. Perhaps the most significant result is the inhibition observed with H2S and HCN. Warburg believes that the biological action of these substances depends, as in the case of CO, on their chemical affinity for the iron-containing respiratory enzyme. His proof for this assertion is far less convincing than that offered for the CO action. In fact, it is largely based on the well-known affinity of H»S and HCN for heavy metal compounds. Warburg ig- nores the possibility that both H»S and HCN are reducing agents, which, as can be easily shown, reduce physiological sulphur compounds. The ob- served inhibition of mitosis therefore may be due (1) to a reducing action upon the organic sulphur system, (2) to an action upon catalytic heavy metals, as copper or iron, or (3) to a com- bination of these actions. As regards the action of COs, Spek and Cham- bers have shown recently the ease of penetra- tion of this substance into Amoeba, so as to cause a decrease in cytoplasmic pH. The inhibition of mitosis observed with COs stands in contrast to the ineffectiveness of HCl, lactic and pyruvic acids, and may be related to alterations in the physico-chemical conditions of the nucleoproteins of chromatin (see Hammersten). The principal results and conclusions herewith reported can be summed up as follows: Growth of cells requires protein synthesis. It is shown that one important step in this synthesis is governed by the intracellular proteinase, whose action—lytic and synthetic—is controlled by the oxygen tension. The function of oxygen in this process can best be explained by the assumption that the enzyme contains a reversibly oxidizable active group, which in its reduced state favors lysis, and in its oxidized state synthesis of pro- tein. The state of oxidation of this active group may be determined by the apparent oxidation-re- duction potential of the system, in the regula- tion of which oxygen, catalytic heavy metals and 196 cat THE COLLECTING NET organic sulphur compounds appear to play an im- portant role. Analysis of the action of glutathione upon Amoeba proteus indicates that this cell compon- ent, under the experimental conditions used, ac- celerates nuclear growth and thus promotes mi- tosis. No evidence was obtained which would in- dicate that glutathione is a mitotic stimulus in the strict sense. Minute amounts of copper salts in- hibit nuclear growth. Within certain limits the ef- fect of copper can be antagonized by glutathione. Work on dividing amoebae indicates that an increase in the external hydrogen ion concentra- tion by HCI does not influence cell division ; lac- tic and pyruvic acids inhibit cytoplasmic division ; CO» has a tendency to inhibit mitosis. Inhibition of mitosis results from exposure to HS, HCN and ether. The inhibition observed with H»S and HCN is reversible. The implications of these re- sults are discussed with regard to the chemical re- actions underlying mitosis. There is a consider- able independence of nuclear and cytoplasmic fis- sion in Amoeba, depending on chemical changes of the environment. DiIscuUSSION Dr. Gudernatsch: Did I understand that no cell division occurs even under glutathione stim- ulation unless both cell and nucleus have reached the proper size? We all have been trained under the dogma of the nucleus-plasma relation, This is a very important point because, under gluta- thione stimulation, nuclear division occurs with- out ensuing cell division, as Dr. Chalkley has pointed out. Nuclear division alone may occur, as we know, in some cells; particularly under pathological conditions (e.g. tuberculosis) giant cells are formed—sometimes with as many as forty or fifty nuclei. Dr. Voegtlin: In amoebae exposed to gluta- thione, nuclear fission occurs when the nucleus of control cells divides. Nuclear division may or may not be followed by cytoplasmic division, in- dicating a certain degree of independence of the two processes. Dr. Gudernatsch: Glutathione contains gly- cine, glutamic acid and cystine. Have you tried the three acids or any two of them in mixture? Dr. Voegtlin: As one control solution we used alanine (in place of cystine) plus glycine and glutamic acid. We have never tried glycine, glu- tamic acid or cystine separately. Dr. Gudernatsch: I asked this question about mixtures of acids because, as I mention in my paper, we have used various two and three acid mixtures. Which of the three acids do you con- sider the important one in the molecule? Dr. Voegtlin: Cystine or cysteine. ently glycine is inactive. Dr. Gudernatsch: Have you tried cysteine? Appar- [ Vor. TX. No. 80 Dr. Woegtlin: No, we have never worked with cysteine for the reason that cysteine is not satisfactory for working in the physiological pH range. Cystine is rather insoluble. Dr. Gudernatsch: Cysteine is much more sol- uble than cystine, but if you use cystine, how much of it remains in the aqueous medium? Are you satisfied that adequate proof of tem- perature effect has been submitted ? Dr, Voegtlin: J place very great importance on the temperature factor in studies on mitosis. However, I question whether the temperature coefficient of mitosis alone is sufficient proof for the assumption that mitosis is a chemical process. Dr. Harris: Do you consider the size of the cell to be significant, merely as indicating the size of the nucleus ? Dr. Chalkley: Under given conditions for a certain cell size, there is a certain nuclear size. It tends to be only very slightly variable. In other words, I wouldn't say it is a fixed thing, but characteristically, without a doubt, you have as- sociated with a volume of cytoplasm a certain ~ volume of nuclei. That holds not only for the mono-nucleate, but for the poly-nucleate cells. If you have a cell with three nuclei in it, it will be approximately three times the volume of a cell with one nucleus. Dr. Harris: Do you consider that cell divi- sion or nuclear division is dependent wholly upon the size of the nucleus? } Dr. Chalkley: In this sense, the size of the nucleus is merely an index of what Dr. Voegtlin referred to as a general chemical condition for! the nucleus. I think you will find that under any normal growth conditions the nucleus must grow’ to a certain size before the cell divides. It is an! important index of the physiological state of the nucleus. You might assume it to be an index of the quantity of chromatin, or more likely, an in- dicator of the osmotically active material within the membrane or an index of the amount of water in the nuclear protoplasm. We have some evi- dence that hydration of the protoplasm is one of the important factors influencing cell division. There is an intake of water into the cytoplasm. The nucleus swells remarkably during the pro- phase. Dr. Harris: Do you consider the action of glutathione to be chiefly concerned with the in- take of water in the nucleus? Dr. Chalkley: J couldn’t say, but it is cer- tainly concerned with the transference of some material essential to the nucleus, from the cyto- plasm to the nucleus. That is fairly reasonable, but is purely hypothetical. To prove this, it would be necessary to identify a given material in the cell without injuring the cell. It is more or less speculative. We haven’t the equipment to do it. Until we have this we can’t prove it, — _ ES rere”’”"—=“‘é’’=~- Aucust 25, 1934 } THE COLLECTING NET 197 Dr. Voegtlin: It is always difficult to explain mechanisms. We spent years trying to find out something about the biochemical mechanism un- derlying the arsenic action. We made progress but there are many points that have to be eluci- dated further. Dr. Schram: Do you conclude, from your ob- servations, that Dr. Hammett’s postulates of the stimulating effect of the sulfhydryl group on mi- tosis is subject to revision? Dr. Voegtlin: This is a question of definition of terms. Glutathione accelerates nuclear growth in Amoeba and, therefore, can be considered as a stimulus of nuclear growth. We haven't any evidence so far to prove that glutathione in any way accelerates mitosis direct- ly, apart from its growth accelerating action upon the nucleus. Dr. Hammett has not worked with isolated cells. He has not differentiated between growth and division of cells, except in his experiments with plant roots. His generalization doesn’t hold for Amoeba. Dr. Riddle: 1 should like to ask whether you have made observations on the relative amount of glutathione in tumor cells as compared with normal cells. Dr. Voegtlin: Yes, we have done what we could do. The methods for the quantitative es- timation of glutathione are only reasonably ac- curate, because all the methods, even the latest modifications, are not specific enough for gluta- thione alone, because in all these methods, oxidizing agents are used. It happens that vitamin C and other reducing agents in tissues also react with these oxidizing agents. The his- tory of blood sugar estimation shows what a tre- mendous amount of work had to be done to make the technique sufficiently specific. Malignant tumors have, roughly speaking, the same gluta- thione content as liver, or perhaps, liver has even more than the tumor, and most of that gluta- thione is in the reduced form. The benign tum- ors which we studied had an exceedingly small glutathione content, much smaller than that of normal tissues. I don’t agree with Waldschmidt- Leitz. He proposed the idea that tumors grow so well in the body because of a shift in the equi- librium between the reduced and oxidized form of glutathione in malignant tissue. There is no shift in this ratio in the malignant tissues as com- pared with that of normal tissues. Miss Hoffman: Wouldn't the amount of glu- tathione in reduced form, in contrast to the amount of the oxidized one, run parallel with the rate of oxidation in the cell? Dr. Voegtlin: Yes. It seems to run parallel, as Hopking and Elliott showed with liver tissue. We have done similar experiments with red blood cells where we estimated the oxidized glutathione by means of a special method and found that there is a shift in the ratio of the two forms of glutathione according to the oxygenation of the blood. Miss Hoffman: On the basis of the amount of oxidized glutathione and the limit of the error by which you could determine that glutathione, could you estimate the amount of proteolysis? Dr. Voegtlin: That is what we did. Dr. Abramson: Dr. Voegtlin has shown that the oxygen tension influences the quantity of amino acid nitrogen produced in a protein digest and that this action is reversible. He has, him- self, pointed out his attempts to show the resyn- thesis of protein. It would be of further inter- est to know if: (1) Experiments are available on crystaliz- able proteins with known molecular weights. (2) There is any evidence beside the nitro- gen and the cystine analyses, that a strictly re- versible reaction was occurring. Dr. Voegtlin: (1) So far, no experiments have been made with crystalline proteins of known molecular weight. We considered using crystalline ovalbumin in place of coagulated egg- white, as the former might be used for molecular weight determinations of the resynthesized pro- tein by Svedberg’s method. However, this meth- od requires great experience. (2) Protein was precipitated with trichlor- acetic acid under carefully standardized condi- tions. Amino acid nitrogen determinations showed a marked decrease during the oxygena- tion of the digests, at a time when protein syn- thesis was proceeding at the highest rate. Dr. Gudernatsch: Do you happen to know if amoebae can live without all three amino acids? Dr. Voegtlin: Not for a very long time. Dr. Rahn: The amoeba has to feed on bac- teria which probably contain a great variety of amino acids, Dr. Chalkley: You can’t tell just what is es- sential. You can’t study the amino-acid nutri- tion of amoebae unless you can devise a synthetic medium. 198 THE COLLECTING NET [ Vor. IX. No. 80 ORIGIN AND CHEMICAL NATURE OF THE ORGANIC MATTER IN SEA WATER AND SEA BOTTOM Dr. SELMAN A. WAKSMAN AND Dr. CorNELIA L, CAREY Professor of Soil Microbiology, Rutgers University, and Assistant Professor of Botany, Barnard College Organic matter is present in the sea in three distinctly different forms, namely, (1) as liv- ing plants and animals, ranging from the micros- copic diatoms and protozoa to the giant kelps and whales; (2) as organic matter in the sea bot- tom; (3) as organic matter in solution. From the economic point of view the first form is no doubt the most essential. Since it is only transitory in nature, however, and of comparatively short duration in the sea, it need not be considered here in detail. Sooner or later it is partly consumed by higher forms of animal life, partly transformed into the simpler (biolog- ically speaking) organic complexes, and _ partly changed into its constituent elements which again become available for plant life. The organic matter in the sea bottom varies considerably in concentration depending on the nature of the bottom material, on the distance from shore, on the time of deposition, ete. Quantitatively, it was found to range from 0.5%, for sand and calcareous bottoms, to as high as 10% for certain mud bottoms. The most im- portant and significant characteristic of this or- ganic matter is its carbon-nitrogen ratio (C:N), which ranges from 8:1 to 12:1, with an average of 10:1. A study of its chemical nature revealed the fact that it consists largely of three definite complexes: (1) lignin-like substances, originat- ing from the algal residues: (2) proteins, large- ly synthesized by bacterial action in the sea as well as by the animal population of the sea; (3) polyuronides, originating from the plant and bac- terial residues in the sea. This type of organic matter may be designated as marine humus, be- cause of its similarity to the humus in land soils. The third form of organic matter, which is present in sea water, in true solution or col- loidal solution, may be designated as water hu- mus. Putter, in his famous hypothesis on the nu- trition of animals in the sea, suggested that the organic mater present in true solution in sea wat- er forms the most essential source of nutrients for marine animals, as compared with the or- ganic matter in suspension. This hypothesis aroused considerable interest and met with much opposition. Krogh, for example, denied the abil- ity of animals to utilize the organic matter pres- ent in the sea in true solution. He went as far as to claim that even hacteria are unable to attack this organic material. In a careful study of the chemical composition of the organic material dissolved in sea water, Krogh established the fact that 1 cubic meter of water contains 0.244 grams of nitrogen and 2.36 grams of carbon. Assuming that the carbon content of the organic matter dissolved in the water is 50%, 1 liter of water is thus found to contain 5 mg. of organic matter. These results pointed to certain definite conclusions: namely, the uniformity of distribu- tion of the organic matter in sea water; its spec- ific chemical nature; and its resistance to decom- position, even to the action of bacteria. The last conception will have to be modified, as a result of the experimental evidence obtained in our in- vestigations. The changes in the transformation of the or- ganic matter in the water can be measured by the following methods: (1) the rate of change ot bacterial numbers in the sea; (2) carbon crans- formation as determined by carbon dioxide evol- ution and oxygen absorption; (3) nitrogen trans- formation, as determined by ammonia evolution. The first method, namely the change in bacterial numbers in the water, may be considered as the most sensitive index of the amount and nature of the available energy material in the water. By eliminating the action of the autotrophic bacteria, one finds that the development of the marine bacteria is limited by the supply of available or- ganic matter. Bacterial multiplication can be con- sidered as a direct index of the amount of avail- able organic matter in the sea. In considering the transformation of the or- ganic matter in sea water, one must distinguish between the organic matter present in suspen- sion, such as the various types of plankton, and the organic matter present in solution. The latter has been reported to be 7 to 65 times as abun- dant as the former. The origin of the organic matter in solution can be traced to several dis- tinct sources: (a) the solution of some of the algal and diatom constituents in the water: (b) formation of substances following bacterial de- composition of the plankton, as well as of the higher plants and animals in the sea; (c) organ- ic matter removed by streams from land. It can easily be shown that the second source is by far the most important. The first, if it is of any sig- nificance at all, can become so only at a period of maximum plankton activity; the third source may become of importance in regions close to land. The fact that dissolved organic matter is uniformly distributed in water even at great depths and at considerable distance from land in- 4 7 ss Aucust 25, 1934 ] THE COLLECTING NET 199 dicates that these two sources may be only of temporary and local significance. Definite evidence can be submitted to prove the theory that the bacteria are largely responsible for the formation of the organic matter dissolved in the sea water. Furthermore, it can be shown that this organic matter is not absolutely resis- tant to bacterial attack, but, under certain condi- tions of temperature and oxygen supply, it can undergo decomposition, with the result that some of the constituent elements become mineralized and are again made available for plant life in the sea. All plant and animal residues are decom- posed in the sea by bacteria; some of the con- stituent substances are rapidly attacked, while others are more resistant to decomposition. These go partly into solution and are partly pre- cipitated, and gradually settle to the sea bottom. The resulting sediment has all the characteristic properties of marine humus. The different forms of organic matter, namely, that which remains in solution and that which is precipitated out of solution, are very similar in chemical nature, and are somewhat resistant alike to bacterial decom- STUDIES ON EVOLUTION IN THE In his lecture on evolution in the South Seas Dr. Henry E. Crampton expressed his inability to cover all the details of the work, which has been in progress for twenty-eight years, but stated that he would present his remarks in four parts: first, the problem of the experiments; sec- ond, the area which was included in their scope; third, the material; and fourth, some of the re- sults. The lecture was illustrated by slides of scenes of the islands where his work was done, and pictures and charts of the snails which formed the material for his work. The problem to be considered is one of biologi- cal evolution, the study of definite groups of or- ganisms in order to understand the phenomena of their evolution, something of the dynamics, and to see if one may, in the open, find results approximating those obtained by scientists work- ing in the laboratory. The area covered lies mostly in the Pacific Oceania, which includes the so-called South Sea Islands (though many of them are north of the equator). Twelve trips altogether were made to these islands, and four other trips, for the pur- pose of comparison to the West Indies and South America. The Pacific Ocean, the field of the study, covers over one-fourth of the globe, a fact which people do not fully realize until they have sailed upon it. Of the thousands of islands distributed throughout Oceania, ten different groups have been personally investigated, of which the Society position. Evidence is presented to prove these facts, by the use of the methods outlined above. On comparing the rate of decomposition of the organic matter in sea water or in the sea bottom with that of organic complexes of known chem- ical composition, using the rate of bacterial mul- tiplication and the course of oxygen consumption as indices of bacterial activities, one can calcu- late the proportion of organic matter which is readily subject to bacterial action. A theory is put forth that the organic matter dissolved in water is in a state of equilibrium be- tween the algal, diatom and animal population of the sea from which it is formed, on the one hand, and the bacterial activities, as a result of which it undergoes further decomposition, on the other hand. This organic matter is further in equilibrium with the bases in the water which tend to precipitate it, and with the organic mate- rial or humus in the sea bottom upon which it settles, gradually becoming an integral part of it. (This article is based on a seminar report pre- sented at the Marine Biological Laboratory on August 7.) ISLANDS OF THE SOUTH SEAS* Islands, with Tahiti as the principal member, are the most important for the present study. The Carolines,the Hawaiian Islands, Samoan group, Fiji Islands and the Mariannes are among the others. They lie within an area extending 4500 miles in length, and 2500 miles in width. The cluster of the Carolines alone has 3000 members. The islands, according to commonly accepted opinion, are the remaining high peaks of a sub- merged land mass. The character of the terrain would seem to bear this out, in prefernce to the alternative theory, which is that they are volcanic masses thrown up from submerged marine vents. Of all the islands, Moorea yielded most satis- factory results, and was headquarters for the in- vestigation. This island is fairly high, with plenty of shade, moisture and vegetation. It is surrounded by low coral atolls, flush with the water, which are very dry, and do not provide any material. Equally relevant to the investigation with the topographical characters of the islands are the ethnographic characters of its inhabitants; these must be taken into consideration as illustrating racial and geographical correlation, and as pos- sible carriers in migrations of the snails. The natives of neighboring islands are most nearly similar in features, habits, etc. Others, farther away show the effects of cumulative changes. *Summary of evening lecture presented at the Marine Biological Laboratory on August 10, pub- lished with the approval of Dr. Crampton. 200 THE COLLECTING NEG The natives of the islands are Polynesian, Me- lonesian, and Micronesian. Of these the Maori, a Polynesian people, are a very excellent example of natural selection, as their migration to New Zealand carried them to a harsher climate and land than that where they came from, and only the more hardy could maintain themselves. The materials used in the investigation are cer- tain land snails of the genus Partula. These snails require moist situations, not that they need the actual moisture themselves, but they subsist upon decomposing vegetation which grows there. Collections were made systematically in each island, valley by valley, and all available speci- mens were taken on those islands where the snails were found. More than 150,000 have been secured. Naturally, some situations could be eliminated from the start from their dry or other- wise unsuitable character. The actual collecting is only a part of the work, as the creatures must be brought into the laboratory, classified, grouped and the results tabulated. An enormous amount of work is involved here. [ Vor. IX. No. 80 Among the numerous results cited were in- stances of biological evolution which can be said to have been actually observed. From 1861-1888 Dr. Garrett collected these snails and made fairly accurate observations on his findings. When the present investigations were first started in 1906- 1909, it was anticipated that results would merely bear out those obtained by Garrett, and in more detail. Re-collecting and tabulating, distinct dif- ferences were found. Some species were found to have changed from predominantly dextral to predominantly sinistral in their shell coil. The sizes of shells ranged from long slender individuals to little stout ones. Colors range from dark through medium, banded, unbanded, to a light orange-pinkish. _ Results of the investigations check with those obtained in the laboratory and the Mendelian theo- ries. Variations are congenital in causation and environmental conditions have nothing to do with their origin. TECHNIQUES FOR EXPERIMENTAL WORK AT THE MARINE BIOLOGICAL LABORATORY Dr. Henry J. Fry In the first number of The Collecting Net this summer we made mention of our ! plans for the publication of a series of articles which afterwards would be assembled to form a manual of methods for investigators working in marine biology. The vol- ume is being designed especially for Woods Hole workers, but it will undoubtedly be useful to biologists at other marine stations. It is a privilege to be able to announce that Dr. Henry J. Fry has consented to edit the volume in question and to print the preliminary plans in the present number. It would be of value in various ways if the available facts concerning the handling of eggs and other materials used at the Marine Biologi- cal Laboratory were assembled in a form con- venient for reference. This suggestion, which has been made by various investigators from time to time, is now practicable because THE CotLEcTING Net will do the secretarial work and publish the results. The purpose of this preliminary note is to pre- sent a tentative scheme for organizing the infor- mation concerning the handling of eggs. The topics included should cover all major points of significance to a diverse group of investigators. Those who have suggestions for additional topics are asked to call them to the attention of the writer. A committee will carry out the project. The workers most familiar with each type of material will be asked to supply what information they have with reference to each topic. As the results are compiled they will be published in Tuer Cot- LECTING Nev; a handbook will eventually result. Considerable information of this type is al- ready at hand—e.g., “Methods for Experimental Embryology with Special Reference to Marine Invertebrates,” by E. E. Just (THE COLLECTING Net, 1929); “Physical and Chemical Constants of the Egg of the Sea Urchin, Arbacia punctula- ta,” by E. Newton Harvey (Biological Bulletin, Vol. LXII); the instruction sheets used in the embryology course; and other sources. However, only in one or two cases is information available concerning most of the proposed topics; in the majority there are wide gaps in our knowledge. Nevertheless, it is desirable to organize and pub- lish the facts now known, in order that such gaps may eventually be filled in. Although the proposed scheme applies only to eggs, it is planned that techniques applicable to other materials will be included later. These are so varied, however, that they can hardly be brought under a common scheme. GENuS SPECIES (ORDER, PHYLUM) I—SEASON When are the gametes usually in optimum condition? Are there lunar or other varia- tions? Aveusr 25, 1934 ] THE COLLECTING NET 201 II—CoLLeEctTING 1. If the organism is not obtainable through the Supply Department, how is it collected ? 2. Under what conditions should the material be kept until used? I1I—Securine Eccs 1. If the organism is bisexual can the sexes be differentiated externally ? 2. Can sexually mature individuals be differen- tiated from immature ones, either externally or internally ? 3. If the eggs are shed naturally, under what conditions are they obtained ? 4. If eggs are secured artificially, how are the gonads removed and the eggs obtained? How are ovaries differentiated from testes ? 5. If the body fluid and tissue extracts are harm- ful, how are their effects avoided ? 6. Are the eggs harmed by ordinary disturbances such as pouring from bowl to bowl? 7. Should the unfertilized eggs be washed? How often? Or does washing impair their capacity to be fertilized? TV—StTRUCTURE OF THE UNFERTILIZED Eccs . Amount of eggs per female. . Diameter and shape of egg. . Transparency or opacity. . Specific gravity. . Indications of polarity: presence of micropyle ; localization of materials; position of nucleus; yolk distribution ; etc. 6. Viscosity. mMbwWD 7. Membranes and jelly; consistency; thickness ; etc. Modes of removal. 8. Conditions when shed, with reference to ma- turation. Does contact with sea water bring about any development? If so, how far does it proceed, and what is the time schedule of events at a given temperature? V—FERTILIZING THE Eccs 1. When should the eggs be fertilized ? 2. How long can they stand without deteriora- tion and under what conditions ? 3. How is seminal fluid obtained? How long can it be kept without deterioration and under what conditions ? 4. What dilution of the seminal fluid is neces- sary? What amount of the sperm suspension is added to what quantities of eggs in what amounts of sea water? Does polyspermy oc- cur easily ? VI—HANDLING THE DEVELOPING Eccs 1. To what extent can the eggs be crowded with- out impairing development ? 2. How frequently should they be washed ? 3. Are they harmed by ordinary disturbance? 4. Structure of the membrane and jelly in con- trast to that of the unfertilized egg. 5. Viscosity in contrast to that of the unfertilized egg. 6. Time schedule of events at a given tempera- ture: Membrane and jelly phenomena Movements of granules and other materials First and second polar body formation History of sperm aster and approach of pronuclei First cleavage, second cleavage, etc. Blastula formation Gastrulation When do embryos begin to swim? 7. What are the effects of temperature upon the usual time schedule? 8. The percentage of normal development to be expected when eggs are handled properly. 9. What is the “spread” of the time of first cleavage in the egg-set of a single female? 10. To what extent does the time when 50% of the eggs have undergone first cleavage, differ from the egg-set of one female to another? 11. The technique of rearing larvae. VII—Oruer Data 1. Fixatives suitable for the various cell com- ponents 2. Methods of artificial parthenogenesis 3. Special techniques VIII—BreviocGRAPHy (List of previous investigations, with mention of subject of the investigation, if that is not ob- vious from the title.) Dr. G. E. Gates who is in this country on his sabbatical year from Judson College, Rangoon, Burma, recently visited the Marine Biological Laboratory in order to make use of the library facilities. He has now returned to the Museum of Comparative Zoology at Harvard University, where he is continuing his investiga- tions on the systematics of the earthworm. Dr. ANN H. Morcann and Dr. EvizapetH AvAms, professor of zoology at Mt. Holyoke College, visited the laboratory on August 22nd. Dr. and Mrs. Ropert STABLER left Woods Hole on August 20th, and after a vacation in Spencerville, Maryland, will return to the Uni- versity of Pennsylvania. 202 THE COLLECTING NET [ Vor. TX. No. 80 The Collecting Net A weekly publication devoted to the scientific work at marine biological laboratories Edited by Ware Cattell with the assistance of Mary Lawless Goodson, Rachel Warner Parker, Barbara Allee, Anne Janney, Martin Bronfenbrenner and Annaleida Snyder Cattell. Printed by The Darwin Press, New Bedford THE BEACH QUESTION IN THE LIGHT OF A CHANGING ENVIRONMENT (A Continuation of the Editorial Contributed by Dr. Henry McE. Knower) It is fortunate indeed that the success of the work of the laboratory proper appears to have remained to date uninjured, not unduly affected by the great increase of complexities in its Er- ternal Environment. The chief centers of meeting at the Mess, the Club and the Beach have attracted always larger numbers, including many outsiders with interests only related in general to the Laboratory. They all joined freely, however, in the varied activities now presented: attendance at lectures, discus- sions, use of libraries, club activities, the Choral Club and other numerous musical occasions, the developments of the Summer School of Science for children, much social interchange among all groups, and in all sorts of out-of-door diversions and athletics. It was surely the adherence of the biological workers to the principles already outlined of “live and let live,” unwillingness to interfere with the individual freedom of thought and ac- tion of their co-workers that kept the Internal Environment so free of harmful outside influ- ences. On the other hand it has already been pointed out that the biological community has been aided from without by an exceptionally permissive and encouraging attitude from the very first. In this, we repeat, the Woods Hole people have set a notable example of hospitality, on the beach for instance; and the town of Falmouth continues to leave these local problems to the residences here, refusing to come in to regulate until forced to it. Let us hope never!—and that agitators for fur- ther town regulation here will not force the issue against long standing precedent. The natives are only rivalled by the continued hospitality of the owners of extensive tracts along our shore line, in the woodlands and on the neighboring islands. They have always kept their property and even their homes open for the enjoyment of our pub- lic and continue to do so, in evident appreciation of the consideration and avoidance of damage and disturbance of our group. There is not space here to list the many contributions from _ its friends which have favored the work of the lab- oratory and added to the attractiveness and help- fulness of its External Environment, but a few must be emphasized. We not only have to thank the Cranes for the land and plant we work in, but for provision of summer homes nearby. The Fay’s cooperation in joining this move on the beach and on the Hill, as also Miss Fay’s special cordiality in sharing her large estate with the local public has furnished an- other delight for those seeking quiet recupera- tion. I do not know what the Woods Hole na- turalists, workers of intense pace who need such restful refuges would do if these fine woodland retreats should be shut off or sold and broken up into lots. Let us hope some power for lasting good to the town preserves it all intact, as a park for the benefit of future generations. Mention must also be made of the kindness of the rest of the Fays at their Nobska Beach, the Forbes in keeping open their island shores, and the Lillies and Warbasses at their piers and in their homes. All of these items of out-of-door background, of course, apply to the beach question, and I re- gret that I shall have to pass on with mere men- (Continued on page 206) To the Editor: Dear Sir, I have noticed your editorials regarding the Woods Hole Beach and I would like to say that I have felt very resentful at the condition in which I find it, so much so that I have given much thought to it besides many hours of actual labor together with some men I have hired at my own expense to try and improve it. Also, I have dug sand and redeposited six or eight small cartloads, and already I feel somewhat rewarded by noticing many small children playing and making castles in the sand as children love to do everywhere. Today the sea was rough and at high tide I went down to see the effect of my efforts—my sand was still there. My feeling is that if a substantial quantity of sand were deposited very high on the beach, possibly twice during the summer, it would answer all requirements. I doubt if this would cost more than $60 or $70. Then if the owner of the bath house would allow someone to run some solid planks well down into the water so that bath- ers could avoid the stones, Woods Hole would have a fine approach to very good bathing. Perhaps it might be necessary to drive a few piles deep into the sand to make a solid support for a runway; I realize that a rough sea does tear up a beach. I suggest that the Town of Falmouth could place not more tha nfour or five large boulders below the water line. If huge enough, they would not bother the bathers but would break up the force of the sea which would in turn help to keep. the beach in good order. The ideal situation would be to have the bath house back some 20 or 30 feet and then cover the upper part of the beach with fine sand, but perhaps this is asking too much from private owners. Yours very truly, BRYANT BAKER, a CaP nS tgp! oP Og a Wo \< ar rp, ea SS Aveusr 25, 1934 ] THE COLLECTING NET 203 REEMS) (OF Dr. C. R. Moore of the University of Chicago has been made chairman of its department of zoology succeeding Dr. C. M. CuiLp, who will become professor emeritus on October Ist. Dr. Moore will be among those present at the con- ference of the Committee on Sex-Research to be held on August 25th at the Marine Biological Laboratory. Dr. ENRIQUE BELTRAN, professor of zoology at the University of Mexico, was appointed last January to the directorship of the newly created Instituto Bioteenico which is devoted to routine and research work in the field of agriculture, for- estry, animal husbandry and fisheries. Profes- sor Beltran was at the Marine Biological Labor- atory in 1932 on a Guggenheim Fellowship, and worked with Professor Gary N. Calkins. He is now engaged in some investigations of biological conditions and fauna of the Lake of Patzcuaro in the State of Michoacan. A card from BoHumiIL Krajnik, Sc.D., M.D., who worked at Woods Hole during August and September, 1931, on the physiology of the mus- cles of Busycon canaliculatum includes the fol- lowing statement: “I am now Privat Docent of the General Biology and Assistant of the Mas- aryk University, Medical School, Brno, Czecho- slovakia. I am now working in the field of gene- tics, especially on the human constitution and heredity of the pathological abnormities, and eu- genics. I have also interest for the genetics of protozoa and for the hydrobiology.” A fellowship carrying a stipend of three hun- dred and fifty dollars and free tuition for gradu- ate work for the year 1934-1935 is available to a graduate (male) student in biology at the Phil- adelphia College of Pharmacy and Science, Phil- adelphia, Pennsylvania. The applicant must be a graduate of some college or university of rec- ognized standing in biology. It is desired that his primary interest be in physiology. Further information can be obtained from Dr. Amo Viehoever through the office of the Marine Bio- logical Laboratory. The 480-pound sea-turtle which has heen the main attraction for several weeks in the Fisheries ‘basinette,/ was removed last week and is to be sent to the aquarium at Washington, D.C. The turtle came from the West Indies in a shipment of tropical fish, and being too large for the aquaria at New York and Boston, it was sent to the Bureau of Fisheries at Woods Hole where it has spent the main part of the summer. INTERES? Dr. ReGinaLp G. Harris, director of the Bio- logical Laboratory at Cold Spring Harbor, visited Woods Hole on Wednesday and Thursday. He has been spending a few days with friends in Cataumet. Dr. FRANK BLAIR HANSON, assistant director of the science division of the Rockefeller Foun- dation, has attended the meeting of the Genetics Society this week and will attend the conference of the Committee on Sex-Research. Those coming to Woods Hole from a distance to attend the meeting of the Trustees held on August 14th, were Dr. W. B. Scorr of Prince- ton University; Dr. H. C. Bumpus, Brown Uni- versity; Dr. Ross G. Harrison, Yale Univer- sity; Dr. D. H. TENNANT of Bryn Mawr Col- lege; Dr. FRANZ SCHRADER, Columbia Univer- sity; Dr. M. J. GREENMAN of the Wistar Insti- tute; Dr. B. M. DuceGar, University of Wiscon- sin; and Dr. H. B. Goopricu of Wesleyan Uni- versity. The whales which formerly were the basis of the famous whaling industry at New Bedford, are now being threatened by extinction, and measures are being taken for their protection by the passage of the Whaling Industry Regulation Bill. Nature reports that this bill is an attempt to secure international action to cut down the destruction of whales in antarctic seas, for the possibility of their extermination imperils the future of the industry. In recent years the use of ‘floating factories’ which can operate outside of territorial waters has rendered individual gov- ernmental control practically impossible. In 1931 the governments represented in the League of Nations at Geneva bound themselves to a sys- tem of licensing all whaling ships connected with their countries to conditions giving complete or partial protection to certain species of whales. This agreement prohibits the capture of ‘Right’ whales and those below certain sizes, as well as females accompanied by their young. The orchestra which has been playing at the last few M. B. L. club dances has been organized by several of the boys in and around the Marine Biological Laboratory. Keith Noble of Oberlin College who plays the guitar and Matthew Pratt, drummer, are members of the collecting crew. Lloyd Deewest plays the violin and is a student at Tufts Medical College. Robert Keltch, pian- ist, formerly played with Red Nichols of Chicago over station WGN; Bud Barstow of Falmouth plays the saxophone. 204 THE COLLECTING NET [ Vor. IX. No. 80 INTERLABORATORY COOPERATION AND SCIENTIFIC EQUIPMENT (Continued from page 189) equipment in research, and by a different treat- ment than by conference-symposia. For the present we may think of the matter as a dual cooperation. The prosecution of a piece of research which, for example, involves the use of a large group of animals, will at some time or other revolve around the equipment which may most effectively effect the completion of the work. If as often happens an incubator is re- quired one would consult colleagues, catalogs of manufacturers, or the scientific literature. If the latter alone were taken as a measure of advance in (or a guide to) incubators and methods of in- cubation, one could easily draw the conclusion that there has been a rapid development of ideas without really establishing a method of control- ling temperature which is uniformly successful. In other words, there are so many types of ther- moregulators that a biologist, at least, finds him- self entirely confused when he attempts to ascer- tain from the literature what devices he should use. Now, one might ask the question, will a con- ference-symposia on incubators and thermo-regu- lators aid an individual to start out with a satis- factory approach to his problem? For every at- tempt that has been made to discuss matters ot this kind in groups, there have been increased difficulties to establish the point that a real ther- mo-regulator for a biologist is not available. As- suming that the method of consideration is at fault—would it not be better for us to resort to the consideration of such equipment by two or three more closely interested individuals get- ting together when the problem is raised, and by thrashing it out free from the difficulties of a formal discussion or symposia ? In apparatus methods generally, whether con- cerning incubators or x-ray equipment, two 1n- terested individuals, or one perhaps more inti- mately acquainted with the subject than the other may reach a solution in less time than in a more formal set-up, as e.g. a group discussion. The factors involved are in part: (1) a knowledge of what has been done for a particular purpose, (2) what equipment is generally or actually available and (3) what, particularly, is required for the problem at hand. By dual cooperation is meant the intensive consideration by two individuals who for the moment may concentrate upon a par- ticular phase of a complex problem. If such “sive and take” methods were extended by small groups in our laboratories, with less bias and pressure, might not a greater advancement in scientific endeavor be counted upon? For, in contrast to the individual treatment of the sub- ject, a group discussion of some data in a prob- lem may easily become involved in personal ex- perience which will not be of interest to the group as a whole; or, if developed to the point where it is of interest to a large portion of any group, the discussion may not aid an individual with a per- plexing problem in his mind. Certainly provision should be made to extend what we here call “dual cooperation.” Discussions where one individual may assist himself to learn a new technique or clear up a problem by conference with another is important. It is also an individual duty to assist another scientist in the advancement of knowl- edge free from elaborate problems. Can a medi- um of exchange or a freedom to do this be pro- vided ? It is suggested that greater opportunity be pro- vided for informal discussions of mutually inter- ested workers. This could be done by less crowd- ing of formal meetings into a gathering of scien- . tists. Perhaps also some friendly means should be taken to encourage those who require help to reach the proper source of information; and to tactfully discourage the over-solicitous but warped individual. Beyond the point of acquainting one’s self with adequate equipment for research, there is the grave problem of securing devices for a par- ticular problem. Such devices may not appear to be available through commercial or other known channels. What should be done about this situa- tion? Should the investigator who needs a de- vice for his problem, do without it because it is not at hand? Should he modify his problem in order to work with what is available in the labor- atory where he wishes to work? Should he write to another laboratory for the loan, or make other arrangements to use the equipment which he would like to have? What cooperative plan can be created to effect a solution of this acute prob- lem? Rather than publish a list of stocks which may often be changed or altered, it would seem to be feasible that an arrangement be made between technically minded individuals in several lapora- tories for the exchange of information concern- ing scientific equipment. At the present time the almost prohibitive cost of some scientific equip- ment may act as a check on further scientific re- search. If arrangements can be made to provide adequate tools for research, through cooperative loans, there might be considerable advancement in this direction. Again this may be best ar- ranged through “dual cooperation,” where indi- viduals of one laboratory act as liaison officers for the purpose. As a suggestion for the correction of the pres- ent difficulties with regard to equipment for re- ————_- Aucust 25, 1934 ] THE COLLECTING NET 205 search, the following is proposed: (1) exchange between institutions of individuals who are ac- quainted with the technical aspects of research as by residence and individual conferences at more than one laboratory during a season; (2) the in- terchange of information concerning equipment developed for a particular problem by sketch and brief but clear discussion, distributed to labora- tories indicating their willingness to cooperate ; (3) an effort to make uniform those types of equipment used more or less generally by labora- tories in résearch. In this category may be in- cluded thermo-regulators, incubators, motor-re- duction gears, refrigerators, etc. Finally, with regard to cooperation, of one form or another, there should be, on the part of those interested in the technical aspects of labor- atory research a heartier and more constructive endeavor to aid the industrial laboratories. There are several causes of misunderstanding in this connection between industrial laboratories and re- search workers in the universities. Liaison work concerning equipment and methods is poorly done by salesmen and others. Printed records and ca- talogs are often inadequate. A more informal and personal (e.g. dual) cooperation may be of benefit, for the reason that much is being devel- oped in industrial laboratories which can not well be published or reported at scientific meetings. Such information sometimes cannot be officially spread before a symposia except in a more or less sterile form. A real provision by research laboratories to advance good work may be ar- ranged by liaison conferees if protection against dishonesty and a check upon the “Priority Seek- er” can effectively be devised. Ordinarily, as for example, at our marine laboratories and at con- ferences, ‘“by-meetings” of pairs of scientists will crop up. Here may be a way for advancement not otherwise practicable. Could a progressive method of arranging for these smaller, intensive group meetings be made at scientific assemblies, and seminar conferences—or must we depend upon chance and individual aggressiveness ? REPORT OF THE MEETING OF THE GENETICS SOCIETY OF AMERICA IN WOODS HOLE Dr. John H. Gerould ,of Dartmouth College, acted as chairman of the meetings since Dr. Sewall Wright and Dr. Donald F. Jones, presi- dent and vice-president of the Society, were not in attendance. . The morning sessions were at- tended by about seventy-five people, thirty-four of whom were members of the Society. The following is a list of papers which were presented in addition to those appearing on the printed program: Morning Sessions The Linear Distances between Genes, Chromo- som Maps and Crossing over in Drosophila melanogaster: Morton D. Schweitzer, Colum- bia University and California Institute of Technology. Effects of varying dosages of x-rays on meiotic chromosomes of Gasteria, treated at various stages of meiosis: Alfred Marshak, Harvard University Biological Laboratories. Thursday Afternoon Session Genetics and the Development of the Gameto- phyte: Dr. J. L. Cartledge, Carnegie Institu- tion, Cold Spring Harbor. Gene Balance and Development: Dr. Whiting, Carnegie Institution, Cold Harbor, Pee Spring Dr. E. G. Conklin acted as chairman of this session. Six additional demonstrations and exhibits were shown on Wednesday afternoon, as follows: Some effects of centrifugal force applied to Dro- sophila eggs: Ruth B. Howland, Harry G. Al- baum and Samuel Kaiser, Washington Square College and Brooklyn College. Exhibit of Opalina spindle fibres: University of Pennsylvania. Hermaphrodites in Drosophila: G. T. Lebedeff, Carnegie Institution, Cold Spring Harbor. Cytological evidence of the elimination of the x- chromosome in the eggs of Drosophila melano- gaster: Morris Rabinowitz, Washington Square College. Possibility of producing haploid Drosophila ex- perimentally (androgenetically): B. P. Son- nenblick, Washington Square College. Slides of sections of styles showing pollen tubes: J. T. Buchholz, Carnegie Institution, Cold Spring Harbor. i. Dee Chenr A clam-bake: sponsored by the Society and prepared by Chief Lewis of the Cayadet- ta, was held Wednesday night at Sippe- wisset. Seventy people were present to enjoy the clams, lobsters and other delicacies of a shore dinner, and later enjoyed a beautiful moonlight evening, 206 THE COLLECTING NET [ Vor. IX. No.-80 THE BEACH QUESTION IN THE LIGHT OF A CHANGING ENVIRONMENT. (Continued from page 202) tion of other stimulating as well as restful con- tributions to the External Environment by others sympathetic with the true inwardness of our work and needs. Music has been an especially delightful enjoy- ment furnished us by lovers of the muse, who also realize the essential need of our particular group of workers for its influences to transport us out of narrow or fatiguing routine and help stimulate the higher imaginative constructive powers. All who heard will long remember Mr. and Mrs. Charles Crane’s open air concerts by the Russian choir and the Balalaika orchestra. The musicales of Mrs. Murray Crane followed and many at the Warbasses, whose Sunday forums added another pleasurable and profitable diver- sion. Then the successful choral society and club concerts, and finally Mr. Frost’s importation of professional musical experts of superior quality giving concerts of high standing. We must men- tion Mr. Persinger’s group and the Washington Quartet. Already one practical result of this generous support of music has been the discovery and encouragement of several outstanding musi- cians from among us. Only to mention those who have appeared here oftenest, we have Nancy Wilson, ‘cellist; Mary Fay, singer; Eric War- basse and Guila Bustabo, violinists. The increase of private social interchange, even more than the Club and general associations, from the Hill to the shore has tended to maintain the desirable cultural atmosphere sought here. Our reminiscences, too brief to call history, since I am obliged to omit important aspects of the situation, and interesting details which might make it a live picture, emphasize, I think, certain underlying principles which have come into con- flict and which we must now line up in a few final remarks and suggestions in regard to their application to the recent revival of the beach question. To begin with the summer people may well be divided into two groups: (1) those trying out the situation and not yet rated as regular members of the community, and (2) the settled members of the M. B. L. Corporation who come regularly and have reached an agreement as to attitude and the desirable atmosphere here, and have be- come accepted as proven “compatibles.” I think we may rate the trustees selected from this group as the best examples. Let us bear in mind that the trustees are in no sense an arbitrary ruling body but rather consti- tute an elected board maintaining a general con- sensus as most effective for the laboratory. We've tried to show, also, how many valuable associations outside have aided these leaders to carry on the work with assurance to individuals of freedom and lack oi dictation in their work and in other relations here. Thus has arisen a unique type of university organization most tay- orable for the carrying on of research. To put it briefly, the biologists, like the natives of Woods Hole, have devoted themselves strictly to their own business, avoiding involvements which might standardize or limit them. This attitude of our permanent group is, of course, in marked contrast to that commonly maintained at summer resorts where activities proceed thoughtlessly, with lack of responsibility for the preservation of any such particular atmosphere as would favor the kind of work cultivated here. So, when questions of :rights and ethics of members in this community are discussed, those who would resent intrusion in laboratory prob- lems should stand with beach-lot-owners against such intrusions on their enjoyment of external privileges and quiet leisure enjoyed by the others. I must say emphatically that the beach-lot- owners have not been uncooperative nor shown the lack of practical Ethics which has been at- tributed to them by some; since they have, for a number of years, followed generous precedent in permitting occupation of their beach fronts by the general public at all hours. After twenty years, only three years ago they were forced in self-protection to withdraw these privileges and: close off their property. They were forced to fence off their water fronts for the same reasons that have brought about restrictions of the privi- leges of the general public, not only in the Gan- sett woods of Woods Hole, but all along the At- lantic Coast, wherever crowds gather. This is on account of the inconsiderate selfish rudeness and misuse of their property and comfort on the part of a small minority, mostly outsiders who have come to the beach in automobiles. These incompatibles with our environment disturbed the beach in every way, left trash, garbage on the private waterfronts, and created noisy, un- bearable conditions. This was at the high tide of visiting strangers from outside. So the fence went up, and champions of the so-called public rights to wide-open beaches everywhere arose, forgetting that the real suffer- ers were the beach-lot-owners, and that rights of such minorities should be equally preserved. The public had been protected already, how- ever, for when lots were sold on the hill and along the beach, Miss Fay and Drs. Strong and Meigs arranged in advance to preserve the privi- ee ee ee he, ee ee —— se Te ee Aucust 25, 1934 } THE COLLECTING NET 207 leges of the town people and the biologists; at the same time granting private waterfronts to the few who should buy there. This was accepted in good faith by the present beach-lot-owners ; and natives and biologists acquiesced in the allot- ments. Two years ago after public meetings and con- siderable discussion as to rights and courtesies in which the majority sentiment seemed to be strongly in favor of giving the beach-lot-owners immunity from interference; Drs. Strong and Meigs (the latter at considerable financial sacri- fice we believe) gave formal deeds for the right to the use of their holdings by our local residents, hoping to settle the matter and exclude outside interference. The town of Falmouth accepted this settlement last Winter and refused to do more, in spite of the urge of some extremists to have all the northern beach-lots taken over as public property. So the beach problem was thrown back on our own group for self regulation through the funda- mental principle of “‘live and let live” which we have repeatedly quoted as being so potent here in controlling mistakes of dictation and maintain- ing freedom throughout. It was decided to leave the beach lot owners undisturbed, and turn atten- tion to other methods of relieving a_ situation which does indeed call for action. This seemed to mean a settlement of discussion which had been proved futile. Last year Nature also stepped in and silenced complaints by, a demonstration of a possible method for improving the public allottment of beach by wiping out the advantages attributed to the lot owners; since the storms of the previous Winter shifted the sand from the lot-owners’ front to the public section, thus giving the sun- bathers the very conditions they had clamored for, and leaving only stones for the private lot fronts. Unfortunately, this year the sand was shifted back, causing a reopening of attacks on the fence and again raising questions of law and ethics. This unexpectedness and illogical habit of Nature is familiar to us, of course; but we must not fol- low leadership which deserts tested and effective precedent; so our only safe reaction is to stand out against attacks on individual rights and pri- vileges, and turn back to history for support in permitting and encouraging individual freedom. History, and the experience of a number of gen- erally accepted leaders then force us into dis- agreement with the editor of THE COLLECTING NET as to the importance of removal of the fence and an unethical attitude of the beach lot-owners. We feel he is attacking sufferers instead of the real enemy. and we see no constructive sugges- tion in his pleas. Right here, however, | must stop to record my appreciation for much constructive work he has done for our community, and say that I, (and others agree with me in this), rate his usual con- duct of THe CoLtLectine NET, together with the Scholarships, as valuable contributions to the Laboratory. Many of us appreciate his untiring devotion in various ways to the best interests of the Laboratory. He has asked for criticism while implying that some of it furnished him has been rather drastic and threatening. And he has offered to defer to fair criticism of his attitude. We are content to leave the case to history and what seems to be the unescapable logic of experience; that persis- tence in further championship of interference in the beach situation is contrary to our best guid- ance for the welfare of the biological work and environment here. So we ask him to desist. On the other hand it seems that his quotations from leaders among us who have seemed to threaten him are probably extreme expressions of individuals. They are certainly too drastic for general acceptance, for there would certainly be general resentment and disagreement with any action to force his withdrawal. Individuals are often hot-headed in personal criticism ; but it seems impossible, I believe, that a majority of those to whom we have delegated regulation here in accord with our traditions will fail to grant him approval of his constructive side, and welcome its free development. We should certainly expect this, even if they are forced at the same time to ask him to refrain, as all of us should, from catalyzing activities which have been shown to be contrary to our accepted guiding principles. So much restriction of an individual’s freedom must be asked and agreed in by all who believe in cooperative methods for the benefit of an institution. Finally, in regard to the solution of the beach problem, we seem to have been offered the only practical alternative for action in the shifting of the sand. I believe with many others that the building of a jetty to direct the sand where it is desired is the best suggested scheme for perma- nent improvement. The property has been en- trusted to the local public, native and biological, and it is now their problem to develop it for ade- quate use. We cannot discuss the details, but would advocate the formation of an operating committee, who shall secure the means to provide the jetty and further improvements needed, this means rafts and supervision. In a word, the problem is to use the property which we have and make it adequate, rather than to fight for further extensions, 20! THE COLLECTING NET [ Vor. IX. No. 80 NY. State Fish Hatcheries - Carnegie Institution > Blackford Hall dormitory + Laboratory buildings - Main Building + Residences THE- BIOLOGICAL | LABORATORY i Sa Co Cold Spring Harbor ~—- THE EFFECTS OF X-RADIATION ON CELL STRUCTURE AND GROWTH A GENERAL SURVEY OF RADIOBIOLOGY GrorGE L. CLARK Department of Chemistry, Massachusetts Institute of Technology (Continued from previous issue) VI. Stimulating Effect of Irradiation For years the legend that roentgen rays or radi- um, under certain conditions of dosage, may in- crease the growth and metabolism of cells has gained wide circulation. This notion has arisen from the attempt to apply to these agents the so- called Arnt-Schulz law, according to which small doses stimulate and large doses depress cellular metabolism. Based on pharmacologic grounds, this doctrine has not been generally accepted, even by pharmacologists. The attempt to apply it to the action of roentgen rays is unwarranted, be- cause the experimental evidence, on which it is based is extremely meager and apparently invalid. That a measure of acceleration in cellular meta- bolism may occur under certain conditions has been shown repeatedly both in animals and plants, but such unusual acceleration is a transient phase of reaction and is invariably followed by more or less pronounced inhibition of function and cellu- lar degeneration. Another factor in the propaga- tion of this notion of a stimulating action of the rays has been the regression of pathologic lesions after exposure to small doses of roentgen rays. Such regression is best explained by the excep- tional radiosensitiveness of certain varieties of cells. As the result of primary degeneration of certain cells a secondary and indirect stimulation may sometimes be observed. Such is the increase in connective tissue cells in certain tissues and or- gans after repeated irradiation; the connective tissue is laid down to replace other cells which the rays have caused to undergo degeneration. Any primary or direct acceleration of cellular meta- bolism must be regarded as an effort of the cell to counteract or compensate for the noxious influ- ence of the rays; in other words, it is purely a defense reaction. Continued acceleration of meta- holism can not be induced by roentgen rays or radium, which always cause degenerative changes or have no effect whatever. Irradiation of cer- tain tissues, such as the skin, repeated over a long period of time may cause hyperplasia of the epi- thelium, and this in turn may lead to malignant transformation. This is not stimulation in the sense here employed, but the alteration of a nor- mal to an aberrant function due to chronic irri- tation. One phase of the problem of stimulation is found in the effect of X-rays on sensory pro- cesses. The experiments in the writer’s labora- tory of auditory effects in dogs are perhaps most extensive. Animals were trained to a conditioned response, associating a standard sound with an electric shock through one paw, and the acuity of hearing measured. Irradiation was made through the skull, with dosage up to 11,000 roent- gens. All animals showed a transient increase in acuity following a latent period of 11 days of 5.5 decibels, continuing over a period of 12 to 36 days. Even though the animal showed deleteri- ous effects from the X-rays such as loss of hair, skin burns, anemia, and extreme restlessness, the acuity of hearing increased. Experiments are now in progress in which irradiation is made over parts of the body entirely removed from the brain and auditory mechanism, in order to ascer- tain whether the effect is a direct or indirect sys- temic one. Indications are that the latter is the true explanation and not some specific stimulating effect on the auditory nerve system. VII. Medical Implications of Cell Reactions to Radiation. X-ray therapy is indicated when it is desirable to produce the following effects: a. Inhibition of the growth or function of glands and cells. Differentiated cells such as those composing glands and hair follicles, physio- logically active cells, young cells, cells about to divide, lymphoid tissue and tissue of embryonic type are all markedly radiosensitive. Skin diseases characterized by hyperactivity of the glands, hy- perthyroidism, diseases which may be cured by oe ee ee ee eee. ee oe ee ee ee ae ee Eee Aucust 25, 1934 ] THE COLLECTING NET 209 checking ovulation, leukemia, and many other conditions are successfully treated by irradiation. b. Solution of hyperplastic connected tissue such as uterine fibroid (microscopic éxamination reveals atrophy of myomatous cells and hyaline sclerosis of the connective tissue with dilatation of blood vessels. c. Reduction of lichenification, by inhibition of overgrowth of epidermal cells, and destruction of fungi. d. Anodyne effect. The relief of itching has been very frequently accomplished. The relief of pain, except when due to a lesion which can be cured by irradiation, is less certain, though many writers have reported an analgesic effect upon neuralgic pain. e. Reduction of inflammation. There is a favorable experience in the treatment of carbun- cles, pneumonia in certain stages, erysipelas, etc. The rate and mode of reaction of inflammatory lesions indicate that the rays act chiefly by de- stroying the infiltrating lymphocytes, the excep- tional sensitiveness of which has already been pointed out. Evidently these cells contain pro- tective substances which enable them to neutral- ize bacterial or other toxic products which give rise to the inflammation; when the cells are de- stroyed by irradiation these protective substances are liberated and become immediately available for defensive purposes. f. Destruction of benign and malignant tissues. While it is not the purpose of this paper to enter upon a discussion of the problem of can- cer, yet a few fundamental facts bearing upon the effects of irradiation are in order. (1) First, the specific sensitiveness of differ- ent kinds of cells is here, as in the case of normal cells, the most important single factor in the treatment of neoplasms. The susceptibility of tumors to irradiation agrees closely with the radiosensitiveness of normal cells of the same kind as those from which the tumors are derived and of which they are largely composed. Desjar- dins shows that the inordinate hyperplasis of lymphoid structures which characterizes Hodg- kin’s disease, lymphosarcoma and lymphatic leu- kemia retrogresses under irradiation at the same rate as normal lymphocytes are known to be de- stroyed by similar exposure. “In fact, so striking is the parallel that irradiation is now being used daily as a means to distinguishing such conditions when their clinical features do not permit abso- lute identification. In some cases, indeed, the radiotherapeutic method of diagnosis is more ac- curate and dependable than microscopic examina- tion.” Ewing has been able to demonstrate strik- ingly how various bone tumors (endothelial mye- loma, benign giant-cell tumor, etc.) can be easily distinguished from other neoplasms which affect the skeleton and are noteworthy for radioresis- tance. Isaacs at the University of Michigan be- lieves that nothing happens to cells during irradia- tion that would not have happened to them if they had lived their normal life, except that X-rays make them go through the process somewhat faster. X-rays act by hurrying the onset of old age and not by killing directly. (2) Splendid progress is being made in the study of chemical effects. McDonald and asso- ciates at the University of Pennsylvania in the past few years have demonstrated that the cancer cell is distinguished from the normal cell by the following : (a) In tumor tissue for every 13 sugar mole- cules attacked, 12 are split into lactic acid and 1 is oxidized, while in normal tissue the ratio is 1:1. (b) In cancer blood plasma the pH is 7.47 or 8.7% more alkaline than normal. (c) Blood glucose is high. (d) Calcium is low. (e) Potassium is high. Irradiation must in whole or part bring about conditions to restore normal energy adaptation, increase acidity (proved), decrease blood glucose, increase calcium and decrease potassium content. The mechanisms involved in these chemical trans- formations we do not know. Carrel has shown that malignant types of cells digest fibrin and feed upon substances or tissues which are not utilized to the same extent by normal cells. (3) Following is the simplest though inade- quate description of cancer which is possible at the present time*. A cancer is an abnormally organized epithelial tissue which usually grows at a greater rate and to a much greater extent than the normal tissue from which it develops. There is good evidence that the malignant transformation depends, as Hauser’ maintained, upon a fundamental change in the biological characters of the cell. Accord- ing to von Hansemann? and Boveri, the cancer cell is an epithelial cell that has undergone an ir- reparable change involving its chromosome com- plex owing to atypical mitosis. The majority of its descendants, however, receive the same new complex by regular mitosis. Cellular abnormalities, including cancerous transformation, are brought about by certain phy- sical, chemical or biological agents acting directly upon the cells, upon their local environment, or upon both. In most instances, a state of chronic irritation is imposed upon the tissues and certain isolated cells instead of undergoing atrophy and death as do most, persist in the altered environ- ment. Subsequently the isolated cells undergo progressive abnormal changes apparently of pre- cancerous nature. Later, because the character- istic structure of a cancer cell is not clearly dis- cernable through the microscope, the pre-cancer- ous cells may be indistinguishable from truly malignant ones. 3 * See Kline, Ohio State Med. J. 1933. 210 THE COLLECTING NET [ Vot. IX. No. 80 Once fully developed, the cancer cells repro- duce cancer cells. As pointed out by Blair Bell*, cancer cells are characterized by as avid a respon- siveness to the chemotaxis of nutriment as is the trophoblast. Like the trophoblast, cancer cells possess an unusual capacity of establishing con- nection with a regional blood supply. Growing rapidly through the tissues, the new cells sur- round and penetrate the blood vessels and the lymphatics. Small masses of cancer cells becom- ing detached in the vessels are transported to local or "distant organs. In the metastatic foci the can- cer cells, just as in the primary site, grow rapidly through the tissues and readily establish a con- nection with the blood supply. By interfering with various organ functions, by their degenera- tion products, by causing hemorrhage and by un- dergoing secondary infection the cancerous growths cause anemia, emaciation, loss of strength and finally the death of the individual. According to Boveri, all differentiation of cells is due to certain alterations in one or more chro- mosomes. Accordingly, metaplastic changes such as the change of columnar cervical or uterine epi- thelium to stratified squamous type follow certain alterations of the chromosome complex of the af- fected cells. The change to a malignant tumor cell then may follow the direct specific action on certain chromosomes of such agents as X-rays, radium, tar, toxins, mutagens (Murphy®) and viruses but more frequently follows after the chronic irritation of tissues by the agents men- tioned and by others such as the friction of rub- bing, heat, aniline, paraffin, bacteria and animal parasites. According to von Hansemann, the cells undergoing certain atypical mitoses, suffer a re- gression in differentiation and acquire a greater capacity for multiplication. Elaborating von Hansemann’s idea of anaplasia, Boveri maintained that a state of chronic irritation gives rise to a great increase in cells and favors abnormalities in cellular division. A suppression of cell division after the division of the centrosome and the nu- cleus followed by a tetrapolar mitosis, he claimed, may result in a malignant tumor cell. This cell, however, transmits its characteristic properties by normal bipolar mitosis and multiple such repro- ductions constitute the cancer. (4) Fundamental principles of X-ray cancer therapy. Modern therapeutic technic not only involves the radiosensitiveness of tumors, but also the relationship of the normal matrix. Tumors proceeding from radiosensitive matrices are radio- sensitive without regard to their degree of differ- entiation ; and tumors proceeding from radioresis- tant matrix tissues are likewise radioresistant in a state of complete differentiation. Osteosarco- mas, fibrosarcomas and adenocarcinomas showing little sensitiveness, imitate the structure of the matrix tissue, and in all such cases, even includ- ing growths in relatively atypical manner, have the same radiobiologic quality as their matrix tis- sue. What in practice divides radiosensitive from radioresistant tumors is not the amount of radia- tions required for destruction of the tumor, but whether this amount is permissible from the standpoint of its effects on other tissues that are necessarily included in the irradiation. The radio- sensitiveness of malignant tumors is not in all cases greater than that of their matrices; other- wise there would be no malignant tumor that could not be made to disappear under irradiation and cancer would be a completely curable disease. Complete disappearance of the tumor can be achieved with certainty only when the matrix tis- sue can be destroyed. However, some tumors have an entirely different structure from their matrices, such as rotnd-cell sarcomas. These are composed of cells not found in mature connective tissue. They possess a radiosensitiveness which makes possible their disappearance, whereas spin- dle-cell sarcoma does not differ in sensitiveness from its matrix. But in most cases therapy must involve an injury to the irradiated matrix whose reaction is a criterion of the reaction of tumor tissue. This gives us a new principle of dosage, based not on the destruction of the tumor, but on disap- pearance of normal tissue of the matrix; the ra- diobiology of the normal tissue becomes the basis of radiotherapy of malignant tumors. (5) A new method of pathological diagnosis. The great value of the microscope in diagnosis is self apparent. And yet it is clear that many problems of cell structure and behavior are be- yond the power of any microscope to solve. For this reason other methods such as specific radio- sensitiveness have been welcomed. In a paper published three years ago* the X-ray diffraction method of structure research was extended to the actual structure of normal and pathological tis- sues, exactly as crystals of metals, or cellulose, or fibrous proteins could be studied. With this “su- permicroscopic’”’ penetration down to molecules of the complex substances of cells, a fundamental molecular change from normal to abnormal was demonstrated with comparable specimens. Hence, though only the slightest beginning has been made, another great application of X-rays has been opened to biology and medicine. It is sing- ular that X-rays will produce subtle changes in tissues and that we can use X-rays, now in a dif- ferent sense, to find out what change has been wrought by the original irradiation. Beyond a doubt in some great clinic with every intellectual and experimental facility, this new method of fine structure analysis can be made to answer many of the unanswered questions of radiobiology, and by improved diagnosis lessen the scourge of cancer on the earth. * Clark, Bucher and Lorenz, Radiology, 17, 482 (1931). yee Aucusr 25, 1934} THE COLLECTING NET 211 for Rapid, Accurate Serial Sectioning The wide popularity of this efficient instrument is due to its many out- standing features of construction and design. Its high degree of accuracy results from its compact rigid construction with its complete freedom from accuracy-destroying vibration. The working mechanism is enclosed for protection from dust and foreign matter by the metal cover, yet is easily accessible when necessary. This Microtome is widely used for rapid serial sectioning, cutting sections AT uae GUE Geatate with accuracy down to one micron in thickness. Built to extreme stand- 5 4. shale eqn ards of precision, it is equipped with an unusually heavy knife block. The PRODUCTION single-piece upper part moves in an arc, with the knife edge as the center of rotation, for setting to the proper cutting angle. Two substantial clamps hold this position and a graduated scale provides for recording it. The knife block is adjustable toward and away from the object. The bal- Oot eee haaal ance wheel is grooved to take a belt when the instrument is motor driven. AND B & L FRAMES While regularly furnished for paraffin sectioning only, this Microtome can be equipped for cutting small celloidin specimens. Write to Bausch & Lomb Optical Company, 671 St. Ba Paul Street, Rochester, } S( : & O New York for particulars. I f L OPTICAL INS RU NPE Nit S- oF OR oT We SCIENCES 212 THE COLLECTING NET [ Vor. IX. No. 80 INTERNATIONAL — ORIGINAL MODEL MINOT ROTARY MICROTOME A precision instrument which retains the desir- able and simple features of the original Minot Model. UPON REQUEST of many pathologists during the past six years, we are again manufacturing the International Minot Rotary Microtome. This is the same mi- crotome found in laboratories to-day after satis- factory service of more than twenty years. Descriptive bulletin on request INTERNATIONAL EQUIPMENT COMPANY 352 WESTERN AVENUE Makers of Fine Centrifuges BOSTON, MASS. COLD SPRING HARBOR | | Books Reduced SYMPOSIA ON 1-3 to'2.3 OEE QUANTITATIVE a) alee BIOLOGY on Water Street Volume I (resulting from conference-sym- THE COLLECTING NET posia of 1933 and dealing largely with surface WOODS HOLE, MASS. phenomena) contains papers by Harold A. Abramson, D. R. Briggs, Robert Chambers, Barnett Cohen, Kenneth S. Cole, Hugo Fricke, : ce Herbert S. Gasser, A. V. Hill, Duncan Mac- || (ER \\ Innes, L. Michaelis, Stuart Mudd, Hans Muel- Can) ler, W. J. V. Osterhout, Eric Ponder, Theodor \a & Svedberg, D. D. Van Slyke. ‘Zz u From a book review: “If this initial high ew 2 KS a ¢ standard (Volume I) is maintained, it is diffi- aa cult to see how a worker in this field can do MICRO SLIDES without these volumes...” The price of Volume I is $3.35. Volume II COVER GLASSES will appear in the autumn of this year. The prepublication price of Volume II, bound in DO NOT FOG f cloth, is $2.90, cash with order. After publi- Ask your dealer — or write cation, the price will be $3.35. Persons pur- (giving dealers name) to s R chasing Volume II may obtain Volume I for $3.00. Address the Biological Laboratory, D : iN Ciay-Apams ComPANy 25 East 26th Street New YORK Cold Spring Harbor, L. I, N. Y. a Ov ?9—0O0—_000 0S om" (wi bo our PART YS THE COLLECTING NET 213 Avucust 25, 1934 } Leitz Roll Film Micro Attachment WE DO OUR FART i. LEITZ, Inc., Dept. 571, 60 East 10th St., New York, N. Y. BRANCHES: Camera “MIFILMCA” This compact device is indispensable for photo- micrography of living objects where short ex- posure is essential. The small negative size is not detrimental to the revelation of detail be- cause special film emulsions and fine grain de- velopers make it possible to enlarge the photos thus obtained to considerable size. This and many other instruments of great value to the Biologist will be displayed at our exhibit at R. G. Thompson's, Main Street, during the month of August, 1934. Washington, D. C., Chicago, Hl., San Francisco, Calif., Los Angeles, Calif. Blakiston Announcement FOR PUBLICATION EARLY IN 1935— MANUAL OF LAND AND FRESH WATER VERTEBRATE ANIMALS Thoroughly Revised Edition By H. S. PRATT, Professor of Biology at Haverford College MOTTIER—Textbook on Botany for College Students. $4.00. ROBBINS—Botany of Crop Plants. $4.00. ROBBINS & RAMALEY—Plants Useful to Man. $3.00. PALLADIN-LIVINGSTON — Plant Physiolo- gy. 3d Am. Ed. $4.00. BARTON-WRIGHT—Recent Advances Plant Physiology. 2d Ed. $4.00. BARTON-WRIGHT— Recent Advances Bot- any. $4.00. HUNTER & LEAKE — Recent Advances Agric. Plant Breeding. $4.00. SANSOME & PHILIP — Recent Advances Plant Genetics. $4.00. GOULD—Pocket Med. Dict. 10th Ed. $2.00, Index $2.50. P. BLAKISTON’S SON & CO., INC. 1012 WALNUT STREET PHILADELPHIA 3d Ed. The Turtox Biological Red Book Two hundred and thirteen pages containing the most complete listing of supplies ever offered for the Biology laboratory. Profusely illustrated with photographs, drawings and color plates, it presents the materials used in the biology laboratory in a well organized and concise form. Ask us to mail a copy to your teaching ardress. GENERAL BIOLOGICAL SUPPLY HOUSE Incorporated 161-763 EAST SIXTY-NINTH PLACE CHICAGO 214 THE COLLECTING NET [ Vor. TX. No. 80 rill DIAPHANE THE IDEAL MOUNTING MEDIUM Diaphane is a synthetic medium made of resins and solvents which is superior in a number of ways to balsam for mounting. DIAPHANE WiLL CORPORATION ROCHESTER N.Y: Diaphane is neutral to all stains. The re- fractive index of 1.483 makes it suitable for the most delicate work, permitting viewing objects which are totally invisible in the usual mount. It is more convenient than balsam. Objects may always be mounted directly from ab- solute alcohol, frequently from 70% alco- hol, and usually from 95% alcohol. It is thus possible to mount specimens without employing dangerously volatile preparations. For the most delicate work, Diaphane Solv- ent may be employed in place of alcohol of high concentration. Diaphane is non-oxidant, works easily, and preserves delicate stains. Mounts made with it keep perfectly without scaling. Diaphane is available in two forms, color- less and green. The latter contains a copper salt which serves to intensify hematoxylin stains. Diaphane, colorless . So opi -60 Diaphane, green, containing a color which intensifies Hema- toxylin stains é F A ora art “75 100 ml. 2.00 500 ml. =7.50 Diaphane Solvent, for thinning Diaphane and for use as a clearing agent before mount- ing with Diaphane. .. . 500 mi. 2.50 1000 ml. 4.50 WILL CORPORATION LABORATORY APPARATUS AND CHEMICALS ROCHESTER, N.Y. THE WISTAR INSTITUTE STYLE BRIEF Containing 170 pages, 23 text figures and 37 plates, published January, 1934 This guide for authors, in preparing manu- scripts and drawings for the most effective and economical method of publishing biologi- cal research, has been prepared by the Staff of The Wistar Institute Press and the codper- ative efforts of more than fifty editors con- cerned in the editing of journals published by The Wistar Institute, and presents the con- sensus of opinion on many points relating to the mechanical preparation of manuscripts and drawings for the printer and engraver. Due attention has been given to the relative costs of various methods of reproducing tables and illustrations with a view to reducing the costs of publishing papers. The work has been revised, rewritten and enlarged since the first copy was prepared and submitted to editors, in order to offer as much information and illustrative material on the subject as is possible within reasonable limits. It will save authors much time and expense in preparing papers for publication and tend to expedite the publication of research. Address Price $2.00 The Wistar Institute of Anatomy and Biology Thirty-sixth Street and Woodland Avenue PHILADELPHIA, PA. Angle Centrifuges (Swedish) Developed by Dr. Ragnar Lundgren of St. Goran Hospital, Stockholm, Sweden during his work at the Syphilitic Clinic of the Karo- linska Institute. Used by a leading N. Y. Research Institute in many of their laboratories, especially for Wasserman and Serum work, The glass tubes are revolved at an angle be- low 50°, which results in a much faster de- position of sediment. As there is very little resistance to air, considerably less power is required than for ordinary centrifuges. Write for bulletin No. 528. EIMER & AMEND Est. 1851 Inc. 1897 Headquarters for Laboratory Apparatus and Chemical Reagents Third Ave., 18th to 19th St., New York, N. Y,. PMS) Aueust 25, 1934 | JHE COLLECTING NET ZEISS [ IKON } The universal camera 24x36 mm. TWELVE INTERCHANGEABLE LENSES TEN SPEEDS Bat 2, eas e ayilos 1/25, 1/50)" 1/00! 1/200, 1/500, 1/1000, Ideal for scientific photography. Com- pact and convenient. Metal focal plane shutter for slow and high speeds. Built- in long base range finder. Convenient two-finger control. Loads like a roll film camera. Daylight loading 36 ex- posure spools. Ask for literature on Contax and accessories eit CARL ZEISS,.INC., “sane Skeleton of Fish in Case Models, Specimens, Charts for Physiology, Zoology, Botany, Anatomy, Embryology, ete. Catalogs will gladly be sent on request. Please mention name of school and Spalteholz j peatetle) Te gine Gees | = | ete aleae Preparations Human Aap I~ = Crav-Anams Company Zoological 25 EAST 26th STREET NEW ae Modeliof Human\Heart Visit our display rooms and museum. The new and original features of this SPENCER No. 8 Research Microscope are described in de- tail in our booklet M-51-S. Prices are listed. Write for your copy today. ae § hive Peceminently a Hosp eT of fine muctescepes, Spencer 3 test welieventent 0 é v ‘ : esearch Mi “s the a: Vs é i egeanch = ze TOSCO Pe. at pe without exception, the finest, mest precise muctescope offered a to the AC tentisl today. NEW YORK BUFFALO, THE COLLECTING NET _ ___[ Vor. IX. No. 80 Vol. IX. No. 10 SATURDAY, SEPTEMBER 1, 1934 Annual Subscription, $2.v0 Single Copies, 25 Cents. THE DEDICATION OF THE MOUNTAIN LAKE BIOLOGICAL STATION Proressor Ivey F. Lewis, Director The buildings of the Mountain Lake Biolog- ical Station of the University of Virginia were formally dedicated on the afternoon of July 21st THE PRELIMINARY DISSECTION OF LIMULUS Dr. ELBert C. CoLe Professor of Biology, Williams College* Information secured from two of the larger biological supply houses indicates that several in the presence of representatives of many of the Southern colleges. The Station is located one mile north of Mountain Lake in Giles County, Virginia, at an elevation of approximately 4000 feet, on the divide be- tween the Mississippi and the Atlantic drainage areas. In the vicinity is a wide variety of biological conditions vary- ing from peat bogs and cran- berry swamps to the dry Al- leghany Mountain tops, in- cluding mountain and lowland streams and the nearby Moun- tain Lake, said to be the only natural lake in the Southern mountains and one of the highest in the East. The buildings are of a rustic type of construction, but pro- vided with electric current and running water obtained by gravity from a_- spring above the Station on Bear Cliff Mountain. These buildings include the John B. Laing Laboratory, (Continued on page 231) containing four class Scholarship Awards THE COLLECTING NET takes pleasure in announcing the award of its Scholarships for 1934 to the following students: Embryology: William Richardson Spofford, II Protozoology: Arlene Johnson De- Lamater Botany: Arthur Allan Cohen Physiology: Will be announced later Zoology: Will be announced later In accepting the award of one hundred dollars the student must spend not less than six weeks in full-time research at Woods Hole or at Cold Spring Harbor during the summer of 1935. The recipi- ents are selected by members of the staff of the courses as being most competent to carry on inde- pendent reaserch as well as being in need of financial assistance. hundred specimens of the horseshoe crab, Limu- lus polyphenius, are sent annually to various in- stitutions in this country. Such figures indicate that the presentation of a new method for preliminary dissection would be of interest. Alee (1922)? gave an ac- count of the method formerly used at the Marine Biological Laboratory. The preparation was made by the instructors and was a long, tedious and somewhat dangerous job. In the summer of 1927 the writer devised a new way of prepar- ing Limulus for student dis- section. This method is rapid and simple. It is also less dangerous than any method involving extensive use of a sharp knife in cutting the tough carapace. Limulus is first bled either by cutting off the legs close to the body with a pair of tinsmith’s shears, or by slitting the thin walls of the vascular (Continued on page 230) TABLE OF CONTENTS The Preliminary Dissection of Limulus, Dr. AS Ol? WN! oocoonsadecondcovopeobenn 227 Id, (Ch COS 4 ctibcnodooeno.ae otto a cor oad 217 , i , : é y Plasmodesma in Plant Tissues, Dr. L. G. Mountain Lake Biological Station. Account Tivineston: .. uncon ee ee ee 228 OfeLhemMedicationmery cre are id) eiste tae elslei= 217 . Effect of Centrifuging on the Alga Griffith- Geter. os ey aigt ale Dr) Vieton, Schechter) ..5 <7. he. 228 An Experimental Study of Striated Muscle The Differentiation of Rat Gonad Primordia in vivo, Dr. C.‘C. Speidel ............«-.- 225 in Normal Adult and Gonadectomized Rat NiditorialsPagebl ri. saisisaie suse cs cteardlevets cis 226 Hosts, Adrian Buyse .................... 229 THE COLLECTING NET [ Vou . IX. No. 81 THE SITE OF THE WOODS HOLE OCEANOGRAPHIC INSTITUTION PIER IN 1870 and brought passengers to Woods Hole, bound A painting from memory by Franklin L. Gif- ford who recently celebrated his eightieth birthday at his home in Woods Hole. The berth of the “Af- lantis” is located just to the right of the stage coach; the site of the engine house and a portion of the yard of the Woods Hole ¢ eeopaonie In- stitution appears in the picture. The painting por- trays the steamer “Monohansett’ landing at Bar Neck Wharf in August, 18703 with over 900 pas- sengers aboard on their way to Vineyard Haven Camp meeting. The vessel often towed whaling vesse's into Woods Hole when they were unable to proceed under their own sails. The building on Bar Neck Wharf was first used as a freight shed by the New Bedford and Martha’s Vineyard Steamboat Company, and was originally on the present site of the library. After the railroad wharf was built, the freight-house and passenger depot on the old Bar Neck Wharf were pur- chased from the Steamboat Company by William Studley who rebuilt it as a residence for himself. The house is now located on North Street. The yellow stage coach on the wharf met all the boats, for New Bedford and the Vineyard. It had a regular route between Falmouth and Woods Hole. The land in the distance is Naushon Island, while the low tide in the foreground exposes the sand bar on Grew's Clam Flats. Walter Lus- combe’s coal dock now covers the flats, and the Penzance Garage is located on the old Bar Neck Wharf. Two lightships can be seen in the dis- tant harbor. The ‘‘square-rigger’’ came from Italy loaded with brimstone for the Pacific Guano Company which was situated on Penzance Point in 1863. This chemical laboratory and manufac- turing plant was for thirty years the principal in- dustry of Woods Hole. Crude guano (excre- ment of sea fowls which was rich in phosphates and nitrogenous matter for plant growth) was shipped to Woods Hole from distant deserted is- lands. At the works at Woods Hole the guano was combined with bone scrap to produce a su- perior type of fertilizer. The Guano Company was in operation from 1863 to 1895, employing regularly from 150 to 200 men. SerteMBER 1, 1934 } THE COLLECTING NET 219 NY. State Fish Hatcheries - Carnegie Institution + Blackford Hall dormitory - Laboratory buildings + Main Building - Residences ~ LABORATORY Harbor —- THE GENE AND ITS ROLE IN ONTOGENY M. DEMEREC Carnegie Institution of Washington, Department of Genetics, Cold Spring Harbor, N. Y. In a paper published recently on “What is a gene?” (Demerec 1934) the gene was defined as a minute organic particle, capable of reproduc- tion, located in a chromosome and_ responsible for the transmission of a hereditary characteris- tic. This definition may well be used as a start- ing point of the present discussion in which it is intended to analyze the definition, to discuss sev- eral other characteristics of the gene, to present evidence which indicates that a gene plays an im- portant part in the biological processes of an or- ganism and to point out the possibilities regard- ing the role played by genes in processes of dif- ferentiation and development. Analysis of gene definition. In the early stages of genetic research it became evident from the similarity m the behavior of chromosomes and the behavior of various characteristics de- termined by genes that there must be a close re- lationship between genes and chromosomes. Evi- dence began to accumulate from sex-linkage, linkage groups, non-disjunctions, and later from translocations, inversions and other chromosomal rearrangements, to prove beyond all reasonable doubt that genes are located in chromosomes. At present there is also sufficient evidence to show that genes are arranged in a linear order within the chromosome and that this order is retained with great regularity. Infrequent changes in the order of genes which do occur under natural con- ditions, the frequency of which can be increased by various radiations, are an abnormal, rather than a regular, occurrence. Each gene therefore has a definite position within a chromosome, has its locus in the gene-string. Genes are extremely minute organic particles. They are probably ultramicroscopic. Estimates hy various workers (Morgan 1922, Muller 1919, Gowen and Gay 1933) place the size of the gene close to the size of a large organic molecule. Genes possess an important property of self re- production. The evidence now available indicates that a given gene complex is the same every- where in the body of an organism; which means that genes regularly reproduce like structures. It is known, however, that changes in genes do oc- cur every once in a while. Among the millions of cells constituting the body of a large organism it may be expected, therefore, that some of the cells would differ in their genic complex but this difference should be taken as an exceptional, rather than a regular, occurrence. In the case of unstable genes, which change with a high fre- quency, it may be expected that cells with unlike genic constitution in regard to these particular genes would be found quite frequently. Such differences in genic constitution of various cells of the body, however, are due to changes similar to those mentioned earlier, and the increased fre- quency of them does not constitute valid reason against the postulate that changes in the genic complex do not occur regularly during growth and differentiation of the organism. The hereditary units, the minute organic par- ticles called genes, must be very powerful agen- cies, since they are responsible for the appearance of various hereditary characteristics of the or- ganism. It is known that the presence of one kind of gene in a certain locus instead of another- kind will produce a marked difference in the ap- pearance of the organism. For example, the eye color of Drosophila will be either white, ivory, tinged, buff, eosin, apricot, cherry, blood, or red, depending on the kind of gene which is present in one particular locus of the X-chromosome. It is known, however, that no single gene has the sole responsibility for the appearance of any one characteristic. The phenotype of an organism is a product of the interaction of the whole comple- ment of genes, although certain of them mav have a greater influence on the expression of cer- tain characteristics than some other genes have. Importance of genes in living functions of the organism. Deficiencies, viz. absences of genes in various loci, occur quite frequently under na- tural conditions. Their frequency, is much _ in- creased in the material treated with X-ray radia- tion, In Drosophila melanogaster, deficiences are 220 THE COLLECTING NET [ Vor. TX. No. 81 known for about thirty regions of the chromo- somal complex. For the most part, they are very small, some probably affecting not more than one locus. All deficiencies discovered so far in Dro- sophila are found to be lethals when homozygous. This indicates that the presence of certain genes, probably a great majority of them, is essential in order that an organism may live, which means that genes play an important role in the vital pro- cesses of an organism. Evidence contradicting this generalization may be found in a case presented by Stadler (1934) describing a deficiency in maize affecting about 1/6 of one chromosome, a deficiency which is viable in the haploid condition (gametophyte ). As pointed out by Stadler himself, however, there is the possibility that maize may be a poly- ploid species and that this may make possible the survival of deficient gametophytes because of gene reduplication. =Hy ie an ae It it it La f iy Hf a fait sity ne reste ai ai aa a an i ae i ee hi i } Hutte i} . ii edie aay i yh if Lo _ veneer 4 iets ie ce - a . i i) ay ; oe . i BSS Bias Spy a ie