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Meade Hh eee itiaetrel we 2 pines ares me Ha ener St ede cae Pt PRA) Frets : : elehegey 1h ee , snail rate ane: ith Me whee dae Hho at 7 ij rE th i » hy ~ 4 _ ; Se iy aius as aie Hiaetguiuanctes : ee fir ieyag ae nH eee i) UP Adar fa We deta ate iil ty nytt a aera) ‘ea to t rina 7 yal uel at et en : ’ eh 808 Aiea " ia asf ae sit faa Cy it i wat Heh FOR THE PEOPLE FOR EDVCATION FOR SCIENCE LIBRARY OF THE AMERICAN MUSEUM OF NATURAL HISTORY i: | 4. nl as aie _ ae ~ © oy, i ne rs, ofjat ed OE PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE INCLUDING THE PACIFIC COAST BRANCH, MINNESOTA BRANCH AND WESTERN NEW YORK BRANCH. VOLUME XIX 1921-1922 EDITED BY THE SECRETARY NEW YORK 1922 4 De Wt Sok einer at Von TaD ANG _ J re - - p > ee 7 > Ww Zz CONTENTS. PAGE SCIENTIFIC PROCEEDINGS: Communications of the one hundred seventeenth meeting............... I Communications of the thirtieth meeting, Pacific Coast branch.......... 48 Communications of the first meeting, Minnesota branch................ 59 Communications of the one hundred eighteenth meeting................ 69 Communications of the one hundred nineteenth meeting................ 103 Communications of the second meeting, Minnesota branch.............. 132 Communications of the one hundred twentieth meeting................. 149 Communications of the one hundred twenty-first meeting............... 195 Communications of the third meeting, Minnesota branch............... 252 Communications of the one hundred twenty-second meeting............. 265 Communications of the thirty-second meeting, Pacific Coast branch...... 302 Communications of the one hundred twenty-third meeting.............. 309 Communications of the thirty-third meeting, Pacific Coast branch....... 345 Communications of the fourth meeting, Minnesota branch.............. 351 Communications of the one hundred twenty-fourth meeting............. 365 Communications of the fifth meeting, Minnesota branch................ 412 Communications of the second meeting, Western New York branch...... 423 RECAPITULATION OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE See oa Shes ea ete ae We Oi snd pee Ea Yap see lee 434 EXECUTIVE PROCEEDINGS (117th-124th meetings). ...........c0cccccevecees 457 REGISTER OF NAMES AND ADDRESSES OF THE MEMBERS..............0000002 464 NTE IS he oe Sree Foe ha Se Ra ay wo os beak ow owe meee 475 SES SS ye ee ee 476 Pee or Ses CAEN TIV IC: PROCEEDINGS. 2 ook. soc oc osc Sew w Sec suc ecw sec’ 480 111 Fae ot } WS SCIENTIFIC PROCEEDINGS. One hundred seventeenth meeting. ABSTRACTS OF COMMUNICATIONS. Cornell University Medical College, October 19, 1921. President Wallace in the chair. I (1748) Further studies on the nature of botulinus toxin. By J. BRONFENBRENNER and M. J. SCHLESINGER. [From the Department of Preventive Medicine and Hygiene, Harvard Medical School, Boston, Mass.| In trying to duplicate in vitro the conditions as they exist when botulinus toxin is taken by the mouth, we have observed that the acidity equal to that of the stomach contents not only leaves the toxicity of botulinus toxin undiminished, but would actually increase its potency. It has been repeatedly stated in the literature that botulinus toxin resists action of acids, but so far as we know nobody has observed the increase in potency of this toxin resulting from the change in its hydrogen ion con- centration. In attempting to establish the extent of this increase in potency we found that under the suitable conditions of the experiment the botulinus toxin which ordinarily kills mice in amounts not smaller than 3 X 107’ cc. can be increased in potency to such an extent that 3 X 10-7! cc. occasionally and 3 X 107! cc. quite regularly kills mice of 18-20 grams in less than 48 hours after the intra- ' peritoneal injection. While the total solids of such a minute dose _ of toxin amount to only 3 X 10-*% grams (this amount including also the inorganic portion of the medium) the toxic product thus obtained, nevertheless, possesses all the essential characteristics of bacterial toxins: it is thermolabile, it acts only after an incuba- tion period, it reproduces in experimental animals typical symp- I 2 SCIENTIFIC PROCEEDINGS (117). toms of the botulinus poisoning and it exhibits strict specificity in its neutralization with the homologous antitoxin. Our studies, thus far, were limited to the toxin produced by a single strain of Bacillus botulinus, but the experiments are in progress to determine whether the observation can be extended to toxins produced by other strains of Bacillus botulinus as well as to toxins of other bacteria. This work is a part of the investigation of food poisoning, con- _ ducted under the direction of Dr. M. J. Rosenau, professor of preventive medicine and hygiene, Medical School of Harvard University. The investigations are made under the auspices of the Advisory Committee on the Toxicity of Preserved Foods of the National Research Council, and under a grant of the National Canners’ Association. 2 (1749) Some plant sources of vitamins B and C. By FRANCISCO O. SANTOS. [From the Sheffield Laboratory of Physiological Chemisiry, Yale University, New Haven, Conn.] Several plant foods were tested for their content of vitamins Band C. Togi (sprouted mongo), okra, and avocado were found to be comparatively high in vitamin B. One half gram of each of them as daily supplement to the standard vitamin B free diet caused the recovery in weight of rats which had been declining because of lack of this accessory food factor. Mongo, sweet potato leaves and duhat (Eugenia jambolana) contain enough vitamin so that one gram of each of them as daily supplement caused the recovery in weight of rats which had been declining due to lack of vitamin B. Artichokes, bilimbi (Averrhoa caram- bola), banana flower bud and bamboo shoots are relatively poor in vitamin B. i The vitamin B in mongo was increased in germination, a fact contrary to the finding of Grijns that the antiberi-beri vitamin is lessened in amount as germination takes place. Mongo is relatively poor in vitamin C. Togi when fresh is relatively rich in vitamin C; but after it is prepared for culinary use, the vitamin C is destroyed. OBSERVATIONS ON PANCREATIC RENNET. 3 The observation of several investigators that vitamin C is in- creased when peas, lentils, and beans are germinated has been verified in the case of mongo. Ten grams of mongo as daily supplement to the scorbutic diet failed to protect guinea pigs from scurvy, while five grams of fresh togi as supplement to the same scorbutic diet cured three guinea pigs of the disease. 3 (1750) Observations on pancreatic rennet. By ALBERT A. EPSTEIN. [From the Department of Physiological Chemistry, Mt. Sinai Hospital, New York City.| Pawlow and Parastschuk,! Vernon? as well as Delezenne* have called attention to the presence of rennet in the pancreatic secre- tion of experimental animals. Wohlgemuth*‘ claims to have found it in human pancreatic secretion, but not without some difficulty. Notwithstanding these observations some doubt seems to exist in the minds of a number of investigators in this field. Textbooks on physiology do not class rennet with the other pancreatic fer- ments. Fresh or well-preserved dried preparations of pancreatic ex- tract ordinarily do not show any milk coagulating ferment. When solutions of such extracts are permitted to deteriorate the rennet function comes into evidence. While studying the pan- creatic ferments I have found that the presence of rennet in ex- tracts of this organ may be demonstrated constantly in a number of different ways. 1. Rennet may be liberated by heating a solution of the extract from 50 to 65° C. for a period of about 10-15 minutes; the most favorable temperature being 60° C. Flocculation usually occurs upon heating, but the ferment remains in solution. 2. The addition of suitable amounts of hydrochloric acids re- veals the presence of rennet. 1 Pawlow, J. P., and Parastschuk, S. W., Zeitschrift fur Physiologische, 1904, xlii, rs. a 2Vernon, H. M., Journal of Physiology, 1903, xxix, 302. 3 Delezenne, Soc. Biol., 1907, Ixiii, 98. 4Wohlgemuth, Biochem. Zeitschrift, 1917, ii, 350. 4 SCIENTIFIC PROCEEDINGS (II7). 3. By treating solutions of pancreatic extract with colloidal iron and other precipitants such as uranium acetate, alcohol, sodium sulphate and others. Calcium chloride solution in con- centration accomplishes the same result. 4. The addition of products of peptic digestion, such as those of gliadin or Witte’s peptone, to solutions of pancreatic extract also liberate the rennet. 5. Serum of a rabbit immunized by intravenous injections of pancreatic extracts, when added to solutions of pancreatic ex- tract, liberate the rennet. Whatever method of activation is used, in every instance, the rennet itself remains in solution, and some substance is precip- itated, which before precipitation conceals the presence of the rennet. I reported some of these results at a Section meeting of the American Chemical Society, held last September, and concluded at the time that the rennet in the pancreatic extract was probably present not as a pro-enzyme, but as an active enzyme mixed with substances which are antagonistic to its action. The conclusion is based on the foregoing experiments, the most significant of which is the one showing the effect of immunized serum on in- active pancreatic extract. Apparently the inactive solution of pancreatic extract is capable of producing in an immunized animal an antibody for the substance in the pancreatic extract, which is antagonistic to the rennet. The antibody thus produced is in the nature of a precipitin. The lack of any specific method of activation seemed to support this view that there is no pro-ferment. However, the proof is indirect, hence not final. Various attempts to recover the antagonistic substance in active form proved futile. The most that can be said about it is that it probably is a substance of protein nature which coagulates at a temperature between 60 and 65° C., is precipitable by sodium sulphate and other precip- jtants, and is capable of producing a precipitin in immunized serum. These facts brought to mind the former controversy concerning the nature or state of the rennet in gastric mucosa. You will recall that rennet is believed to exist in two states, that of an active enzyme (or rennet) and as a pro-enzyme (or OBSERVATIONS ON PANCREATIC RENNET. 5 pro-rennet), and that under the influence of very small quantities of acid at the optimum temperature, the pro-rennet is rapidly transformed into the active rennet. This result is regarded as the product of true activation. Hedin*, however, interprets the facts in another manner. He assumes that the pro-rennet is merely a combination of rennet with a substance antagonistic to it, and on the following grounds. If the pro-rennet be treated with dilute HCl, the rennet is set free and the antagonistic sub- stance destroyed, hence, its inhibitory action is lost. On the contrary, a solution of pro-rennet, treated with very dilute am- monia at 37° C., loses all its rennet already free, while the an- tagonistic substance remains unchanged; so that, by adding active rennet to this treated liquor, the rennet is at once rendered in- active. I applied Hedin’s method of proof to the rennet in the pan- creatic extract and found that the results were in accord with his. On closer analysis it became evident that Hedin’s proof was in- sufficient and the conclusion erroneous. Before proceeding to the evidence in substantiation of this, permit me to note the following concerning the pancreatic rennet. The content of this enzyme in pancreatic extract is very large, and goes absolutely hand in hand with the quantity of trypsin present. Means have not yet been found to separate rennet from trypsin, The two appear to be intimately associated functionally and chemically. A method has been devised for the. quantitative recovery of the rennet-trypsin enzyme and for its purification. This will be presented at another time. Suffice it to say for the present that the trypsin-rennet combination constitutes about I-2 per cent. of the total substance of dried pancreatic extract. It is of protein nature, is not precipitated by colloidal iron, is coagulated by heat (at 82—85° C.), is extremely hygroscopic, and of an acid character. It is active only in the presence of Ca, which, however, must be available in ionizable form. In this fact seems to lie the fault in ~ Hedin’s proof. I have stated that fresh or well-preserved pancreatic extract has no milk coagulating properties, but when a solution of it is 5 Hedin, S. G.,Harvey Lectures, 1914, p. 162. 6 SCIENTIFIC PROCEEDINGS (117). treated by heating (up to 60° C.) or by means of colloidal iron and other precipitants, the rennet is set free. Now if some of the original extract is added to the activated preparation, no inhibition of the rennet action occurs. In other words, none of the substance which hinders the action of rennet is present in a freé state in the pancreatic extract. When alkalinized with ammonia the original extract acquires the power to inhibit the action of active rennet preparations. The result, however, is not due to the setting free of the antagonistic substance (as Hedin believes), but to the fact that the calcium ion, which is essential for coagulation, is rendered inert by the procedure. On the other hand, if a solution of active pancreatic rennet is alkalinized with the hydrate or carbonate of ammonia or soda, the enzyme solu- tion is rendered inactive. If, however, CaCl. in sufficient amounts is added to this liquor, the rennet is immediately reactivated. Neutralization or alkalinization of the active rennet solution by means of disodic phosphate, or calcium hydrate, does not in- activate the rennet. It would appear from this that Hedin’s result was not due to inactivation of the rennet by means of the antagonistic substance, but merely to the removal of the calcium ion from the sphere of action. The experiments made thus far seem to indicate that the enzyme substance forms a chemical combination with calcium, in the nature of a salt, and only as such exerts its action. There appears to be some ground for the belief that rennet and trypsin reside in a single chemical unit of the pancreatic substance, and possibly represent two phases of one and the same ferment. I might add in conclusion that, whereas the evidence is in favor of the view that the rennet in pancreatic substance exists as an active enzyme and not as a pro-rennet, definite proof for this opinion is still lacking. EmETIC ACTION OF THE DIGITALIS BODIEs. 7 4 (1751) Further observations on the seat of the emetic action of the digitalis bodies. By ROBERT A. HATCHER and SOMA WEISS. [From the Department of Pharmacology, Cornell University Medical College, New York City.] The application of digitalis bodies directly to the vomiting center, described by Thumas, does not cause emesis. When a digitalis body is injected into a cat in which the spinal cord has been cut at about the level of the second thoracic vertebra, vomiting does not usually occur, but when the cord is severed at the level of the 5th thoracic vertebra vomiting is not prevented. Removal of the stellate ganglia frequently prevents this emesis, and removal of the stellate ganglia with cutting of both vagi prevents the emesis in nearly every case. Removal of the celiac plexuses does not interfere with the emesis following the injection of digitalis bodies. When the nerve supply to the heart is intact the injection of a digitalis body causes emesis, if the animal is in good condition. When all nerve supply to the heart is severed, digitalis does not cause emesis, but mercuric chloride still causes. vomiting in the usual way. Impulses appear to pass up from the heart to the vomiting center chiefly by the way of the sympathetic, and to a less, though probably variable, extent by way of the vagus. When the sympathetic is cut the administration of ouabain usually fails to induce emesis. This may be due to the fact that the impulses passing up the vagus are usually insufficient to set up the codrdi- nated reflex, or it may be that in those cases where vomiting is not elicited by this drug after the sympathetic is cut the vagus does not contain any fibers concerned in this reflex. Evidence is presented to show that digitalis bodies induce emesis by reflex action due to irritation of the heart or its ap- pendages. This is almost certainly a protective mechanism for the heart such as is recognized in the case of other organs. 8 SCIENTIFIC PROCEEDINGS (II7). We wish to offer the suggestion that impulses constantly pass from various organs to the vomiting center and that apomorphin promotes the codrdinated vomiting reflex to such a degree that these normal impulses give rise to vomiting. That the action of apomorphin on the vomiting center is strictly analogous to that of strychnin on the cord whereby con- vulsions—apparently spontaneous, but in reality of reflex char- acter—are induced. Vomiting requires powerful—almost convulsive—contractions — of the abdominal muscles and diaphragm, and the weak stimuli are incapable of setting up the reflex in the unpoisoned animal. It is significant, too, that morphin produces strychnin-like convulsions through its action on the cord (in the frog), and apomorphin-like emesis through its action on the medulla. 5 (1752) III. Experimental rickets. The prevention of rickets in rats by exposure to sunlight.! By ALFRED F. HESS, L. J. UNGER and A. W. PAPPENHEIMER. [From the Department of Pathology, College of Physicians and Surgeons, Columbia University, New York City.] In recent papers it was shown by Hess and Unger that rickets in infants could be cured by frequent short exposures to the sun’s rays.’ 2 By this means and without any alteration whatsoever of the dietary, the characteristic signs of this disorder begin to dis- appear in three to four weeks as noted by clinical examination and by the x-ray. Asa result of favorable experiences of this nature, it was concluded in a study of the seasonal incidence of rickets that ‘“‘hygienic factors, especially sunlight, and not dietetic fac- tors play the dominant réle in the marked seasonal variations of this disorder.”” It seems probable that the ultra-violet rays play a large part in this curative power of the sun, judging from the work of Huldschinsky ‘ and others who recently have shown that 1Hess, A. F., and Unger, L. J., Proc. Soc. Exper. BIoL. AND MED., 1921, xviii, 298. 2 Hess, A. F., and Unger, L. J., J. A. M. A., 1921, Ixxvii. ® Hess, A. F., and Unger, L. J., Amer. J. Dis. Child., 1921, xxii, 186. * Huldschinsky, K., Zeitschr. f. orthop. Chir., 1920, xxxix. THE PREVENTION OF RICKETS IN Rats. 9 infantile rickets can be cured by means of the rays produced by the mercury-vapor lamp. In 1918 we tried the curative effect of rays from this source, but, lacking the aid of x-ray examinations, could not convince ourselves of their efficacy; since then we have succeeded in curing rickets by this means. Having found sunlight efficacious in the rickets of infants, we proceeded to test its value in the prevention of rickets in rats. To this end a series of white rats were placed on the diet (No. 84) described by Sherman and Pappenheimer,® consisting of patent flour 95.0 per cent., calcium lactate 2.97 per cent., sodium chloride 2.0 per cent. and ferric citrate 0.1 per cent. It has been the ex- perience of the investigators in this laboratory that such a diet invariably leads to the development in rats of lesions which are anatomically identical with those of infantile rickets. In carrying out experiments on rats our practice has been to keep the colony in a semi-dark room, the yellow shades being drawn at all times. In testing the effect of sunlight, the rats (weighing at the outset about 40 grams) were kept in absolute darkness, one series being taken out of the room and exposed to the direct sunlight for a period of 15 or 30 minutes. There was no difference whatsoever in the diets of these two groups. After a period of about three weeks the animals were x-rayed in order to observe early lesions of the epiphyses, and after thirty to forty days were killed and autopsied. These experiments were begun in April when the weather permitted four to five exposures a week. It was found for the first time in our experience that diet No. 84, the “‘rachitic dietary,’”’ did not lead to rickets—that the rats which received sun treatment did not show signs of rickets either by x-ray or by histological examination of the bones. It is un- necessary to discuss in detail the histological criteria which we consider characteristic of rickets, as this question has been fully considered in a previous paper.’ It may be stated briefly that they consist of increased width and irregularity of the prolifera- tive cartilage, absence of calcium deposition and great excess of 5 Sherman, H. C., and Pappenheimer, A. M., J. Exper. Med., 1921, xxxiv, 189. 6 Hess, A. F.. McCann, G. F., and Pappenheimer, A. M., J. Biol. Chem., 1921, xlvii, 395. 10 SCIENTIFIC PROCEEDINGS (117). osteoid in the region of the metaphysis and along the shafts of the bones. In the paper previously referred to it was shown that the intro- duction of 0.4 per cent. secondary potassium phosphate (K,HPO,) in place of an equal weight (replacing about one seventh) of the calcium lactate contained in the rickets-producing diet, completely prevented the development of rachitic lesions; this constitutes an addition of 75 mg. of phosphorus per 100 gm. of the diet. In order to test the counterbalancing effect of phosphate and dark- ness, a series of tests were carried out in the dark with additions of small and increasing amounts of potassium phosphate to the stand- ard dietary (No. 84); to one series 25 mg. were added, to another 75 mg. (constituting dietary No. 85). The rats on these diets were kept in the dark but, to serve as control, half of each series were exposed to sunlight for thirty minutes daily when this was possible. As was to be expected in view of our previous experience and the fact that phosphate tends to protect against rickets, none of the rats which were treated with sunlight developed rachitic lesions. Among the group, however, which were kept at all times in the dark, active rickets developed in spite of an addition of 25 mg. of phosphorus. The addition of 75 mg. was found to be sufficient to prevent the development of this disorder in some of the rats. This amount contituted some-_ what less than the minimum protective supplement to diet No. 84, which in itself contains about 86 mg. of phosphorus. Thus it will be noted that a short exposure to sunlight was equivalent to almost doubling the protective dose of phosphate. If the phos- phate content of the diet is adequate, rats do not develop rickets in spite of being kept in the dark throughout the experiment. The effect of sunlight with other dietaries was also studied, and is being continued. _ DISCUSSION. As sunlight has a marked effect on the bony development of rats, it is evident that in future in similar nutritional investiga- tions, the light factor will have to be controlled and standardized. It seems probable that some of the irregularities and lack of con- formity observed by investigators in this field may be attributed to keeping the experimental animals under dissimilar intensities of THE PREVENTION OF RICKETS IN RATs. PI A. DARKNESS. Duration Microscopic Diet.1 (Days). Rat No. X-Ray. Examina- tion. ERNE aot Poe, eae aed 34 246 R. R. 23 247 — R. 22 248 _— B — 436 R. —_ a 437 R. = 30 438 R. R. No. 84 + 25 mg. P....... 39 262 R. R. 39 263 R. i: a 39 264 a. R. 28 443 R. — 28 444 R. R. (slight) 28 445 R. —_ No. 84 +75 mg. P....... 38 121 neg. neg. 38 122 neg neg 38 123 neg. neg. B. SUNLIGHT. Duration Microscopic Diet. (Days). Rat No. X-Ray. Examina- tion. LP en gn a EL 34 249 neg neg 32 250 sé sé 35 251 Q x 33 439 e os 33 440 si i 33 441 _ - 33 442 ‘s x No. 84 +25 mg. P....... 39 259 s 3 39 260 sé «sé 39 261 ee as No. 84 +75 mg. P....... 38 124 re = 38 I25 ac sé R. = rickets. light. The most interesting aspect of the question, however, is the phenomenon that the sun’s rays are able to stimulate a deposition of inorganic salts where these are lacking. The damaging effect of darkness emphasizes the fact that sunlight is of great impor- 1Diet No. 84 as originally constituted, contained 86 mg. per cent. of phos- phorus. In the fall, however, owing to a variation in the phosphorus content of the flour, this diet was found by analysis to contain only 72 mg. per cent. Rats numbered our 400 (in this table) were fed on the ration having the lower phos- phorus content. I2 SCIENTIFIC PROCEEDINGS (117). tance, not merely for the vegetable world but also for the higher animals. Furthermore, the fact that sunlight is efficacious in the rickets of both human beings and rats, serves to show the similarity of this disorder in these two species. These results indi- cate that in the prevention and causation of rickets at least one hygienic factor plays an important réle which will have to be carefully considered in future studies of this disorder. 6 (1753) Identical twins in pigeons arise from ova of markedly aberrant size. By OSCAR RIDDLE. [From the Carnegie Station for Experimental Evolution, Cold Spring Harbor, N. Y.] During Io years data have been accumulated for yolk size and total egg size in 15,000 to 18,000 eggs of doves and pigeons. Such measurements of these two associated structures permit us, within certain limits, to know some definite things concerning the size of either structure if the weight of the other is known. Another group of 15,000 to 20,000 eggs have been weighed, incubated, and later observations made upon the embryos and young. Incidental to these latter observations 7 instances of identical twins have been found. Such twins other than the seven listed here have almost certainly not appeared; or, if present, they attained a stage of less than 2-day embryos. The figures of Table I make it clear that at least most of the particular eggs which gave rise to twins were of markedly different size from all other eggs then being produced. This is particularly well shown in the first four instances—given in the upper eight rows of figures—since the twin-bearing egg was in these four cases by far the largest egg produced by its parent during one entire year,—and so much larger as to indicate, in all probability, that it contained the largest ovum produced during the year. The seventh case was likewise of aberrant size—being the smallest of a group of undersized eggs. However, the weights of all eggs ob- tained in connection with this seventh case, as also with cases 5 IDENTICAL TWINS IN PIGEONS. 13 and 6, are known to be rather unreliable indices of the weights of the enclosed yolks because the parent birds (K469, P843) suffered from special reproductive disorders which involved the production of irregular and inadequate quantities of shell and albumen, un- paired eggs and embryos often incapable of hatching. The early death of these three pairs of twins is probably to be associated with this circumstance. The significance of the egg weights of the four cases listed at the top of the table is wholly clear since differences of 15 per cent. in egg size (between the two eggs of a clutch) have been found in normal birds to reliably indicate that differences of yolk size lie in the same direction. The two cases listed at the top of the table have been earlier fully described! and the data given there will likewise demonstrate the abnormally large yolk size which must have been present in cases 3 and 4 of the present tabulation. Apparently the known facts concerning these cases of twins do not well accord with a strict application of Stockard’s? conclusions as to the cause of twinning and ‘‘double monsters,” particularly as he has described it in relation to birds, since in the present cases we learn that the twin-producing ovum of the pigeon is ‘‘marked”’ for twinning even before it leaves the ovary. However, it seems possible that even these instances may fall within the range of his generdl explanation. We have learned that extraordinary yolk-size means a low oxidizing level of the ovum. Since this level is lowest in the largest ovum of the given bird this excep- tionally low level may account for the first 4 cases of the list. In the last three cases the size of the contained ova is questionable but in these cases a disorder of the reproductive organs—already known to involve the abnormal functioning of some of the en- docrine glands—may conceivably effect a retardation of develop- ment previous to gastrulation as Stockard’s theory demands. The early death of many of these particular embryos, as well as an apparent excess of twins derived from the meager amount of this material, may afford evidence for such retardation. It should also be stated that in case 3 the embryo was subjected to ice-box temperatures (13°-16° C.) during the first 23 hours after laying; and that the parent in case 4 was a generic hybrid. 1 Riddle, O., Jour. Exp. Zoél., 1918, xxvi, 227. 2 Stockard, C. R., Amer. Jour. Anat., 1921, xxviii, I15. 14 SCIENTIFIC PROCEEDINGS (117). The sex of the twins from the large twin-yielding ova (sex known for first 3 cases only) is of real importance; for, much earlier work by the author has shown that in pigeons the females arise from larger ova and males from smaller ova. Each of the three present cases supplies a rigorous test of the validity of that conclusion,—and each affords a confirmation. If either of these extraordinarily large ova had produced male twins, it would have directly contradicted the conclusions drawn from related lines of study conducted over a period of several years. If prospectively male twins were present in this series they were necessarily con- fined to origins from yolks of relatively small size. TABLE I. SIZE OF EGGs YIELDING TWINS COMPARED WITH OTHER EGGS FROM SAME BIRD. Data on Eggs of ae ee — Twin-bearing Clu tch Weight of Minimum Weights ; : 5 Eggs for Other Than vs Laid by Twin-bearing No. of | 3 Same Female Eggs Female | 6 ; Per | Sex (or | Immediately: (Same Year). Parent. Date. W at Cent. of Stage). Pras 59 Tas Mast] Mil coe : e- - |Maxi- - eg fore.! | ter.1 |mum. | mum. Total A248 Kr; 4/7 7.43 fot 9:62.| 6.22 |>-4.33,1.27.10 23 K2| 4/9 | 10.63 | +43.1 2g 8.75 | 8.92] 9.17] 8.46] 22 60. Fi | 3/5 8.07 of 8.21 | 7.58 | 8.68 |. 6.72 22 F2| 3/7 | 10.08 | +24.9| 99 8.13 | 7.84] 8.65 | 6.15] a1 V49? D1 | 10/16 | 15.77? rou 14.80 | 17.05 | 17.40 | 14.80 8 D2 | 10/18 | 20.60 | +30.6 ee 15.33 | 18.06 | 18.23 | 15.33 7 P450 Dr1| 5/30! 8.92 of 8.76 | 9.02] 9.21 | 7.82 17 D2| 6/1 | 10.39 | +16.5 | (4—5d.) 8.00 | 9.17] 9.75 | 8.12 14 K465 | Er | 3/4 8.00 (1.0d.) | 7.59 | 7.05 | 8.87} 6.87] 17 E2| 3/6 8.05 | + 0.6 | (2.5d.) 8.24 | 8.50] 9.29) 7.82 13 K465 | K | 5/14 | 8.47 (4.5d.) | 7.905 | 8.18 | 8.87] 6.87] 17 8.50 | 8.64) 9.29 7.82 13 P843...| Cr | 3/15 | 6.40-| — 3.7 | (3.0d.) 7.51 | 7.04] (?) (?) 2 | C2] 3/17| 6.64 (2.0d.) 7.10 | 6.90] (?) (?) 3 + In some instances the twin-bearing clutch was preceded or followed by fewer than 5 eggs (see second and last columns). * Common pigeon; all other groups are ring-doves. Note.—The weights of twin-bearing eggs are set in italic type. First and second eggs of the clutch are kept separate throughout the table. THE VITAMINES OF .YEAST. I5 7 (1754) The vitamines of yeast and their réle in animal nutrition. By CASIMIR FUNK and HARRY E. DUBIN. [From the Research Laboratory of H. A. Metz, New York City.] The question whether pigeons and rats require for their well- being the same vitamine B has been discussed at length by Mitchell and Emmett a few years ago with the conclusion that vitamine B must be different from the antineuritic substance. Funk and Macallum have tested the phosphotungstate precipitate obtained from yeast and have found that while it was strongly curative for avian beriberi, it induced only moderate growth in rats. The present writers were able to show recently that by fractional ad- sorption with fuller’s earth or norit it is possible in most cases to effect an almost quantitative separation of the B-vitamine, cura- tive for avian beriberi from another substance, which we pro- visionally have called vitamine D and which acts on yeast and certain bacteria. In practice the separation is effected as follows: One liter of autolyzed yeast is shaken with 50 g. of fuller’s earth; the filtrate which in the majority of cases was found in- active for avian beriberi was treated twice with the double amount of fuller’s earth, the combined precipitates carrying down quantita- tively the vitamine D, the last filtrate being devoid of the two above-mentioned substances. Having succeeded in this separation (the procedure varying somewhat with different samples of autolyzed yeast) we thought it worth while to test out the fractions obtained on animals, making simultanous tests on pigeons, rats, yeast and streptococci. The experiments carried out with six rats and four pigeons in every case will be repeated and extended and the present com- munication is only of a preliminary character. While the pigeons were found to need only the vitamine B when fed on a vitamine-free diet, the rats exhibited a somewhat different behavior. They _were fed the usual so-called synthetic diet with cod liver oil as source of vitamine A. The rats receiving the vitamine B or D fraction as an addition grew only for a few weeks at a slow rate and started to die out after two months. While increasing 16 SCIENTIFIC PROCEEDINGS (117). the amount of one vitamine did not have any effect, the addition of the missing component in both of the above cases caused a prompt resumption of growth. The rats given both B and D from the start together with the last filtrate, which contains neither B nor D, showed a normal behavior both in regard to growth and appearance. The influence of the last filtrate does not seem to be very important but has to be investigated. The results suggest that the rats and possibly other mammals require, besides the vitamine A, at least two vitamines of the B type, namely the B and D vitamine, for their well-being and growth. 8 (1755) Comparative buffering value of American peptones. By J. BRONFENBRENNER, G. G. DE BORD and P. F. ORR. [From the Department of Preventive Medicine and Hygiene, Harvard Medical School, Boston, Mass.] Some time ago one of us! reported before this Society the results of the inquiry into the effect of the composition of the medium as affecting the reliability of the cultural methods of identification of bacteria, and has particularly insisted on the role of the buffer and on necessity of quantitative adjustment af media in respect to its buffer content. . In the present investigation we have attempted to determine the buffer content of a few of the commercial peptones with the view of determining the limits of possible variation in the buffer content in the media prepared in different laboratories as due to the choice of peptone alone. The method used was that of de- termining electrometrically the hydrogen ion concentration of the various peptone solutions before and after the addition to them of measured amounts of acid and alkali respectively. The study demonstrated the fact that initial reaction of different peptones varies within fairly broad limits, that due to complexity of com- position the buffering action of any given peptone varies at dif- ferent zones of hydrogen ion concentration, and that buffering action of one peptone at a given hydrogen ion concentration may 1 Bronfenbrenner and Schlesinger, Proc. Soc. Exp. Brot. AND MEeEDb., 1918, xvi, 44. BUFFERING VALUE OF AMERICAN PEPTONES. 17 exhibit as much as five times more buffering action than another peptone at the same hydrogen ion concentration. In general, the peptones tested showed the highest degree of variation in buffering effect in the zone of the hydrogen ion concentration limited be- tween Py = 9 and Py = 8, and the lowest degree of variation in the zone between Py = 4 and Py = 5. As to the absolute con- centration of buffering.salts, these were found in most peptones to be the highest at the zone of the lowest concentration of the hydrogen ions and not in the zone of neutrality or of high hydrogen ion concentration where the buffering action would be most de- sirable for the use in media for identification of bacteria. Below is a table showing the relative buffering action of pep- tones at various Py levels. | Peptone. on Pe PH | PH PH PH | 9-8. Beg et Fo, SG ee Re wn as oes Arce Sek se | 9 5 1 5 LE PROGR rt LO Acs avies Soee II 8 4 5 I5 Mpa 2c 5 TAD. a. Bes | 6 6 5 4.5 10 P-L i alee I ale ac ae a / 34 II 7 6 14 ES S05 eagle ee ne | 12 8 9 ” 14 TSE ee oe ee 13 8 5 4 10 NGC a oy a air sas, | 20 II 9 a 12 This work is a part of the investigation of food poisoning, con- ducted under the direction of Dr. M. J. Rosenau, Professor of Preventive Medicine and Hygiene, Medical School of Harvard University. The investigations are made under the auspices of the Advisory Committee on the Toxicity of Preserved Foods of the National Research Council, and under a grant of the National Canners’ Association. 9 (1756) Some mathematical relations in the Wassermann reaction. By STERNE MORSE. [From the Psychiatric Institute, Ward’s Island, New York City.] Von Krogh’s! equation, y = x”/(x" + k), has not received the the consideration by immunologists which its very close statement of the facts in several immune mechanisms capable of numerical 1Von Krogh, Journal of Infectious Diseases, 1916, xix, 452. 18 SCIENTIFIC PROCEEDINGS (II7). expression would warrant. It will in general, for instance, closely state the amount of hemolysis in a system where complement is the only independent variable, x, y, is the proportion of hemolysis read colorimetrically, and m and k are constants. It is often more convenient to use it in the form of x” = k[y/(1 — y)]. If this expression is put into logarithmic form, ’ n log x = log k + rps OEE 4 - the expression is linear when expressed graphically, that is, if plotted on logarithmic paper, the data will fall more or less ac- curately on a straight line whose slope numerically expressed will equal 7, and whose intercept on the axis y will be the reciprocal of k. Moreover, if two complements are compared, the intercept of their graphs on the axis of x will be reciprocals of their con- centration referred to any unit in which we may choose to ex- press such concentrations. In theory and this is to a large extent borne out in practice, this intercept on the axis of « is independent of the value of n. n varies in the case of blood cells with the individual from which the blood is drawn, with the age of the blood cells, and with the treatment which they have experienced. It is low when the cells are suspended in Ringer’s solution, high when they are suspended in salt solution, is increased with the age of cells and in general with harmful conditions, such as the presence of antiseptics in small concentration and the like. It decreases as the concentra- tion of cells is increased. It varies under the conditions and WASSERMANN REACTION. 19 technic used in this laboratory', from 2 to 6, generally around 3.5. In order to calculate the constants of the above equation from numerical data, I have devised the nomogram illustrated in Fig. I. This enables the value of 1 to be directly ascertained from any pair of values of x and Yr (x1, ¥1,) and (xe, ye), where as before x is as THEE AUNT Hatt a lil TEETH aos Huw apo HE aa HH HET ET TT TTT PEED UIT MRT AMA ae Ht Lf HH HBleaewn HH ales a} CPE TET Te TTT TT ose REN ED RARSi Mees fed pate pee ft Bue Sentara E Golttceamectlscccmmmcn t+} 5 Besheee rH HH HH os B Ht HoH Spaeaetssscceseae HH es a oa H44 C288 RPRRD = Biles rniie SsosBestllsdiEas 0.80 SU ESRRTE Hit Sr Eleni caeteee i SitssssssSee = E ss 0.76 233586535 =35S=s5 i szcs ; Pt ad === H scs==s aHiSE Eee! Sintssee 060 ay PEST TE biases 0.55 == Pty eee = il rarecerseese: BESRRa esheets it 38 tI 8 6 Sarg i Mmwecaca Coot Roe 0.10 & sBalutittsaai s@eve esa Se ie an ir ~- leg = Fic. 2. Codédrdinate paper — log y/(1 — y) vs. log x. 1 Morse, Psychiatric Bulletin, 1916, i, 47. 20 SCIENTIFIC PROCEEDINGS (117). the independent variable, complement, and y is the proportion of cells which are hemolyzed. Possibly a more convenient method still is the use of the codrdinate paper shown in Fig. 2, where log [y/(1 — y)] is directly compared with log x.. In the case of the nomogram a straight edge is laid between the values of y; and yo on the lines AB and CD respectively and the point where it crosses the line EF marked. A line is then drawn between the values of x; and x. on EF and CD respectively and this line is ~ moved parallel to itself until it passes through the point where the - first line crosses EF. It will then intersect the line OP at the value of ” required to satisfy this pair of values. In the use of the codrdinate paper the values are plotted directly and the slope measured. This second method has the advantage over the first of being dependent on all values of x and y and not merely upon a pair of values. The Wassermann reaction is in the last analysis an estimation of complement after certain procedures are performed. This method is, it is believed, the nearest to an absolute measure of complement which has yet been devised and has a precision under favorable conditions of 3 per cent. or better. The reaction between syphilitic antibody-antigen complex and complement appears to follow the same law, wherein the logarithm of the proportion between the amount of complement absorbed to that unabsorbed varies linearly with the logarithm of the amount of antigen antibody complex present, the slope of the graph in this case ranging round 1 or a little higher. If these considerations are valid, one can make certain state- ments as to the Wassermann procedure which are at variance with the theory of the reaction as ordinarily conceived. In the first place, the estimation of complement should be performed under such conditions as to bring the amount of hemolysis in the neighborhood of 50 per cent., which corresponds to the value log |y/(1 — y)] = o. *The precision of measurement of comple- ment by using this point can be calculated to be and is in fact at least 10 times as great as the precision obtainable by the common methods. In the second place, a true measure of the amount of syphilitic antibody antigen complex is given, not by the absolute amount of complement absorbed, but by the proportion which TYPING STRAINS OF BACILLUS BOTULINUS. 21 the amount absorbed bears to that unabsorbed. Thirdly, it follows from the second conclusion that the actual amount of complement which is used in the reaction is not important within limits, except as it affects the slope of the plotted logarithmic curve. This gives a method susceptible of considerable accuracy for the comparison of any unknown syphilitic serum or spinal fluid with a standard syphilitic serum which has previously! been shown to be indefinitely preservable by appropriate technic. 10 (1757) Experiment in new method of therapy of paralysis agitans. By M. H. WEINBERG and T. SCHUBB. [Pitisburgh, Pa.| Starting out from the premise that paralysis agitans is due to hyperparathyroidemia, as advocated by several observers, we proceeded to prepare a parathyroidectin substance for the treat- ment of this condition. Experiments were conducted on rabbits and on goats. The two external parathyroid glands of the goat were removed, and after forty days the blood of the goat was withdrawn and glycerinized. The administration of this blood to Parkinsonian patients seems to show promising results. Further study of this method of therapy is now under way. II (1758) Typing of different strains of Bacillus botulinus by immunologic methods. By J. BRONFENBRENNER, M. J. SCHLESINGER and S. C. CALAZANS. [From the Department of Preventive Medicine and Hygiene, Harvard Medical School, Boston, Mass.] A number of strains of Bacillus botulinus isolated both abroad and in this country represent a fairly uniform group in so far as their cultural characteristics and the symptoms produced by their toxin are concerned. However, in respect to neutralization of ‘toxin by antitoxin there exist two sharply distinct groups of this organism, thus suggesting that in fact we are dealing with two distinct antigens. lloc. cit. 22 SCIENTIFIC PROCEEDINGS (117). In this attempt to find a method by which the existence of two antigenic varieties within the group of Bacillus botulinus could be established without the recurrence to the toxin-antitoxin test we found that neither the complement fixation nor the precipitation tests give satisfactory results. The agglutination test, however, offered a ready means for grouping as the results obtained with this test were in accord with those obtained by toxin-antitoxin tests. The agglutination test has permitted us to classify also such strains of Bacillus botulinus which have lost their toxicity — under the conditions of test tube cultivation. As a control in all the above experiments we included a strain of Bacillus sporogenes and found that contrary to the statement in the literature all but one of the strains obtained by us from different laboratories in this country are free from Bacillus sporogenes contamination as judged by the above serologic tests. 12 (1759) The antiscorbutic potency of strawberries. By CLARENCE A. SMITH, OLAF BERGEIM, and PHILIP B. HAWKE. {From the Laboratory of Physiological Chemistry of Jefferson Medical College, Philadelphia, Pa.] Several guinea pigs were fed a diet of oats, milk, and hay until they were decidedly scorbutic. They were then given ex- pressed strawberry juice, either fresh juice or juice previously boiled for five minutes. The symptoms of scurvy were overcome within seven days by the administration of ten c.c. per day of either boiled or unboiled juice. Strawberries, therefore, appear to be relatively rich in water-soluble C, and their content of this vitamine is not seriously decreased by five minutes boiling. 13 (1760) A modified anaphylactic reaction induced by X-rays. By R. G. HUSSEY (by invitation). [From the Rockefeller Institute for Medical Research, New York City.] The following observations are of interest in connection with a theoretical consideration of the mechanism of classical serum anaphylaxis. ANAPHYLACTIC REACTION INDUCED By X-Rays. 23 We have found it possible, as has v. Heinrich also, to modify the manifestations of anaphylaxis in guinea pigs by exposing them to X-rays. Guinea pigs weighing about 250 grams were given an intraperitoneal inoculation of 0.1 c.c. of horse serum (I c.c. of a I — 10 dilution) for sensitization. Half of the number of sen- sitized animals were radiated immediately and then each day thereafter for lIodays. The X-rays were delivered from a Coolidge tube governed by a spark gap of 3 inches with 10 milliamperes of current. The distance from the anode to the surface of the animal’s body was 6 inches and the total time of exposure was 10 minutes. 14 days after sensitization 0.1 c.c. or 0.01 c.c. of horse serum was inoculated into the jugular vein as an intoxicating dose. In the animals sensitized but not X-rayed, typical anaphylactic manifestations and usually acute death followed the inoculation with either amount of antigen. The X-rayed animals, on the other hand, showed either very slight or no objective anaphylactic manifestations. If, however, 4 weeks were allowed to elapse from the time of sensitization, then a similar amount of antigen inoc- ulated intravenously, there was no difference in the behavior of X-rayed animals as compared with the controls. Further, it was found that radiation at any time other than during the incuba- tion period did not induce a modified reaction. With these facts established, we directed our attention to a study of the anaphylactic state of isolated tissue. It may be said that many investigators describe the anaphylactic reaction of isolated smooth muscle as an index of the reaction of the animal body as a whole. Indeed, this phenomenon is regarded by them as the most important evidence which indicates that the locus of antigen-antibody union is intracellular. Female guinea pigs of about 225 grams were sensitized by an intrapleural inoculation of 0.1 c.c. of horse serum and subsequently treated as described in the original experiment. At intervals of 14 and 30 days following sensitization, the horns of the uteri were re- moved and segments of these treated in accordance with the principles of the well-known Dale method for studying the physi- ological behavior of isolated tissues. For each tissue preparation, we employed a suspension bath of 250 c.c. of oxygenated Locke’s 24 SCIENTIFIC PROCEEDINGS (117). solution kept at a constant temperature of 38° C. When the_ muscle developed a satisfactory tone and rhythm, 0.2 c.c. or 0.5 c.c. of horse serum was added to the bath at a point which per- mitted uniform diffusion throughout the fluid before coming in contact with the tissue. The uteri of the sensitized and X-rayed animals reacted typically with maximal response just as did the uteri of the sensitized animals not X-rayed. The tracings of the uteri removed at both intervals show no essential differences. The information furnished by the data presented we believe to have a direct bearing on the controversial point regarding the locus of antigen-antibody union which results in anaphylactic shock. The results of our experiments indicate that the anaphy- lactic reaction of isolated smooth muscle is not an index of the reaction of the animal as a whole. Also it is indicated that other factors than the reaction of sensitized smooth muscle should be taken into account in the statement of a theory concerning the mechanism of anaphylactic shock. We have now in process an investigation in which we are de- termining the relation between the existence of free antigen and the presence of precipitins in the circulation of animals X-rayed and not X-rayed. The results of our experiments to date indicate that free antigen remains in the serum of X-rayed animals for a much longer period than is found in animals not X-rayed. A full report of these results, together with studies on passive anaphyl- axis, will be published later. 14 (1761) Contribution to study of diphtheria toxin. By P. J. MOLONEY and L. HANNA (by invitation). [From the Research Division, Connaught Antitoxin Laboratories, University of Toronto, Toronto, Canada.] The results reported are those of experiments planned tc throw further light on the mechanism of toxin production by B. diphtheria. An extended series of test-tube experiments was carried ou in which the Park 8 strain of the diphtheria bacillus was grov in broth and daily counts made of the number of viable organis od athe CONTRIBUTION TO STUDY OF DIPHTHERIA TOXIN. 25 (as estimated by the poured plate method), and at the same time the toxicity of the broth, free from organisms, was estimated. The growth curve had the characteristics of those reported at various times in the literature for other organisms; 1.e., a period of logarithmic growth, and then a continued rise which in general reached a maximum in 48 hours, a sudden falling off of the number of viable organisms and then a more gradual decrease, with some- times small increases in the number of viable organisms during the period of decrease similar to the observations of Graham-Smith! for staphylococcus. It is suspected that these small secondary increases may only be apparent, and are due to the method of counting. The toxicity of the broth was very low at the end of 24 hours (approx. 1 M.L.D. per 0.5 c.c.), with a small increase at the end of 48 hours and then a rapid increase (if the increase and decrease of the cells was characteristic), and after a maximum was reached a falling off in toxicity which was sometimes very sudden. This sudden falling off in toxicity has also been reported by Bunker.? The hydrogen-ion concentration of the broth during growth showed an initial increase and then a gradual falling off; this agrees with the findings of Bunker? and Davis.? Bunker in his paper gives the limits of maximum toxicity at Py 7.8 — 8.25; contrary to this, good toxin has continued to be formed at Py 8.7 _ There are several ways in which the above results may be interpreted: I. Reproduction and toxin production do not go on at the same time; a cell produces toxin when it is incapable of division; apparently it is not accumulation of toxin which inhibits repro- duction since toxin—200 M.L.D’s per c.c.—will support growth when replanted with B. diphtherie. 2. A non-toxic substance may be produced during the period of cell division which is transformed into toxin; or this non-toxic substance acts on some constituent of the broth and produces toxin. Non-toxic germ-free broths (after organisms had grown ‘for less than 24 hours) which were sterilized in various ways— 1 Graham-Smith, Journal of Hygiene, 1920 (10). , 2 Bunker, Journal of Bacteriology, 1919, iv. 3 Davis, Journal of Bacteriology. 1920, v. we 26 SCIENTIFIC PROCEEDINGS (I17). heating at 56° C., berkefelding, and by adding various disinfectants, phenol, gentian violet, etc——and then allowed to incubate at 37° C. for 5-6 days, failed to show any toxin production. Some experiments by Walbum! are interesting in this connection. Walbum deduced the existence of what he called pro-toxin; he injected a mixture of toxin plus peptone into a guinea pig in such amount that the pig died in five days; the same amount of peptone and toxin given separately to two other guinea pigs failed to kill. The experiments hardly seem conclusive. The peptone might. conceivably injure the animal so that the toxin would kill more easily. He tried no experiments with other reagents than peptone, to check this point; and no experiments were undertaken to test the specificity of any toxin which may have been formed by this mixture. 3. There may be some sort of autolytic disintegration of the cells—so far there has been no evidence of this sort of mechanism. Organisms at different stages of life activity—after growing I-2-4- 6 days—were incubated at 37° C. for 6 days with saline and with distilled water. The clear sterile liquid in these several cases showed no toxin whatever. A possible objection to these ex- periments is that the digestion was not carried on in a colloidal substrate. Appropos of this were the attempts made to prevent growth by planting a very large number of organisms, but these were unsuccessful, for no matter how large a number of organisms was planted there was always some growth. However, in every case so far tried the broth planted with a very large number of organisms ‘showed less toxin than the same broth planted with a loopful. In some cases the difference was very marked. This difference in toxicity was apparently not due to absorption of the toxin by the organism; toxin allowed to stand in the ice chest with large numbers of organisms at different stages of life activity did not show any change in toxicity. . 4. During the period of cell division a substance may be pro- duced which acting on the non-viable cells produces toxin. The following results bear on this. After growing the organisms for less than 24 hours the whole was sterilized, in some cases by heating to 1Walbum, Zeit. Immunitatsforschung, 1909 (3), originale. ~ THE CHANGE IN REACTION OF DYING TISSUE. 27 56° C. and in others by adding disinfectants, and in each case allowed to incubate at 37° C. for 6 days. Sterilized thus by heat, by phenol and by gentian violet there is no evidence of toxin production. Experiments to test more thoroughly these various hypotheses are being planned. Details will be published in full elsewhere. 15 (1762) The change in reaction of dying tissue. By WITHROW MORSE and H. C. VAN DER HEYDE. [From the Department of Physiological Chemistry, School of Medicine, Morgantown, W. Va.] In the studies of tissue enzyme action which the senior writer has been making since 1910, there has always been the question of the stages immediately following the death of the tissue and also of the conditions of reaction of medium, which have been shown to regulate the character of the process, that is, the rate and equilibrium. An attempt was made by Strauss and Morse! to determine the reaction of medium in the kidney during hematog- enous infarction brought on by ligation of the blood vessels and at the same time to determine whether autolysis proceeded or not. The former collaborator (D. C. S$.) being called for service rendered it impossible to complete this series of studies. Earlier still? the Sgrensen colorimetric method was employed in similar work, but the obvious difficulty of the time element involved in the dialysis inhibited very critical conclusions. Recently, Dernby? applied the S¢grensen solutions with the Clark-Lubs indicators to the study of the problem, but the critical point regarding the inception of autolysis and the state of reaction of medium in the earliest stages was not investigated. In his third paper in the “Studies of Autolysis’’* Bradley and collaborator found ‘‘soon 1917, Xiv, I7I. . 2 Morse, M., J. Biol. Chem., 1916, xxiv, 163. 3 Dernby, K. G., J. Biol. Chem., 1918, xxxv, 179. 4 Bradley, H. C., and Taylor, J., J. Biol. Chem., 1916, xxv, 261. 28 SCIENTIFIC PROCEEDINGS (117). after death”’ a reaction of Py = 7.00 in normal liver, but inasmuch as beef and pig livers were used, it is probable that the source of supply was slaughter-house material as in previous work in the series, while there is nothing to indicate that the experiment with horse liver involved the incipient stages, so that no data seem to have been given which would permit one to judge how soon post mortem the experiments were conducted. Here, as in the studies of Dernby, the colorimetric method involving dialysis was em- ployed (p. 263, l.c.). The writers are unable to find in bio- chemical literature any other studies of this nature and the follow- ing results of their work are presented with the view of interesting investigators in the problem where facilities are available for furthur work. Method.—Guinea pigs were used, the pig being struck on the head with an iron mallet, laparotomy rapidly performed, the liver exposed and frozen im situ by means of an ethyl chloride spray. Then the liver was excised while the heart still beat, transferred to a cold mortar in an ice-bath at — 5° C., wherein it was ground to a snow. ‘The temperature of the liver mass, however, varied but little from zero Centigrade; in this connection it is well to recall the findings of Foster and Moyle! in studies on muscle, where ex- posure to temperatures of from — 5° C. to — 8° C. led to relatively great development of acidity (lactic), the low temperature acting similarly to mechanical injury. The snow obtained in this way was transferred to the electrode vessel of the gas chain apparatus? and the temperature of the mass within the vessel was brought rapidly to about 20° C. by means of the warm hand. Potenti- ometer readings were made at frequent intervals and the readings followed for thirty-six hours. The contents of the vessel were agitated, moderately, by means of a stirrer, operated by a small motor. In order to check the apparatus, controls were run on S¢érensen NaOH — KH-2PO, buffer mixtures, the variation from the expected being but slight in any one case. By this means, likewise, the time for reaching equilibrium was established as far * The writers were permitted to use the apparatus belonging to the Department of Soils, West Virginia University. I II. THE CHANGE IN REACTION OF DyING TISSUE. 29 PROTOCOL. Time. Temperature. . 3:45 (Pig killed) 3:51 (Transferred to electrode vessel) acum spciatls .i3% rg" Bs Ms aie aa 20 rie ded elas ae wre 20 POD ais. sided pin’ 20 MPN dee Sa Ee aes 20 ee. Ca 21 7 SRE 21 os: Sipe alee araeas 20 Mess 5 te): SE 3% 20 Ce ee ee one 20 Area te se shoes 20 Ve: Sees oe pone 20 ea Nosians alts Sim 21 Bim us ikh. £5 21 Re es 5a eal o's) a. vce 21 eric ee ie ae 2I SIR weet ice So's 21 Be tone atinlt: «aye 2I 7:39 (Toluene added) 21 G05 (a.01.) ... 19 Dd OS aoe eae 22 7245 RES. wo - 21 110 fee se: 21 Ot. Mw sss 22 Discontinued. 70°34 Killed.. ..-. Millivolts. Calculated Pa (Saturated electrode) 420 510 §30 535 540 542 550 615 635 645 660 664 665 660 657 652 648 646 630 640 590 587 Sap 485 10:20 Transferred to electrode vessel; heart in cadaver still beating. Tie a 20 Pees eek = 26 (Vessel warmed) ee a: ORC 20 (Vessel cooled) BO ee oh 21 POS is ete 21 ic Se en 2% EO:Gaey wae: 3 ge Bisel ahs oes 22 PROM eS. eth: 22 CHAOS Sos a 22 EM a oe bs 23 a ee ae ae 4 22 DR oe hehe 8 eos ae aie eteck sss ve 23 BL ae Ata & 23 AA ee Rte oe 22 a) at a 29 S245 A. ta ot 20 8 a ae Bt 20 OOS 8 ok or et 20 Discontinued. 3/0 495 490 497 500 500 502 497 496 495 622 641 643 626 620 605 590 520 527 528 6.80 6.11 6.20 4.67 30 SCIENTIFIC PROCEEDINGS (II7). being necessary. This figure is taken as a basis for the tissue work. The writers are unable to determine any factor in the tissue which may prolong the period of reaching equilibrium and while it is possible to explain the results obtained, as having to do with inequilibrium, the burden of proof is rather upon this aspect of the question, for one must show why liver tissue should demand more time for reaching equilibrium than the buffers. _ The protocols following are those of two experiments. A third was conducted with practically identical results: The results are striking, the reaction of the tissue being de- cidedly acid at the first reading, taken within five minutes after the time of excision of the liver. Then there is a slow fall to neutrality, which is reached within about 45 minutes. A rise ensues, which continues for a considerable length of time, over 24 hours at least. The meaning of these findings is not clear, but they may be due to the fact that acid is produced at first in an explosive way, a con- clusion which is justified by the studies of Fletcher,! who found that one fifth of the CO, produced by an excised muscle arose in the earliest stages; by the studies of Fletcher and Hopkins,? who found, always, in dying tissues lactic acid; and by the investiga- tions of Foster and Moyle,’ who found 0.218 per cent. lactic acid developed in injured muscle (minced) as compared to uninjured muscle 12 days at 0° C., 0.017 per cent. Secondly, the buffer action of the proteins, etc., in the tissue may exert its effect, caus- ing a “‘fixing’’ of the free acid, but finally this effect is nullified by a saturation of the buffers and a rise in free acid begins. If these results are free from criticism, a more substantial basis for the conception of how autolysis proceeds is available. Bradley showed in his first series of studies that the proteins of the sub- strate in autolysis became altered in some way whereby they be- came more digestible in tissue hydrolysis under the influence of the tissue protease and Dernby virtually substantiates these findings. The older work of Dochez, of Hedin and of Rowland point to this conclusion and the interpretation of relation of reac- tion to substrate is in keeping with the recent studies of Northrup, 1 Fletcher, W. M., J. Physiol., 1902, xxviii, 354. 2 Fletcher, W. M., and Hopkins, F. G., J. Physiol., 1907, xxxv, 247. 3 loc. cit. CURE OF INFANTILE RICKETS BY SUNLIGHT. 31 Falk and others upon different material. If we assume that the alkalinity of the tissue is slowly changed to acid reaction, it is _ difficult to see how low hydrogen ion concentration can operate to render the tissues more digestible, whereas a high degree of acid, such as we have found developed in the liver in the present study, may well be imagined to exert a profound influence upon the character of the proteins of the liver, for this concentration re- sembles that of gastric juice, especially that of the young subject,! where proteins are digested rapidly. Since the above statements were written, the electrometric method has been checked by the Sorensen colorimetric method supplemented by the indicators of Clark and Lubs. Practically identical results have been obtained with both liver and kidney. The details of the method, with results and discussion, will be given in another place under the following title: ‘Further Studies on the Reaction of Dying Tissues,’”’ by Withrow Morse and R. Goldberg. The question will be raised therein, whether the sug- gestion made by Paul Erlich (‘‘Die Aenaemie’’) that the reaction of the nucleus is acid, is applicable here. 16 (1763) The cure of infantile rickets by sunlight as demonstrated by a chemical alteration of the blood. By ALFRED F. HESS and P. GUTMAN. [From the Department of Pathology, College of Physicians and Surgeons, Columbia University, New York City.] It has been shown by one of us (A. F. H.) that the rickets of infants can be cured merely by frequent exposures to the sun’s rays.” Animal experiments carried out in this laboratory con- firmed these clinical observations. They clearly demonstrated that rickets could be either prevented or brought about in rats fed a standard diet, according to whether they were subjected for 1 McClendon, J. F., Amer. J. Physiol., 1915, xxxviii, 191. 2 Hess, A. F., and Unger, L. J., Proc. Soc. ExPpER. BIOL. AND MED., 1921, Xvili, 298. Hess, A. F., and Unger, L. J., J. A. M. A., 1921, lxxvii, 30. 32 SCIENTIFIC PROCEEDINGS (II7). short periods to the sun’s rays or were kept at all times in the dark. The present report adds substantiation from the chemical side to the clinical and anatomical evidence brought forward in the previous papers. In a recent article Howland and Kramer! have shown that the inorganic phosphorus of the serum of infants suffering from active rickets is reduced, and that during the process of healing, especially upon the administration of cod liver oil, the phosphorus content gradually rises to normal. In view of the marked clinical im- provement following sun treatment, it seemed of interest to ascer- tain whether this procedure was accompanied by a chemical altera- tion of the blood. For this purpose the rapid colorimetric method of Bell and Doisy? was used, in which the color is developed in protein-free filtrates through the reduction of phosphomolybdic acid by hydroquinone in alkaline sulphite solution. Special attention was paid to the inorganic phosphorus of the blood, although in many instances the so-called acid-soluble and _ total phosphorus was also estimated. It will be seen from the accom- panying chart that the normal figures for inorganic phosphorus are sufficiently constant to render this test of clinical value (Table I). TABLE I. BLoop PHOSPHORUS IN NORMAL INFANTS. Inor- Acid Inor- Acid Age, | ganic. |Soluble.| Total. Age, | ganic. |Soluble.| Total. Name.| Mos. Name. | Mos. Mg. P. per 100 c.c. Blood. Mg. P. per 100 c.c. Blood, M.K. 6 4.40 14.5 } ee 10 4:48 18.5 45.8 B.S. 4.33 16.2 38.3 S.D. 10 4.31 57:2 46.3 F.M. 15 4.60 17.8 41.6 BH, 12 4.60 18.1 53-5 B.S. 4.42 17.2 56.8 A.M. II 4.76 13.0 37.8 H.H. II 4.80 25-9 43.0 H.M. 7 4.10 16.0 68.2 G.H. 8 4.69 16.4 34.3 B.F. 6 4.00 18.9 M.D. 4 4.65 17.2 2955 A.R. 13 4.44 18.7 52.5 j¥; 18 4.44 16.0 49.6 D.B. 9 4.17 18.5 39.4 B.R. II 4.39 16.8 43-6 H.R. 13 4.61 19.6 40.5 W.L. - 4.10 | 14.9 67.1 | S.F. 4.00 if # 6 4.10 15-3 |. 63-5 H. B. 4.40 B.B. 6 4.05 24.5 — L. S. 4.00 A.S. 6 4.25 15.0 54.7 242, 4.80 A.A. 2 4.17 15.9 35.0 M. G. 4.34 M.C. II 4.14 18.7 — TS. 10 4.20 18.3 41.4 1 Howland, J., and Kramer, B., Am. J. Dis. Child., 1921, xxii, 105. 2 Bell, A. F., and Doisy, E. A., J. Biol. Chem., 1920, xliv, 55. CuRE OF INFANTILE RICKETS BY SUNLIGHT. ea, The infants were placed in the direct sunlight for a half hour to several hours, the period varying according to the intensity of the sun and the sensitiveness of the skin. Previous to treatment the majority of infants showed the usual clinical symptoms of mild rickets and the characteristic signs on x-ray examination. Such, however, was not invariably the case; it has been our experience that infants may manifest the classical signs of rickets, accompa- nied by a low inorganic phosphate content of the blood, and, nevertheless, show apparently normal epiphyses at the wrists and other joints. In the course of the sun treatment the babies be- came markedly tanned, the rachitic signs diminished or disap- peared, and the general condition improved. The accompanying table, which shows successive examinations of the blood, requires little interpretation (Table II). It will be TABLE II. BLoop PHOSPHORUS OF RACHITIC INFANTS TREATED WITH SUNLIGHT. Inorganic P. Acid Name. Age Total.! (mos.). | 6/22 | 7/21 8/11 | 9/16 | 10/18 | Sol.} ) gn: ae 7 2.80 2.75 4.14 4.13 5.7 43.6 | i eae 5 au3 3-4 4.16 4.22 23.0 41.0 EAut. tees 13 2.77 275 3.53 4. L5.2 43.6 Wats. s « 8 3.1 3.18 Beg 4.28 15.0 36.0 Bee ys ete DS 3.0 3.02 2.16 3.87 14.5 44.6 CM... 37 3-4 | EP Ir | 4.3 19.3 52.0 a a 7 3.0 3.9 4.0 16.6 38.5 Se 18 4.0 3.977 To.g 56.8 ee RS I5 4.6 3.067 7. 15.0 39.0 seen that the inorganic phosphorus of the rachitic infants steadily increased from month to month, starting generally below 3.5 mg. per 100 c.c. of blood and gradually being restored to the normal level which must be considered about 4.0 mg. On the other hand, the results of the determinations of ‘“‘acid-soluble’”’ and of total phosphorus are not sufficiently definite to warrant deductions. There were no alterations in the diet throughout these periods, the infants receiving the usual milk formulas; all were given orange juice daily, the older children getting cereal in addition. It is evident that sunlight not only brings about a cure of the rachitic lesions, but in so doing occasions chemical changes in the 1 Tests made with blood used for the first inorganic P. determination. 2 Previous to treatment. 34 SCIENTIFIC PROCEEDINGS (117). blood similar to those noted when the cure is effected by cod liver oil. This is of interest as affording testimony that the curative process occasioned by these divergent therapeutic agents is funda- mentally the same. These observations establish a chemical basis for heliotherapy in rickets. They furnish also, as far as we know, the first definite evidence of metabolic change in the animal body brought about by the solar rays. 17 (1764) Dissociation of microbic species. II. Mutation in pure-line strains of the bacillus of rabbit septicemia. By PAUL H. DE KRUIF. [From the Laboratories of the Rockefeller Institute for Medical Research, New York City.] The coexistence of two distinctly different types of microbe in cultures of the rabbit septicemia bacillus has been reported in a previous paper.1 These varieties, once separated, appear to breed true to type for many passages. The organisms have been designated as types Dand G. Type D is very virulent for rabbits, grows diffusely in liquid media, and yields highly fluorescent, rather opaque colonies on serum agar. Type G is of extremely low virulence, exhibits a granular sedimenting growth in fluid media, and grows in the form of translucent, non-fluorescing colonies on serum agar. The two types show no noticeable dif- ferences in morphology or in fermentation reactions. Immuniza- tion and agglutination reactions indicate their antigenic com- munity. It seemed necessary to determine whether the two varieties coexist in cultures isolated from infected rabbits or whether one variety arises from the other. Type D (virulent) is the microbe invariably obtained from the naturally infected rabbit. Type G has only been found after artificial cultivation has been carried on for some time. But since the primary isolations were made from colonies which conceivably might arise from two or more 1 Jour. Exper. Med., 1921, xxxiii, 773. DISSOCIATION OF MICROBIC SPECIES. 35 organisms, it would be unjustifiable to conclude that the original type D had changed into the microbe of the G variety. Con- sequently, 8 pure-line strains were isolated from a D culture by the Barber method. Single cells, removed from three-hour cul- tures, were planted in undiluted rabbit serum. The percentage of positive cultures obtained by performing the entire operation in serum was much higher than when broth was employed. The resulting pure-line strains were planted daily in undiluted rabbit serum. Tests were carried out to determine the conditions under which the low-virulent type G makes its appearance. The method of detection of this type consisted in streaking the test material upon the surface of 10 per cent. rabbit serum agar. The G type colonies can be readily distinguished from those of the strongly fluorescent type D. With proper attention to the technique of streaking, quantitative estimates of the propor- tionality between the D and G varieties can be made. In undiluted rabbit serum, with daily transplant, type D breeds true for long periods of time. The G variety has seldom been observed to arise under these conditions. In plain broth, transplanted daily, a few G colonies have been detected after 25 passages. On the other hand, when 3 or 4 days are allowed to elapse between transplants in this. medium, many colonies of this type make their appearance on the serum agar sub-plates. This observation led to the following experiment. 0.05 c.c. of a pure-line strain, type D, was seeded into tubes of plain broth and of undiluted rabbit serum. The tubes were placed at 37° C. and a loopful of the material from each tube was streaked at 12-hour intervals on rabbit serum agar plates. In the sub-plates from un- diluted rabbit serum no G colonies were detected during incuba- tion for 109 hours at 37° C. In the plain broth, G colonies began to appear at 48 hours, and had reached a concentration of 50 per cent. of the total organisms in 109 hours. These G colonies, fished from serum agar plates, remained true to type for over 50 passages, showing no tendency to revert to the parent D form, even when returned to undiluted rabbit serum. All of the pure- line strains under study have been found to undergo this mutation when allowed to stand in plain broth, but do so with varying degrees of rapidity and completeness. 36 SCIENTIFIC PROCEEDINGS (II7). It was considered probable that filtrates from D cultures might hasten the D—G ttransformation. Accordingly, cultures of 6, 24, 48, and 72 hours were filtered through Berkefeld candles. After sterility had been proved, 0.05 c.c. of pure-line strain B—D, was seeded into 10 c.c. of each of the above filtrates, into controls of sterile broth, and into undiluted rabbit serum. The tubes were incubated at 37° C. and streaked on serum agar plates at intervals up to176 hours. Contrary to expectation, the number of _G colonies arising in the 6- and 24-hour filtrates was extremely © small, and comparatively few appeared in that of 48 hours. In the 72-hour filtrate G colonies appeared at a rate and in a con- centration approximately parallel to that of the control broth. The mutation had reached 50 per cent. in 176 hours. In the undiluted rabbit serum no G colonies appeared at any time during the experiment. It would seem, then, that early filtrates from D cultures are antagonistic to the D — G mutation. 7 The Cy+ of the broth seems, within limits, to have no effect upon the rapidity of the mutation. If anything, an acidity > Py = 7.0 retards the process. Tests were made down to Py = 6.0, beyond which point it is difficult to obtain growth. An effort was made to discover the constituents of plain broth that encourage the tendency of type D to change to the G variety. Pure-line strains of the former were planted in beef infusion, and in various concentrations of peptone (Fairchild). The Py of all the media was adjusted to 7.4. It was found that little or no mutation occurred in the beef infusion up to 200 hours at 37° C. In 0.5 to I.0 per cent. concentrations of peptone some D—~G change was noted.. But when higher concentrations, up to 20 per cent., were employed, a very rapid mutation set in, reaching go per cent. of the total organisms in 96 hours. This was true even when the peptone solutions were made up to volume with beef infusion. Control tubes of undiluted rabbit serum and of beef infusion showed one or two G colonies at 120 hours, but none after 144 hours or after 8 days. This experiment indicates that peptone in suitable concentrations accelerates the D — G process. The G colonies arising in these experiments, and sub-cultured to undiluted rabbit serum, were frequently tested for their dis- tinguishing characters, 7.e., low virulence and granular growth in DISSOCIATION OF MICROBIC SPECIES. 34 fluid media. They were found in every case to satisfy these criteria. What is more, the acid agglutination point is distinctly different to that of the D variety. It is in the nature of a physical constant for each type, and is an important differential criterion. All of these characters persist throughout many passages in un- diluted serum, a medium markedly antagonistic to the original change. It cannot be said that the presence of the peptone causes the mutation D—G, since the change occasionally occurs, though very rarely and in small amount, in undiluted rabbit serum. On the other hand, the presence of peptone in suitable concentra- tion greatly accelerates a reaction toward which a tendency already exists. It is of interest to note that four pure-line strains, kept on ice in undiluted rabbit serum for three months without passage, showed no evidence of the appearance of G colonies. 18 (1765) Dissociation of microbic species. III. Differentiation of microbes D and G by acid agglutination. By PAUL H. DE KRUIF. [From the Laboratories of the Rockefeller Institute for Medical Research, New York City.] Granular sedimenting growth in liquid medium is one of the principal characters differentiating microbe G (bacillus of rabbit septicemia) fromits parent D form. Type G exhibits the granular appearance not only in plain broth, but in serum broth and in undiluted serum as well. This fact led to the examination of the comparative acid flocculation points of the two types. The method used was that of Michaelis, later described in full by Beniasch. The suspensions of types G and D were prepared by washing the sediments from 5 per cent. serum broth cultures in large volumes of distilled water. After this procedure had been re- peated four times, the final suspensions were carefully brought to equal turbidity. Prepared in this way, the G type suspension shows a stability equal to that of D. The tests for acid agglutinability were carried out with mix- 38 SCIENTIFIC PROCEEDINGS (I1I7). tures of Na lactate-lactic acid, range Py = 4.7 to Py = 2.4, and with Na acetate-acetic acid, range Py = 5.6 to Py = 3.2. The mixtures of these buffer series with the microbic suspensions were incubated at 43° C. for 16 hours. Readings were taken at the end of this time. A distinct difference in acid agglutination optimum for the two types was observed. The optimum for type G in general occurs at a range between Py = 4.7 and Py = 4.0. Type D, on the other hand, shows complete sedimenta- tion between Py = 3.5 and Py = 3.0. Many strains of the two types have been examined with invariably the same result. This observation furnishes an important differential criterion for the two varieties. The constancy of the acid agglutination optimum for type D is very strict. That for type G is slightly less so, but the variation is never so great as to cause it to be confused with D; 19 (1766) Dissociation of microbic species. IV. Factors influencing the acid agglutination optimum of types D and G. By PAUL H. DE KRUIF. [From the Laboratories of the Rockefeller Institute for Medical Research, New York City.] It is generally supposed that the acid flocculation optimum of bacteria is referable only to the Cy+ and is not influenced by the character of the buffer salts or the anion of the acid. This in- terpretation is questionable in the light of the following facts. Microbes D and G were tested against a glycocol-HCl buffer series, range Py = 3.0 to Py = 1.2. The same suspensions were tested simultaneously with the Na lactate-lactic acid and the Na acetate-acetic acid series employed in the experiments de- scribed in the preceding paper. ‘The results are presented in the following table. This experiment indicates that other factors besides the Cy + are important in the interpretation of the acid agglutination point of the organisms in question. For example, complete flocculation of type G occurs at Py = 3.0 in the glycocol HCI series, while no DISSOCIATION OF MICROBIC SPECIES. 39 flocculation whatever was observed at the same Py in the Na. lactate-lactic acid series. It would appear that either the Cl~ of the acid, or the glycocol possessed the property of broadening the optimum zone or of shifting it toward a higher Cy. Acip AGGLUTINATION OF TYPES G AND D IN VARIOUS BUFFER SERIES. Complete Flocculation. Buffer.Series. PH Range. G. D. Ua US ist a ea 3.0 to 1.2 3.0, ++ at 2.8 3.0 to 2.4 Na lactate-lactic acid........ 4.7 to 2.4 4.7 to 4.1 3-5 to 3.3 Na acetate-acetic acid........ 5-6 to 3.2 4.7 to 3.8 Ls ee ee This and other considerations led to experiments which suggest an explanation for the granular growth of microbe G in plain broth. Washed suspensions of this organism are strongly aggluti- nated by beef infusion between Py = 7.4 and 6.8. This range represents the Cy4 occurring during the growth of type G in broth. On the other hand, peptone (Fairchild) and Naz:HPOQu, the other constituents of broth, agglutinate type G very little or not at all in this acidity. Types G and D were next subjected to tests with varying amounts of beef infusion, which were adjusted to varying acidities, from Py = 7.5 to Py = 2.0. The dilutions of beef infusion were varied from 1-2 to 1-40. Each dilution was tested over the range of acidity just mentioned. Incubation at 43° C. for 16 hours. It was found for type G that as the acidity increases, down to Py = 4.5, the amount of beef infusion necessary to cause complete agglutination becomes less. At Py = 4.5 to Py = 4.0, complete flocculation occurs with traces of beef in- fusion or with none at all. But as the acidity is increased beyond this point, that is, at Py =4.0, increasing amounts of the beef in- fusion are necessary to produce this result. The same phenom- enon is observed for type D, the only difference being that the complete flocculation of this variety by a given concentration of beef infusion demands a higher Cy than in the case of type G. For type D, the optimum zone lies between Py = 4.0 and Py = 3.0. Beyond this, in the direction of greater acidity, more and more beef infusion is necessary to produce complete flocculation. 1 No flocculation at 2.7 and 2.4. 40 SCIENTIFIC PROCEEDINGS (II7). It will be observed that the range of Cy4 at which the smallest amount of beef infusion is required is for each type precisely the zone of the acid agglutination recorded in the preceding paper. This experiment indicates that the beef infusion, per se, does not cause the agglutination. It merely widens the acid agglutina- tion zone. This would seem to throw light upon the mechanism of the granular growth character of type G in plain broth. Suspensions of types D and G were similarly tested against decreasing concentrations of peptone at varying Cy. In these experiments the results were of a different nature, as might have been expected from the failure of peptone to agglutinate type G at Py = 7.5 to Py = 6.8. In the case of peptone, the optimum for type G lies at a range between Py = 3.0 and Py = 2.5. That for D, at Py = 2.5. Peptone, therefore, seems to shift the optimum zone in the direction of a higher Cy 4, an effect analogous to that observed in the glycocol-HCl buffer mixtures. In the case of microbe D, strong concentrations of peptone (1-2 and 1-4) actually suppress flocculation completely at Py = 3.0. This effect is analogous to the pre-zone phenomenon in immune reac- tions, since for the higher dilutions of peptone at this Cy4, com- plete agglutination readily occurs. It would appear from the foregoing that while the flocculation in all cases under consideration is due to H-ions, at the same time other factors, such as glycocol, peptone or beef infusion, either shift or broaden the acid agglutination optimum. 20 (1767) Dissociation of microbic species. V. Further considerations in regard to the virulence of microbes D and G. By PAUL H. DE KRUIF. [From the Laboratories of the Rockefeller Institute for Medical Research, New York City.] The wide variations in virulence between microbes D and G, bacillus of rabbit septicemia, has been demonstrated in the first paper of this series... Microbe D, the type found in natural in- 1 Jour. Exper. Med., 1921, xxxiii, 773. DISSOCIATION OF MICROBIC SPECIES. 41 fections, is possessed of powerful invasive properties, while its mutant form G is characterized by very low virulence. The fixity of the character of high virulence for type D is demonstrated by the following experiment. Strain R-19, type D, was tested a few days after its isolation from a rabbit dead of broncho-pneumonia. The test was carried out by injecting high dilutions of a six-hour serum broth culture intrapleurally into young rabbits of 600 grams weight. The strain proved itself fatal in 10-® c.c. of the serum broth culture. This culture was transplanted every seven days on serum agar. Tests made one and three months after the first experiment indicated its virulence to be still of the same titer. The individuals of a given strain of type D appear to differ very little in the characteristic of virulence. Six pure-line strains, isolated by the Barber method from stock strain R-15, were tested for virulence by the method just described. All were fatal in dose of 107* c.c. The virulence of type D not only remains constant during passage on serum agar, but persists under conditions that may be considered as distinctly unfavorable. For example, a pure-line strain of type D was planted in plain broth. It was allowed to remain at 37° C. for 9 days and 12 hours without further trans- plantation. At the end of this time a culture was streaked on a serum agar plate. Marked D — G mutation had occurred, counts showing D = 40, G = 60. A colony of each type was fished into serum broth tubes. These were incubated for the usual time, diluted appropriately, and injected into two series of rabbits of 600 grams weight. The D culture, injected over a range from 1o~’ to 107! c.c., proved fatal in every case. The G culture, on the other hand, failed to provoke a noticeable effect, even when 0.5 c.c. of whole culture was injected. It has been remarked in the second paper of this series that pure-line strains of microbe D may mutate during daily passage in plain broth. A pure culture of type D, virulence 10-*, was transplanted daily in plain broth for 25 passages. Its virulence remained constant during this time. At the 30th passage a few G colonies were observed on the serum agar sub-plates. In two months, sub-cultures on serum showed a large preponderance of G 42 SCIENTIFIC PROCEEDINGS (117). colonies. The virulence had fallen to 10-* c.c. Twenty-five days later, no D colonies could be demonstrated. The mutation D — G was complete, or type G had completely outgrown type D. o.1 c.c. of culture failed to produce a fatal effect. The at- tenuation of this culture is to be referred to the gradual replace- ment of the primordial D by the mutant G form. It is possible to predict the virulence from the relative preponderance of the two types, as evidenced by colonies on serum agar plates. It is _ possible to procure sub-cultures of very high or of very low vir- » ulence by selection of one type or the other, so long as any of the D type remain. While the virulence of microbe G is very low, 1.0 to 2.0 c.c. of whole culture may occasionally produce fatal infections, especially in young rabbits. The organisms recovered from such animals at necropsy retain their granular growth character, but may gain perceptibly in virulence. After three animal passages, a type G culture has been observed to reach a virulence of 10~ c.c. But despite this increase in invasive power, the non-fluorescence of its colonies persisted, its granular growth character intensified, and its acid agglutination optimum rose to > Py = 5.6. It is apparent from this experiment that it is unsafe to state that low virulence goes invariably hand in hand with the other characters gained in mutation. Experiments are under way to determine whether this artificially produced virulence of type G is permanent or evanescent. The route of infection is important in determining the ability of the low-virulent type G to gain a foothold in the animal body. A culture of microbe G which produced no perceptible effect when injected into rabbits intrapleurally in dose of 1.0 c.c. gave rise to abscesses when injected subcutaneously in 0.1 c.c. These lesions remained sharply circumscribed, but the type G organism could be recovered from them for several weeks after the appearance of the abscess. The phenomenon of vicariously greater susceptibility to sub- cutaneous injection would seem to be due to the rapidity with which phagocytes are mobilized against type G when this organism is injected into the pleural cavity. Within 6 hours after intra- pleural injection of 1.0 c.c. of serum broth culture of type G, no = ™ eae PREVENTION OF RICKETS IN RATS BY SUNLIGHT. 43 free organisms could be demonstrated in the aspirated fluid. Polymorphonuclear cells were present in large numbers and phagocytosis was intense. The virulent type D, on the other hand, gains its foothold primarily by reason of the late appearance of phagocytes following its intrapleural injection. The occurrence of the low-virulent type G would seem to afford an excellent opportunity for the investigation of the prop- erties or products of secretion which give the parent D type its characteristic of high virulence. 21 (1768) II. The prevention of the development of rickets in rats by sun- light. By P. G. SHIPLEY, E. A. PARK, G. F. POWERS, E. V. McCOLLUM and NINA SIMMONDS. [From the Department of Pediatrics, Johns Hopkins University, Baltimore, Md., Dept. of Pediatrics, Yale University, New Haven, Conn. and School of Hygiene, Johns Hopkins University, Baltimore, Md.| In June, 1919, Huldschinsky! reported that the ultraviolet ray exerted a curative action in rickets. Thecriterion on which he relied was the evidences furnished by the X-ray of calcium deposi- tion at the ends of the long bones. He found that there were definite signs of calcium deposition after four weeks of treatment and that at the end of eight weeks healing was almost complete. In May, 1920, Huldschinsky * again reported the curative effects of treatment with the ultraviolet ray in rickets in a series of thirty children, aged between one and one half and six and one half years, who exhibited all clinical manifestations of the disease. In all, healing was accomplished after twenty-two to twenty-six treatments covering a period of two months. In April, 1920, Putzig* corroborated the findings of Huldschinsky. He obtained 1 Huldschinsky, K., Deutsch. Wcehnschr., 1919, xlv, 712. 2Huldschinsky, K., Zischr. f. orthop. Chir., 1920, Ixxxix, 426. 3 Putzig, H., Therap. Holbmonatschr., 1920, viii, 234. 44 SCIENTIFIC PROCEE DINGS (117). cures by means of the quartz lamp in premature infants suffering from rickets. In July, 1920, Riedel! further confirmed Huld- schinsky’s findings in a series of one hundred children suffering from rickets. In June, 1921, Hess? confirmed Huldschinsky’s findings in a series of six cases. The favorable influence of sunlight in rickets has been recog- nized by some students of the disease for a long time, notably by Palm® (1890), and experimental evidence of its beneficial effect on mineral metabolism in the puppy has been furnished by Rac- | zynsky‘ in 1912. Huldschinsky made use of sunlight together with the ultraviolet ray in two cases of his series and Riedel relied on treatment with sunlight in some of his cases, supplementing with the quartz lamp ray only on sunless days. Hess* was the first, so far as we are aware, to demonstrate, by means of the radiograph, that sunlight alone exerts the same curative action as the ultraviolet ray. All the investigations which have been made up to the present time in regard to the curative effects of both the ultraviolet ray and sunlight in rickets have been made on human subjects of the disease and all the evidence has been fur- nished by means of the radiograph. In order to satisfy ourselves concerning the action of light in rickets as well as actually to see the changes produced in the bones we performed the following experiments. Eighteen rats about six weeks old and weighing between forty and fifty grams were placed on diet 3,143 which, as previous ex- perience’ has shown, produces rickets comparable in every re- spect to the rickets manifesting itself in human beings. The ration has the following composition: WURCAE ois Asre CSA 2 ce we Cae os ie an Sees oa eras Pe. See 33.0% RAI aig Ra ie oe eA ah Oe Zhe thas ev es Rid Div Se ee See Oe 33.0 Cede t ie 5 apa Sip te 5169 weve 4 Ka Te ULES b A ben eas a 15.0 Wheat gitee oi.:. . Siw wep bees eran vats} dp glee ee ee eee 15.0 rs a's as Sth oats dice p coed ie ae ae aah one ale a 1.0 CMC Ss ocak + Ev baie bee es SSE en 2 I ees ale viol» Sets gee 3.0 1 Riedel, G., Miinchen. med. Wchnschr., 1920, Ixvii, 838. 2Hess, A. E., and Unger, L. J., Am. J. Dis. Child., 1921, xxii, 186. ’Palm, T. A., The Practitioner, 1890, xlv, 270-279 and 321-342. 4Raczynski, J., Compt. rend. de L’Association Internationale De Pédiatrie, Paris, 1913, p. 308. 5 Hess, A. F., and Unger, L. J., J. A. M. A., 1921, Ixxvii, 39. 6 McCollum, E. V., Simmonds, Nina, Shipley, P. G., and Park, E. A. J. Biol. Chem., 1921, xlvii, 507. PREVENTION OF RICKETS IN RATS BY SUNLIGHT. 45 It contains nearly twice the optimal content of calcium, and is decidedly below the optimum in its content of phosphorus and in fat soluble A. Otherwise, it is well constituted. Twelve rats placed upon this diet were sent to New Haven, there to be exposed to sunlight. The remaining six rats were retained in Baltimore to be kept as control animals under ordinary laboratory conditions in a large, well-ventilated room completely screened with windows of ordinary glass. The animals treated with the sunlight were divided into two groups and placed in fairly large wire mesh cages. Each clear day the cages were carried out of doors and placed in the sunlight. At first, the weather being warm, the rats were exposed to the sunlight for two short periods of twenty minutes each. Soon, however, the periods were lengthened to six or even more hours. During the experi- mental period, which covered between sixty-two and sixty-seven days, the rats were exposed to the sunlight on every day except nine. The total exposure to sunlight during the experimental period varied between two hundred and forty-two and two hundred and seventy-three hours. The average daily exposure was four hours. When first exposed to sunlight, the albinos developed con- junctivitis; the ears of all, in particular the albinos, began to peel; the skin of the tails became sunburned and rough; the hair of some of the albinos acquired a yellowish tint. Long before the experiments were completed it became evident that the animals treated with sunlight were not developing rickets. Though they did not grow normally, they remained extremely active, climbing and darting about the cages. Toward the end of the experiments the males became sexually active; one of the females became pregnant. The control rats, killed at the expiration of two months, showed all the gross and microscopic evidences of rickets, the characteristic deformities of the thorax, enlargement and distortion of the costochondral junctions, fractures of the shafts, enlargements at the wrists, ankles and knees, and the ends of all the long bones. The bones cut with diminished resistance. On section a deep rachitic metaphysis entirely free from calcium was exposed. Into it the proliferative cartilage extended in irregular prolongations. The trabeculz were surrounded with broad zones of osteoid. 46 SCIENTIFIC PROCEEDINGS (117). The rats exposed to the sunlight, on the other hand, showed none of the evidences of rickets. The thorax was not deformed; the costochondral junctions were normal. There were no frac- tures of the ribs. The ends of the long bones were not enlarged. The long bones cut with great resistance. On microscopic ex- amination the cartilage was normal. The proliferative zone was completely calcified. The trabeculae were completely calcified. The condition found was normal except that both microscopically and grossly the bone was more delicate than in the rat of corre- sponding age reared on satisfactory diets. Though the sunshine completely prevented the development of rickets, it did not entirely compensate for the deficiency of phosphorus in the diet, either as regards the growth and development of the rat as a whole or of the skeleton. There were some noteworthy findings outside the skeleton. An abundance of fat was present. In the control rats the fat was scant. The thymus was only partially involuted. In the control rats it was completely involuted. The spleen was not enlarged. DISCUSSION. Sunlight effectually prevents the development of rickets in the rat. We have already shown,! as has also Pappenheimer, that cod-liver oil prevents the development of rickets in the rat. As nearly as we can judge from the radiographs furnished by Huld- schinsky and others the mode of healing at the cartilage-shaft junction induced by the ultraviolet ray (sunlight) is exactly analogous to that which occurs after the administration of cod- liver oil, as determined by Howland and Park.?. The time rela- tions are also similar. Huldschinsky found that the ultraviolet ray produced definite evidences of healing at the end of four weeks, and at the end of two months almost complete healing. Howland and Park found that cod-liver oil first gave rise to evidences of healing at the junctions of the cartilage and shaft of the long bones three weeks after the administration was begun and that at the end of about two months the calcification of the diseased ends of 1Shipley, P. G., Park, E. A., McCollum, E. V., and Simmonds, Nina. Proc. Soc, Exper. Biot. AND MED., xviii, 227, 1921. 2 Howland, J., and Park, E. A., Arch. Pediat., 1920, xvxvii, 411. Howland, J.,and Park, E. A., Bull. Johns Hopkins Hosp., November, 1921. (To be published.) -__ —_—eor PREVENTION OF RICKETS IN RATS BY SUNLIGHT. 47 the shafts seemed to be complete. Moreover, as the result of the gross and histological examinations made on the rats fed the rickets-producing diet 3,143 but exposed to sunlight it is possible to say that the changes produced by sunlight in the skeleton do not differ in any important respect from the changes produced when the animals are kept in room light but on a diet supplemented by cod-liver oil. Cod-liver oil contains something which is essential for optimal cellular function. Light also contains some- thing which is essential for optimal cellular function. Cod-liver oil or light when made available to an organism previously de- prived of either permits the organism to put into successful opera- tion adaptations or defense mechanisms which otherwise would have been ineffectual. Neither cod-liver oil nor light meets the defects in the composition of the diet directly by supplying to the body either calcium or phosphorus but meets them indirectly by so raising the potential of cellular activity as to secure the most efficient utilization possible of those substances available in the body which are directly or indirectly concerned with ossifica- tion and calcification. 48 SCIENTIFIC PROCEEDINGS (117). ABSTRACTS OF THE COMMUNICATIONS PACIFIC COAST BRANCH. Thirtieth meeting. Berkeley, California, October 15, 1921. 22 (1769) The production of tyrosine by a putrefactive anaérobe. By IVAN C. HALL and FLORENCE FINNERUD. [From the Department of Bacteriology and Experimental Pathology, University of California, Berkeley, California.| In caring for our collection of anaérobic cultures which now numbers 69 strains distributed among 15 clearly recognizable species and 4 strains as yet unidentified, we have observed that certain ones habitually form white crystalline products in the deep brain medium that is used for preserving the stock cultures; namely 3 strains of B. bifermentans, 4 of B. centrosporogenes (a new species shortly to be described for the first time), 1 of B. histolyticus, and especially an unplaced culture, herewith designated No. 106, that resembles B. sporogenes in certain properties but differs in its striking crystal formation. All of these are actively putre- factive anaérobes. We have failed to observe such crystals in 6 strains of B. Welchii, 3 of B. Novyt, 2 of B. butyricus, 19 serologically homolo- gous strains of B. sporogenes, 5 heterologous strains of B. sporo- genes, 2 strains of B. botulinus Type A, 3 of B. botulinus Type B, 7 of Vibrion septique, 7 of B. tetant, 3 of B. putrificus, and 1 each of B. Chauveaun, B. sphenoides, B. tertius, and B. tetanomorphus. Of this list B. sporogenes and B. botulinus are actively putrefactive, B. tetani and B. putrificus less strikingly so. References to crystals supposed on microscopic grounds to be tyrosine in cultures of putrefactive anaérobes are scattered through the literature, a review of which is reserved for a more detailed report. So far as we are aware, no one else has recovered tyrosin in a state of high purity from a pure culture of any single bacterium. Our studies to date show that culture No. 106 produces crystals, macro- and microscopically resembling tyrosin, in ground meat, TYROSINE BY A PUTREFACTIVE ANAEROBE. 49 brain, salmon, milk and suspended casein mediums not containing fermentable carbohydrates, 7.e., monosaccharides in excess. The early stages of incubation are marked by clouding and vigorous gas production. The crystals begin to appear in 4-6 days at 37° C. along with a visible liquefaction of the protein as well as odorous evidence of putrefaction. Meat and brain mediums are blackened presumably owing to the precipitation of iron sulfide by the action of sulfuretted hydrogen upon the iron freed by proteolysis. Milk, salmon and casein mediums are not blackened, although sulfuretted hydrogen is produced, except upon the addi- tion of iron ions, as by the inclusion of an iron nail. These facts fail to support a suggestion that the blackening of certain proteins by putrefactive anaérobes parallels the supposed action of a tyrosinase in transforming tyrosin into melanin in the animal body. The recovery of the crystals in a pure form involves the removal of the water-soluble constituents by washing the partially digested culture mediums with cold water, extraction of the crystals by boiling water, with or without the addition of ammonia, followed by hot filtration to remove undissolved proteins, concentration of the filtrate by boiling, crystallization from the concentrated filtrate by cooling, removal of the crystals by cold filtration and repeated clarification with animal charcoal in boiling water alter- nated with crystallization in the cooled filtrate. Impurities soluble in cold water are removed by washing at each cold filtra- tion and the crystals are dried after rinsing with 95 per cent. alcohol followed by ether. Quantitative methods of extraction are yet to be devised; there is considerable loss at each step excepting in the treatment with ether. The sample displayed, about 0.8 gram, represents the purified product from several liters of culture. The crystals are identified as tyrosine by their physical and chemical properties. To the naked eye they appear as snowy ‘white flakes with a silken sheen, the individual needles barely visible. These flakes may be readily suspended in alcohol or distilled water; in the latter particularly the characteristic crystals may be seen with a hand lens as colorless double-pointed needles. When allowed to crystallize slowly from somewhat dilute hot 50 SCIENTIFIC PROCEEDINGS (II7). water or ammonia solutions, they readily form the sheaf-like bundles so characteristic of tyrosin. They are soluble in boiling water, N/10 ammonia, and dilute mineral acids, slightly soluble in dilute acetic acid and relatively insoluble in cold water, cold and hot absolute alcohol, ether, toluene, acetone, benzine, carbon disulphide, glycerine and chloroform. They are not decomposed in aqueous solution by heating in the Arnold sterilizer at 100° C. or in the autoclave at 15 lbs. pressure for 1 hour. They give Pirie’s, Hoffmann’s and Denige’s tests.! The senior author is now engaged in perfecting the method of extraction and in studying the crystal formation of other an- aérobes. 23 (1770) A method for the preparation of cystin. By CARL L. A. SCHMIDT. [From the Department of Biochemistry and Pharmacology of the University of California, Berkeley, California.] A number of years ago Folin? described an improved method for the preparation of cystin which has come into general use. It is based on the fact that the solubility of cystin is a minimum in solutions possessing an acidity between Py 4-5. To obtain the optimum acidity for the precipitation of cystin, the HCl used to hydrolyze the protein is neutralized by the addition of sodium acetate. Although good yields of the amino acid are obtained by this method it nevertheless is not economical for the produc- tion of cystin in quantity since large amounts of relatively ex- pensive materials are required. Neutralization of HCl with sodium acetate results in the simultaneous precipitation of humin which later necessitates the repeated use of large quantities of charcoal to effect its removal. In the method described below the HCI is in large part re- covered by vacuum distillation. Use is then made of commercial finishing lime to neutralize the remaining HCl, to precipitate 1 Hammerstein-Mandel, ‘‘A Text-book of Physiological Chemistry,”’ J. Wiley & Sons, N. Y., 1912, p. 150. 2¥Folin, O., J. Biol. Chem., 1910, viii, 9-10. A METHOD FOR THE PREPARATION OF CYSTIN. 51 the humin! and to hold the cystin in solution. The advantage in the use of lime lies in the fact that it is comparatively insoluble, gives a solution of low alkalinity, thus minimizing the destruction of cystin, and is cheap. Human hair or wool which has been freed from oil by extraction with gasoline is hydrolyzed by heating at 100° C. with twice its weight of concentrated HCl. It requires about 12 hours to effect complete hydrolysis. The mixture must not be heated for any length of time beyond the point at which the biuret test is either negative or feebly positive since, as Van Slyke? has shown, cystin is destroyed during the process of hydrolyzing the protein. The greater part of the protein is removed by distilling in vacuo at a temperature between 60-70° C. and the original volume of the solution is restored by the addition of water. A thick aqueous suspension of commercial finishing lime is now slowly added, care being taken to avoid any considerable rise in temperature, until the mixture has acquired a chocolate color. It is then fil- tered by suction through a Buchner funnel and the residue washed a number of times with distilled water. The filtrate should be clear and possess a light brown color. Hydrochloric acid is now added to partially neutralize the alkaline solution and it is finally acidified by addition of acetic acid. On standing over night in the ice chest, sedimentation of the crude cystin takes place. This is filtered off and is dissolved in a minimum quantity of 5 per cent. HCl. The solution is decolorized by boiling for several minutes with a small quantity of charcoal which has been pre- viously boiled with HCl to remove the calcium phosphate, and the cystin is precipitated by the addition of sodium acetate to the hot solution until a drop of the solution ceases to turn congo red paper blue. The mixture is filtered at once and the cystin is washed a number of times with hot water to completely remove the last traces of tyrosin. Typical hexagonal plates of cystin are obtained. To compare the relative yields of cystin by the method of Folin and the above-described method, 1.3 kilos of human _ hair ~ were hydrolyzed and divided into two equal parts. The yield of 1 Hanke, M. T., and Koessler, K. K., J. Biol. Chem., 1920, xliii, 521-526. *Van Slyke, D. D., J. Biol. Chem., 1911, x, 38. 52 SCIENTIFIC PROCEEDINGS (I1I7). cystin isolated by the Folin method .was 7 per cent., while the lime method gave a yield of 6.3 per cent. 24 (1771) A globulin as the principal protein of the pecan nut: Its chemical and nutritive properties. By F. A. CAJORI (by invitation). [From the Department of Chemistry, Stanford University, California.] Pecan meal, prepared by removing the oil from the whole shelled nut, was extracted with 10 per cent. sodium chloride solution. This extract containing the proteins of the meal was subjected to fractional precipitation with ammonium sulfate and fractional coagulation by heat. The results indicate that the large part of the protein of the pecan nut is a globulin. This globulin has been isolated and purified and the distribution of its nitrogen determined by the Van Slyke method. Large amounts of basic amino-acids were found to be present in this globulin. It gives a strongly positive test for tryptophane. In general the analysis agrees fairly well with the recently published results of Dowell and Menaul} on mixed pecan proteins. Normal growth has been observed in young rats on diets in which the protein of the ration was derived from the pecan nut, | indicating that this nut furnishes adequate quantities of those nitrogenous complexes necessary for growth. In order to render pecan nut diets suitable as rations for rats, it was found necessary to remove the outer layer of the nut since this layer contains large amounts of tannin. Previous failure to observe normal growth in rats on pecan nut diets may be ascribed to the injurious or distasteful effect of the tannins that were present in those diets, and not to an inadequacy of amino-acid yield of the protein of this nut. 1 Dowell, C. T., and Menaul, P., J. Biol. Chem., 1921, xlvi, 437. Du Bots HEIGHT-WEIGHT FORMULA. 53 25 (1772) A modification of the Du Bois height-weight formula for surface areas of newborn infants. By HAROLD K. FABER and MARGARET S. MELCHER. [From the Subdivision of Pediatrics, Stanford University Medical School, San Francisco, Cal.] In 1916 Sawyer, Stone and Du Bois announced a series of measurements by which the surface area of adults and children could be estimated with an average error of 1.3 per cent. Ina subsequent paper of the same series (1916) by Du Bois and Du Bois, a new formula based on height and weight alone was pre- sented which was stated to have an error of + 5 percent. This formula is as follows: A=W* x H:5 x C, or log A = log W X .425 + log H X .725 X log C. The constant C was found to be 71.84 (log 1.857). It was not known whether the formula held for children under two years. Since the first method is based on a separate estimation of the surfaces of the extremities, head and trunk there seems to be no reason why this method should not be applicable at any age. The height-weight formula should however apparently be checked for young infants. In a series of 100 newborn babies, none over 12 days old, ranging in weight from 2,140 to 4,520 grams and in height from 45.2 to 56.9 cm., the surface area was measured by the Sawyer, Stone and Du Bois method and compared with the results obtained by the height-weight formula of Du Bois and Du Bois. Taking the former as the correct measure, we found that the latter showed a constant deviation below the former which averaged IgI sq. cm. or a mean error of — 8.6 per cent. Correcting the constant, it was found that the surface area could be computed in these infants by the height-weight formula with an average error of + 2.5 per cent. 54 SCIENTIFIC PROCEEDINGS (117). For newborn infants the corrected formula is as follows: A=W x H5 x 78.50, or log A = log W X .425 + log H X .725 + 1.895. 26 (1773) On the relation of blood-volume to the nutrition of the tissues. I. The effects of hemorrhage and intravenous injections of gum- saline on the response to the administration of a mixture of carbon dioxide and room air, and of room air alone. By ROBERT GESELL, CHARLES S. CAPP and FREDERICK S. FOOTE. [From the Department of Physiology, University of California, Berkeley, California.]} At the last meeting of the American Physiological Society some experiments were reported on the effects of hemorrhage on the response to a gradual reduction in the percentage of oxygen in the respired air. These experiments were performed on the normal unanesthetized dog connected by means of a mask with a rebreathing apparatus arranged to absorb the carbon dioxide of the expired air as the oxygen was consumed. The purpose of the experiments was to determine the detrimental effects of hemorrhage and the subsequent effects of replacing the lost blood with a gum-saline solution. | We reasoned that if a normal percentage of hemoglobin and a normal flow of blood are essential for a normal gaseous exchange, that the response of an animal to a reduction in the percentage of oxygen in the respired air would be altered by hemorrhage; and further that if the intravenous injection of gum-saline accelerated the volume-flow of blood out of proportion to the accompanying dilution, the reduced tolerance to low pressures of oxygen would be improved. To our surprise we were unable to detect, with the methods employed, any decrease in tolerance after hemorrhage amounting to 3 per cent. of the body weight. Wedo not attempt to definitely explain these results as yet, but wish to point out a striking effect — RELATION OF BLOOD-VOLUME TO NUTRITION. 55 of hemorrhage. It invariably produced a quieting effect upon the dogs accompanied by a softness of muscle indicative of a marked reduction in muscular tonus. It is possible that the decreased demand for oxygen on the part of the tissues partly explains our data. When carbon dioxide was allowed to accumulate in the re- breathing apparatus as the oxygen percentage of the respired air decreased, hemorrhage produced a definite decrease in tolerance to the combined changes in carbon dioxide and oxygen. But the restlessness of the animals under these conditions contraindicated an effort at quantitative results without anesthesia. We have recently conducted a series of experiments on the dog under morphine-ether, and morphine-urethane anesthesia. In these experiments we compared the ventilation during the administration of a mixture of carbon dioxide and room air with that of room air—first with a normal blood volume, second after hemorrhage, and third after the replacement of the blood with gum-saline. Consistent results were obtained in I0 experiments. The administration of a 5 per cent. mixture of carbon dioxide in room air invariably increased the ventilation over that of room air alone. The increase in ventilation was markedly augmented by hemorrhage. In some cases the augmentation was excessive. Subsequent injections of gum-saline reduced the response almost to that obtaining before hemorrhage. Striking augmentation of response to carbon dioxide mixtures resulted from hemorrhages amounting to I per cent. of the body weight and less. In some experiments changes in respiration were associated with the ad- ministration of carbon dioxide alone. In other experiments the ventilation of room air was increased by hemorrhage, and de- creased by injection as well. These experiments, we believe, demonstrate that hemorrhage amounting to I per cent. of the body weight or less may, under the conditions employed in these experiments, produce a detrimental effect upon the volume flow of blood, and that the injection of an inert solution improves the circulation in such a way as to enhance the gaseous exchange. . 56 SCIENTIFIC PROCEEDINGS (117). 27 (1774) . On the relation of blood-volume to the nutrition of the tissues. II. The effects of hemorrhage and subsequent injections of gum- saline upon the volume-flow of blood through the striated muscle of the dog. By ROBERT GESELL. [From the Department of Physiology, University of California, Berkeley, Caltfornia.] The effects of blood-volume upon the volume-flow of blood were studied by means of the drop method. While in some animals hemorrhage amounting to I per cent. of the body weight had a relatively small effect, in many animals the same or smaller hemorrhage markedly decreased the volume-flow of blood. Sub- sequent injections of a 6 per cent. suspension of gum in a 0.9 per cent. sodium chloride solution augmented the flow out of all proportion to the dilution entailed. 28 (1775) On the relation of blood-volume to the nutrition of the tissues. III. The effects of hemorrhage and subsequent injection of gum- saline upon the response of the sartorius muscle of the dog to rapid electrical stimulation. By ROBERT GESELL. [From the Department of Physiology, University of California, Berkeley, California.]} By comparing fatigue curves elicited by equal periods of stimulation of the sartorius muscle, we found that hemorrhage had a detrimental effect upon the endurance of the muscle, that hemorrhage amounting to I per cent. of the body weight or less may decrease the response of the muscle to stimulation. Sub- sequent injection of gum-saline decidedly improved the response of the muscle to further stimulation. WAVES OF BLOOD PRESSURE. 57 29 (1776) On the relation of blood-volume to the nutrition of the tissues. IV. The effects of hemorrhage and subsequent injection of gum- saline on total oxygen consumption. By ROBERT GESELL, CHARLES S. CAPP and FREDERICK S. FOOTE. [From the Department of Physiology, University of California, Berkeley, California.] The effects of progressive hemorrhage were studied on the dog under morphine-urethane anesthesia. We found that the greater the hemorrhage the greater the reduction in the amount of oxygen consumed and that a hemorrhage amounting to 1/2 per cent. of the body weight may elicit a decided reduction. Sub- sequent injection of gum-saline, bringing the blood volume back to normal, increased the amount of oxygen consumed. The amount of oxygen consumed immediately after an injection was greater than the consumption a few minutes later. We believe this, along with the decreased amount of oxygen consumed, points to an oxygen hunger during a period of decreased blood-volume. The results here reported are in agreement with those recently published by Doi.! 30 (1777) A comparison of the waves of blood pressure produced by slow and by rapid breathing. By ROBERT TROTTER, PHILIP EDSON and ROBERT GESELL. [From the Department of Physiology, University of California, Berkeley, California.] The effects of rapid breathing were compared with those of more normal breathing upon the systolic blood pressure in man. Supplementary data were also obtained on the dog and cat. For the well-known changes of blood pressure that occur during a single respiration, and which are more or less synchronous with ‘the changing respiratory phases, we have proposed the name of simple cardio-respiratory waves to distinguish them from those waves produced by rapid breathing. 1 Doi, Y., Journal of Physiology, 1921, lv, 249. 58 SCIENTIFIC PROCEEDINGS (117). The oscillations of pressure elicited during rapid breathing by the interference method we have designated as cardio-respiratory interference waves. The most striking difference in the respiratory relations of the simple and interference waves is that in the simple waves the blood pressure changes are complete within the period of a single respiration, while in the interference waves the gamut of the blood pressure changes is run through in the interval of several respirations. The production of interference waves of blood pressure is de- pendent upon the establishment of cardio-respiratory cycles in which the number of respirations is greater by one or lesser by one than the number of heart beats making up the waves and occurring in the same time interval. When these conditions are fulfilled we may conceive of the heart beats as moving through respiration, the direction of the movement being determined by the relative rates of the heart and respiration, that is, whether the respiratory rate is slower or faster than the heart rate. A cardio-respiratory cycle is complete when two beats (the first and last of the interference wave) fall at ap- proximately the same time in respiration. Whereas in the simple respiratory waves we found the biahinet pressure to be associated with approximately the beginning of inspiration, in the cardio-respiratory interference waves we found the highest pressure to obtain at approximately the beginning of expiration. Without definitely assigning the responsibility for the pro- duction of interference waves to any particular respiratory factor, we are inclined to favor the hypothesis that they are primarily due to the changing intra-thoracic pressure accompanying respira- tion. It is possible by breathing slightly slower or slightly faster than half the heart rate to produce double interference waves of blood pressure, that is, under these conditions, two waves of blood pressure may be in progress simultaneously. Each of the double waves is formed by alternate heart beats, one being made up of the even numbered and the other of the odd numbered beats. Double cardio-respiratory interference waves are to be explained in the same manner as the single waves. EXPERIMENTAL RICKETS. 59 The results of these experiments on the dog and cat are in ~ agreement with those obtained in man. Cardio-respiratory interference waves particularly of the double type with alternating beats occur simultaneously in sacri- fice experiments in dogs and cats. We therefore point to our work as occasionally explaining pulsus-alternans and blood pressure waves of the third order. ABSTRACTS OF THE COMMUNICATIONS, MINNESOTA BRANCH. First meeting. Minneapolis, Minnesota, October 12, 1921. 31 (1778) Experimental rickets. By J. F. McCLENDON and HARRY BAUGUESS (by invitation). [From the Laboratory of Physiological Chemistry, University of Minnesota, Minneapolis, Minnesota.| In a large number of experiments on feeding albino rats in which white wheat flour was used as the main part of the ration, bone abnormalities were absent or transitory. Since Sherman showed that rickets could be inhibited by simply adding phosphate to the diet we concluded that the normal condition of the bones of our rats was due to the phosphoric acid content of the casein used in the diet. After substituting casein by lactalbumin or edestin, bone abnormalities appeared in one hundred per cent. of our rats. If casein was fed to the extent of 6 per cent. of the ration bone abnormalities were reduced and apparently the dis- turbance was transitory, since the bones became hard and cast dense shadows with the x-rays without changing the diet, yet some of the deformities were preserved. With a basic ration of white flour containing 6 per cent. sea salt, the addition of edestin or lactalbumin to improve the protein did not in any way decrease the abnormalities. The edestin carries vitamine B and if wheat germ extract is added to the lactalbumin in order to furnish vitamine B, the abnormalities still persist. The addition of spinach up to 5 per cent. of the ration did not decrease the ab- 60 SCIENTIFIC PROCEEDINGS (117). normalities, whereas 0.2 per cent. furnished sufficient vitamine A to keep the animal alive for 3 or more months. The addition of 0.5 gram of butter fat per day did not lessen the abnormalities. In order to determine the growth of the bones x-ray plates were made by photographing a large number of animals on the same plate and comparing the density of the shadows of the bones. In addition, determinations of calcium intake and output and calculations of the calcium retention were made. Considerable individual variation was found, but when long metabolism periods (about a week) were used and all our animals with abnormal bones were — averaged we found that rats which were weaned and placed on the diet at the age of twenty-one days and left on this diet at least three days before commencing the metabolism study showed a re- tention of 2.7 milligrams of calcium per rat per day for the first two weeks, 1.7 for the third week and 1.5 for the fourth week. In some individual rats shortly before death we obtained negative calcium balances. It would indicate, therefore, that the dis- turbance of calcium metabolism is increasing in severity from the first to the fourth week. We do not know the average calcium retention of the normal rat but we assume that it is close to 5 milligrams per day for the ages corresponding to our rats. This is based on calcium content of whole rats. It seems probable, therefore, that calcium retention can be used throughout the course of the disorder as an index of the severity of the disease, provided adequate methods are used to determine the calcium retention. In order to avoid errors due to transfer of the excreta we have made round cages with quarter-inch-mesh wire screen bottoms that sit in silica dishes six inches in diameter and have two bird-feed cups attached, one for water and one for food. Any food spilled from the container into the dish does not cause an error because it does not affect the difference between the intake and the outgo of calcium. At the end of the metabolism period the cage is lifted from the dish and the excreta ashed in the dish. In order to separate the calcium from the dissolved ash as calcium oxalate we have used a bromphenol blue as an indicator for hydrogen ion concentration. If the solution which is acid is neutralized until it reaches Py, = 4 the calcium oxalate will not be appreciably soluble and yet no calcium will precipitate as EXPERIMENTAL RICKETS. 61 phosphate. In case of adding too much alkali it is best to use acetic acid to bring the solution back to Py = 4, because the danger of overstepping the end point is very small, since the mixture of sodium acetate and acetic acid can vary considerably in composition with little change in the hydrogen ion concentra- tion. This is McCrudden’s method in principle, but the indicator makes us more certain of the Py. The calcium oxalate was titrated with potassium permanganate. We do not wish to discuss the diagnosis of rickets, although our animals showed the same appearances as those described in the papers of Sherman, McCollum and Hess. Dr. C. M. Jackson has very kindly offered to work out the morphological changes in great detail and publish them so that they will be available. From a practical standpoint, however, types of non-rachitic osteoporosis caused by calcium deficiency do not seem to be very common among human beings. Bone abnormalities which are possible on human diets at present in use deserve considerable study whether they are called rickets or not and we use the word rickets merely for convenience. Some of our rats getting more phosphoric acid than that contained in the wheat flour seemed to recover from the disturbance of metabolism just as infants may recover from rickets, with the reservation that the diets of our rats were unchanged in percentage-composition and only changed in the quantity eaten per day, whereas very little exact data is to be had on the diets of human beings. In case the abnormalities are not great enough to cause permanent deformities we do not know of any means of detecting the previous history of rickets without diagnosing it at the time of its occurrence. The x-rays and calcium balances, however, may be used for diagnostic purposes on rats without killing the animals. Rats die easily under ether and it is difficult to get them absolutely quiet without danger of killing them unless they are held mechanically. They may be stretched out by tying their feet to a stiff ring of suitable size after slightly etherizing them. 62 SCIENTIFIC PROCEEDINGS (117). 32 (1779) Rapid determination of surface tension. By ROBERT G. GREEN (by invitation). [From the Department of Bacteriology, University of Minnesota, Minneapolis, Minn.| An apparatus was demonstrated by means of which the surface tension of a liquid is rapidly determined by the drop-weight method. From one to six drops of the liquid to be measured is | required. The apparatus consists essentially of a delicate torsion wire balance and an adjustable scale on which the surface tension is read in dynes per centimeter. 33 (1780) The influence of the surface tension of the culture medium on bacterial growth. By W. P. LARSON. [From the Department of Bacteriology, University of Minnesota, Minneapolis, Minn.] Pellicle-forming bacteria such as the B. tuberculosis, B. subtilis and others of that group which habitually grow upon the surface of liquid medium, will grow throughout the body of the medium by depressing its surface tension from 59 dynes, the S. tension of ordinary broth, to 40-45 dynes. By analogy with the floating needle experiment it may be assumed that when the pellicle-form- ing bacteria are properly wetted they no longer grow upon the surface of the medium but throughout the body of the broth or even at the bottom of the flask. The further observation has been made that the B. subtilis and B. anthracis, when grown in media of low S. tension, finally be- come asporogenous. Cultures of B. anthracis grown under such conditions and sterilized by heat at 60° for 30 minutes protect guinea pigs. The enhanced wetting of the bacteria brought about by the addition of soap probably creates better nutritive condi- tions which cause the organisms to grow without forming spores. Castor oil soap when in aqueous solution is perfectly clear, does DETERMINATION OF DISSOLVED OXYGEN. 63 not hydrolyze as readily as most other soaps, and has therefore been used extensively in our experiments. It is more toxic to some bacteria than potassium or sodium stearate. ‘This is prob- ably due to the fact that castor oil soap is dialyzable and probably dialyzes into the cell and disturbs the salt balance by precipitating the calcium, magnesium and salts of the heavy metals. Bacteria such as the pneumococcus and streptococcus will not grow on low tension media, while the organisms which inhabit the intestinal tract grow abundantly on media of low tension. This is not surprising since the contents of the intestines have a low S. tension due to the presence of bile, soaps and other S. tension depressants. It is well known that many of the intestinal bacteria when inoculated in broth grow near the surface of the medium. This is particularly true of the cholera vibrio. This selective localization is probably due to the fact that the S. tension reducing substances concentrate at the surface of the medium thus creating a favorable environment for these bacteria. Incidentally it may be pointed out that the bacteria which grow well in low tension media are better antigens than the streptococcus, pneumococcus and others which refuse to grow under such conditions. 34 (1781) A micro-Winkler method for the quantitative determination of dissolved oxygen. By E. J. LUND. [From the Department of Animal Biology, University of Minnesota, Minneapolis, Minn.| Winkler’s method for quantitative determination of dissolved oxygen may be applied to Io c.c. or even 5 c.c. samples of water in the following way. One tenth of a cubic centimeter of each of the two solutions MnCl, and NaOH — KI are added from 1 c.c. burettes graduated to 0.1 c.c. or less. The thiosulfate solution of the usual concentration is diluted to ten times its volume. The iodine is titrated in a tall dish using a 5 c.c. burette. The end joint is just as definite as that in the ordinary procedure. The percentage error is also the same, about I per cent. The distinct 64 SCIENTIFIC PROCEEDINGS (II7). advantage in the micro method lies in the possibility of greatly shortening the duration of the tests, thus making it possible to follow the time course of respiratory exchange over relatively short periods of time. Owing to the small volumes used, temperature adjustment is rapid. With good manipulation the maximum error is less than 0.005 c.c. of O2 gas. The method is being used in studies on oxygen consumption by small organisms such as protozoa, eggs and certain kinds of tissues. 35 (1782) Does the introduction of an ethoxy group into aromatic com- pounds increase their bactericidal action upon the pneumococcus and the gonococcus? ! By ARTHUR D. HIRSCHFELDER and L. J. PANKOW. [From the Department of Pharmacology of the University of : Minnesota, Minneapolis, Minn.| Morgenroth and his collaborators have shown that when an ethoxy group is substituted for the methoxy group in quinine derivatives and ethylhydrocuprein is produced, the substance takes on markedly increased pneumococcicidal action in vitro and in vivo. Solis Cohen, Kolmer and Heist found that ethylhydrocu- prein hydrochloride was from eight to twenty times as strong an antiseptic for the pneumococcus as quinine hydrochloride. Mor- genroth and Levy had shown that no such difference between quinine and ethyl hydrocuprein could be observed in the case of the streptococcus. We find that when cultures of gonococcus are exposed to starch bouillon containing quinine hydrochloride or ethylhydrocuprein in 1/10,000 dilution and then transferred to plates of rabbit’s blood agar, growth occurs if the exposure to the drug has lasted only ten minutes but the bacteria are killed if the exposure has lasted thirty minutes. Ethylhydrocuprein has therefore no specific action against the gonococcus. However, as ethylhydrocuprein is too toxic for successful use in the chem- 1 The investigations recorded in this paper were rendered possible by a grant of funds granted by the United States Interdepartmental Social Hygiene Board, for the discovery of better medical measures for the prevention and treatment of the venereal diseases. BACTERIAL ACTION OF AROMATIC COMPOUNDS. 65 otherapy of lobar pnuemonia, substances which are less toxic must be sought for. The first question to be determined is whether pneumo- coccicidal properties are common to ethoxy compounds in general or whether this property is peculiar to ethylhydrocuprein and its closely related compounds. We have accordingly tested the bac- tericidal action of the ethyl ethers of various aromatic compounds, and compared them with the corresponding hydroxy compounds. In making the tests a suspension of pneumococcus type I. from the Rockefeller Institute was suspended for the desired interval in a broth solution of 0.9 per cent. NaCl containing the substance whose action was to be determined; and after the desired interval a loopful of this stroked across a rabbit’s blood agar plate and in- cubated 24 hours. In the experiments with the gonococcus a strain was used which had been isolated from a case of typical anterior gonorrhoeal urethritis in the Outpatient Service of the Genito Urinary Division of the University of Minnesota, and which had been grown in Vedder’s starch bouillon and on starch bouillon agar and rabbits’ blood agar. Tests were made in the same way as for pneumococcus except that the drug was mixed with starch bouillon. The cultures were then transferred to rabbits’ blood agar and incubated 24 hours. The following substances were tested, + indicating growth of the cocci after being exposed to the drug for the period in- dicated ,—indicating that the cocci did not grow. OH /~\ COONa . 1. Sodium salycilate 5% the Pneumococcus { + after 10 min. — after 60 min. \f OC2H; \, COONa 2. Sodium ethylsalicylate! 5% Pneumo + after 60 min. 1 Substances Nos. 2, 4, 10, II, 12, 13 were prepared by Mr. Merrill C. Hart, chemical assistant in the department of pharmacology, University of Minnesota, and No. 9 was kindly furnished by Prof. Roger Adams, University of Illinois, to both of whom we extend our thanks. 66 Io. Il. SCIENTIFIC PROCEEDINGS (II7). OH Me Sodium phenolsulphonate 5% Pneumo + after 2 hours. 1% Gono + after 30 min. SO:0Na OC:H; . Potassium phenetolsulphonate 5% Pneumo + after 2 hours. 1% Gono + after 30 min. SO:0K OH . Para aminophenol 1% Pneumo + sometimes — sometimes after 30 min. I:1000 Gono — in 10 min. NH: OC2H; . Para phenetidin 1% Pneumo + sometimes — sometimes after 30 min. 1% Gono — after 10 min. NH; ~ OH . Para nitrophenol 1% Pneumo + after 10 min. — after 30 min. 1: 500 Gono + after 10 min. — after 30 min. NOs OC:Hs I: 1000 Gono + after 60 min. . Para nitrophenetol ) 1: 1000 (sat. sol.) Pneumo + after 60 min. NO: OC:Hs5 . Phenetidinethylalcohol Pneumo + after 30 min. I: 250 Gono + after 30 min. NHCH:CH:0H OH \.CH;0H Saligenin 2% (in serum) Pneumo -— after 60 min. 2% Gono + after 30 min. a! — after 60 min. OC:Hs CH:0H Saligenin ethyl ether Sat. sol. in 0.9% NaCl (about 1: 10,000). Pneumo. Results vary. 8 repetitions — after 5 min. 16 repetitions + after 30 min. BACTERIAL ACTION OF AROMATIC COMPOUNDS. 67 OC;Hs / \ CH:0H 12. Saligenin iso amylether | | Sat. sol. less than 1: 10000. 2. Pneumo — after 5 min. ra Gono — after 10 min. Staphylococcus and Bacillus coli both + after 30 min. OOCCH; CH:20H 13. Acetyl saligenin I: 200 Pneumo — after 5 min. : Gono — after 10 min. From the above-recorded experiments it is evident that in the simpler aromatic substances, predominantly water-soluble like sodium phenolsulphonate and salicylate or predominantly lipoid- soluble like para amino phenol and para nitrophenol, whether nitrogen-free or containing nitrogen, the introduction of an ethyl group upon the ring does not confer pneumococcicidal or gonococci- cidal powers. Whenever any difference is noted the hydroxy compound is a somewhat stronger antiseptic than the ethoxy. There is therefore no analogy in this regard between the simpler aromatic compounds and the quinine derivatives. SCIENTIFIC PROCEEDINGS. ABSTRACTS OF COMMUNICATIONS. One hundred eighteenth meeting. New York Post-Graduate Medical School, November 16, 1921. President Wallace in the chair. 36 (1783) Alcohol and white rats: a study of fertility. By E. CARLETON MACDOWELL. [From the Station for Experimental Evolution, Cold Spring Harbor, oe a This paper deals with the effect of alcohol fumes upon the size and number of litters produced by white rats and their descendants. Details of the administration of the alcohol have been published. The treatment was given from the age of 28 days through the lives of the rats, with the exception of the females on the 28 days fol- lowing the birth of a litter. After mild initial doses, each rat was left in the fume tank each day until it was completely anesthetized. Brothers and sisters of the treated males and females were used as controls. The matings were all between treated males and treated females or their descendants, and between the controls. Each group of test matings in each generation had its own control group raised at the same time and under the same conditions of environ- ment. The data came from four main groups of rats: those treated, their treated children, their untreated children, and their untreated grand children from the untreated children. Size of Litters —The average size of all the litters produced by the treated rats was 10 per cent. less than the control average. Nine pairs of treated offspring from these treated rats gave litters ‘that were 10.3 per cent. smaller than their control litters. Ten pairs of untreated rats from the treated parents gave litters that were II.2 per cent. smaller than the controls. And eleven pairs 1 MacDowell and Vicari, Jour. Exp. Zo6l., 1921, xxxiii, 209. 69 70 SCIENTIFIC PROCEEDINGS (118). of untreated rats from untreated parents and treated grandparents gave litters that were 13.3 per cent. smaller than their control litters. So there appears to be a reduction in the average litter size in each generation that is about ten per cent. of the respective controls. However, in no case is the reduction statistically signifi- cant. This is probably due to the small size of each sample, since the combination of all the generations gives a difference that is 3.6 times its probable error and so is to be considered significant. Alcohol appears to have caused a modification in litter size that persists for two generations after the original treatment and is not increased by a second generation of treatment. This is the result to be expected from a definite germinal modification. Number of Litters —The numbers of litters are based upon the production during equal periods of the test pairs and their own particular control pairs. In some cases the period was longer than in others, but opportunities for the tests and controls to produce litters were equal in each case and therefore equal in the totals. The number of litters is, accordingly, purely relative. 44 treated pairs produced 32 litters, whereas on the basis of their controls 91 litters were expected. This was a reduction of 65 per cent. + 3.37 which is 19.2 times its probable error and so, significant beyond all question. Treated rats from treated parents produced 14 litters when 22 were expected on the basis of their controls. This is a reduction of 35 per cent. + 6.91 and is 5 times its prob- able error. Untreated rats from treated parents produced 33 per cent. + 8.20 more litters than the controls, and the untreated rats from untreated parents and treated grandparents 55 per cent. + 8.4 more litters than the expectation. Both these differences are fully significant. The number of litters was strongly reduced when the rats themselves were treated, but, just as soon as the alcohol was further away, the number of litters at once increased and the test animals produced significantly more litters than their controls. The obvious interpretation of this result is that alcohol has acted as a selective agent by preventing those females from having litters that bore weaker determinants for the production of litters. This accounts for the apparently odd fact that two generations of treatment made less difference than a single genera- tion of treatment. The offspring of treated animals are a selected EXPERIMENTS WITH B. ENTERITIDIS (MURIUM). 71 lot. Genetically they have higher litter-producing powers than the controls; when alcohol is given to them it causes a reduction in the number of their litters, but this reduction is half as great as the reduction caused by the treatment of their parents which were genetically equal to the controls. The alcohol has sorted out differences already present. This is a very different result from that given by litter size, which demands the assumption of alcohol modifications. It is to be concluded, then, that alcohol works upon the size of litters and the number of litters through different channels. This leads to two generalizations: first, that fertility is a complicated character whose different measures are not all manifestations of the same factors; second, that the action of alcohol upon animals is very complicated; it may act through different channels and in differ- ent ways, so that the end results in any special case are due to the interaction of different tendencies. Students of experimental alcoholism must recognize the complex nature of their problem, and, leaving behind the familiar method of generalizing from end results, focus attention upon the problem of the channels through which alcohol may work. 37 (1784) Experiments with B. enteritidis (murium)! on normal. and immune mice. By LESLIE T. WEBSTER. [From the Laboratories of the Rockefeller Institute for Medical Research, New York City.] These experiments were undertaken to ascertain varieties and degrees of resistance of normal and immune mice to fixed doses of B. enteritidis (murium). 1. If live cultures of this organism are injected intrapleurally or intraperitoneally into normal mice, there occurs an initial lag in the rate of bacterial multiplication lasting a few hours followed by a rapid and continued acceleration of growth until the death of the animal. If live cultures of this organism are given per os to normal mice, there occurs an incubation period of 5-6 days, after which the 1A serological and cultural description of this organism will appear in the Journal of Experimental Medicine. 72 SCIENTIFIC PROCEEDINGS (118). animal usually develops symptoms of disease and succumbs. A small percentage of mice, however, prove refractory to infection by this route. | 2. If live cultures of this organism are injected intrapleurally or intraperitoneally into mice previously ‘‘ vaccinated”’ intrapleurally or intraperitoneally, they are partially destroyed and held in check by the protective mechanisms of the animal body for two or three days. Subsequently, the rate of bacterial multiplication increases gradually until the death of the animal. The partial protection following this type of vaccination is entirely of a general nature; no evidence of a ‘‘local immunity”’ has been obtained. Mice given I,2, or 3 subcutaneous doses of this organism vaccine show a similar relative increase in resistance to the subsequent injection of live organisms er os as intraperitoneally. 3. Feeding mice live or killed cultures of this organism induces a definite protection against subsequent intrastomachal and intra- peritoneal injections of live organisms. ‘The immunity developed in this way is also of a “‘general’’ as opposed to a “‘local’’ nature. 38 (1785) Therapeutic application of Bacillus acidophilus. By LEO F. RETTGER and HARRY A. CHEPLIN. [From the Bacteriological Department, Yale University, New Haven, Conn.]} In previous communications to the Society (1920 and 1921) we stated that Bacillus acidophilus may be implanted in the human intestine by the oral administration of (1) minimum quantities of lactose or dextrin, (2) whey broth cultures of B. acidophilus, or (3) a combination of lactose and the acidophilus culture in which the amounts of each are cut in half. Early in 1921 the milk culture of this aciduric organism was substituted for the lactose- and whey broth cultures and subsequent implantation experiments have been carried on with the acidophilus milk. In the work on pathological cases we received the friendly coéperation of practicing physicians, who not only supplied us with many of the most interesting subjects, but who furnished us THERAPEUTIC APPLICATION OF BACILLUS ACIDOPHILUS. 73 with a history of the individual cases, and in two instances with X-ray photographs. The subjects comprising the first group under observation may be divided for convenience into the follow- ing classes: Chronic constipation with the symptoms of so-called autointoxication and other pathological conditions, some of them acute............... ee cee eevee 20 Chronic diarrhea following an attack of bacillary dysentery... 0d. ecu 2 Colitis, at times DGG Y, ARG Mote OF TESS MUCOUS i. case ct fc oe pula ees cee Bence 2 ae Pace Setar pelt tele jain flats sata ollie rad i ark pea ae a 2 RIN a CEPA CI: boule fae cs Pe ert cers wb wellnahe ae Rb se bles desea Fn ees 3 These 29 cases are exclusive of those which have come under our observation within the past two months; nor do they include those which have been and are being studied in other institutions through our codperation. Brief reference will be made to only a few of these 29 cases. The subjects were instructed to bring to the laboratory one or two specimens of stool before taking the acidophilus treatment, and for a while daily samples, when procurable, after the first use of the acidophilus milk or of the milk plus stated amounts of lactose. Bacteriological examinations were made of these speci- mens and the results correlated with the clinical findings. Persons who had been afflicted with chronic constipation usually received one quart of the acidophilus milk plus 100 grams of lactose daily, the powdered lactose being added to the milk in the flask and the contents thoroughly shaken. The subjects were instructed to take the daily supply in three equal portions, and as nearly as possible two hours before or after meals. This schedule was at times: varied to suit the particular needs of the cases. There were no restrictions as to diet, but the subjects were urged to refrain from the use of food which by experience or training they knew to be harmful. In all of the diarrheal cases (including colitis and sprue) the treatment consisted in the daily administration of from 500 to 1,000 c.c. of the acidophilus milk without added milk sugar. The milk was well tolerated by the patients. CHRONIC CONSTIPATION. The first two cases, which had a long history of most obstinate constipation and in whom the symptoms usually accompanying 74 SCIENTIFIC PROCEEDINGS (118). such condition were of the most aggravated type, responded within the course of less than a week to the use of one quart of acidophilus milk daily without any added lactose. A very close correlation between the clinical and bacteriological results was established. Two other subjects who for many years had had marked enteroptosis and a condition approaching at times intes- tinal stasis yielded readily to the administration of one quart of the milk plus 100 grams of lactose, and after the first few days required only 500 c.c. of the milk with as little as 25 to 50 grams of added milk sugar daily. wha ae iN 22.8 16.8 13.8 21.2 Second month......... are ee ore 45.1 31.6 26.7 19.4 Es WHOMEVER Gn een ea Dee 24.5 20.4 15-5 14.6 PeSIO Ry: SONI sh icned es aeienaes 21.1 17.2 13.5 II.4 UREDD MOU sinc ac os oh c Cod baa sea 16.2 13.6 — 8.9 ESR SOOOER Fi iis tal Sale pce eee 14.4 _ —_ 5.8 Sevemetl Mant. 5s i Sicecle eees'ce vas 11.6 —_ — 5.9 ee WOE ses 4 PG SO rer G eee 7.4 _ —r 3-9 ee EN |, ca be A tariein ete dita 2 5.1 —_— _ 4.0 * Average of ten large published series of observations. WEIGHT INCREMENTS OF PREMATURE INFANTS. 135 The increments thus determined were next compared with those of fetuses of the same gestation age and with those of full- term newborn children. The norm for fetal growth in weight was estimated from the following empirical formule: (1) Y = 0.24 X*? + 400, where Y is the body-weight in grams and X is the body length in cm., and (2) Y = 8.9 (X — 1) — 0.27 (X — 1)? — 6.5, where Y is the body length in cm. and X is the age in fetal or lunar months. The norm for weight increment in the first nine postnatal months was determined by calculation from 1o large series of published aver- ages on the increase in body weight in the first year. When the monthly weight increment rates of the premature infants are compared with these norms it is found that they fall much closer to the calculated rates of growth of fetuses of the same size and age than to those of newborn children. This is shown particularly well by group A of the smallest prematures which were approxi- mately 7 fetal months old when born. The comparison is shown graphically in the chart below. 80 % TO 60 50 Ist 2nd 3rd Ath 5th 6th Tih 8th 9th mo. Fic. 1. A graph showing monthly percentage increment in weight of a group of premature infants ranging from 1.0 to 1.5 kg. in birth weight. The solid line represents the rate of increment of the premature children in the first 9 months after birth. The upper broken line represents the calculated rate of increment of the fetuses of the same size and age as the prematures. The lower broken line represents the rate of increment of full-term children. 136 SCIENTIFIC PROCEEDINGS (119). These results indicate that premature children, after a short period of retarded growth incident to the adjustment to the extrauterine environment, tend to regain the fetal rate of growth and to follow this course of growth until some time in the latter part of the first year when the rates of fetal and postnatal growth approximate one another. In other words the growth tendency of prematures is in general that of fetuses of the same size and age rather than that of full-term children. These results are in agreement with those of Hammett! on growth capacity and body weight in the first two weeks of postnatal life, and with certain findings of Cammerer,? but seem to be in opposition to some of the conclusions of Schwarz and Kohn? and of Ylppé.4 That this opposition is apparent rather than real will be shown in a later communication. 68 (1815) An undetermined principle obtained from poison ivy. By E. D. BROWN. [From the Department of Pharmacology, University of Minnesota, Minneapolis, Minn.] An undetermined principle has been obtained from poison ivy which so far as I have been able to find differs in its behavior from that of any substance previously described. It came down as a precipitate after long standing of a filtrate after precipitating with lead acetate. No work has as yet been done with the substance except to determine a few of its properties. It is non-irritant when applied to the skin, neutral to litmus and is bitter to the taste. It has a melting point of 190° when heated slowly, insoluble in cold water and fairly soluble on boiling, imparting a lemon yellow color to the solution. It is soluble in ammonia water, acetic acid and hot alcohol. Insoluble in alcohol in the cold, ether, chloroform, petroleum ether and acetone. It 1 Hammett, F. S., Amer. Jour. Physiol., 1919, xlv, 396. 2 Cammerer, W., Jahrb. f. Kinderheilk., 1900, liii, 381. * Schwarz and Kohn, J. L., Amer. Jour. Dis. Children, 1921, 296. ‘Yippd, Zeitschr. f. Kinderheilk., 1919, xxiv, 179. EFFECT OF HEAT ON Cow’s MILK. 137 gives striking color reactions with H:SO,, HNO;, KOH, CaOH and other reagents. f A solution to which a few drops of silver nitrate are added gives a pink color changing quickly to a dull green. Iodine as Lugol’s solution gives a pink color which quickly fades. | With ferric chloride a very dark blue or black changing to brown. On the addition of Fehling’s solution it turns green and on boiling a slight reduction occurs. It does not reduce Fehling’s solution in the cold on long stand- ing even after it had been previously boiled with acid and again rendered neutral. This leads to doubt as to its being a glucoside. With Millon’s reagent it turns a port-wine color rapidly be- coming darker which suggest the possibility of its belonging in the group of the phenols. 69 (1816) The effect of heat on the calcium salts and rennet coagulability of cow’s milk.* By LEROY S. PALMER. [From the Section of Dairy Chemistry, Division of Agricultural Biochemistry, University of Minnesota, St. Paul, Minn.] When milk is boiled a precipitation of a portion of the calcium phosphates occurs, the amount of fixation being proportional in general to the amount and duration of heat applied. Soldner? first called attention to this fact and his observations have been confirmed by numerous investigators, among whom may be men- tioned Boekhout and de Vries,? Purvis, Brehaut and McHattie,? and Grosser.‘ It has been commonly believed, also, that some fixation of the * Published with the approval of the Director as Paper No. 288, Journal Series, Minnesota Agricultural Experiment Station. ~ 1S6ldner, F., Landw. Versuchs., 1888, xxxv, 351. 2 Boekhout, F. W. J., and de Vries, J. J. O., Landw. Versuchs., 1901, lv, 221. ’ Purvis, J. E., Brehaut, A. H., and McHattie, A. C. N., Roy. Sanit. Inst. Journ. Trans., 1912, xxxiii, 154. 4 Grosser, Paul, Biochem. Zeiischr., 1913, xlviii, 427. 138 SCIENTIFIC PROCEEDINGS (I19Q). calcium phosphates takes place during the holding process of pasteurization.” The fact that pasteurization of milk retards the coagulability of the casein by rennet and the fact that this prop- erty can be restored by the addition of calcium chloride to the milk have been presented in support of the view that heat changes some of the soluble calcium salts to an insoluble form. The experi- mental evidence for such a change is, however, contradictory. Solomin! noted that a little phosphorus falls out of milk when the temperature is raised to 80° C. Diffloth? found a decrease of 26 per cent. in the soluble phosphates when milk was held at 60° C. for 30 minutes. Rupp,’ however, approached the problem by filtering raw and pasteurized milk through a clay filter and analyzing the filtrate for calcium and phosphorus. He found no change in the calcium and phosphorus after holding the milk for 30 minutes at 68.3° C. Milroy* held the fresh milk at a tempera- ture just below the boiling point for one hour and, after filtering through an ordinary filter, noted a decline in calcium. He ex- plained this result on the basis of a transformation of dicalcium phosphate into basic calcium phosphate. Grosser made similar studies on samples of milk which had been boiled for 5, 10 and I5 minutes, respectively, and noted a negligible loss in phosphor- us in the filtrate, but a slight loss in calcium. Daniels and Loughlin® have recently obtained qualitative evi- dence that calcium phosphates are thrown down when milk is pasteurized by the holding process, in that they have noted a nutritional calcium and phosphorus deficiency of such milk which could be prevented by feeding the washings from the walls of the vessel in which the milk was pasteurized or by addition of calcium phosphate to the rations of the animals (rats) in the experiments. The explanation commonly held for the effect of heat on the calcium phosphates of milk originated with Sdldner who believed that the calcium of milk is present as mono- and dicalcium phos- 1Solomin, P., Arch. f. Hyg., 1897, xxviii, 43. 2 Diffloth, Paul, Bull. d. Sci. Pharm., 1904, x, 273; Zeit. Nahr. Genussm., 1906, xi, 455. ’ Rupp, Philip, U. S. Dept. of Agr., Bureau of Animal Ind. Bull., 1913, clxvi, I-16. 4Milroy, T. H., Biochem. Jr., 1915, ix, 221. 5 Daniels, A. L. and Loughlin, R. J., Biol. Chem., 1920, xliv, 381. EFFECT OF HEAT ON Cow’s MILK. 139 phates which, on heating, pass to tricalcium phosphate and are thus precipitated. This explanation has apparently never been submitted to critical examination. Monocalcium phosphate is readily soluble in water and its solutions decompose on boiling giv- ing rise to dicalcium phosphate whose solubility is so low that a heavy precipitation of phosphates occurs. The solubility of di- calcium phosphate is, however, only 0.135 to 0.561 part per 1000 of water, depending on the saturation of the water with CO. The more highly concentrated solution naturally gives up some of its calcium phosphate on heating, due to the loss of COz. The solution in pure water also clouds up on boiling. The facts just cited seem to support, in general, Sdldner’s theory. However, the experimental results of Grosser* and Rupp,’ cited above, show that there is actually little if any decrease in the calcium phosphates dissolved in milk when the milk is held at pasteurization temperatures or boiled for some minutes. At the same time there is abundant evidence, as indicated, that heat does precipitate calcium phosphates from milk. How are these divergencies in results to be explained? It occurred to the writer that a simple explanation of these divergencies is afforded by the experimental evidence brought forth by Van Slyke and Bosworth! that the calcium phosphate of cow’s milk is wholly in the form of dicalcium phosphate, amount- ing to about 1.75 parts per 1000, on the average. These figures are greatly in excess of the maximum solubility of dicalcium phosphate, even in water saturated with CO,. These investi- gators found, moreover, that the dicalcium phosphate of cow’s milk was retained on the Pasteur-Chamberland filter when milk is filtered through this medium. The natural conclusion to be drawn from these results is that the calcium phosphate of cow’s milk, which appears to be wholly in the form of CaHP0Os, is present in colloidal solution, and that the aggregates of particles are sufficiently large that they do not pass through the Pasteur- Chamberland filter, or even through the Bechloidt filter used by Grosser, or the clay filter used by Rupp. This conclusion coin- cides with the results of Grosser and Rupp who obviously were 1'Van Slyke, L. L., and Bosworth, A. W., N. Y. Agr. Exp. Sta. Tech. Bull., IQ14, xxxix, I-17. 140 SCIENTIFIC PROCEEDINGS (119). not dealing with the colloidal matter of milk in their analyses of filtrates from the clay filters which they used. The experiments which have shown a gross decline in calcium phosphates or in which the precipitated phosphates have been seen, when milk is heated, are to be explained, therefore, solely by the effects of heat on colloidal solutions of dicalcium phosphate. As a matter of fact the loss of calcium and phosphorus from milk on boiling observed by Séldner showed a ratio of one molecule of calcium to one of phosphorus such as exists in dicalcium phosphate. EXPERIMENTAL. In order to determine what the effect of heat is on colloidal CaHPO, solutions, such a solution was prepared by grinding CaHPO,, which had been washed free from electrolytes, to an impalpable powder in a porcelain ball mill. This powder was then ground further in the mill in the presence of a 0.6 per cent. gelatin solution. After settling, the supernatant fluid presented a very satisfactory colloidal suspension of CaHPO,. It was dis- tinctly milky and showed the usual Tyndall effects in a striking manner. ‘The concentration of CaHPO, was not, however, as high as had been expected, the solution being found to contain only 0.542 gram per 1000. Possibly a higher concentration would have been obtained if a stronger solution of gelatin had been employed or a better colloid stabilizer used. Gelatin was chosen, however, because it is not coagulated by heat. The effect of heat on this colloidal solution of CaHPO, was determined qualitatively only by heating a portion of it in a water bath at 63° C. for 30 minutes. A heavy precipitation of CaHPO, resulted and the filtrate showed much less evidence of a colooidal suspension. This simple experiment shows rather conclusively that it is not necessary to assume any transformation in the composition of the calcium phosphates of milk during heating to account for the partial fixation of these salts. The phenomenon is readily accounted for by the effect of heat on a colloidal solution of CaHPO, which renders such a solution much less stable and causes the aggregates to pass, in part at least, to the crystalloid form. EFFECT OF HEAT ON Cow’s MILK. 141 The results secured in this experiment have a bearing, also, on the alleged effect of heat on the calcium salts of milk as affecting the coagulability of milk by rennet. It seems evident that the only calcium salts affected by heat are colloidal calcium salts and the question is therefore raised as to the possibility of the colloidal CaHPO, of milk playing a part in the rennet coagulability. In order to determine whether this is true or not, two 200 c.c. portions of fresh whole milk were dialyzed in collodion bags against running distilled water for 48 hours, using I per cent. toluene as preservative. When rennet was added to this milk there was no coagulation even after several hours. One drop of 4 molar CaCl, solution added to 100 c.c. of the rennet treated milk caused instant coagulation. The same result followed the addition of 2 or 3 drops of dilute HCl solution. The addition of 10 c.c. of the colloidal gelatin solution of CaHPOx..to 100 c.c. of the rennet treatment milk was, however, without any effect. It is apparent that the colloidal CaHPOs, of the milk does not play any part in the rennet coagulation. The indications are, also, that the effects of heat on rennet coagulation which can apparently be overcome by the addition of soluble calcium salts are not to be explained on the grounds of an effect of the heat on the calcium salts of the milk but rather on the grounds of an effect of heat on the casein itself.. Just what this may be is not definitely clear, as yet. The author has this problem under investigation and hopes to be able to present definite data on it at a later date. It will be sufficient to point out at this time that the explanation of this phenomenon involves the fact that rennet coagulation is unquestionably both a chemical and a colloidal reaction. The calcium caseinate of milk is in colloidal solution. Rennet appears to hydrolyze the calcium caseinate into two molecules of calcium paracaseinate. The clotting of the calcium paracaseinate is a secondary phenomenon which is a gellation, perhaps of the nature of a crystallization of colloid—in this case a hydrophyllic colloid in a state of hydration. The conditions which govern what is regarded as a normal clotting of the calcium paracaseinate are evidently disturbed by the application of heat to the colloidal calcium caseinate of the milk. Zoller! has recently shown how 1 Zoller, H. F., J. Ind. Eng. Chem., 1921, xiii, 510. 142 SCIENTIFIC PROCEEDINGS (119). the properties of the casein of milk are affected by heat so as to have a marked influence on the precipitation of casein by acids and on the hydrophyllic properties of the casein thus precipitated. Apparently the addition of soluble calcium salts to milk helps to restore the conditions existing in raw milk which govern the normal rennet clot. All the interrelations of calcium and rennet coagulation have obviously not been determined. A study of these relations is at present in progress in this laboratory. SUMMARY. It is shown that the partial fixation of the calcium salts of milk by pasteurization or boiling is readily explained simply on the grounds of the effect of heat on colloidal solutions of CaHPOu, the calcium phosphate natural to cow’s milk. It is shown further that the effect of heat in retarding the rennet coagulability of milk is not related directly to the loss of colloidal CaHPO, because the addition of colloidal CaHPO, to dialyzed milk does not restore its coagulation by rennet, while the addition of CaCle or HCI does restore this property. The phenomenon of rennet coagulation is discussed briefly from the standpoint of the chemical and physico-chemical reactions involved, and also from the standpoint of the possible bearing which the addition of calcium salts to heated milk has on this phenomenon. 70 (1817) The velocity of development of the demarcation current in the frog’s sartorius. - By GEORGE EDMESTON FABR. [From the Department of Medicine, University of Minnesota, Minneapolis, Minn.] Urano and Fahr have experimentally established the fact that the potassium ion is almost exclusively the only cation within the muscle cell of the frog’s sartorius. Overton has shown that the demarcation current of the frog’s sartorius may be inhibited or have its sign reversed by replacing the lymph fluid surrounding the muscle cell by a fluid containing K ions in place of the normally DEMARCATION CURRENT IN FROG’s SARTORIUS. 143 Na ions. This action is reversible. The author has repeated Overton’s experiments and in addition to confirming them found that the relation of K in the muscle cell bathing fluid to the potential developed by injury of the muscle is a quantitative one and also that it is possible to get not only a change of direction for the demarcation current by replacing Na in the cell bathing fluid by sufficient K but also to get a pseudo action current under these circumstances. This pseudo action current only travels a short distance along the muscle, is of slow rate of progression and is accompanied by a small, slow contraction. From the above experiments it was concluded that a displace- ment of K ions across a semi-permeable cell boundary was respon- sible for the demarcation current of muscle. At the moment of cutting or injuring a muscle cell there is opportunity for ion equilibrium at the cut surface, whereas there is ion and thus electric strain at the uncut surface. If this hypothesis is true the velocity with which the demarcation current develops to its maximum intensity is of the order of 1/100,000 of seconds because of the speed of ions and the distance to be travelled by them. Garten has cut the surface of the frog’s sartorius and determined the speed with which the demarcation current rises to its maxintum value by means of the capillary electrometer. He believes that his experiments show that more than 1/1,000 of second is necessary for the demarcation current to develop. The analysis of Garten’s capillary electrometer curves is based on the formula D(ds/dt) + Ks = C1. This formula neglects the mass factor because usually the mass is so small in relation to the friction that it may be neglected. d*s ds pe OR ante = ee M+ D_+Ks=Ci is the equation which accurately describes the forces acting on the capillary when a potential difference is applied to it. Curves analyzed according to the first formula do not give an accurate picture of the development of potential difference during the first 1/1,000 of a second. Therefore we do not believe that Garten has proved that it takes more than 1/1,000 of a second to fully establish the current of injury of a frog’s muscle. 144 SCIENTIFIC PROCEEDINGS (119). In the year 1913 the author attempted to solve this question in the physiological laboratory of the University of Giessen under Professor Garten. To establish the current of injury a frog’s sartorius muscle was cut by a rifle bullet. This bullet cut a copper wire just before entering the muscle substance. The copper wire was shunted across a string galvanometer through which a constant current was passing. This galvonometer immediately responded with an excursion which was used as time marker. After passing the muscle the bullet cut another copper wire shunted across a second galvanometer. This galvanometer was connected to one electrode at the end of the muscle farthest from the injured portion. The other electrode lay beneath the injured portion of the muscle. It was possible to cut all but a few fibers of the muscle by means of the bullet and keep the muscle firmly attached to the electrodes. The moment the rifle trigger was pulled a hymographion carrying film was shot at the speed of 4,000 mms. per second. The galvanometer excursions were recorded on this. It was possible to get uniform velocity over a large portion of the film. A time marker recorded the time and ordinates were established as well as abscisse by mechanical devises. After re- cording a current of injury the two copper wires were replaced and a bullet shot through them as before. In this experiment a constant current equal to the previously measured demarcation current was thrown through the galvanometer at the moment the second copper wire was cut. We thus had a picture of the excur- sion of the string under the influence of a constant current to compare with our demarcation current. It was possible to meas- ure time with an error of less than 1/10,000 of a second because of the speed of the film and the uniformity of the velocity of the hymographion. Apparently all the demarcation current excur- sions of the string reproduced the constant current excursions within the limits ef errors of the method. That is the curves covered one another so closely that one could infer that the demarcation current is established with its maximum value within 1/10,000 of a second after injury. The strongest objection to the ion theory of the demarcation current is therefore removed. SoME ETHERS AND ESTERS OF SALIGENIN. 145 71 (1818) The pharmacological action of some ethers and esters of saligenin.' By ARTHUR D. HIRSCHFELDER and HERMAN H. JENSEN. [From the Department of Pharmacology, University of Minnesota, Minneapolis, Minn.] Benzyl alcohol saligenin and other aromatic alcohols have been shown to possess local anesthetic and antispasmodic action (Macht; Hirschfelder; Hjort). Hirschfelder and Quigley have also demonstrated that the local anesthetic action of benzyl alcohol and its derivatives is diminished when one of the inactive hydrogens (i.e., in the CH»2 of the CH,OH carbinol group) is substituted by another radical; or, in other words, that the secondary aromatic alcohols are not as good local anesthetics as the primary, and that substitutions for both the CH2 hydrogens (tertiary alcohols) causes complete loss of local anesthetic action. Although Hirschfelder, Lundholm and Norrgard had demon- strated that methyl and ethyl substitutions on the phenolic hydroxy] of saligenin rendered the substances more irritating than saligenin, a more extensive study of this type of substitution products was desirable, particularly on account of the fact that they furnished some alcohols homologous with acetyl-salicylic acid. The substances studied may be devided into three groups: I. Ethers of saligenin with substitution on the phenolic hydroxyl: 1.e., the ethyl, -butyl, iso-amyl and benzyl ethers. A Cut OCH.2CH2CH:2CHs3 CH,OH ene \ V/ |i EF: Ethyl saligenin. N-Buty! saligenin. * 1 The substances used in this research were prepared by Merrill C. Hart as by- products of an investigation of the phenolic alcohols and their derivatives as anti- septics and for the chemotherapy of the venereal diseases, with the aid of funds furnished by the United States Interdepartmental Social Hygiene Board. Their preparation has already been described elsewhere (M. C. Hart and A. D. Hirsch- felder, Jour. Am. Chem. Soc., 1921, July). 146 . SCIENTIFIC PROCEEDINGS (119). /CH OCH:CH:CH OCH2CeH; Y™ CHOH ‘absthics--( ) CHIOM 7 III. IV. Iso-amy] saligenin. Benzyl saligenin. II. Esters of saligenin with substitution on the phenolic hydroxyl (acetyl and monobenzoy] saligenin). OOCCH; OOCC,.H; (~ CH.OH CS GE al V. Vi. Acetyl saligenin. Benzoy] saligenin. III. An ester with substitution on both Levee (dibenzoyl saligenin). OOCC,H; ( CH,OOCC.Hs SS. VI. Dibenzoyl saligenin. All of these esters and ethers except the dibenzoyl compound are oily liquids, and the latter is a solid; and all are practically insoluble in water but soluble in the usual organic solvents and in olive oil. Their pharmacological properties were therefore studied by dissolving the substances in olive oil and emulsifying this with acacia by the Continental method. Toxicity tests, made by determining the dose which was lethal for frogs in twenty-four hours, gave the following results: For the ethers, ethyl saligenin 0.5 mg. per gram frog, n-butyl 0.25 mg., iso-amyl 0.12, benzyl 0.36 to 4 mg.; for the esters, acetyl saligenin 0.9 mg., benzoyl 1.0 mg., dibenzoyl 2.0 to 3.0 mg. All of these compounds are therefore more toxic than saligenin itself. The local anesthetic action was tested by dipping the frog’s foot into the emulsion and then into one per cent. sulphuric acid. Two per cent. emulsions were used. Anesthetic action set in from two to five minutes after exposure to the drug and lasted SOME ETHERS AND ESTERS OF SALIGENIN. 147 from ten to twenty-five minutes, except in the case-of the normal butyl ether which was more prolonged and lasted from one to two hours, thus being more prolonged than that of saligenin. With the dibenzoyl ester no anesthesia whatever was obtained, even after thirty minutes’ exposure to a five per cent. emulsion. This corresponds to the results which have been obtained with benzyl benzoate and other benzyl esters by Macht and others. All these substances are, however, very irritating. On the tongues of human beings they give rise to a bitter taste and an intense burning sensation which is most marked with the normal butyl, and least marked with the dibenzoy] ester. Upon the contractions of excised segments of rabbits’ duode- num in 400 mils of aérated Ringer-Langendorff solution at 38-39 the addition of 2.5 mils of ethyl saligenin decreased the amplitude, slowed the rate, and finally caused complete inhibition. 1.5 mils of n-butyl, 0.5 mil of iso-amyl, 1.0 mil of benzoyl ester, and 10.0 mils of the dibenzoyl ester produced the same effects; but only 0.8 mil of the benzyl ether was required to produce complete inhibition, accompanied by a very marked lowering of tone. This lowering of tone was also striking with the iso-amyl but not with the other ethers. In the case of the acetyl ester there was at first an increased amplitude (probably due to acetic acid from hydroly- sis) with slowing of the rate, gradually followed by inhibition. When injected intravenously into starved rabbits, anaesthe- tized with ether, no visible effect upon the contractions of the exposed small intestine could be observed through a glass window in the abdominal wall after the ethyl and the iso-amy] ethers, but the z-butyl, the benzyl, the benzoyl and the dibenzoyl compounds all caused a definite inhibition of peristalsis and a well-marked dilatation of the intestine. Contrary to the findings of Mason and Piek, and in accord with the experiments of Macht, we also observed this inhibition after the injection of benzyl benzoate. When applied locally to the rabbit’s intestine, all our compounds, with one exception, caused inhibition and dilatation. The acetic ester, however, on intravenous injection augmented the peri- stalsis, and when applied locally produced spastic contractions. All the emulsions, on intravenous injection, caused a fall of blood pressure, which varied from a sudden transitory fall in the 148 SCIENTIFIC PROCEEDINGS (119). case of the ethyl, 2-butyl and iso-amyl ethers to a more gradual and more prolonged fall after the acetyl, benzoyl and dibenzoyl esters. The benzyl ether caused a sudden and more prolonged fall, which lasted four to five minutes. Control emulsions, injected at the same slow rate, gave no effect whatever. Perfusion of the frog’s circulatory system gave a marked vaso constrictor effect, in striking contrast to the vasodilation obtained with emulsions of saligenin and benzyl benzoate. This constriction is probably due to irritation of the arterial walls. SCIENTIFIC PROCEEDINGS. ABSTRACTS OF COMMUNICATIONS. One hundred twentieth meeting. College of Physicians and Surgeons, January 18, 1922. President Wallace in the chair. 72 (1819) Is there more than one kind of rickets?! By E. A. PARK, P. G. SHIPLEY, E. V. MCCOLLUM and NINA SIMMONDS. [From the Department of Pediatrics, Yale University, New Haven, Conn., the Department of Pediatrics, Johns Hopkins University, and the Department of Chemical Hygiene, Johns Hopkins University, Baltimore, Md.] TI. CLINICAL OBSERVATIONS SUGGESTING THE EXISTENCE OF MORE THAN ONE KIND OF RICKETS. For at least two years our attention has been attracted to the possibility that there might be more than one kind of rickets. We were led to think of this possibility as the result of the con- sideration of certain peculiar manifestations of the disease and associations with other diseases. The facts are as follows: Rickets occurs with great frequency in premature children, even when breast fed. It seems to affect the head more than the extremities orribs. Ina group of cases of rickets the disease shows an especial tendency to involve the shafts of the long bones declaring itself clinically by the occurrence of multiple fractures from trivial causes. There is a curious association between rickets and certain forms of secondary anemia (the so-called alimentary anemias and 1 Abstract presented at the one hundred nineteenth meeting of the Society for Experimental Biology and Medicine. 149 150 SCIENTIFIC PROCEEDINGS (120). the anemias of the von Jaksch type). The rachitic involvement of the head in the children suffering from the combined conditions appears to be out of all proportion to the involvement of the extremities. Rickets is associated with the most severe forms of chronic interstitial nephritis or developmental defects of the kid- ney in which there is the most extreme degree of disturbance in renal function. Curiously, tetany sometimes occurs with rickets and sometimes not. Since the conception that there might be more than one kind of rickets was supported solely by clinical ob- servations of at most a suggestive character, we did not feel at liberty to express it as a definite hypothesis. Recently, however, our experiments have yielded results which indicate that there may be two distinct forms of the disease. Il. THE EXPERIMENTAL EVIDENCE. In previous publications! we have described certain defective diets which, when fed to the young rat, produced marked dis- turbances in the growth and calcification of the skeleton. The diets in question were all insufficiently supplied with a factor or factors present in cod liver oil. They differed from each other considerably, however, in the composition of their mineral fraction, chiefly, however, as regards the calcium and phosphorus. They may be divided into two groups according to the relative amounts of those two elements present. In one group, the phosphorus was at a low level but the calcium at or above the optimal level; in the other group, on the other hand, the calcium was at an extremely low level but the phosphorus was not far from the optimal. When the diets of the first group (the phosphorus being deficient and the calcium-phosphate ratio high) were fed to young rats living under ordinary laboratory conditions (room light), there developed a diseased condition of the skeleton which was identical in all essential particulars with that seen in the rickets 1 McCollum, E. V., Simmonds, Nina, Parsons, H. T., Shipley, P. G., and Park, E. A. Jour. Biol. Chem., 1921, xlv, 333. Shipley, P. G., Park, E. A., McCollum, E. V., and Simmonds, Nina. The Johns Hopkins Bulletin, 1921, xxxii, 160. McCollum, E. V., Simmonds, Nina, Shipley, P. G., and Park, E. A. Amer. Jour. of Hygiene, 1921, i, 492. McCollum, E. V., Simmonds, Nina, Shipley, P. G., and Park, E. A. Jour. Biol. Chem., 1921, xvii, 507. Is THERE MORE THAN ONE KIND OF RICKETS? 151 of human beings. The costo-chondral junctions were greatly en- larged; in some animals the thoracic wall was sunken at the sites of the costo-chondral junctions and the shafts of the ribs were fractured. The long bones of the extremities were enlarged at the ends; they could be cut and broken easily. Between shaft and cartilage lay a yellowish zone two to three mm. deep, the rachitic metaphysis. The proliferative cartilage extended in ir- regular prolongations toward the shaft. Calcium deposition in the cartilage was entirely lacking or extremely defective. The intermediate zone between cartilage and shaft presented the picture typical of the metaphysis in the bones of rachitic children. It was composed of cartilage in all stages of metaplasia or de- generation into a material indistinguishable from the osteoid, trabeculz consisting of osteoid, blood vessels bordered by marrow elements, scattered, irregular deposits of calcified material incased in osteoid, and connective tissue. All were intermingled in a disorderly manner. The trabecule of the shaft were surrounded by broad investments of osteoid. When the diets of the second group (the calcium being deficient, the phosphorus at a level not far from the optimal and the calcium- phosphate ratio low) were fed to rats kept under ordinary labora- tory conditions (room light), there developed a diseased condition of the skeleton which also bore marked resemblances to the lesions found in the rickets of human beings. The gross deformities caused by the second group of diets were as great or greater than those caused by the first group and corresponded exactly to the deformities found in rachitic children. The thorax was even more deformed than in the rats fed the diets of the first group; it was flattened from side to side and marked at the sides by deep grooves following the costo-chondral junctions; the angular deformities produced by the costal cartilages and the shafts projected into its interior; the costo-chondral junctions were enlarged and greatly distorted; fractures in the shafts of the ribs were especially numerous. The lower ends of radius and ulna were enlarged as were also the ends of all the long bones of the extremities. The bones were extremely soft and weak. Between the cartilage and the shaft was a white intermediate zone one to three mm. deep. Microscopic examination showed that the cartilage was entirely 152 SCIENTIFIC PROCEEDINGS (120). free from calcium or nearly free from it and was invaded in an irregular manner by the vascular elements of the shaft. In con- sequence the cartilage extended toward the shaft in irregular prolongations. The cells of the cartilage in proximity to the shaft showed evidences of degeneration and metaplasia. The inter- mediate zone was composed of cartilage in a more or less degen- erated state, osteoid trabeculz, blood vessels surrounded by marrow elements, a few deposits of calcium for the most part situated near the periphery and connective tissue. The trabeculz of the shaft were bordered by rather broad zones of osteoid. A loosely arranged fibrous tissue invested many of the trabecule. In those places in which it filled in the spaces between them, it gave rise to histological pictures which closely resembled those presented by the fibrous marrow in the rickets of human beings. The pathological condition induced in the bones by the diets of the second group did not, however, exactly correspond at all points to that usually found in the human subjects of the disease. The cartilage was invaded and its columnar arrangement was disrupted to a less extent than is commonly the case in the rickets of human beings. The metaphysis was composed in larger part by osteoid trabeculz. Though these osteoid trabecule were free from calcium deposition, they, nevertheless, retained a certain semblance of orderly arrangement. The osteoid zones about the trabecule were not so broad as in the rats on the diets of the first group, though they were quite as broad as the osteoid borders in the bones of rachitic children. Cells evidently derived from the fixed tissues with large basophilic granulations were numerous in the immediate vicinity of the trabeculae. Resorptive activity was exceedingly marked. In the fundamental respects, however, in particular, degeneration and metaplastic changes in the cartilage, defective calcification of the cartilage and trabeculz and the con- sequent osteoid production, the irregular invasion of the cartilage and the production of a rachitic intermediary zone, the patho- logical conditions produced by the faulty diets of the second group corresponded to the rickets of human beings. While one of the diets in question, diet 2638, did not give constant results, in some animals it produced a pathological condition corresponding to that found in human rickets even in its minor details. Is THERE MORE THAN ONE KIND OF RICKETS? 153 III. Discussion. Our experiments make it clear that in the rat, when deprived of certain active light rays and an unidentified factor contained in cod liver oil, a pathological condition corresponding in all fundamental respects to the rickets of the human being can be produced through the diet in two ways; it can be produced (1) by diminishing the phosphorus in the diet and supplying calcium in excess of the optimal or at the optimal concentration, or (2) by reducing the calcium but maintaining the phosphorus at a con- centration somewhere near the optimal. In the former case the calcium-phosphate ratio in the diet is large, in the latter case it is small. We have not the slightest doubt that in the human being similarly deprived of active light and the unidentified factor it would be possible to produce true rickets through a manipulation of the calcium and phosphorus of the diet in the two ways men- tioned. As the result of our experiments we are led to believe, therefore, that there are two main forms of rickets, one character- ized by a normal or nearly normal blood calcium and a low phos- phorus, the calcium-phosphate ratio being high, the other by a normal or nearly normal blood phosphorus but a low calcium, the calcium-phosphate ratio being low. Between the two forms there are probably innumerable intermediary forms marked by calcium- phosphate ratios which are less effective in preventing calcium- phosphate deposition. Diets with these intermediary ratios _manifest themselves pathologically as rickets in various stages of healing or of healing and relapse or as slight disturbances of calcification of the skeleton which have more or less remote resemblance to rickets. At one time we were under the impression that tetany sharply marked off one form from the other, 7.e., that the two main divi- sions of rickets were ordinary rickets and the rickets of tetany. The investigations of Howland and Kramer! have shown, however, that tetany in a latent form, at least, may be associated with the low phosphorus form of the disease. Though tetany may occur with the low phosphorus form, however, it is probably regularly associated with the low calcium form and may be regarded as a 1 Howland, John, and Kramer, Benjamin. Amer. Jour. of Dis. of Child., 1921, xxii, 105. 154 SCIENTIFIC PROCEEDINGS (120). symptom of the latter. The low calcium form of rickets is, generally speaking, the rickets of tetany. Tetany may of course occur independently of rickets. If tetany persists long enough, however, evidence of defective calcification of the skeleton would almost certainly develop. . On theoretical grounds entirely, we think it possible that the rickets which develops in the youthful subjects of the severe functional derangements of the kidney may belong to the low calcium form of the disease; in other words, that the rickets in these patients may be truly endogenous in origin depending primarily on the inability of the kidney to excrete phosphorus. If this view proves to be correct ‘‘renal dwarfism”’? is in reality renal rickets and ought so to be called and regarded. 73, (1820) Variations in aliquot fractions of gastric contents. By NICHOLAS KOPELOFF. [From the Department of Bacteriology, Psychiatric Institute, Ward’s Island, New York City.] Aliquot fractions obtained by the Rehfuss method of fractional gastric analysis do not accurately represent the total gastric contents, as indicated by the results of the following experiments on subjects showing no clinical evidence of gastric disease: 1. Instead of the usual periodic aspiration, the total gastric contents were removed after three quarters of an hour by with- drawing 10 c.c. fractions in rapid succession. A wide variation was found in the acidity of these fractions, indicating that the total gastric contents are not a homogeneous mixture and that a single 10 c.c. sample is not a valid aliquot. This was more noticeable in subjects having a high rather than low gastric acidity. 2. (a) By inserting three Rehfuss tubes in one individual and aspirating the fractiens simultaneously at fifteen minute intervals, it was found that there was considerable variation of acidity in different parts of the stomach at the same moment. X-ray pictures established the relative position of the tubes. 1 Barber, Hugh. Quart. Jour. of Med., 1921, xiv, No. 55. EFFECT OF COOKING UPON VITAMIN IN CABBAGE. 155 (6) From these data it is shown that widely divergent curves of acidity may be plotted which depend entirely upon the experi- mental error of the method and not upon the subject’s gastric condition. (c) The inadequacy of the titration method and the importance of hydrogen ions and buffer salts in measuring gastric acidity is indicated. 3. A subject who could regurgitate his total gastric contents at will, was given the test meal on different days and the total contents were regurgitated at different intervals from the time of ingestion to the test meal. The curve of acidity plotted in this manner differed radically from the curve obtained in the usual manner. Furthermore, discrepancies were noted between the acidity of an aliquot removed immediately prior to regurgitation and the acidity of the total gastric contents. Results obtained by the Rehfuss method may be more validly interpreted if: (a) the analysis is repeated until a satisfactory agreement in curves is obtained; (0) the tube is kept at a constant level; (c) aliquot fractions are large; (d) little saliva is swallowed; (e) acidity is measured in terms of hydrogen ions and buffer salts. 74 (1821) The effect of cooking upon the antiscorbutic vitamin in cabbage. By WALTER H. EDDY, E. SHELOW and R. A. PEASE. [From the Department of Physiological Chemistry and the Department of Foods and Cookery, Teachers College, Columbia University, New York City.| The present report is one of a series of studies undertaken to determine the effect of the new cooking implement known as the pressure cooker upon the vitamin content of such foods as are adapted to preparation in that device. The cooker also affords in its manipulation an opportunity to throw light on the response of vitamins to certain combinations of destructive influences that are absent or different from those met in older methods of cookery. The present study is confined to a comparison of the effects of pressure versus open kettle cooking on the vitamin C content of cabbage. 156 SCIENTIFIC PROCEEDINGS (120). In open kettle cooking the routine involves immersing a given wt. of cabbage in a kettle containing one cup of boiling water and continuing the boiling until the cabbage is in a condition suitable for the table. This required from 45 to 90 minutes according to the weight of cabbage used. In bringing the cabbage to the same condition with the pressure cooker the following procedure was necessary: One cup of water was placed in the cooker and brought to boiling. Cabbage was then added, the cover clamped on and the cooker placed over the flame. ‘The valve was left open until dry steam issued, then closed and heating continued until a pressure of 15 lbs. was reached and a temperature of 121° C. From the time of adding the cabbage to this point required 3-5 minutes. The flame was then regulated to maintain this pressure and temperature for thirteen minutes. At the end of that time the cooker was removed from the flame and allowed to cool until the pressure had dropped to 5 lbs. The valve was then opened, cover removed, and the cabbage drained. The contrasts in cook- ing conditions are summarized as follows (the Py was determined on the drained-off liquor): COOKING CONDITIONS. Pressure Cooker. Open Kettle. Time of Cooking 2.55. 6.2.35 20 minutes in all 45-90 minutes total I3 minutes at 15 lbs. Temperatures maintained. ...3-5 minutes at 100-121° C. 45-90 min. at 100°C. I3 minutes at 121° C. 2 minutes at 121-100° C. Reaction: Exp. IIandI..... Py 5.6-5.8 Py 5.6-5.8 Yap. BE, CofE Oe Ss Py 4.6-4.8 Contact with atmos. oxygen..In atmosphere of steam Practically immersed throughout cooking in boiling water period. throughout cooking period. Guinea pigs were used as experimental animals throughout the experiments and in all cases the pigs were fed for a preliminary period of at least two weeks on the basal diet plus raw cabbage ad lib. Only those -which gained consistently on this diet were selected for the experiment and those which approximated 350 gms. so far as possible. For basal diet the LaMer-Sherman combination was selected: EFFECT OF COOKING UPON VITAMIN IN CABBAGE. 157 LAMER-SHERMAN BASAL DIET. Per Cent. Skim milk powder (Krystallak) heated two hours at 107° C.................. 30 ee TE a Pa ns 5, hoe Sb das dic 'p owe tals Ae oles bine sla wale « oe ea ys 10 aC are Me salNrti te WP ee LS yi, Alcon gener ig hk ede fend mi gge kb edd pts orally ees 59 ne ee Ct eR Re ne na era io aedia Ph cage) oP a aia Wp eve toa ele 2 alt Eee I 100 The cabbage was fed separately by hand and complete consumption observed of the daily allowance. The amounts used were I gram raw to control pigs and 5 or 10 grams cooked to experimental! pigs. 5 grams cooked cabbage is equivalent to 5.4 grams raw. 10 grams pressure cooked is equivalent to 10.7 gms. raw and 10 grams open kettle cooked to 10.8 gms. raw. Three experiments are reported herewith. Expeiiment I was a qualitative expe1iment over a twenty-day period. During this period 4 pigs were fed the basal diet plus I gram raw cabbage daily and two were put on basal diet without supplement. Two others were given basal diet plus 5 grams pressure cooked cabbage daily. At the end of 20 days the animals were chloroformed and autopsied. The result seemed to indicate that 5 grams cooked cabbage was very little protective as the animals showed marked scurvy symptoms. It also demonstrated the efficiency of the control diets. (See Chart, Exp. I.) In the second experiment the control diets were repeated and four groups of experimental animals followed. These received 5 grams or ten grams daily of pressure cooked or open kettle cooked cabbage. They were kept on this diet until death ensued from scurvy and were then autopsied and the symptoms of scurvy verified. The controls on I gram raw cabbage per day were continued 82 days to make sure of the protection and then chloro- formed and autopsied to confirm this point. (See Chart, Exp. II.) In the third experiment three animals were placed on 10 grams pressure cooked cabbage per day plus basal diet but in this case the water in which cooking took place was acidified by diluting 40 c.c. of vinegar to 234 c.c. with water. The liquor drained from the cabbage at the end of the cooking period registered a Py of 4.6—-4.8. All of these animals died of scurvy. (See Chart, Exp. III.) A summary of the significant data connected with these three experiments is given in the following table. The growth curves are shown in the chart. 158 SCIENTIFIC PROCEEDINGS (120). TABLE. Diets (Symbols). A. Basal diet plus one gram raw cabbage daily. ’ B. Basal diet without cabbage. C. Basal diet plus 5 grams pressure cooked cabbage daily. D. Basal diet plus 5 grams open kettle cooked cabbage daily. E. Basal diet plus Io grams pressure cooked cabbage daily. F. Basal diet plus 10 grams open kettle cooked cabbage daily. G. Basal diet plus 10 grams pressure cooked acidified cabbage daily. Experiment I. fui it Diet. | aoe. A, B. | ee Pee is Ss Ps eee Ave. of 4 Ave. of 2 Ave. of 2 Bays. On dicts 25 s..0 tae ce 20 20 20 Wt. at beginning............ 250 225 225 fet ey ) ER AM co eR tee 350 185 220 Autopsy findings. ........... No scurvy Scurvy Scurvy i Experiment IT. Diet. AS B. CG dD. | Te, | EK; ' | | | eee Pig New, Dart 6 | 12 Bic Sao A POSS 7 Te ae & 2) 9 Days on diet...| 82| 821! 32] 32| 46] 41 | 44] 48) 45] 59| 52 | 48 Wt. at beg. ....| 400 358 | 378 | 387 | 364 | 376 | 411 | 339 | 362 | 396 | 351 |384 Wt. atend..... | 300 | 498 | 220 | 207 212| 272 235 | 235 | 252 | 207 | 200 244 Gain or loss. ...| 100 4140) 158 180 | 152 | 104 176/104 | 110| 189 | 151 |140 igniter... s2 5. Nia No} SE) S Ss S$ Ss Ss Ss S$. [ss Adrenals, >. 2... — | 640 aes (453 439 | 302 509 | 481 | 350 | —— | 389 324 mgms. | Experiment III. | Diet G Ple Ness eux ears Ce Re 23 24 25 Da G OM TAG. bnoa 02k tp a ee / AI 26 37 Wt. Ot-Dem. 2h e ean ee 469 315 395 WC: t CA 57.) us a 7h eid ae 300 230 225 Pe ey ei es Ye 169 85 170 AUEOOEY 6 68 bs. a ee oe Ss S Ss PATACOTOIG 21 oo teas) o.0 pe te 697 412 496 mgms. SUMMARY. 1. The LaMer-Sherman basal diet proved quantitatively ac- curate in producing death by scurvy within the periods predicted by the authors. 1S” sign. scurvy. ‘‘N” sign. no scurvy. EFFECT OF COOKING UPON VITAMIN IN CABBAGE. 159 2. One gram raw cabbage daily, added to the basal diet, proved completely protective against scurvy; no symptoms appearing in 82 days on the diet and the absence of symptoms in this case was confirmed by autopsy on the chloroformed survivor. 3. The methods of cookery were such as to indicate that in spite of acid reaction and exclusion of oxygen the temperatures used were sufficient to destroy enough vitamin C to make an intake of 10 grams daily inadequate for protection against the disease. In this respect the cabbage vitamin seems to be much Nuss ince Ey J Ce NS s- Be S Bee. Growth curves expressed in grams. D indicates death by scurvy and the numeral following indicates days elapsed on diet. less resistant than the orange juice or tomato juice carried form. It is of course possible that the intra-cellular oxygen of the cabbage is a factor in this difference. The results however harmonize with those of Ellis, Steenbock and Hart,! who showed that drying for 35 hours in an atmosphere of COz at a temperature of 65° C. proved extremely destructive to the cabbage vitamin. iT biol. Chem., 5021, xlvi, 367; 160 SCIENTIFIC PROCEEDINGS (120). 4. At the maximum intake used, no difference was observable between the pressure cooker and the open kettle as a destructive agent. X-rays of excised ribs of the animals of Experiment II. The numerals on the I gram raw cabbage daily; 0, pigs 3 and 8, no cabbage in diet; 4, 5 grams pressure cooked cabbage and 3, 5 grams open kettle cooked cabbage; 7, 10 grams pressure photograph have the following significance: 5 is pig 12, 82 days on basal diet plus cooked cabbage and 8, 10 grams open kettle cooked cabbage daily. : —— oe | A New SULPHUR-CONTAINING AMINO ACID. IOI 75 (1822) A new sulphur-containing amino acid isolated from casein. By J. HOWARD MUELLER. [From the Department of Bacteriology, College of Physicians and Surgeons, Columbia University, New York City.| In a report on a study of the cultural requirements of strep- tococci made last year before this society, the writer stated that a compound containing sulphur had been isolated from casein, which was apparently not related to cystine, and which seemed to be required for the growth of the test organisms. Although subse- quent work has shown that this sulphur compound, when pure, is apparently not concerned in the growth of streptococci, it seemed desirable to make a study of the substance, both because of the uncertainty of the nature of non-cystine protein sulphur, and also in order to be able to effect a separation of this compound from the bacterial growth inducing factor in the amino acid fraction under investigation. While there are still many points to be cleared up in connection with the substance, perhaps enough information has been obtained to warrant a preliminary report. There have been a number of difficulties met with in the work. The yield is very small, and probably not by any means quartita- tive, and further, no insoluble compounds suitable for separation have so far been found, so that purification has consisted largely in methods for the removal of impurities. In order to obtain sufficient material for analyses, thirty pounds of commercial Argentine casein were used. Briefly, the method consists in hydrolysis with sulphuric acid, neutralization with sodium carbonate, and precipitation with mercuric sulphate solu- tion. From the washed precipitate, freed from electrolytes, a considerable quantity of other material is removed by a second precipitation with mercuric sulphate, the sulphur compound this time remaining in the filtrate. Further purification is effected by precipitation of the filtrate, after removing electrolytes, by silver sulphate and barium hydroxide, and the compound itself is ob- tained from the silver filtrate, freed, of course, from Ag and Ba, by fractional crystallization, finally from dilute acetone. The yield from thirty pounds was about Io grams. 162 SCIENTIFIC PROCEEDINGS (120). The compound crystallizes from water, dilute alcohol and dilute acetone, in shining white microscopic plates or rosettes of rather indefinite crystal form. As a criterion of pulity, amino nitrogen determinations were carried out afte1 successive crystalli- zations until constant values were obtained. At this time the proportion of N to S was 2:1, and it was assumed that the prep- aration was pure. Analyses of this product indicated the formula C,,;He35NeQO,4. Unfortunately, when most of the amino acid had been used up, it was found that the remaining material contained as an impurity a substance forming a hydrochloride relatively insoluble in concentrated HCl. Since the entire prep- aration had not been crystallized at the same time, and only the first portions, used for quantitative analyses, had been checked up by amino nitrogen and sulphur determinations, it is possible that the impurity was not present in the material which was analyzed. From the few tenths of a gram remaining, the insoluble hydrochlo1ide was removed as completely as possible by dissolving in boiling HCI and allowing to crystallize on ice for several days. The filtrate, containing the sulphur compound, was freed from HCl by evaporation and by AgsSO,, and the sparingly soluble copper salt prepared by boiling the solution with Cu(QH)». and filtering. The salt separates as microscopic, pale blue platelets. About 0.25 gram of this salt was obtained, and a single combustion and amino nitrogen determination gave the formula Cy,;He:- SN.O.Cu, corresponding exactly with that obtained fo1 the amino acid in the earlier analyses. The combustion was done by the Dennstedt method, permitting the simultaneous determination of carbon, hydrogen, sulphur and copper. The nitrogens are both in the form of amino groups, and since the solution of the compound in water is practically neutral, it is probable that two COOH groups are present. A single formol titration gave results which were slightly low, but in fair con- formity with this supposition. The sulphur is not in the lead- blackening form, and is not readily split off as sulphate by boiling with acids or alkalis. The number of hydrogen atoms present suggests a straight chain compound, but no direct evidence of the structure has been obtained. In order to rule out the possibility of the mateiial used for analyses having been impure, it is planned to prepare another lot INHERITANCE OF SUSCEPTIBILITY. 163 of about the same quantity, after which complete analytical data and a detailed account of the preparation will be published elsew here. When the composition and properties of the amino acid are definitely established, it will be necessary to show that it is really ‘a primary component of protein. It is possible that the sulphur has been introduced into the molecule either from the sulphuric acid used in hydrolysis, or from the HS used throughout the preparation. The possibility of introducing sulphur from sul- phuric acid is rather remote, since the amino acid has not the properties of a sulphonic acid. However, it can be excluded only by the use of enzyme digestion or alkali hydrolysis in the primary breaking down of the protein, and this has not yet been attempted. In the preparation as outlined above, it is apparently not possible to avoid the use of HS, and this factor can be ruled out only by the elaboration of a method based on quite different principles, or by careful quantitative determinations of total sulphur throughout the various fractions, which is not likely to prove very satisfactory. It is not apparent, however, just how sulphur from H.S could be introduced into any of the known amino acids to give a compound of the above formula. Should it be possible to exclude these sources of extraneous sulphur, this compound will probably account for a part at least of the non lead-blackening sulphur known to be present in certain proteins. The amount present may well be considerably in excess of the present yield, since the method is obviously not quantitative. 76 (1823) The inheritance of susceptibility to implants of splenic tissue in mice. 1. Japanese waltzing mice, albinos, and their F, generation hybrids. By C.-C. LITTLE and B. W. JOHNSON. [From the Carnegie Institution of Washington, Cold Spring Harbor, N. Y,] The use of the terms ‘‘auto,” ‘‘homio,” and “hetero’’ trans- plantation has been general and of great value in the long series 164 SCIENTIFIC PROCEEDINGS (120). of experiments which have dealt with the transplantation of normal and of neoplastic tissues in vertebrates. Following a number of experiments in this field, covering a wide range of material, it has become generally recognized by biological investigators that the closer the genetic relationship between the host and the donor of the graft tissue, the greater is - the likelihood of persistent and progressive growth of an implant of tissue from one to the other. Similarly it has been found that in the ordinary “‘laboratory”’ races of mammals, inbreeding has not been intensive enough to have produced a close degree of genetic resemblance between individuals within the race. Without this resemblance the con- tinued growth of tissue transplants made from one animal to another is impossible. When close relatives such as parent and offspring or litter mates are picked for this interchange of implants, there is, as Loeb and others have pointed out, more chance of persistence of the implants than when unrelated animals are used. Loeb! has proposed the term “‘syngenesio-plastic transplantation ”’ for experiments involving the close relatives referred to. When, however, closely inbred races of known genetic constitu- tion are used, results are obtained which show that the distinctions between “homio,”’ “‘syngenesio,”’ and “‘auto”’ transplantations are only relative and may be deliberately broken down by picking animals of certain definite genetic constitutions for experimenta- tion. Thus in animals of a closely inbred and genetically homoge- neous strain of Japanese waltzing mice [already described in con- nection with experiments on the inheritance of susceptibility to transplanted tumors’], the general reactions of an individual to subcutaneous transplants of bits of its own spleen (autotrans- plants), or to bits of the spleen of another Japanese waltzing mouse of the same inbred race (homiotransplants) were the same. ‘ . ° . ' Both implants ‘‘auto’’ and ‘‘homio”’ persisted successfully, estab- lished a blood supply, and remained healthy. 1 Loeb, Leo, Journ. Med. Research, 1918, xxxix, 39-57. 2 Little, C. C., and Tyzzer, E. E., Journ. Med. Research, 1916, xxxili, 393-453- INHERITANCE OF SUSCEPTIBILITY. 165 TABLE I. Autotrans- Homiotrans- plants. plants. Cross No. Races. + PSD Ws A te . a _ os = I Japanese waltzing X Japanese waltzing 22 1*¢ 22 1*a 2 Japanese waltzing 14 1*b oO 15*b X albino 16 1*¢ oO 17*c 3 Japanese waltzing 23 (s) 9) 23 X Fi hydrids 33 oO 33 fe) -++ means persistence of the implant in a healthy condition. — means disintegration of the implant. *a—Mouse sick and probably unable to provide adequate nourishment for either implant. *b—One mouse negative to both auto and homio, probably due to poor operative technique or to mouse being in poor physical condition. *c—One mouse negative to both auto and homio, probably due to poor operative technique or to mouse being in poor physical condition. These results are tabulated in the top two lines of Table I. It should be noted that in one animal neither the autotransplant nor the homiotransplant persisted. Records show that this ani- mal was in markedly poor physical condition and this without doubt accounts for the elimination of both implants. When interchange of splenic implants was made between Japanese waltzing mice and unrelated albino non-waltzers, the results shown in lines 3 and 4 of Table I were obtained. One Japanese waltzing mouse and one albino failed to support either the auto or the homio implant. This was probably due to poor technique involving infection after the operation. If these animals are subtracted from the totals we find that the auto- transplants in either the Japanese waltzers or the albinos are successful, while the homiotransplants from albino to waltzer or vice versa are uniformly unsuccessful. This, it will be remembered, is the result ordinarily obtained in homiotransplantation and adds another piece of evidence to our belief that similarity in genetic constitution is essential for success- ful implantation of splenic tissue. The interesting and crucial test of the correctness of the hypothesis on which the experiments were planned is found in the 166 SCIENTIFIC PROCEEDINGS (120). case of reciprocal transplants between Japanese waltzers and F; generation hybrids, formed by crossing together the waltzers and the albinos. In this case, analogy with the tumor work and knowledge of the genetic constitution of the animals lead us to suspect that implants of waltzing mouse spleen should grow in the F, hybrids, while implants of splenic tissue from the hybrids should not persist in the waltzing mice. This was actually found to be the case, as can be seen from the two bottom lines of Table I. The gametes of the Japanese mice were, by hypothesis, essen- tially equal in respect to their genetic factors. Each F, hybrid had therefore received from its waltzing mouse parent approxi- mately the same genetic contribution. This was of such a nature as to make possible the persistence of implants coming from the Japanese mouse. (The same was found to hold true in the case of tumors of the Japanese mouse.) The F, tissue which was implanted in the Japanese waltzers comes from animals approximately half of whose genetic consti- tution is determined by their albino parent. Since the Japanese mice had not the genetic factors which made up the albino complex, we should not expect that they could support the hybrid implants. The results, therefore, in all three series are in complete harmony with the hypothesis advanced by Little and Tyzzer (1916), namely: that the susceptibility of any mouse to implants of foreign tumor tissue depends upon the genetic constitution of the host in its relation to the genetic constitution of the animal from which the implant was taker. The value of the term ‘‘syngenestotransplaniation”’ seems to be greatly impaired for: (a) Parents (waltzers) failed uniformly to support implants of the splenic tissue of their progeny. (b) Progeny (F, hybrids) grew regularly the splenic tissue of their parents. There is in this case an absolute difference in the results obtained, depending upon which race is used as the host. The fact that the ‘genetic’ relationship changes while the ‘‘pedigree’’ relationship remains the same, shows that the former is the important factor in determining the nature of the result. OBSERVATIONS ON COD-LIVER OIL AND RICKETS. 167 The evidence obtained from the Japanese waltzing mice shows that within a closely inbred race, homiotransplants of splenic tissue may be quite as successful as autotransplants. The experiments further show that 7m all probability suscepti- bility to transplants of splenic tissue depends upon the same general principles of heredity found to apply in the case of tumor tissue— namely, multiple mendelizing factors. 77 (1824) Observations on cod-liver oil and rickets. By T. F. ZUCKER, A. M. PAPPENHEIMER, and MARION BARNETT. [From the Department of Pathology, College of Physicians and Surgeons, Columbia University, New York City.] In view of the nearly specific action that cod-liver oil has on rickets, it is of interest to inquire into the nature of the substance conferring on it the therapeutic properties. Several attempts have been made to isolate from it materials that could be made responsible for its action. Gautier and Morgues? isolated the organic bases contained in the oil and separated from them two alkaloid-like substances besides the simpler aliphatic amines. Funk? also worked with this mixture of bases which he frac- tionated in various ways. None of these observers, however, have published any data on the action of the isolated material. Stéltzner* claims, without giving any details of his evidence, except the statement that he cured even the worst cases of rickets, that hydroxy acids confer upon cod-liver oil its pharmacological properties. Freudenberg and Klocmann# had expressed similar ideas and prepared calcium salts of the unsaturated acids of cod-liver oil which they used in the treatment of spasmophilia. Wacker and Beck® believe that ‘‘besides other chemically not yet well characterized substances, cholesterol plays a significant role in the antirachitic fat soluble factor A.”’ 1 Gautier and Morgues, C. R. Acad. Sci., 1888, cvii, 110 and 626. * 2Funk, Biochem. Bull., 1915, iv, 365. * Stdltzner, Miinch. med. Wochenschr., 1921, \xviii, 272. 4 Freudenberg and Klocmann, Jahresb. f. Kinderh., 1913, Ixxviii, 47; 1914, xxix, 700. 5 Wacker and Beck, Berl. klin. Wochenschr., 1921, \xxxv, 453. 168 SCIENTIFIC PROCEEDINGS (120). With a good test object now available in the rat made rachitic on the phosphorus low diet described by Sherman and Pappen- heimer, the problem of determining the point in question is much easier. The rickets of children and the experimental rickets in rats both respond in the same manner to treatment with cod-liver oil! and there is no reason to believe that the substance active in the two cases should not be the same. We first isolated the crude bases according to Gautier and Morgues and found them inactive. Next the oil was hydrolyzed with sodium hydroxide and the fatty acids separated. The fatty acids, when reasonably purified were entirely inactive, although in one of the first experiments a rather impure fatty acid fraction did slightly promote calcification. The residue of unsaponifiable matter gave a marked curative action. From this the bases were again isolated and these bases obtained after hydrolysis were also inactive. From the unsaponifiable matter in solution in alcohol a goodly portion of the cholesterol was crystallized out. This cholesterol fraction also was inactive. The material freed from most of the cholesterol was now more active than before. In this manner, we obtained fractions which on being diluted with ninety parts of cotton-seed oil, which had been found to be inactive, gave a curative effect a little stronger than the original cod-liver oil. The results were controlled by both X-ray of tibia and histological examination of rib sections. This material also contains the fat soluble factor A, as we have been able to cure with it ophthalmia produced by deficiency of fat soluble. Very recently, Steenbock, Nelson and Hart? have made similar other extracts of saponified cod-liver oil and reported curative effects on ophthalmia in dogs. Although there is no longer any good reason to believe that rickets is a simple fat soluble deficiency,’ the relation of the fat soluble A factor to the curative property of cod-liver oil remains to be worked out. To sum up, we may say that the antirachitic substance of cod- liver oil can be demonstrated in the ether soluble “‘ unsaponifiable”’ 1 Shipley, Park, McCollum, Simmonds and Parsons, Jour. Biol. Chem., 1920- 21, xIv, 343. 2 Steenbock, Nelson and Hart, Am. Jour. Physiol., 1921, lvii, 14. * Pappenheimer, McCann and Hess, Jour. Biol. Chem., 1921, xlvii, 395. 4Shipley, McCollum and Simmonds, Jour. Biol. Chem., 1921, xlix, 399. DISTRIBUTION OF INORGANIC PHOSPHATE OF BLOOD. 169 fraction after alkaline hydrolysis. It is not an organic base of the type described as occurring in cod-liver oil. It is not choles- terol, but similar to cholesterol in its solubilities. The suggestion is made that it may be a sterol related to cholesterol or a choles- terol derivative. The fatty acids of cod-liver oil are entirely inactive in curing rickets. 78 (1825) The distribution of inorganic phosphate of the blood between plasma and cells. By T. F. ZUCKER and MARGARET B. GUTMAN. [From the Department of Pathology, College of Physicians and Surgeons, Columbia University, New York City.] Since Greenwald’s work on the organic acid soluble phosphorus our knowledge of the various phosphorus compounds in the blood is steadily increasing and acquiring significance. The determi- nation of the inorganic blood phosphate, however, has been very questionable, particularly in corpuscles, due to the ease with which the organic acid soluble phosphate is hydrolyzed. Any method which requires considerable time or in which the phosphate has to be precipitated, or in which the red cells are washed or otherwise manipulated, comes very gravely under suspicion of having allowed a significant amount of hydrolysis to take place. When working only with plasma these precautions are not so necessary. Bloor’s! figures for inorganic phosphate in the cor- puscles are admittedly high. A method very well suited to esti- mation of inorganic phosphate is that of Bell and Doisy? in which the color of the blue reduction product of phosphomolybdic acid is measured as in Folin’s uric acid and phenol determintion, the limiting factor, however, being the phosphate. Our results have been briefly as follows: When working rapidly with the Bell and Doisy method, the inorganic phosphate in the plasma and the whole blood is the same within the limit of error of the method. The few exceptions to this were traced to improper handling of the blood or too long a ‘time elapsing before the determination. Even here the whole 1 Bloor, Jour. Biol. Chem., 1918, xxxvi, 49. 2 Bell and Doisy, Jour. Biol. Chem., 1920, xliv, 55. 170 SCIENTIFIC PROCEEDINGS (120). blood gave figures much lower than Bloor’s. This shows that the phosphate ion in its relation to cells and plasma behaves differently from all the other ions studied in this respect. The chloride, for instance, is never present in cells and plasma in the same concen- tration. This exceptional role of the phosphate ion is, however, not so surprising when we consider the organic acid soluble phosphate, in a sense the counterpart of the chloride combined with protein. TABLE I. DISTRIBUTION OF INORGANIC PHOSPHATE BETWEEN WHOLE BLOOD AND PLASMA. Adults. Children. Subject. Whole Blood. Plasma. Subject. Whole Blood. Plasma. P.G. I 3.5 3.4 ce: le ae 3.0 3.01 OS Rae 3.52 3.58 BOO. So, vs 3.02 2.78 G.M. 1 4.0 | 3-78 M.E... 4.0 4.01 GA 2 3.98 3:94 Ot: Ae ek 4.61 4.63 bo Sats as: 3.1 2.8 DB... eres 4.17 4.26 ) ok ERTS rare 3.38 3.36 R.M 4.10 4.08 Gute Peteten ast 3.50 F.B 3.75 3.49 MUL bck 3.16 3.18 Piet ai atose 4.17 4.12 Do ee Eyes 4.25 4.35 A conclusion of practical import is that when inorganic phos- phate is to be determined in blood, it is immaterial whether it is done on whole blood or plasma, providing it is done immediately by the method of Bell and Doisy. | If the colorimetric reading is made within half an hour after the blood is drawn, whole blood is no higher than plasma, but if an hour elapses the whole blood is higher by about 0.5"mg.; after several hours the difference is a milligram or more. It seems reasonable enough to assume that if at the time the blood is drawn the phosphate ion is always present in equal concentration inside and outside of the cell, it is freely diffusible into and out of the cell when the blood circulates. Iverson! has shown that phosphate in organic acid soluble form can accumu- late in.the corpuscle at a good rate if inorganic phosphate is injected or added to blood. We would, therefore, expect that the phosphate formed by hydrolysis would diffuse out of the cells into 1 Iverson, Biochem. Zeil., 1921, cxiv, 297. INORGANIC PHOSPHATE OF BLOOD. I7I the plasma. This seems not to be the case in the blood after it is taken from the body. TABLE II. To ILLUSTRATE EFFECT OF TIME AFTER DRAWING BLOOD ON THE INORGANIC PHOSPHATE. Within % Hr.| Within rt Hr. | After 5 Hrs. | After 20 Hrs. TS 5) 2 a a 2.4 2.5 WET Soe ts sk ess 2.5 2.8 3:4 Bi ASIA OS ses ss 2.55 2.53 Ce EP Re eee 2.6 3.08 4.02 These results lead to the following conception of the rdle of inorganic and organic acid soluble phosphorus in the blood: The réd cell is totally permeable to the phosphate ion, 7.e., no ‘‘osmotic influences’ control the distribution of the phosphate ion inside and outside of the cell. Phosphate ions can be taken up by the cell and stored as organic acid soluble phosphate (Iverson). This organic phosphate is hydrolyzed very easily when there is need for phosphate ion in the plasma, similar to the liver glycogen yielding blood sugar. The diffusion of the phosphate out of the cell, however (at least in vitro), is slower than its rate of forma- tion by hydrolysis. ‘To substantiate this view we will still have to show under what conditions inorganic phosphate can diffuse out of the cell. We have no indication so far that phosphate distribution is influenced by the COs, tension. We are now col- lecting data on this point, as well as on the whole subject from the point of view of the acid soluble phosphate. 79 (1826) Observations on the inorganic phosphate of blood in experimental rickets of rats. By M. B. GUTMAN and V. KNEELAND FRANZ (by invitation), [From the Department of Pathology, College of Physicians and Surgeons, Columbia University, New York City.] _ The work of Howland and Kramer! on the level of the inor- ganic phosphate in the blood in human rickets, and some con- 1 Howland, J., and Kramer, B., Amer. Jour. Dis. of Child., 1921, xxii, 105. 172 SCIENTIFIC PROCEEDINGS (120). firmatory experiments undertaken by Hess! led to the conclusion that during the period of active rickets in children, the inorganic phosphate of the blood is reduced, and that during the process of cure by either sunlight or cod-liver oil, the phosphate rises again to its normal level. Since the experimental rickets produced in rats is comparable in most important respects to human rickets, it was thought of interest to determine whether the same changes in blood phosphate could be demonstrated in rats. In applying the work of Howland and Kramer to experimental rat rickets, we have obtained results which on the whole agree very well with those of theirs reported at a recent meeting of the Society of Biological Chemists. Because of the small quantities of blood which can be obtained from the animals, it seemed advisable to make the determinations on whole blood rather than plasma if possible. Experiments undertaken to show the relative distribution of the inorganic phosphate in plasma and whole blood indicate that the level of the phosphate is practically the same inside and outside the cells and is maintained at a constant level from day to day. Therefore, as far as the inorganic phosphate is concerned, it is immaterial whether the determinations are made on whole blood or plasma. (By the colorimetric method of Bell and Doisy.?) Table I shows average figures for the inorganic phosphate in theb lood of rats on rickets-producing, normal and high phosphorus diets. TABLE I. INORGANIC PHOSPHATE OF RAT’s BLOOD. Inorganic Phosphate as Mgs. P per 100 c.c. Whole Blood. Blood Diet. ita: ete: Phosphate. ie Ce Eva Me mina- Rickets. Raa TERIA ot AES Te No. % P. | % Ca.| Rats. | tions. Max.| Min.| Av. Norm..| ? ? 55 43 None in 95 % 8.2 | 5.1 | 6.2 Bas vas 86 550 40 18 Marked in 100% 4.9 | 2.0 | 3.2 2 ee ee 72 380 10 y] Marked in 100% 5.0 | 2.3 | 3.5 RE Be 550 7 2 Sl. osteoporosis 5-5 | 5-3 | 5-4 er ssn 120 380 16 |+1%4 Sl. rickets in 25% 7:4 | 3.2 | 6.2 85c 596 020 16 6 Mod. atypical ric. 7.6 | 6.0 | 6.6 | ee 596 O15 12 9 Mod. atypical ric. 9.8 | 6.5 | 8.4 cs, Hous 310 550 3 3 None 9.8 | 9.4 | 9.6 1 Hess, A. F., and Gutman, P., Proc. Soc. Exper. Biot. AND MED., 1921, xix, 31. ? Bell and Doisy, J. Biol. Chem. 1920, xliv, 55. DISTRIBUTION OF INORGANIC PHOSPHATE OF BLOOD. 173 We find that in general a reduction in the inorganic phos- phate of the blood runs parallel to the degree of severity of the rachitic lesions. It will also be seen that the blood phosphate of rats (on these rather specialized diets) may be greatly influenced by the level of phosphate intake. On normal diets the range is from 5.0 to 8.2, averaging 6.2, and on high phosphorus intake a very wide range from 6.0 to 9.8, averaging about 8.5 (mg. P per 100 €.c.). | The dividing line between rickets-producing diets is however sharp. Rats on diets containing 86 mg. per cent. phosphorus all develop rickets and the range of blood phosphate from 2.0 to 5.0 averages usually around 3.2. When as little as 75 mg. per cent. phosphorus is added to the diet, the rachitic lesions fail to appear and the blood phosphorus averages between 5.5 and 6.0. The study of the blood phosphate on rats under light or cod- liver oil therapy brings up several interesting points. TABLE. Il, PREVENTION AND CURE OF RICKETS. Inorganic Phosphate as Mgs. P per 100 c.c. Whole Blood. No. of Blood Phosphate. Diet. Deter- Treatment. Rickets. minations. ee | Ee [ 84 18 Untreated Beading 4.9 2.0 22 + to +-++ 84 10 Mercury vapor lamp Beading 5.4 2.9 4.1 +to + 84 8 Cod-liver oil prepara- Calcification 5.9 2.4 3.95 tions +to +++ As shown in Table II one group of rats on diet 84 (containing 86 mg. per cent. P) were treated with the light from the mercury vapor lamp as a preventive measure. In almost every case complete prevention was secured but the blood phosphorus, while distinctly above that of the controls, was nevertheless in the upper range of rachitic blood. This would seem to indicate that the rats can produce non-rachitic bone at a lower level of phosphorus intake, under the influence of light, than is possible without its ‘presence. Curative experiments with cod-liver oil preparations on another group of rats on the same diet show active calcification of cartilage ’ 74 SCIENTIFIC PROCEEDINGS (120). going on while the blood phosphate is still in the rachitic range. We therefore conclude that a definite deposition of calcium salts may occur before the blood phosphorus regains its normal level. We have as yet no experiments in which the rats were carried through till the healing process was complete, but if we may draw analogies from the work of Howland and Kramer on human rick- ets, we cught to find that when healing is complete the blood phosphate will regain and maintain its normal level. It seems therefore that calcification is not directly controlled by the level of the blood phosphate. Experiments are, however, in progress to determine the nature of the relation between these two factors. 80 (1827) Experiments with quinidine on conduction and on the refractory period in the dog’s heart. By ALFRED E. COHN and ROBERT L. LEVY. [From the Hospital of the Rockefeller Institute for Medical Research, New York City.] These experiments are based on the theory of fibrillation developed by Garrey and Mines on the suggestion of A. G. Mayer and recently elaborated by T. Lewis. The theory, although it illustrates best the condition known as flutter, is directly applicable also to the state of fibrillation. In the normal heart the stimulus for contraction arises at the sinus node and passes in a radial fashion to the rest of the muscle of the auricle. In flutter, and in fibrillation, there is apparently no fixed point at which the stimulus originates. It appears to be dislocated from the node but it is not yet known what activity goes on in this structure when the abnormal rhythm prevails. Instead of the usual arrangement, an excitation wave courses continuously through the muscle of the auricle, usually over a circular path about the open- ings of the great veins, as Lewis’s experiments show. In order that a continuous circuit may be maintained it appears to be necessary (1) that the path shall have a sufficient length; (2) that the rate of passage shall be sufficiently slow, and (3) that when the stimulus returns to its starting point, the muscle is ready to receive it. It is clear, on this plan, that: (1) if the muscle EXPERIMENTS WITH QUINIDINE. 175 mass involved is too small, a path long enough to permit the development of a circus becomes impossible; (2) if the rate of conduction is too fast, the stimulus returns to its starting point before the muscle at that point is ready for its reception; (3) if the refractory period of the muscle at the starting point is pro- longed beyond the time consumed by the stimulus to make its circuit, the muscle cannot be reéxcited. The establishment and maintenance of circus movement depends then on (1) a large mass of muscle; (2) a slow rate of conduction, and (3) a brief duration of the refractory period. When it became clear to us after trial in patients that with quinidine one could terminate the state of fibrillation in about half the individuals afflicted, we began early in 1921 to examine the nature of the activity of this drug. Certain effects which we found the drug to possess on the heart and circulation we reported to this Society at its meeting on May 18, 1921. At that time we reported that the study of its effect on the rate of conduction of impulses through the muscle was under way. We have since added to these studies an investigation of its effect on the duration of the refractory period. On these two phases of the subject we report now. During the course of our work, Lewis’s report dealing with the same phases of the subject has been published. Although the general viewpoints of the two researches are the same, they show a certain difference in that we have attempted to maintain the conditions of the experiments as nearly natural as possible and have, perhaps, in consequence of this difference in plan, arrived at somewhat different results. These experiments were carried out on dogs, anesthetized with ether only. Artificial respiration, at constant pressure and volume, was maintained by the method of Meltzer and Auer. The chest was split and the right auricle exposed by an incision of the pericardium. A wick was sewed to the heart near the base and another near the apex of the auricular appendix. The two wicks led to non-polarizable electrodes. These electrodes formed parts -of two separate galvanometer circuits which were completed by indifferent electrodes inserted in the muscle of the chest wall. Electrodes were also placed on the right fore and left hind limbs. 176 SCIENTIFIC PROCEEDINGS (120). Records at suitable times and for different purposes were then obtainable of the usual lead 2, and of excitation at the base and at the apex of the auricle. A combination of any two records could be taken. Electrodes for sending break shocks into the auricles were permanently fixed in the auricle near the sulcus terminalis. The vagi remained undisturbed. The rate of the heart was natural. By taking simultaneous records of the excita- tion at the base and apex of the auricle, the distance between the electrodes being known, the rate of conduction could be calculated. The refractory period could be known exactly by obtaining and arranging in series the duration of the periods after the waves of auricular activity which the break shock terminated (Table I). These periods fall into two groups: those in which the break shocks fail to elicit a response, and those which bring about a response. The longest of the periods after which a shock fails to elicit a response is the refractory period. The break shocks were signalled by a special device to the camera and were photographed. They were likewise signalled and inscribed on the smoked record of the blood pressure. Records were made before and after injecting quinidine. We report now on six experiments (Table II). The refractory period rose in four: 0.0042 sec. (Exp. 39), .0502 sec. (Exp. 47), 0.0336 sec. (Exp. 48), 0.0120 sec. (Exp. 46), and fell in two: 0.0101 sec. (Exp. 41) and 0.0084 sec. (Exp. 42). In two the change, one upward and one downward, was insignificant (Exps. 39 and 42). The rate of conduction fell in four (Exps. 39, 42, 46 and 47), and remained practically unchanged in one (Exp. 48). In experi- ment 41, the change in this rate is unknown. It is important to point out that after injecting quinidine the heart rate rose 4 times (Exps. 39, 46, 47 and 48). In experiment 46, the rate fell after subsequent injections. In two experiments (Exps. 41 and 42) the rate fell though the change was slight. In the cases in which the rate rose there was an increase in the duration of the refractory period. This result is contrary to what is anticipated, for with a rise in rate, Lewis’s experiments lead one to expect a fall in this measurement. In cases in which the rate fell, there was a fall also in the length of the refractory period. If the circus movement, as it is now believed, underlies the state of fibrillation, and if it depends on the factors which have EXPERIMENTS WITH QUINIDINE. 177 TABLE I. EXPERIMENT 48. Male Dog—8.o kgm. Refractory Periods. : After Before Slee Quinidine. 0.02 see cara: Wo responses. . < sd «5 334s .0825 .0844 .0560 .0876 .0601 -0904 .0605 .OOIS -0607 .09017 .0653 -0917 .0655 -O94I1 .0663 .0950 .0666 -0985 .0678 .0989 .0695 -1000 -0699 -I061 .0700 -1066 .0708 -1078 -0754 -1079 Mer. period .. <.\ 0.626 0763 .1099 TIGSBGHBES S05 Soe eb sms .0766 -1106 .0770 -Irr4! -0792! ok 123" 0812 -I129 .0827 -1135! .0844 .1138 .0855 -II45 -0859 1161! -o861 -II9Q4 .0868 -I200 .0892! .1207 -0960 , eee 38 95 49 21 15 46 27 71 | 38 22 15 47 27 72 39 23 | 16 48 27 73 39 24 16 49 28 74 40 25 17 50 28 75 40 26 17 51 29 76 40 27 18 52 29 77 41 28 18 53 30 78 41 29 19 54 30 79 42 30 ~—CO 19 55 31 80 / 42 31 20 56 31 81 43 32 20 57 32 82 43 33 20 58 32 83 44 34 21 59 33 84 44 . DETERMINATION OF UREA. : 197 five volumes of water are added and, after mixing, a comparison is again made, the result this time being found by consulting Table II. These tables cover a range from Io to 100 mg. of urea N per 100 c.c. blood. TABLE II. DILUTION,—1 VOL. SOLUTION : 7 VOLS. WATER: 1 VOL. NESSLER’S SOLUTION. | Mg. N Mg. N Mg. N Reading. per Reading. per Reading. per | 100 c.c 100 c.c 100 c.c 20 | 31 ra 32 46 62 71 92 se 33 47 64 72 94 23 34 48 65 73 95 24 36 49 66 74 96 25 oe 50 67 75 97 26 38 51 68 76 98 27 39 52 7° 77 99 28 41 53 71 78 100 29 42 54 72 39 43 55 73 31 44 - 56 74 32 46 57 76 33 47 58 77 34 48 59 78 35 49 60 79 36 50 61 80 37 51 62 82 38 53 63 83 39 54 64 84 40 55 65 85 41 56 66 86 42 58 67 88 43 59 68 89 44 60 69 90 45 61 79° gt The tables represent the averages of many determinations made on sunny and on cloudy days. This minimizes the slight differences due to variations in intensity of light. Results obtained with this method agree closely with analyses of the same specimens by the aeration method in which 2 c.c. of ‘blood were used. The micro method may be performed easily in 20-30 minutes with sufficient accuracy for clinical purposes. 198 SCIENTIFIC PROCEEDINGS (121). 92 (1839) The vitamine requirements of certain yeasts and bacteria. By LOUIS FREEDMAN! and CASIMIR FUNK. [From the Laboratory of Biochemistry, College of Physicians and Surgeons, Columbia University, and the Research Laboratory of H. A. Metz, New York City.] In discussing the nutritional requirements of the microérgan- isms, we cannot overlook the important réle that the vitamines play. The identity of the vitamine that influences the growth of the lower organisms is still an open question. The bulk of evi- dence, however, points strongly to the conclusion that this sub- stance is distinct from vitamine B, although it is closely related to it; and our results lead us to draw the same conclusion. Preliminary experiments with beef-heart infusions, peptone, and autolyzed yeast solutions, have shown us that these media contain substances which have a comparable growth-stimulating action on hemolytic streptococci and yeast cells. Thus a beef- heart infusion gives a profuse growth when inoculated with streptococci, whereas this medium, when decolorized by boiling with 2 per cent. of its weight of norit charcoal, loses its growth- stimulating activity, even on addition of a glucose-salt solution. This confirms some of the results obtained by Mueller. When I per cent. peptone or autolyzed yeast solutions are added to the decolorized infusion, the medium again becomes favorable for the growth of streptococci. Analogous results were obtained with these substances on the growth of yeast cells. As it was more desirable to separate these activating substances from the bulk of impurities with which they are associated in their natural media, we subjected beef-heart infusions and autolyzed yeast solutions to fractional adsorption by means of fuller’s earth and norit according to the method of Funk and Dubin. ‘These authors have shown that at least two different substances can be separated from autolyzed yeast by means of fractional adsorption 1 The data in this paper was taken from a dissertation to be presented by Louis Freedman in partial fulfillment for the degree of Doctor of Philosophy, in the Faculty of Pure Science, Columbia University. VITAMINE REQUIREMENTS OF YEASTS. 199 with fuller’s earth. By this method it is now possible to separate the vitamine active for yeast growth, which has been provisionally called ‘‘vitamine D,’’ from that of the anti-beriberi or B vitamine. TABLE I. EFFECT ON GROWTH OF YEAST CELLS AND STREPTOCOCCI OF SHAKING OF AUTOLYZED YEAST WITH FULLER’S EARTH AND NOrIT. Yeast Bacterial No. Growth. | Growth. Fuller’s Earth: I. Autolyzed yeast (5% solution). .......00..c.5000. 12.5 sep a. a ‘* shaken with 50 grams per liter.... 9.5 ++ 2(a). Baryta extract of fuller’s earth (from 2).......... oe ++ Z- Autolyzed yeast (filtrate from 2) shaken with 100 PPI EOE oe cans inn ee sles eee ee hue co 5.0 - 3(a). Baryta extract of fuller’s earth (from 3).......... 4.0 ++ 4. Autolyzed yeast (filtrate from 3) shaken with 100 emmeR PeMeRg eh co2k ievinas ak ek Ave & eb eee Ge 0.5 _ 4(a). Baryta extract of fuller’s earth (from 4).......... 0.0 = Norit: S: Autolyzed yeast shaken with 50 grams per liter.... 10.5 + 5(a). Acetic-acid extract of norit (from 5)............. 4.0 ++ 6. Autolyzed yeast (filtrate from 5) shaken with 100 NS OP IE seo os sia one Ak Bay Oia nie bee ae = 3.0 _ 6(a). Acetic-acid extract of norit (from 6)............. 3.0 + * Autolyzed yeast (filtrate from 6) shaken with roo RE SOE TIEOINS Se PS aged ie eee aia e Seale So 0.0 _ 7(a). Acetic-acid extract of norit (from 7)............. 0.0 | - TABLE, If. SHOWING EFFECT ON GROWTH OF YEAST CELLS AND STREPTOCOCCI OF FRACTIONAL SHAKING OF BEEF-HEART INFUSIONS WITH FULLER’S EARTH AND Norit. Yeast Bacterial No. Growth. | Growth. Fuller’s Earth: I. Beef-heart infusion 1 c.c. [equiv. to (? gm.) beef- WE RETR a oS ow ia ee eae fe aes hs lee Bae ae 12.0 +a 2: Beef-heart infusion shaken with 50 grams per liter. 3.0! ++ 2(a). Baryta extract of fuller’s earth (from 2).......... 1.0 ~ ve Beef-heart infusion (filtrate from 2) shaken with 100 PeM CE MEET 2 5 iat 5 o's’: dina 6 a eee eee 0.0 _ 3(a). Baryta extract of fuller’s earth (from 3).......... 1.0 — Norit: 4. Beef-heart infusion shaken with 20 grams (2%) per eS Se ea 2 gSSas Wap wh Wiehe GENET pia ne Eee ark Si OD + 4(a). Acetic-acid extract of norit (from 4)............. 7.5} +-+ \5. Beef-heart infusion (filtrate from 4) shaken with 50 peas pee lifer =... sis fo ne WR 0.0 _ 5(a). Acetic-acid extract of norit (from 5)............. 0.0 = 1 Average result of several extractions. 200 SCIENTIFIC PROCEEDINGS (121). The activated adsorbents were extracted with baryta and glacial acetic acid respectively, and the influence of these ex- tracts were tested on the growth of yeast cells and streptococci. The results which we obtained and which are embodied in Tables I and II, show that the substances which stimulate the growth of streptococci and yeast cells, as extracted from beef-heart and autolyzed yeast solutions, apparently belong to the class of vitamines of the water-soluble B type, but are not identical with B vitamine. They are comparable in activity and show similar © properties in that they are easily extracted from their natural sources by the same adsorbents, and are again recovered from the adsorbents without appreciable loss in activity. ACTION OF PROTEIN HYDROLYSATES ON BACTERIA AND YEAST CELLS. It is very well known that protein hydrolysates stimulate the growth of certain bacteria, and this stimulating action has been attributed at various times to the presence of unknown substances in the protein molecule. To test out this theory, we subjected to acid hydrolysis twelve animal and ten vegetable proteins, which were prepared and purified by the usual methods, particular care TABLE III. QUANTITATIVE ACTION OF PROTEIN HYDROLYSATES ON STREPTOCOCCI. Pu. of Standard Culture Medium = 7.3. Se ES SATE SS © SRR OE ES SO SO A ES SS No. Hydrolysates of the Proteins. Crowih: Change (1 c.c. used in each test.) in Du. : | Casein (purified) FIC) beydrolgaatet yes. ivi didtadues + 5.8 pecs (sterile conten)... ES _ 7.3 Casein (purified) H,SO, hydrolygates. 6 fees + 5-3 2) (sterile control)............ — 7.0 Casein (technical) HCl hydrolyaate. 0.0010, ie oe + 5.8 3(a). (sterile control)............. - 7.3 ; Gelatin (commercial) «6 vicigiwix’ 5-04 toe oy ive amaen + 6.0 4(a). © {eterile control) 614 fe iit tee ib his _ 7.2 z. Gelatin (prepared and purified in laboratory)........ _ 7.2 (9): 2 (sterile COmErO). 51625 0265'S OSE Laas Pe eae _ 7.3 Edleetitas gua ei: 3s sleine ee Palanan Nadie a dada evans + 6.5 sb ¢ (sterile control).< «'-¢.lveGn MeV rte laces _ 7.3 4. Yeast protele .a85 i.7-F BOR eS eee We td Th ete etceald se + 4.9 7(a). ‘6° (UCT CORI 5 boc ret en ad in cok - 7.3 Notre.—Hydrolysates of 18 other proteins found to be inactive. DIFFERENT IMMUNOLOGICAL TYPES OF B. PERTUSSIS. 201 being taken to have them free of vitamines. These hydrolysates were tested on the growth of streptococci and yeast cells. The bacterial growth was measured by the increase in the acidity of the medium, by means of the Sérensen Indicator method. The action on yeast was not constant, and in most cases showed a growth inhibition due to the known inhibiting action of certain amino acids. | The results on streptococci, which are summarized in Table III, strongly suggest that the growth-stimulating action of protein hydrolysates is not due to a constituent part of the protein mole- cule, but to a vitamine-like substance, probably similar to vita- mine D, which is present as an impurity, and which cannot be removed by the known methods of protein purification. 93 (1840) The existence of different immunological types of B. pertussis. By CHARLES KRUMWIEDE and LUCY MISHULOW. [From the Bureau of Laboratories, Depariment of Health, New York City.] Twenty-two strains having the typical morphological and cultural characteristics of B. pertussis have been studied by means of the agglutination and agglutinin absorption reactions. These tests have demonstrated that the cultures studied fall into two serological groups. If the two groups are tentatively designated as “‘a’”’ and ‘“‘b”’ the results may be briefly described as follows: Anti-serums for group “b”’ agglutinate the strains of group ‘‘b”’ but agglutinate the strains of group ‘‘a’’ very slightly or not at all. The absorption of group ‘‘b” serum by group ‘‘a”’ strains does not appreciably reduce the agglutinins for group ‘‘b.”’ Group “‘a’”’ serum, however, agglutinates group ‘‘b”’ strains to a considerable extent. The absorption of group ‘‘a’’ serum by group “‘b”’ strains results in a reduction of the agglutinins for strain ‘‘a.’’ The serological differences, therefore, are sharply - defined in one direction, but group relationship is shown in the reverse direction. These findings are of immediate interest because of their possible bearing on the use of pertussis vaccines. 202 SCIENTIFIC PROCEEDINGS (121). 94 (1841) The applicability of the precipitin reaction in determining the infectivity of discharges from gonorrheal infections. By MARGARET F. KELLEY. [From the Department of Bacteriology, University and Bellevue Hosptial Medical College, New York City.] The demonstration of gonococci by either culture or smear method is difficult after the subsidence of acute gonorrheal symp- toms. Complement fixation tests on the blood of treated or untreated cases may give negative results. There is need there- fore of a method to determine the persistence of infectiousness. Robinson and Meader! reported encouraging results with the application of the precipitin reaction to discharges of gonorrheal origin. We have attempted to verify their results, working under a grant from the U. S. Interdepartmental Social Hygiene Board. Selected rabbits were immunized with live gonococci to pro- duce the immune serum used for the tests. Specific gonococcus antigen was prepared by autolyzing the gonococcus in salt solution for several days and centrifugalizing to obtain a clear antigen. “Discharge extracts’ from cases were prepared by adding to 2 c.c. of salt solution the secretions obtained from the cervix or vagina. The mixture was allowed to stand over night and then centrifugalized until clear. 0.2 c.c. of clarified extract was then added to 0.2 c.c. of diluted immune serum and to 0.2 c.c. of diluted normal serum as a control. A positive result was shown by the development of a ring of varying thickness and opacity at the point of contact of extract and serum or by the development of a precipitate. The reaction appeared usually from two hours to eighteen hours. With 92 specimens, smears positive, 82 per cent. gave reactions with gonococcus serum while 21 per cent. gave reactions with normal serum. With 49 specimens, smears negative, 61 per cent. gave reactions with gonococcus serum, while 51 per cent. gave reactions with normal serum. The relatively frequent reactions 1G. H. Robinson and P. D, Meader, The Journal of Urology, 1920, iv, 551. ~~ ee PRECIPITIN REACTION. : 203 with normal serum indicated the presence of a non-specific factor and raised the question as to whether the reactions occurring with immune serum alone could be considered specific. Of 17 non-gonorrheal vaginal specimens from children 100 per cent. reacted with immune serum and 94 per cent. with normal serum. Nose and throat specimens and a miscellaneous group of sputums, pus due to infection by bacteria other than gonococci and peritoneal washings from normal mice or mice inoculated with exudates due to pneumococcus or streptococcus, gave similar non-specific reactions. Various methods have been employed in the attempt to elimi nate or lessen the non-specific reactions, so that a specific reaction could be recognized if it occurred. The standardization of the opacity of “‘discharge extracts,” the dilution of the serums or of the extracts, or of both, and finally the heating of the extracts have failed to be of help. . With gonococcus serums, precipitates were most frequently encountered with antigens prepared from the staphylococcus and meningococcus. Absorption of gonococcus serums by _ these heterologous types did not reduce appreciably the reactions ob- tained with extracts from non-gonorrheal sources. That the reactions obtained with gonococcus case extracts could not be considered as specific was most conclusively shown by the persistence of reactions after the gonococcus serum was absorbed by the gonococcus. That is, when gonococci were added to the serum to the point where it no longer reacted with a known gonococcus antigen, it still gave a precipitate with extracts from gonococcus cases. Although a specific reaction might have oc- curred at times, the presence of this non-specific factor would have obscured it. The precipitin reaction, therefore, as recommended by Robin- son and Meader is not applicable for the determination of the presence of the gonococcus in discharges from the cervix, urethra, etc. 204 SCIENTIFIC PROCEEDINGS (121). 95 (1842) A study of oxalic-acid poisoning. By SAMUEL A. BROWN and ALEXANDER O. GETTLER. [From the Chemical Laboratory of The Uniwersity and Bellevue _ Hospital Medical College and of the Pathological Department, Bellevue Hospital, New York City.] The earliest case of oxalic-acid poisoning, reported by Royston, occurred in England, in 1814. Since then, the number of deaths due to oxalic acid and its soluble salts has so increased that today it ranks among the first three poisons in the number of fatalities. A. W. Blyth states that in the five years between 1912 and I916, there were 448 deaths in England and Wales due to oxalic acid. The duration of a case of oxalic-acid poisoning is usually be- tween 2 and 14 days. There is one case on record by Ogilvie (Lancet, 1845) however where death occurred within 3 minutes. Oxalic acid acts locally as a corrosive and also as a systemic poison. Locally it is more or less destructive to the mucous membrane with which it comes in contact. The lips, tongue, pharynx and esophagus are discolored yellowish white, sometimes marked with patches of a reddish hue. The mucous surface of the stomach is coarsely corrugated and presents a bright red color | both in the elevations and depressions; this may change to brown or even black by postmortem action. In some cases the mucous surface is in part or in whole pale, opaque or translucent, and marked by a coarse ramiform vascularity of the submucous tissue. The mucous membrane is soft, pulpy, eroded in patches, thrown into folds, and is easily detached. Perforation is rare. The systematic effects are attested by falling of the blood pressure, arhythmia and retardation of the pulse, slow breathing, paralytic symptoms and fibrillary muscular contractions. Some consider it a poison acting on the extracardiac ganglia. The red blood corpuscles are destroyed, with consequent fatty degen- eration of the tissues. The activity of the muscles is diminished consequent upon loss of irritability. The respiratory muscles are paralyzed. A Stupy OF OxALIc ACID POISONING. 205 Symptoms.—Although more than 1,000 cases of oxalic-acid poisoning have occurred since Christison wrote his treatise, his description still holds good. “If a person immediately after swallowing a solution of a crystalline salt, which tasted strongly acid, is attacked with burning in the throat, then with a burning in the stomach, vomiting, particularly of bloody matter, imper- ceptible pulse, and excessive languor, and dies in half an hour, or still more, in twenty, fifteen, or ten minutes, I do not know of any fallacy which can interfere with the conclusion that oxalic acid was the cause of death. No parallel disease begins so ab- ruptly, and terminates so soon; and no other crystalline poison has the same effect.’’ There may also occur headache, cold extremities, numbness and tingling, loss of voice, cramps, con- vulsions, delirium, coma, etc. Prognosis.—Out of 242 reported cases, there were 132 deaths, a mortality of 54.5 per cent. This ratio, of course, depends upon the manner and speed of treatment. Nor is the outlook for complete recovery favorable if the initial degree of poisoning was severe. There have been a few cases reported in which patients returned after some months suffering from gastric irritability, dyspepsia and symptoms of constriction of the esophagus, the latter due apparently to destruction and subsequent repair of the mucous membrane. Postmoritem.—Aside from the local corrosive action, there are no typical pathological lesions with one exception, namely, in the kidneys, where the cortical substance may present a definite whitish appearance due to the presence in the tissues of crystals of calcium oxalate. None is deposited in the glomeruli. Calcium- oxalate crystals have also been found in the blood, bile, aqueous humor, and pleural and pericardial fluids. Elimination.—Is mainly through the urine. The reported analyses show that from 80 to go per cent. is excreted through this channel. The urine also contains albumin, a reducing substance and hyaline casts. | Treatment.—The oxalic acid should be neutralized and pre- cipitated as quickly as possible, by giving plenty of syrup of lime or a suspension of calcium carbonate. After a few minutes the stomach should be washed out with lime water and, lastly, with 206 SCIENTIFIC PROCEEDINGS (121). plenty of plain water. Stimulants and warmth should be ad- ministered to avoid collapse. Diuretics and an abundance of liquid should be given to combat nephritic conditions, and alkalies should be administered to prevent the tendency toward acidosis. I purpose to present to you to-night a case that came for treatment on the Third Medical Division of Bellevue Hospital, under the direction of Dr. S. A. Brown. This case is of interest because the patient recovered, although antidotal treatment was delayed, and also because of the complete blood study throughout the patient’s stay in the hospital. | J. M., a building superintendent, age 44. Present History—The patient had no bowel movement for 2 days and decided to take what he thought to be a dose of epsom salts. He immediately noticed a peculiar sour taste, with a burning sensation along the esophageal tract, followed presently by severe pains in the epigastrium, which gradually increased. An ambulance was summoned and when the surgeon attempted to examine the patient’s throat he vomited. Thesurgeon, diagnosing it as a case of indigestion, administered sedatives and left. Shortly thereafter the patient developed pain in the lumbar region, the ambulance was summoned again and he was taken to the hospital on October I, 1921, at 4:30 A.M. In the course of the succeeding hours he complained of burning pains in the stomach, first localized, then radiating through the abdomen, and of pain and tenderness in the lumbar region. The next morning, after taking a glass of milk, he again vomited. On physical examination, he was found to be well developed and well nourished. ‘The pupils were equal and regular, reacting readily to light and accommodation. The tongue was clean, the teeth poor, the throat congested. There were no adventitious sounds in the lungs. The heart sounds were of fair quality, no murmurs, rhythm regular, rate 90. The abdomen showed no rigidity, no masses, no tenderness. The spleen was palpable just below the costal margin. The liver edge was also palpable. On deep pressure in the epigastrium, there was pain. There was no edema. Reflexes—no Babinski, no clonus, abdominal cremasteric and knee jerk active. Treatment consisted of fluid diet, with hot packs, colon irriga- A Stupy OF OxALIc ACID POISONING. 207 tions and the following medication: At first bromides and chloral and magnesium sulphate, followed by luminal; later Tr. nux vomica, sodium bicarbonate and veronal. Toward the latter part of his stay in the hospital he was given bismuth subgallate and triple phosphates. It should be noted that no antidotes for oxalic acid were given, as the nature of the poison, if any, was unknown. Upon analysis the salts of which patient had partaken were found to contain 73 per cent. Mg. SO.zand 27 per cent. oxalic acid. The patient stated that he took what is estimated to be I5 to 18 grams of the salt. This means that he obtained 4 to 4.9 grams of oxalic acid. The most common fatal doses are 7 to 15 grams, although there is one case on record in which 3.88 grams proved fatal. The urine was analyzed for oxalates and found to contain 9.2 mg. of oxalic acid in 100 c.c. of urine. It also contained al- bumin, white blood cells, red blood cells, hyaline and granular casts. | The blood was analyzed throughout the patient’s stay in the hospital, at intervals of a few days, with the following results: BLoop ANALYSIS. Time Interval in Days. 4 ime) 14 I9 25 28 31 33 mg mg mg mg. mg mg. mg mg Non-protein nitrogen.| 85 270 200 60 73 37 Ao 37 Urea nitrogen......:. 59 211 I49 39 5r: > 26 ce) 17 Creatinine. ......... 4.3 By 2.3 1.6 Ez I.0 r 5 1.6 Wie Beth ee es 4.1 6.3 2 £5 1.0 | — T.2 1.5 oe ele a 174 98 — 75 70 — 85 —- Alkaline reserve, per SIRES Be eles ees | 48 | 41 46 55 — 59 — 55 The study of the chemistry of the blood shows a gradual increase in the excretory products. This finding may be explained by the mechanical effects of the calcium-oxalate deposition in the kidneys, seen at autopsy in similar cases. The highest point was reached on the tenth day. In connection with the accepted view that cases in which creatinine is higher than 5 gm. do not recover, it is interesting to notice that the creatinine in this case was 5.1 mg.on thetenthday. A possible explanation for recovery 208 — SCIENTIFIC PROCEEDINGS (121). in cases of oxalic-acid poisoning with creatinine over 5 mg. may be that the kidney changes are temporary. The alkaline reserve was below normal throughout the retention period. This evidently indicates that in poisoning by oxalic acid the oxidation processes are subnormal. On the twenty-eighth day all values were back to normal. . The urine output during the first few days was much sup- pressed; the amount voided per day was 30 to 50 c.c. On about the twelfth day, the urine output suddenly increased to 1,900 to 2,200 c.c. It contained little albumin, few hyaline and granular casts, a reducing substance which did not ferment, together with red blood cells, white blood cells and an increased amount of oxalates. In view of the fact that over 4 grams of oxalic acid had been taken and that treatment was delayed (in fact, no antidote was given at all), it is interesting to note that recovery occurred. The most plausible explanation for this is the simultaneous taking of magnesium sulphate, which probably hastened the elimination of the poison. About one month later the patient returned to the hospital complaining of gnawing pain in the epigastric region. This pain usually started a half hour after meals and continued until the next meal. It was aggravated by any kind of food, more so by hot food. He had eructations that were sour in character, was constipated, but not anureous. Fluoroscopic examination showed no ulcer or new growth. The x-ray plates showed a calcareous area in the region of the gall-bladder. This condition is evidently a direct result of the corrosive action of the oxalic acid on the mucous membrane of the stomach. i LIGATION OF THE THYROID ARTERIES. 209 96 (1843) Further studies on ligation of the thyroid arteries in depancreatized diabetic dogs. By G. A. FRIEDMAN and J. GOTTESMAN. [From the Department of Clinical Pathology, College of Physicians and Surgeons, Columbia Unwwersity, New York City.] The experiments described in this and in the following paper were conducted during the year of 1921. 37 dogs were depan- creatized; 9 did not develop glycosuria although in some of them repeated pancreatectomies were performed at various intervals. 3 of the animals showed a transitory glycosuria. 25 dogs became persistently glycosuric; 9 of these dogs died of various causes. Ligations of all of the thyroid arteries were done in 8 of 16 gly- cosuric dogs who survived the pancreatectomies, in about 4 to 7 days after complete or almost complete removal of the pancreas. In one dog the thyroid arteries were ligated in two sittings: first both arteries on the right side and seven days later the arteries on the opposite side. This dog died from tetany on the day following the second ligation. 3 of those who were completely ligated died without demonstrable causes from I to 2 days after ligation. Thus in only 5 dogs could the effect of complete ligation be studied and only in one the effect of partial ligation. The great losses in body weight which usually follow de- pancreatizations were not checked by the ligations. Some interesting points, however, were brought out from the study sufficient to warrant a report. As far as we know there is no literature pertaining to ligations of thyroid vessels in depan- creatized dogs. In a previous paper! we referred to an experiment with dog No. 106 who was completely depancreatized. His urine on daily examinations showed from 5 per cent. to6 per cent. sugar. On the fourth day all thyroid arteries of the animal were ligated. He became sugar-free five days after the operation, and he remained 1 Friedman, G. A., and Gottesman, J., PROCEED. Soc. Exp. BIOL. AND MED., 1921, xviii, 281. 210 SCIENTIFIC PROCEEDINGS (121). without a trace of sugar in the urine for ten days, when he de- veloped distemper and died from pneumonia. Dog 120.—Male. Weight 9.6 kilos. Complete pancreatec- tomy June 22. Glycosuria from June 23 to June 28. Ligation of all thyroid arteries June 28. Sugar positive June 29. Sugar negative on daily examinations of passed and of catheterized specimens from June 30 to July 13. Dog was found dead July 14. Autopsy same day. No demonstrable lesions. Course: good appetite until June 12. No signs of tetany. Sections of the thyroid show many alveoli devoid of colloid (Fig. 1.) Dog 108.—Female. Weight 8.7 kilos. Complete pancreatec- tomy March 16. Glycosuria from March 17 to March 24. Liga- ad ss Siete od . “ + Fic. 1. From dog 123. tion of all thyroid arteries March 24. Glycosuria persisted until April 1. Died April 2 from emaciation. Autopsy on same day. No demonstrable lesions. Course: poor appetite two days after removal of pancreas. Refused food completely from the day following ligation. No acute attack of tetany, but twitchings of the musculature of the back noted two days following ligation. LIGATION OF THE THYROID ARTERIES. 211 Sections of thyroid show extensive hemorrhagic infiltration, marked atrophy. Few remnants of thyroid seen underneath capsule (Fig. 2). Fic. 2. From normal dog. Dog 121—Female. Weight 20 kilos. Complete pancreatec- tomy June 29. Glycosuria from June 30 to July 5. Ligation of all thyroid arteries July 5. Sugar strongly positive July6. Sugar negative July 7. Sugar positive from July 8 to July 14. Course: slight attack of tetany from July 8 to July Io. July II severe attack. July 12 infection noted at neck. Mild attack of tetany. Killed July 14 with chloroform. Large pocket of pus at neck. Dog 128.—Female. Weight 13.8 kilos. Almost complete pan- createctomy July 25. Glycosuria from July 27 to July 30. Liga- tion of all thyroid arteries July 30. Glycosuria persisted until August 6, when she died. No autopsy. Course: No tetany. Infection at neck noted August 3, four days after ligation. Dog 111.—Male. Weight 6.66 kilos. Almost complete pan- createctomy April 3. Glycosuria from April 4 to April 1o. Liga- tion of superior and inferior arteries on the right side April Io. SCIENTIFIC PROCEEDINGS (121). 212 TABLE Tf. S. BLOODSUGAR IN NORMAL DoG Megrm. Sugar per Megrm. Sugar per 100 c.c. Blood. No. 100 c.c. Blood. Ito III I2I 122. 123 124 133 TABLE II. BLOODSUGAR IN DIABETIC DOGS FOLLOWING PANCREATECTOMY. Megrm. Sugar per Megrm. Sugar per 100 c.c. Blood. No. 100 c.c. Blood. oO. I2I 123 124 P26 42 oe See 136 137 138 203 7 £50 12 104 105 106 BOTS or Soe tee oe 108 109 III mk yi ~~ gE Pa Bich From dog 120. FIG. 3. LIGATION OF THE THYROID ARTERIES. 213 Glycosuria from April 11 to April 17. Ligation of superior and inferior arteries on the left side April 17. Sugar positive April 18. Dog died on the following day from a severe attack of tetany. No autopsy. Course: animal was in perfectly good condition until the day following the second ligation. There was a strong ferric-chlorid reaction in his urine on the third day after pan- createctomy. There was no diacetic-acid reaction on succeeding days. Table I—Bloodsugar estimations were made in these animals before the pancreatectomies. They were starved at least 24 hours preceding the operation. The highest figure obtained 104 mmgr. per 100 c.c. blood in 3 dogs in whom estimations were made during the hot summer months July and August. Table II —Dog 124 was glycosuric throughout after pancrea- tectomy, but there was no increase in the contents of bloodsugar. Dog 123 after almost complete pancreatectomy (a minute rem- nant of pancreas was found at autopsy) did not develop glycosuria while under observation for 26 days. His original weight was 11.2 kilos. On the last day of observation his weight was 9.7 kilos, when he was disposed of with chloroform (loss of approxi- Fic. 4. From dog 108. 214 SCIENTIFIC PROCEDINGS (121). mately 60 grm. per day). Such a loss in dogs after almost com- plete pancreatectomy may be considered as a slight one. Sections of his pantreatic fragment showed very few Langerhans islands and sections from his thyroid (Fig. 3) showed unusually large alveoli rich in colloid. Compare this thyroid with one of a normal dog (Fig. 4) who weighed 13 kilos. We emphasize this point because Cohen? and Pariser reported changes in the thyroid in various organic diseases of the pancreas in man whose urine was negative for sugar. TABLE Tilt; MmMcGrR. SUGAR PER I00 C.c. BLOOD AND DATES WHEN BLOOD WAS TAKEN FOR ESTIMATION. Before After Ailey No. Pancrea- Pancrea- Li 4 Remarks. ps igation. tectomy. tectomy. [SS eee) ae ee eee Se Sas Cee ee Se ee ee ee ee 106 | 76 March 2 | 250 March 5 192 March 7 Animal became sugar-free. 90 a 10 50 108 | 67 March 16} 222 March 18 180 March 25 | Bloodsugar diminished af- oe ae 293 225). 4A 30 ter ligation. Increased with tetany. 111 | 78 April 3 154 April 7 244 April 17 Glycemia increased after F909)" eG partial ligation. 120 238 June 28 200 July 1 Animal became sugar-free. 172 oe 5 I2r | 98 June 29 238 July 5 | 213 July 14 | Sugar free 1 day after ligation. Reappearance of sugar with tetany. 128 228 July 30 Did not become sugar-free. Tetany absent. Infec- tion present. Table III.—From this table it becomes evident that partial ligation did not only diminish the diabetic glycemia, but made it more intense. Table IV.—According to Allen*® the weight of the pancreas in dogs is approximately 2 grm. per kilogram of body weight. This table shows our figures come very close to his in completely de- pancreatized dogs. 2 Cohen, Moritz, and Pariser, Hans, Disch. Med. Woch., 1912, 38', 60. 8 Allen, Frederick M., ‘‘Studies Concerning Glycosuria and Diabetes,’’ Boston, W. M. Leonard, publishers, 1913, p. 716. PANCREATIC DIABETES IN DoGs. 215 TABLE IV. Weight of |Approximate Weight of Removed Weight of Body Removed Pancreas Removed No. | Weight Pancreas per I Kilo. Gland Remarks. in Kilos. te of Body According Grams. Weight in to Grams. Allen. 106 gi52 18.2 | 2.4 15.04 No pancreatic remnant at autopsy. 108 8.7 | 21 2.4 17.4 Complete pancreatec- tomy. III 6.56 T7 | 2.4 $5232 Remnant weight at autopsy 4 grams. 120 9.6 21 ' ea | 19.2 No remnant at autopsy. 121 20 ! 44 | 2.2 40 Complete removal. | 128 13.8 20 ‘ 1.45 27.6 Almost complete _ re- | moval. No autopsy. CONCLUSIONS. 1. The mortality of completely depancreatized dogs after liga- tion of the thyroid arteries is high. 2. Glycosuria in depancreatized diabetic dogs was checked after complete ligation of the thyroid arteries. 3. Tetany or infection, or both, seem to interfere with the disappearance of the glycosuria. 4. Partial ligation of the thyroid arteries apparently intensifies the diabetes produced by pancreatectomy. 97 (1844) The relation of the thyroid and parathyroids to pancreatic diabetes in dogs. By G. A. FRIEDMAN and J. GOTTESMAN. [From the Department of Clinical Pathology, College of Physicians and Surgeons, Columbia University, New York City.] Lorand ! (1904) and McCallum? (1909) performed complete thyroidectomies in depancreatized diabetic dogs. The former 1Lorand, A., Compt. rend. Soc. Biol., 1904, lvi, 488. ? McCallum, William George, Johns Hopkins Hosp. Bull., 1909, Sept. 216 SCIENTIFIC PROCEEDINGS (121). worked with 3 dogs and the latter with two. In Allen’s! experi- ments (1913) with diabetic dogs the thyroidectomies were in- complete. Eppinger,? Falta and Rudinger (1908) removed first the thyroid and later the pancreas in two dogs and they did a simultaneous thyroidectomy and pancreatectomy in one dog. Eppinger® and his associates (1909) also studied the relation of parathyroid insufficiency to the metabolism in diabetic dogs by removing simultaneously the pancreas with three parathyroids in two animals. - Lorand has asserted that removal of the thyroid sparing the parathyroids is followed by the disappearance of sugar from the urine in two days in depancreatized dogs. In one of McCallum’s dogs the glycosuria ceased after removal of the thyroid; in the other one it greatly diminished. In one of the experiments of McCallum two parathyroids were spared; in the other all were left in situ. A marked diminution of sugar after thyroidectomy was also reported by Eppinger. In all these experiments, blood- sugar estimates were not made. The duration of life in the animals of Lorand and McCallum was from one to three days after com- plete removal of the thyroid even if the parathyroids were left in situ. Three of our diabetic dogs in whom thyroidectomy with partial parathyroidectomy was performed 3 to 4 days after pan- createctomy, died from 1 to 3 days after the operation. From two of these dogs no urine was obtained, and in one dog the gly- cosuria persisted on the day following the removal of the thyroid. We may mention here two clinical cases cited by Rohdenburg.! One patient was diabetic and later developed exophthalmic goitre. A portion of his thyroid was removed and he remained perma- nently sugar-free. The other patient had exophthalmic goitre for which a portion of the thyroid was removed. Several years later he developed glycosuria. The glycosuria in this case disappeared after removal of more of the thyroid gland. We decided that it would be a better procedure to first partially ligate the thyroid arteries and follow this operation on a later date by partial thyroidectomy, or if possible by thyroidectomy 1 Allen, Frederick M., 1913, p. 848. * Eppinger, Falta and Rudinger, Zischr. f. Klin: Med., 1908, Ixvi, 1. * Eppinger, Falta and Rudinger, Zischr. f. Klin. Med., 1909, 380. * Rohdenburg, G. L., ‘‘Endocrinology,"’ 1920, iv, 63. PANCREATIC DIABETES IN DOGs. 217 alone. One may occasionally succeed in sparing all parathyroids while removing the thyroid especially in larger dogs. In a previous! paper we referred to an experiment with dog No. 100, who became diabetic after removal of a little over one half of his pancreas. Such rare results are occasionally reported in the literature. As the external secretory apparatus of the pancreas was not much affected, but continued functioning, we believe that truly by chance, we produced in this animal a condi- tion which came very close to human diabetes. Inasmuch as this experiment is a singular one in literature we shall briefly refer 10 ite Seven days after the dog had from 2 to 3 per cent. sugar in his urine, both inferior thyroid arteries were ligated. The glycosuria persisted on daily examinations. Seven days later both lobes of the thyroid were removed and we succeeded in sparing all of his parathyroids. There was not a trace of sugar in his urine on the day following thyroidectomy and the urine remained sugar-free for 108 days, although eleven days after removal of the thyroid, additional pancreatic tissue was removed, and ninety-three days after the second pancreatectomy the last remnant of the gland was taken out. The animal was sugar-free four days after the third pancreatectomy. He died on the fifth day from prolapse of the intestines, which was probably brought about by the three laparotomies. ; The dog’s condition was excellent; ate well until the day of the accident. He did not show any signs of myxedema during the time of observation and while his original weight was 14 kilos his weight the day before death was 15.9 kilos, or a gain of 1.9 kilos. His blood sugar remained normal and 14 days after re- moval of the thyroid his sugar tolerance was about Io grams per kilogram of body weight. Fig. 1 shows a photograph of the dog 93 days after removal of his thyroid. Conditions become quite different when one does not succeed in imitating human diabetes and when sparing all the parathyroids -in doing a parathyroidectomy is impossible. Infection also changes the situation. It is our impression that diabetic dogs 1 Friedman, G. A., and Gottesman, J., PROCEED. Soc. Exp. BIoL. AND MED., 1921, xviii, 281. 218 SCIENTIFIC PROCEEDINGS (121). are more susceptible to tetany after thyroidectomy and partial parathyroidectomy than normal dogs. Although in the experi- ments by one of us! in studying the influence of thyroidectomy and partial thyroidectomy on the gastric mucosa, tetany never oc- curred while leaving in situ 2 or 3 parathyroids; the occurrence Fic. 1. Dog 100, 93 days after thyroidectomy. of mild or severe attacks after this procedure was the rule in our diabetic dogs. We were able to observe in one diabetic dog a palliative effect on tetany from calcium lactate injections as pro- posed by McCallum? .and Voegtlin with parathyroidectomy in non-diabetic dogs. The following experiments will show the in- fluence of tetany upon the glycosuria in diabetic dogs and the opposite effect of the thyroid. ‘Friedman, G. A., Jour. Med. Research, 1918, xxxviii, 69- * McCallum, W. G., and Voegtlin, Carl, Jour. Exp. Med., 1909, xi, 118. PANCREATIC DIABETES IN DoGs. 219 Dog 136.—Male. Weight 13 kilos. Partial pancreatectomy Nov. 9: Glycosuria from Nov. 10 to Nov. 16. Ligation of inferior thyroid arteries Nov. 16. Glycosuria from Nov. 17 to Nov. 30. Thyroidectomy Nov. 30. Superior parathyroids left in situ. Thyroid lobes unusually small. Dec. I urine sugar free. Dec. 2 glycosuria. Dog developed a severe attack of tetany early in the morning and died in the afternoon. Autopsy: No demonstrable lesions. Dog 138.—Male. Weight 17.71 kilos. Partial pancreatec- tomy Dec. 3. Glycosuria from Dec. 5 to Dec. 7. Ligation of inferior thyroid arteries Dec. 7. Glycosuria persisted from Dec. 8 to Dec. 10. Thyroidectomy preceded by intravenous injection of 10 c.c. of 5 per cent. solution calcium lactate. Three para- thyroids left in situ. Dec. 12 and 13 sugar positive. Twitchings of the musculature of the back. 50 c.c. calcium chloride 5 per cent. solution by stomach tube daily. Dog developed a severe attack of tetany early in the morning Dec. 14. At 11 A.M. intravenous injection of Io c.c. calcium-lactate solution 5 per cent. Recovered from the attack; but twitchings persisted. At 2 P.M. second attack of tetany. Dog very low, in a dying condition. At 3 P.M. another injection of calcium lactate, same dosage. Dog was catheterized twice and sugar found in the urine by adding 2 drops of Benedict’s reagent. Dec. 15 and 16, no tetany or twitching. Dog had an excellent appetite. Not a trace of sugar in catheterized or passed specimens with either Benedict’s or Nylander’s reagents. Dec. 17, 18 and 19 twitchings of the musculature of back. No actual attack of tetany. Sugar in urine strongly positive. Mild attack of tetany though the dog had received subcutaneously Io c.c. of calcium lactate 5 per cent. solution daily. Dec. 20, attack of tetany. Sugar strongly posi- tive in the urine. Intravenous injection of calcium lactate in the morning and in the afternoon. Dec. 21 and 22, notetany. Urine sugar-free. Subcutaneous injections of calcium lactate were given ‘for three days. Dec. 23 dog was bitten in the back by another dog. On account of the large open wound he was killed. Dog 139.—Female. Weight 12 kilos. Pancreatectomy and ligation of inferior thyroid arteries Dec. 17. Glycosuria from Dec. 18 to Dec. 31. Thyroidectomy Dec. 21. Two superior para- 220 SCIENTIFIC PROCEEDINGS (121). thyroids left in situ. Glycosuria persisted from Dec. 23 to Dec. 25; twitchings of musculature of the back were noted the following day. The twitchings persisted though daily subcutaneous injec- tions of calcium lactate were given. Dog was found dead Dec. 26. Autopsy: No pneumonia. Stomach filled with blood. No food present. Mucosa of the pylorus covered with numerous hemor- rhagic erosions. TABLE V. MnMcr. OF SUGAR IN 100 C.c. BLOOD AND DATES WHEN BLOOD WAS TAKEN FOR ESTIMATION. After After After No. Pancrea- Partial Thyroi- Remarks. tectomy. Ligation. dectomy. 100 95 Feb. 15 Sugar-free 108 days after 66 "5 thyroidectomy. 66 April 15 136 | 250 Nov. 16/ 285 Nov.30} 145 Dec. 2 Increased bloodsugar after partial ligation. Sugar-free when tetany was absent. 138 | 290 Dec. 7 | 310 Dec. 10} 238 Dec. 17 Increased bloodsugar after 100. "(a partial ligation. Glycosuria absent on days when tetany free. | 139 | 228 Dec. 21 | 228 Dec. 21 Pancreatectomy and partial ligation in one sitting. Did not become sugar-free. Mild | attacks tetany after removal of thyroid. Table V.—Note the increase of bloodsugar in dogs 136 and 138 after partial ligation. A similar increase in dog III, previously reported, after the same procedure. In three dogs the diabetes caused by pancreatectomy was not checked by partial ligation, but became even more intense. Compare these figures for blood- sugar with the normal figures in dog 100 after thyroidectomy with the diminished amounts after thyroidectomy with partial para- thyroidectomy in the other dogs. The bloodsugar of dog 138 which amounted to 100 mgr. was found on a day when he was free of urinary sugar and free from tetany. We did not succeed in obtaining blood from dog 139 after thyroidectomy as his veins at the neck collapsed so that it was impossible to introduce a needle. PANCREATIC DIABETES IN DoGs. 221 TABLE VI. Weight of Weight of Approximate N Body Weight Removed Removed Pancreas Weight of > in Kilos. Pancreas in per 1 Kilo of Gland According Grams. Body Weight. to Allen. 100 14 ? ? 28 3-4 10 136 13 17 | fee 26 138 15.71 26 1.65 31.42 139 12 23 I.Q1 24 Table VI indicates that it is possible to obtain a persistent glycosuria after removal of I to 1.3 grams of pancreas per 1 kilo- gram of body weight. Allen! figures that the pancreas of a dog weighs approximately 2 grm. per kilogram of body weight. This makes us believe that we probably removed 14.6 grams in dog No. 100 at the first operation. CONCLUSIONS. 1. Diabetic dogs are more susceptible to tetany after partial parathyroidectomy and thyroidectomy than non-diabetic dogs after the same procedure. 2. The removal of the thyroid in diabetic dogs seems to check the glycosuria provided tetany does not occur. 3. If tetany does occur intravenous injections of calcium lactate may act as a palliative in checking temporarily both the tetanic seizures and glycosuria. 10 Allen, l.c., 716. 222 SCIENTIFIC PROCEEDINGS (121). 98 (1845) Concerning the amount and distribution of stainable lipoid mate- rial in renal epithelium in normal and acutely nephropathic animals, with observations on the functional re- sponse of the kidney. By WM. DEB. MACNIDER. [From the Laboratory of Pharmacology of the Unwersity of North Carolina, Chapel Hill, North Carolina.] The following observations are based upon a study of twenty- six dogs. The animals were under two years old. The dogs were placed in metabolism cages and studied for eight days prior to any experimental interference. During this period, as well as during the period of the experiments, the animals were given 250 c.c. of water twice a day by stomach tube. The diet consisted of scraps of bread and cooked meat. The urine was collected once a day and examined qualitatively for albumin and glucose. The phenolsulphonephthalein test for renal function was made every other day according to the technique of Rowntree and Geraghty. Daily dete1minations of the alkali reserve of the blood (R.p.H.) were made by the method of Marriott. Blood-urea determinations were made by the method of Marshall as modified by Van Slyke and Cullen. At the end of the eight-day period allowed for normal observa- tions, seven of the animals were killed without the use of an anesthetic and used as control experiments. The remaining nine- teen animals were given one subcutaneous injection of 6 mgs. of uranium nitrate per kilogram. Following the use of uranium, observations similar to those previously outlined were continued. The animals that were given uranium were killed without the use of an anesthetic 6 hours, 12 hours, 24 hours and 48 hours following the commencement of the intoxication. At the termination of the observations on both the normal control animals and the acutely nephropathic animals, kidney tissue was at once obtained for microscopic study. Sections of tissue extending through each lateral half of both kidneys were placed in isotonic salt solution Lireoip MATERIAL IN RENAL EPITHELIUM. 223 and without any fixation were frozen, sections made, and stained for lipoid material by Herxheimer’s Scharlach R. method. Such sections were counterstained with Mayer’s Haemalum. Other tissue from both kidneys was fixed with formaline, Zenker’s fluid, and in corrosive-acetic, imbedded in either paraffin or celloidin, and used for a general histological study. NORMAL CONTROL ANIMALS. During the eight days of observation, urine formation by the seven normal control animals has varied from a minimum output of 385 c.c. to a maximum output of 621 c.c. The urine was free from both albumin and glucose. The elimination of phenolsul- phonephthalein by the respective animals in a two-hour period varied from 65 per cent. to 80 per cent. The blood urea varied from 12 to 18 mgs. per 100 c.c. of blood. The reserve alkali of the blood was normal and gave readings between 8.0 to 8.1. When such animals were killed without the use of an anesthetic and kidney tissue studied for the amount and distribution of stainable lipoid material by the use of Scharlach R., the following observations were made. The endothelium of the glomerular capillaries and other vascular tissue of the kidney failed to show the presence of stainable lipoid. The convoluted tubule epi- thelium in young animals such as have been used in this study does not show the presence of stainable lipoid with Scharlach R. In old normal animals as has been previously noted,! stainable lipoid may appear in the epithelium of this portion of the tubule in the form of fine dust-like particles. All of the normal control animals show stainable lipoid in both the descending and ascending limbs of Henle’s loops. In this portion of the tubule such material appears as small particles or fused droplets. The study of the normal control group of animals indicates that stainable lipoid as demonstrated by Scharlach R. with Herx- heimer’s technique of staining is confined to the epithelial cells of the loops of Henle. Lipoid material in this location has no harmful effect on the functional capacity of the kidney and does not interfere with that function of the kidney which is concerned with maintaining a normal acid-base equilibrium of the blood. 1 MacNider, Wm. deB., Jour. Pharm. and Exp. Therap., 1921, xvii, 280. 224 SCIENTIFIC PROCEEDINGS (121). ACUTELY NEPHROPATHIC ANIMALS. The experiments on the animals acutely nephropathic from uranium were terminated as follows. Four animals were killed six hours after the administration of uranium, five at the end of twelve hours, five at the end of twenty-four hours and the remain- ing five animals at the end of forty-eight hours. The four animals killed six hours following the administration of uranium showed no change in the normal functional response of the kidney. During this period the animal of Experiment 4. formed 129 c.c. of urine. The urine was free from both albumin and glucose. Casts were not present. The elimination of phenol- sulphonephthalein during the last two hours of the intoxication was 75 per cent. as compared with the output of 72 per cent. of the dye prior to the use of uranium. The blood urea remained unchanged from the normal reading of 18 mg. per 100 c.c. of blood. The reserve alkali of the blood was unaffected and remained at the normal reading of 8.1. A study of the kidneys of the four animals killed at this early period of the intoxication shows the glomeruli to be normal. Stainable lipoid is not present in the endothelium of the capil- laries. The stainable lipoid in the cells of the loops of Henle, especially in the cells of the ascending limb of the loop, shows an increase in amount when compared with the amount of such ma- terial that can be demonstrated in this location in the normal con- trol animals. The lipoid is in the form of granules and well- defined droplets. The convoluted tubule epithelium failed to show stainable lipoid with Scharlach R. at this early stage of the intoxication. ‘These cells appear normal. The five animals killed at the end of twelve hours following the use of uranium have all shown some change from the normal in the functional response of the kidney. In the urine from three of the animals both albumin and glucose were present. The urine of the two remaining animals contained a trace of glucose but no albumin. The results obtained in Experiment 7 are characteristic for this group. During the twelve-hour period of the experiment the animal formed 322 c.c. of urine. -Heavy traces of both albumin and glucose were present. The elimination of phenolsulpho- nephthalein was reduced from the normal of 70 per cent. to 55 Lirpoip MATERIAL IN RENAL EPITHELIUM. 225 per cent. Blood urea was unchanged. The reserve alkali of the blood was reduced from 8.05 to 7.95. Frozen sections from the kidneys of this group of animals when stained with Scharlach R. show an increase in the amount of stainable lipoid in the cells of the loops of Henle and, furthermore, at this stage of the uranium intoxication, stainable lipoid in the form of dust-like particles appears in the convoluted tubule epithelium. The granules are more marked in the periphery of the cells than in the area immediately around the nuclei. Other than these changes in the stainable lipoid content of the tubular epithelium the kidney tissue appears normal. A study of the course of the intoxication to this point shows the first evidence of an injury to the kidney from uranium to consist of inducing such a disturbance in the cells of the loops of Henle that an increase over the normal of stainable lipoid can be demonstrated in these cells. Such a change in the stainable lipoid content of these cells is not associated with any functional disturbance on the part of the kidney. Ata later period in the intoxication, after twelve hours, lipoid material stainable with Scharlach R. appears in the convoluted tubule epithelium. With this evidence of injury to these cells the elimination of phenolsulphonephhtalein is reduced; there is a beginning depletion in the alkali reserve of the blood, and albumin and glucose or glucose alone appear inthe urine. The total output of urine in such animals is apparently unaffected. Five animals were killed at the end of the twenty-four-hour period of the intoxication. The formation of urine at this stage of the experiments shows an increase over the normal daily output for the respective animals. The urine from all of the animals shows. albumin and glucose. Granular casts are present. The elimination of phenolsulphonephthalein is further reduced. There is no retention of blood urea. The reserve alkali of the blood was depleted in all of the animals. Experiment I0 is representative of the group. The average daily output of urine for this animal before the commencement of the intoxication was 410 c.c. The urine increased to 618 c.c. on the first day following the use of uranium. The urine contained 1.8 gm. of albumin per liter and I.1 per cent. glucose. The elimination of phenolsulphonephthalein was reduced from the 226 SCIENTIFIC PROCEEDINGS (121). normal of 75 per cent. to 30 per cent. There was no retention of blood urea. The reserve alkali of the blood was reduced from 8.05 to 7.9. Frozen sections from the kidneys of the animals at this stage of the intoxication after staining with Scharlach R. show very little change in the amount of stainable lipoid in the cells of the loops of Henle. Such material is abundant and in the form of droplets and fused masses. There is a marked increase in the amount of stainable lipoid in the cells of the convoluted tubules. In this portion of the tubule the small particles that have been described as appearing in this location at an earlier period of the intoxication have fused so as to form small droplets which are numerous. In addition to this change in the convoluted tubule epithelium, these cells show marked cloudy swelling and a com- mencing vacuolation. The capillaries of the glomeruli are en- gorged with blood. They fail to show the presence of stainable lipoid or other evidence of injury. The remaining five animals were killed at the end of forty- eight hours of the intoxication. In two of the animals the forma- tion of urine was in excess of the normal daily output. In three of the animals there was a reduction in urine formation. Urine from all of the animals has shown an increase in albumin and glucose over that observed at the end of twenty-four hours of the intoxication. The elimination of phenosulphonephthalein has — shown a progressive decrease. Only two of the animals show a retention of blood urea. The reserve alkali of the blood shows a progessive depletion. The results obtained in Experiment 16 are representative of the group. The formation of urine was reduced from the average normal daily output of 421 c.c. to 248 c.c. The urine contained 4.7 gm. of albumin per liter and 2.08 per cent. glucose. The elimination of phenolsulphonephthalein was reduced from the normal output of 68 per cent. to10 percent. Blood urea had increased from 14 to 42 mg. per 100 c.c. of blood. The reserve alkali of the blood was reduced from 8.1 to 7.85. Frozen sections from the kidneys of these animals when stained with Scharlach R. show little if any increase in the amount of stainable lipoid material in the cells of the‘loops of Henle or in the convoluted tubule epithelium. Other changes of degeneration in Liporp MATERIAL IN RENAL EPITHELIUM. 227 these cells that have been preceded by the appearance of stainable lipoid have become more marked. The cells show an advanced swelling, which frequently obliterates the lumen of the tubules. Vacuolation and necrosis are well advanced in many of the cells, especially in those of the convoluted tubules. The glomerular vessels are engorged with blood. Occasionally a slight exudate is seen in the subcapsular space. The endothelium of the capillaries has failed to show stainable lipoid. CONCLUSIONS. 1. Lipoid material stainable with Scharlach R. is constantly found in the cells of the loops of Henle in normal dogs. The presence of such material in this location is not indicative of a pathological kidney. The functional capacity of such a kidney is normal. 2. When animals are given one subcutaneous injection of 6 mg. of uranium nitrate per kilogram, the earliest evidence of injury to the kidney consists of an increase in the amount of stain- able lipoid in the cells of the loops of Henle. At this stage of the intoxication there is no evidence of a functional disturbance on the part of the kidney and no change takes place in the acid-base equilibrium of the blood. 3. At a later stage of such an intoxication (12 hours) stainable lipoid material appears in the convoluted tubule epithelium. The vascular tissue of the kidney is uninjured. Associated with such a disturbance in the metabolism of these cells that leads to the appearance of stainable lipoid in the cell there occurs a reduction in the elimination of phenolsulphonephthalein, a depletion in the alkali reserve of the blood and the appearance of albumin and glucose, or glucose alone in the urine. 4. Following this initial injury to the tubular epithelium changes of a more distinctly degenerative type appear in these cells and the functional capacity of the kidney is more severely impaired. 228 SCIENTIFIC PROCEEDINGS (121). 99 (1846) Preliminary report on the effects of vagus stimulation on the dog’s stomach and the influence of asphyxia on these effects. By Z. BERCOVITZ. [From the Hull Physiological Laboratory of the Unwersity of Chicago and Physiological Laboratory of Baylor Medical College, Dallas, Texas.| In a previous report! attention was called to the fact that in the turtle repeated vagus stimulation was followed by a progres- sive decrease in the gastric response to each stimulation. It was further pointed out in another report? that the stomach could not be tetanized by prolonged vagus stimulation, also that reduc- tion in the temperature of the turtle was followed by a decreased gastric response to vagus stimulation. These facts seemed to indicate that in the turtle a complex neuro-muscular mechanism controlled the response of the stomach to vagus stimulation. The object of this study therefore was to determine if the dog’s stomach would respond to vagus stimulation in the same manner as the turtle’s stomach. METHODs. The dogs used in this study were decerebrated in order to avoid the depressing influence of an anesthetic. The balloon method was used for recording gastric contractions. The balloon was introduced into the stomach through (a) healed gastric fistula; (b) through slit in the anterior wall of the stomach near the pylorus; (c) through the mouth and esophagus; (d) through the duodenum. The results were the same in all cases. In most cases a simultaneous blood-pressure tracing was made from the carotid artery and in others the chest was opened, artificial respiration given and the heart observed directly. To produce asphyxia. of the stomach a lifting ligature was placed under the thoracic aorta. The animals were cooled by packing them in cracked ice until the rectal temperature dropped to the desired point. As a rule 14 to 24 hours were required to cool the decerebrated dog to 23° C. 1 Bercovitz and Rogers, Amer. Journ. Physiol., 1921, lv, 323. 2 Rogers and Bercovitz, Amer. Journ. Physiol., 1921, lvi, 257. EFFECTS OF VAGUS STIMULATION ON DoG’s STOMACH. 229 RESULTS. Effect of Repeated Vagus Stimulation.—In the dog stimulations of the vagus with a tetanizing current repeated at short intervals with a given strength of current are followed by contractions of the stomach of approximately uniform amplitude. Complete cardiac inhibition was observed during each stimulation. The apparent ability to produce an artificial rhythm of the dog’s stomach by repeated stimulations of the vagus is in striking con- trast to the rapid failure of gastric contractions on repeated vagus stimulation in the turtle. Effect of Prolonged Vagus Stimulation.—Prolonged stimulation of the vagus with a tetanizing current of moderate strength and also a strong tetanizing current is followed by only a single con- traction of the stomach and its usual subsequent relaxation in spite of continued stimulation. Weak peristalses in the pyloric region may occur after the one contraction during the balance of the stimulation but usually stops shortly after the stimulation has ceased. There is no indication of a tetanus or increased tone of the stomach. These results are similar to those obtained in the turtle. It was noted from the blood-pressure tracings and also from direct observation of the heart that complete vagus inhibition of the heart failed at about the same time as the relaxation of the stomach began. It would seem that the mechanism operating to prevent gastric tetany is the same as that which prevents prolonged complete cardiac inhibition from vagus stimulation. Influence of Cooling—tIn dogs cooled to 25° C. to 23° C. it was found that repeated stimulations of the vagus were followed by no indications of either failure of gastric response or increased or permanent tone as the result of the stimulations. Prolonged vagus stimulation at this temperature is not followed by tetanus of the stomach. It was further noted that at a temperature of 21° C. there was a failure to obtain a gastric response from vagus stimulation but stimulation of the stomach wall was followed by a contraction. No cardiac inhibition was noted at this temperature in response to vagus stimulation. As the temperature was raised there was a corresponding increase in gastric response to stimulation of the vagus. 230 SCIENTIFIC PROCEEDINGS (121). The heart seemed to be more easily thrown into fibrillation in the animals which had been cooled than in correspondingly prepared animals at normal body temperature. Death in most of the experiments on cooling was due to fibrillation of the heart. Influence of Temporary Asphyxia.—In dogs with the thoracic aorta ligated repeated stimulations of the vagus are followed by rapid failure of gastric response to stimulation. Allowing the blood to pass again to the stomach is followed by recovery of the gastric response to vagus stimulation. These experiments would seem to indicate that in the dog as well as in the turtle a complex neuro-muscular mechanism in- fluences the effects of vagus stimulation on the stomach. This mechanism in the dog is very sensitive to partial asphyxia. It may be in the turtle that the rapid failure of gastric response to vagus stimulation is dependent on circulatory changes causing an asphyxia of the complex neuro-muscular mechanism. SUMMARY. 1. In the dog stimulations of the vagus repeated at short intervals are followed by contractions of the stomach of approxi- mately uniform amplitude. 2. Prolonged stimulation of the vagus is followed by a single contraction of the stomach and its usual subsequent relaxation in spite of continued stimulation. Failure of cardiac inhibition occurs at about the same time as relaxation of the stomach begins. 3. Reduction of the body temperature to 25° C. to 23° C. is followed by decreased response to vagus stimulation but no indica- tions of either failure of gastric response to repeated vagus stimula- tion or tetanus. At 21° C. vagus stimulation is without effect on the stomach but stimulation of the stomach wall is followed by a contraction. 4. Partial asphyxia of the stomach by shutting off the blood supply to the part is followed by rapid failure of gastric response to repeated vagus stimulations similar to that observed in the turtle. Removal of the asphyxia is followed by recovery of the gastric response to stimulation. DIPHTHERIA ANTITOXIN. 231 100 (1847) Agglutination phenomena with diphtheria antitoxin. By P. J. MOLONEY and L. O. HANNA. [From the Research Division, Connaught Antitoxin Laboratories, University of Toronto, Toronto, Canada.]| In search for an in vitro test for diphtheria antitoxin the following observation was made: (a) When an emulsion of the diphtheria bacillus, Park 8, is mixed with diphtheria antitoxin, allowed to stand at 37° C. for 1 hour, centrifuged, washed with saline and re-suspended, it is no longer agglutinated by diphtheria- agglutinating serum. (0) The organisms sensitized in this way are inagglutinable by acid agglutination. In this test for the inhibition of acid agglutination the cells are suspended in a buffer solution which gives a maximum agglutina- tion with unsensitized cells. The point of maximum agglutination for acids varies somewhat depending on the culture and the buffer mixture, but for phthalate mixtures (Clark and Lubs) diluted I-I with distilled water it is about Py 4.2 for a three or four-day broth culture of the Park 8 strain. To determine the specificity of the test for diphtheria antitoxin, experiments were carried out along four different lines. 1. The following sera were used in place of diphtheria antitoxin and the test carried through: normal human serum, positive T.B. human serum, normal guinea pig, concentrated tetanus antitoxin, fresh antitoxic serum, normal horse, normal sheep, normal rabbit. In the above experiment there was inhibition of agglutination when the cells had been sensitized with antitoxic serum and with tetanus antitoxin; and when the other sera had been used there was agglutination. A guinea-pig test with the tetanus antitoxin showed that it contained diphtheria antitoxin, and it was subse- quently discovered that this tetanus antitoxin was from a horse which had previously been used for the production of diphtheria antitoxin. When diphtheria bacilli are sensitized with diphtheria agglutin- ating serum instead of antitoxin there is inhibition of agglutination 232 SCIENTIFIC PROCEEDINGS (121). when the cells are subsequently subjected to serum agglutination but acid agglutination is not inhibited. A possibility which was kept in mind was that the effect with antitoxin might be due to the presence of agglutinoids in antitoxic serum. In an attempt to check this, agglutinating serum diluted (1-10) was heated 75-80° C. for 1 hour and subsequently used in the test; both with acid and serum agglutination there was no inhibition. One might conclude from this experiment that the agglutinins were destroyed by the high temperature and that no agglutinoids were produced. Up to the present attempts to pro- duce agglutinoids from agglutinins have not been successful. 2. An emulsion of diphtheria organisms incubated with a mixture of antitoxic serum and diphtheria toxin in suitable quan- tities is subsequently agglutinated both by acid and by diphtheria- agglutinating serum, whereas if the toxin is replaced by an equal volume of broth the cells are not agglutinated. The conditions for this experiment are limited by two factors: there must be sufficient antitoxic serum to sensitize the cells, and an excess of toxin. Diphtheria organisms sensitized with antitoxin are rendered agglutinable by mixing with diphtheria toxin, centrifuging, wash- ing with saline and re-suspending. Diphtheria toxin has this same neutralizing effect on diptheria- agglutinating serum. From this one might conclude that diph- theria toxin contains agglutinogens besides true toxin. 3. In an attempt to apply the test quantitatively two general methods were used: Organisms were sensitized with progressive dilutions of antitoxic sera and the limit of inhibition read as the end point; and mixtures of serum and toxin after standing 1 hour at room temperature were added to sensitized cells directly, and after dialyzing—it had been found that toxin dialyzes through parchment paper—and the dialysate mixed with sensitized cells which were subsequently tested for agglutinability. Neither method gave results which were uniformly consistent with guinea-pig experiments. Using the first method some results were obtained which were parallel to the guinea-pig tests, in other cases the results were reversed, 7.e., in, some cases two sera which by Ehrlich’s method gave, e.g., 300 A.U. and 150 ALU. per c.c. DIPHTHERIA ANTITOXIN. 233 respectively showed by this test that the second contained more antitoxin than the first. In this connection it is well to recall that according to the work of Roux,! Danysz,! Momont! and Cruveilhier,? the results ob- tained with antitoxic sera by Ehrlich’s method do not always parallel those obtained by the French method—a method which has at least the merit that its results are based on animal experi- ments, the conditions of which correspond in a measure to those which obtain in the actual treatment of the disease. These workers claim that in some cases the results are not only not parallel but may even be the reverse of each other. On the other hand according to the work of Marx? the two methods give parallel results. No comparison between the test described here and the French method has yet been carried out. 4. It would be of interest to know whether washed diphtheria organisms do actually take up antitoxin. A carefully controlled experiment to test this was carried out. Organisms which had been washed several times with saline were mixed with a known amount of antitoxin, allowed to stand for 1 hour at room tempera- ture, centrifuged and the supernatant liquid drawnoff. A guinea- pig test with an appropriate amount of toxin showed a fall in antitoxic content of the mixture. The difficulty with this experi- ment is that it is not known when the cells are sufficiently washed. It is proposed to repeat this experiment in the following way: Wash diphtheria cells several times and then wash again batches of these cells and test the first and progressively washed lots to see whether a point is reached where the drop in antitoxin reaches a constant. SUMMARY. Both acid and serum agglutination of Park 8 strain of B. diphtherie are inhibited when the organisms are first sensitized with diphtheria antitoxin; when diphtheria-agglutinating serum is used instead of antitoxin, serum agglutination is inhibited but not acid agglutination. . This inhibition phenomenon with antitoxin is specific, at least for the Park 8 strain. ; Abstract in the ‘‘ Bacteriology of Diphtheria’’; Nuttall, Graham, Smith, 1913, 525. ? Cruveilhier, Ann. Inst. Pasteur, xix, 1905, 249. 3 Marx, Zeit. f. Hygiene, 1901, xxxviii, 372. 234 SCIENTIFIC PROCEEDINGS (121). It is not definitely established whether it is a test for anti- toxin; it is possible that the phenomenon may be due to agglutin- oids, or to a diphtheria antibody not noted heretofore. Nicolle, Debains and Césari* have described a qualitative test for toxin and antitoxin which is based on a precipitin reaction: It is evident from the results above that while the test may be specific for the diphtheria bacillus and for the other organisms used by them, it should be subjected to further investigation before it can be accepted as specific for toxin and antitoxin as defined by Ehrlich’s guinea-pig test. IOI (1848) Action of some purin derivatives on the isolated bronchus. By DAVID I. MACHT and GUI CHING TING. [From the Pharmacological Laboratory, Johns Hopkins Unversity, Baltimore, Md.| In connection with a study of the effects of various drugs on the isolated bronchi of pigs the authors studied a number of purin derivatives. The effects of caffein or trimethyl xanthin in doses of 1-20 mgm. in 25 c.c. of Locke’s solution gave the following results; small doses produced no effect on bronchial muscle or occasionally a very slight constriction. After large doses of caffein a little relaxation of the normal bronchial preparations was noted. When, however, such bronchial preparations were first brought into a state of high tonus or contraction, as for instance on treat- ment with muscarin, the relaxing effect of a subsequent dose of caffein was much more marked. On the whole, however, the results obtained indicated that caffein has a very weak dilator effect on the bronchus. Following experiments with caffein, observations were made on the effects of theobromin or 1-3 dimethyl-xanthin and theocin or 3-7 dimethyl-xanthin. ‘It was found that both dimethyl-xanthins produced much greater broncho-dilatation than trimethyl-xanthin or caffein. The authors were unable to obtain a mono-methyl xanthin but they did study the effects of xanthin itself. Although 1 Nicolle, Debains and Césari, Comp. rend., 1919, clxix, 1433. EFFECT OF COCAINE ON LUPINUS ALBA. 235 xanthin is very slightly soluble nevertheless even very minute quantities of the substance (1 c.c. of I-200,000 solution) introduced into 25 c.c. of Locke’s solutions in which the preparation was suspended were found to produce a very marked relaxation. Hypoxanthin acted in the same way. Going a step further experiments were made with minute quantities of guanidin and adenin and both of these were found to produce relaxation of the bronchus and seemed to be comparatively more potent even than xanthin. Passing to the nucleosid guanosin, the pharmacological action became different. Guanosin produced no effect. A few experiments with adenin nucleotid showed that it also was in- active. Finally tests made with solutions of thymus nucleic acid and yeast nucleic acid gave also no effect on the bronchial muscle. 102 (1849) Effect of cocaine on the growth of lupinus alba: a contribution to ‘“‘phytopharmacology.” By DAVID I. MACHT and MARGUERITE LIVINGSTON | From the Pharmacological and Plant Phystology Laboratories, Johns Hopkins Unierstty, Baltimore, Md.] The effects of cocaine and its decomposition products were studied on the growth of seedlings of the plant Lupinus alba. The seeds were soaked in water and allowed to sprout in a suitable medium, following which the length of the straight roots grown by this plant was measured and the effects of cocaine and other chemicals on the growth of the roots were investigated. The plants were placed in solutions of nutrient salts (Shive solution) and the various drugs were added to such solutions. Controls were made on seedlings suspended in Shive solution diluted one half with distilled water. It was found that the effect of cocaine and its decomposition products on the growth of lupinus was very different from the effect of the same substances on animal tissues. Whereas cocaine is very toxic for animal tissues such as smooth and skeletal muscle, nerves, etc., it required strong solutions of this alkaloid, namely 2 per cent. of cocaine hydrochloride to inhibit 236 SCIENTIFIC PROCEEDINGS (121). the growth of the seedlings. LEcgonin hydrochloride inhibited growth in concentrations of .0055 per cent., while benzoyl ecgonin was much less toxic, requiring 23 per cent. concentration to affect the growth. Methyl alcohol was found to be very little toxic to the roots of the lupinus, requiring 4.8 per cent. to produce an inhibition of growth. Contrary to expectation the most toxic decomposition product of cocaine was found to be sodium ben- zoate, a compound which is practically non-toxic for animal tissues. Sodium benzoate was found to be deleterious to the lupinus root in concentrations of 0.007 per cent.; while the ester methyl-benzoate was found to produce inhibition in concentra- tions of 0.014 per cent. Various simple mixtures of ecgonin, sodium benzoate, methyl alcohol, benzoyl ecgonin, etc., were also studied and the effects of these will be described in the complete paper in the Journal of General Physiology. 103, (1850) Spontaneous cure of rickets in rats. By ALFRED F. HESS, LESTER J. UNGER and A. M. PAPPENHEIMER. [From the Department of Pathology, College of Physicians and Surgeons, New York City.] It has long been believed that in infants healing of rickets occurs in spite of the diet and environment remaining unchanged. Some time ago in the course of an experiment on the curative effect of sunlight in the rickets of rats, it was observed that rickets healed in two of the control animals. One of these rats was on the Sherman and Pappenheimer diet plus 25 mg. P. per cent. and the other on the same diet with an addition of 50 mg. P. per cent. The rats weighed 34 g. at the outset, and after 30 days showed rickets by x-ray. After 62 days the radiograph was negative in one instance and demonstrated healing in the other. Autopsies of both revealed no gross rachitic changes; microscopic examina- tion showed healed rickets at the costo-chondral junctions in one instance, in the other no rickets was found but some autolysis had taken place. 237 SPONTANEOUS CURE OF RICKETS IN RAtTs. "A Suryeay pareve paeory paeaH paeayy 89 v9 "YW payreyy "Y Sureay "Y suryeayy "y Sureazy 'W paeory ol vs 8}2HOY ‘a ‘pow | vo "UW WSS | of ‘UW ‘pow | vs ‘dou JSOWY "A WITS “a "a WSIS ve ge ‘Op “op “op ‘Op *qu99 19d | | | TL | | | | | LS (SISAJOINY) "WON | WON} oS SISOI0d09}SO | ‘YT ON| PS SL°E "YW Ssurpeazy | “wy ON] 09 "a ON SIS9UIS09}SO o’e BATPOCUT | YON] OS "WI snorasid ove | Surmoyjoy siso1odo0a3soO |*y{ *ON] OS of, “Bur *SOIDIPT SSOIL) 4 *d ‘310uy ak *AZ0foyIeg 7 Se Te ee ee Se ee ee ee Se See Ea ee ee ee | eerie i 238 SCIENTIFIC PROCEEDINGS (121). In view of this experiment another series of rats were put on the same diet (Sherman and Pappenheimer plus 25 mg. P. per cent., in the form of secondary potassium phosphate). This addition furnishes a level of phosphorus which is still inadequate for the rat, and leads to rickets. Of the four receiving this diet all showed signs of rickets according to x-ray after 32 days. After 48 days, however, the radiographs showed healing. During this period the rats had made a total gain of 92 g. After 74 days the x-ray showed the lesions apparently healed, and the rats were killed. No gross evidences of rickets were found at necropsy. Micro- scopic examination of the costo-chondral junctions also showed no evidences of active rickets; the rickets was either healing or there was merely osteoporosis. The explanation suggested for this spontaneous healing is that, with a practically stationary weight over a long period, the phosphorus requirement for the building up of new tissue is greatly reduced. The small addition of P. under such circumstances, to the standard rickets-producing diet, suffices to enable the bone to recalcify. 104 (1851) A further report on the prevention of rickets in rats by light rays. By ALFRED F. HESS, LESTER J. UNGER, and A. M. PAPPENHEIMER. [From the Department of Pathology, College of Physicians and Surgeons, New York Cuty.] In a previous communication we showed that the development of rickets in rats fed the standard rickets-producing diet of Sher- man and Pappenheimer can be prevented by daily exposures to direct sunlight for fifteen minutes.! A similar result has been reported by others.? If the rats are placed in a box having flint glass windows, it was found that the sun’s rays which had traversed the glass had lost their protective power. Rays which were re- flected to the rats from a white surface retained some of their 1 Hess, A. F., Unger, L. J., Pappenheimer, A. W., Proc. Soc. Exp. BIOL. AND MED., 1921, xix, 8. 2Shipley, P. G., Park, E. A., Powers, G. F., McCollum, E. V., Proc. Soc: Exp. BIOL. AND MED., 1921, xix, 43. PREVENTION OF RICKETS BY LIGHT RAys. 239 efficacy. Rickets can be prevented in rats by daily exposures of about 2 minutes to the rays of the mercury vapor quartz lamp, or by 4 minutes or less exposure, at a distance of three feet, to the rays from acarbon arclamp. X-rays were found ineffective. In order to test the effect of the pigment of the skin on the protective action of light, a group of white and another group of black rats (the melanic form of the Norway rat) were exposed to the radiation from the mercury lamp. In the first experiment both sets of animals were protected as the dosage was excessive. In the second experiment, when one and one-and-one-half minute exposures were employed, all the white but none of the black rats were protected. The black rats showed rickets by x-ray and by ULTRA-VIOLET RADIATION—WHITE AND BLACK RATS. | Inorg. P. Wet. U.-V. Ray Diet X-Ray Path. Mg. | per cent. White 70-70 I min 843 Neg Neg sé 58-60 sé “eé “se ‘ “é 60-70 sé sé sé sé 46 sc 60-70 13 sé ‘s ‘6 ‘6 “é 60-80 ‘ sé sé ce ‘ 5-45 “es dete 64-70 se ss ‘6 sé 4.44 Black....| 50-60 abs <3 R R oe : 50-60 “é sé “ce “ce sé “ce 60-58 sé sé sé sé sé ¥ ».e.] 50-50 iz“ . e % 2.92 a7 hie bee _ . A 3.00 pathological examination, and their blood contained a less per- centage of inorganic phosphate. This experiment shows that pigment retards the rays which are effective in rickets, and indi- cates one factor in the exceptional susceptibility of negro infants to this disorder. Prolonged exposure to direct sunlight failed to prevent or to delay the onset of scurvy in giunea-pigs. 240 SCIENTIFIC PROCEEDINGS (121). 105 (1852) A note on the preparation of anti-colon streptococcus serum. By JOHN F. ANDERSON. [From the Squibbs’ Laboratory, New Brunswick, N. J.] About two years ago, Dr. H. A. Cotton requested that I should undertake for him the preparation of a combined Anti-Colon Streptococcus Serum prepared from horses treated with cultures of B. Coli and Streptococci isolated from cases under treatment at the New Jersey State Hospital, Trenton, N. J. A certain number of cases under treatment at that institution have been found to yield pure cultures of streptococci of diverse types and also strains of B. Coli. These organisms are obtained from material removed at operation such as tonsils, extracted teeth, lymph glands, especially those found in the mesentery of the colon, and other material from the abdominal cavity. The treatment of the horse was with eleven strains of B. Colt and eight strains of streptococci, most of which belonged to the hemolytic group. The immunization was begun with small doses of the killed organisms, but after several weeks the use of live cultures was adopted. It was found that the injections of the mixed antigens of the two bacteria were followed in some instances by severe reactions in the horse and that when the antigens were.given on different days that the colon antigens were not tolerated as well as the streptococcus antigens. The injection of the colon bacilli induced such violent reactions in some instances that recourse was finally had to the method of preparing the antigen suggested by the author for the preparation of anti-meningococcic and anti-pneumo- coccic sera. The plan of treatment as finally decided upon was the injection of washed broth cultures of streptococci and washed agar cultures of colon bacilli. The two antigens were suspended in salt solution and given intravenously for four injections on successive days followed by a rest period of three days when the injections with gradually increasing amounts were repeated. Vt A orate} ANTI-COLON STREPTOCOCCUS SERUM. 241 After the horse had been under treatment for about 8 weeks, a trial bleeding was made and the titre of the serum determined by the use of the agglutination test as used by the Hygienic Labora- tory for the testing of Anti-Meningococcic Serum. Based on the results of the tests of the trial bleedings, an adjustment was made in the proportion of the individual components of the mixed antigen in order that the serum should be as well balanced as possible. It was found, however, that in spite of all efforts to promote the formation of antibodies to each strain of bacteria used in the injection of the horse that agglutinins, at least, were never pro- duced in measurable amounts to a few strains. This of course is not unique as many have found that some strains of bacteria do not induce antibody formation. A few weeks after the trial bleeding the horse was bled for production by taking 6 liters of blood six days after the last injec- tion and 6 liters 48 hours later. The serum from the two bleedings was combined, four tenths of one per cent. tricresol added, filtered through Berkefeld filters, tested for sterility and for certain antibodies. It was found that the serum agglutinized all of the strains of B. Coli except two in dilutions from I-100 to I-400 and that all of the strains of streptococci were agglutinated in dilutions from I to 200 to I to 400. | In addition complement fixation tests with the serum gave fixation but not to a high degree with both the hemolytic and non- hemolytic groups of streptococci. Protection tests in animals have not been made but it is believed that sufficient data has been accumulated in the literature to justify the advance of the opinion that a properly prepared anti-streptococcic serum will afford a considerable measure of protection against streptococci if given 12 to 18 hours before the bacteria are injected. There is not the same data for an anti- colon serum as but few attempts have been made to make and ~ use such a serum. While most are agreed as to the possibility of giving protection to animals by the previous administration of an anti-streptococcic serum, there is considerable difference of opinion as to the curative value of such a serum, although the weight of opinion is in its favor: 242 SCIENTIFIC PROCEEDINGS (121). Some of the failures of anti-streptococcus serum may have been due to the fact that the particular strain of streptococcus causing the infection was not among those groups used for the immuniza- tion of the horse. The importance of this was not realized until comparatively recently and it is possible that further work may show that a potent anti-streptococcic serum can be prepared for each strain. The pathogenicity of the colon bacillus for laboratory animals shows much variation as the intravenous administration of some strains in small amounts sometimes quickly results in death, while large amounts of other strains are well borne. The symptoms and fatal results are probably due to the action of toxins or poisons contained in the body of the bacteria. The colon bacillus has been claimed to be the cause of a variety of conditions in various parts of the body, particularly in the region of the abdominal cavity, but some observers have thought it questionable if the colon bacillus was the primary cause of the lesions due chiefly to the observation that it is not always found in pure culture. The growth of the colon bacillus is not attended with the production of a soluble toxin or poison of a high degree and there- fore the injection of filtrates into animals of broth cultures is not followed by marked antibody formation, but if animals such as the rabbit or horse are given gradually increasing doses of killed. or live cultures there quickly appear in their blood antibodies such as bacteriolysins, agglutinins, precipitins, and complement- fixing substances. The fact that antibodies are produced in high degree in im- munized animals gives support to the opinion of Dr. Cotton that a combined Anti-Colon Streptococcus Serum is of value in pre- venting post-operation infections due to colon bacilli or strepto- cocci and perhaps may be of value in the treatment of infections by those organisms already developed. COLON-STREPTOCOCCUS ANTI-SERUM. 243 106 (1853) The use of a colon-streptococcus anti-serum as a pre-operation measure. By JOHN WILLIAM DRAPER. [From the State Hospital, Trenton, N. J.] The work here presented is a part of the elaborate research into the causation and treatment of the so-called functional psy- choses, made at the State Hospital at Trenton, New Jersey, under the direction of Dr. Henry A. Cotton. More than thirty years ago, while performing autopsies upon the bodies of patients dying in the hospital for the insane at Chicago, Dr. Albert J. Ochsner! noted that there was present in an unusually large number very marked pathological changes in the colon, and occasionally in the other viscera. He called the attention of the authorities to these findings but was told that even if present, these lesions had nothing whatever to do with the psychosis, which was a personality or psychic disorder, entirely separate and bearing no relation whatsoever to any physical defects which might be present. In spite of his protestations this opinion prevailed. Undoubtedly many other pathologists and surgeons, both here and abroad, have noted the striking frequency with which extensive pathological changes in the abdominal portion of the alimentary canal and elsewhere are to be noted among patients suffering with the so-called ‘‘functional’’ psy- choses. Impressed with the very definite clinical improvements which he had obtained by removing dental and tonsillar foci of infection among these patients, and believing that there must be additional sources for the very evident toxemia among those who made little or no improvement after the removal of these oral infections, Dr. Cotton invited me to conduct a surgical research which should furnish evidence as to the presence or absence of such abdominal infection. This work is now entering upon its fourth year. The pathology is present and the favorable clinical results following 1 Ochsner, Albert J., personal communication. 244 SCIENTIFIC PROCEEDINGS (121). its removal are already rather widely known. Suffice it here to say, that as a result of the application of the usual surgical prin- ciples of detoxication by elimination of all foci within reach, the hospital discharge rate has risen from thirty-five to seventy-six per cent.! Over ninety per cent. of all patients classified as “functional”’ psychotics have marked oral infection. Twenty per cent. present marked evidence of gastro-intestinal disease. Until some better form of therapeusis or early prevention can be found, there seems no better method at hand, as stated by Dr. James Ewing, than surgical removal. Continuing in a report of sixteen specimens of colon and ileum from this series, Dr. Ewing says: “The most marked and constant lesion is pigmentation of the mucosa which has rendered the inner lining brownish or at times dark chocolate in color. This change is most marked in the cecum, diminishing toward sigmoid, but often present throughout the specimen. Sections show the pigment to be lodged in large polyhedral cells lying in the mucosa and at times in the epithelium. Pigmentation of the colon is fully recognized as a sign of chronic intestinal stasis and intoxication. It is sometimes associated with anemia and at times with severe and even fatal dystrophies of nervous and muscular systems.” ‘‘Pouching of the intestinal wall amounting almost to hernial protrusions was observed in most of the cases. These pouches were from one to two cm. in depth. The wall of the pouches was generally thinned, sometimes very much thinned, and the mu- cosa at the bottom was generally eroded, sometimes ulcerated. Through such erosions and ulcerations it is obvious that absorption of fluids and bacteria readily occurred.” “In general, the impression gained from the study of these specimens was that the clinicians were dealing with extensive and somewhat unusual grades of chronic intestinal stasis and catarrhal inflammation with its sequels.”’ In 1919, thirty-four partial resections of the colon were made with thirteen deaths or 40 per cent. mortality. In 1920, fifty-nine partial resections were done with eighteen deaths or 30 per cent. mortality. In 1921, forty-six partial and thirty-one total re- ' Cotton, H. A., ‘‘The Defective Delinquent and Insane,” r92r. COLON-STREPTOCOCCUS ANTI-SERUM. 245 sections were done with ten deaths or 12 per cent. mortality. It is a very simple matter to account for the lowering of the mor- tality rate from 40 to 30 percent. It embraced the training of the staff and the development of an improved surgical technique. The fall from 30 per cent. to 12 per cent. followed immediately upon the introduction of the serum treatment, all other factors remaining as before. Coincident with the 30 per cent. mortality among the insane, the writer’s mortality among private cases was 17.7 per cent., showing that psychotic patients are not good surgical risks. They are all physically sick. Their systolic blood pressure is abnormally low, and, as is well known, they often have an abnormally high small lymphocyte count coupled with a very small number of polymorphonuclear cells, the former sometimes exceeding the latter. Aside from the extensive pathological lesions in the mucous membrane and the wall of the colon, which were found in the speci- mens removed at operation, it was noted that the mesenteric lymph nodes were very much enlarged. In the very beginning of the wo1k these lymph nodes were cultured and various strains of streptococci and colon bacilli were isolated. This finding was of the utmost importance as it clearly indicated that these bacteria were passing through the wall of the intestine and were in all probability the cause of the lesion in the intestinal wall. When these enlarged nodes were found in the mesentery of the colon alone, resection was clearly indicated. In many cases, however, it was found that the adenitis was not limited to the mesentery of the colon, but extended throughout the whole mesentery of the small intestine. In such cases it was evident that removal of the colon would not correct or eliminate the evident lesion throughout the whole of the small intestinal tract. It was necessary, therefore, to devise some other method where- by these infections could be mitigated or eliminated. Autogenous vaccines made from the streptococci and colon bacilli isolated from the lymph nodes, were tried, but without success, probably because of the extent and severity of the infection existing in the intestinal wall, a condition analogous to that found in typhoid fever. As is well known, typhoid vaccine will immunize a patient against typhoid fever, but once the disease is established the 246 SCIENTIFIC PROCEEDINGS (121). vaccines have no value in the treatment, because of the over- whelming number of typhoid bacilli in the intestinal tract. When it was found that the autogenous vaccines were not effective another method had to be devised. As no colon serum had been previously made, Dr. Cotton consulted with Dr. John F. Anderson, who agreed to attempt to immunize a horse with strains isolated in the laboratory at the State Hospital at Trenton. It was found that the colon bacilli, even in small doses, were extremely toxic for the horse, but finally a serum was produced which was very potent. As many of the patients were suffering from combined infection of the streptococci and colon bacilli, it was decided to combine these organisms in one serum, merely for the sake of convenience of administering the serum. As noted below, this serum has proved entirely satisfactory, not only in reducing the surgical mortality, but, combined with the surgical procedure of removing the infected colon, it has apparently hastened the disappearance of the mental symptoms immediately following the operation. Hence, the colon-streptococcus anti-serum, prepared from strains furnished by Dr. Cotton, has been given as a pre-operative treatment, in a series of from eight to ten doses, extending over about a month. Without any other change being made in the surgical technique or post-operative care, the mortality dropped to 12 per cent. Autopsy notes show that this decrease was primarily due to a great reduction in the number of perforations which formerly had occurred in the operation area. It has been noted, clinically, that the character of theimprovementistransient, — probably due to reinfection. This series of one hundred and seventy cases of colectomy is too small to permit of any definite deductions as to the value of this serum, and this report is merely a provisional one, the en- couraging nature of which may not be supported by subsequent studies. BLoop PRESSURES IN GIRLS DURING ADOLESCENCE. 247 107 (1854) Spontaneous agglutinability of bacteria in relation to the antagonistic action of certain cations. By RALPH R. MELLON. [From the Department of Laboratories, Highland Hospital, Rochester, N. Y.] Spontaneous agglutinability of five separate, single-celled (pure line) cultures of diphtheroid bacilli was shown to be a function of growth-cycle developments. The bacillary phase, growing at 37°, was immediately and completely agglutinable by any solution tried. The coccus phase, growing at 20°, formed stable emulsions in NaCl and other salt solutions. By reversing the growth tem- perature, even on the same media, the agglutinability and mor- phology were reversed. Certain of the cultures, still completely ageglutinable by NaCl, formed stable emulsions in Tyrode’s and other equilibrated solutions. The mutual antagonism of the Na, K and Ca ions is believed to explain the phenomenon. The Mg ion was especially beneficial. With various cultures of different age and environment, all possible variations in agglutinability were observed. The amplification of these observations, now in prog- ress, promises to explain some of the paradoxes of the bacterial agglutination, and the observations in themselves constitute the first systematic application of these principles to agglutination. 108 (1855) Blood pressures and heart rate, in girls, during adolescence. A preliminary study of 1,'700 cases. By STANLEY ROSS BURLAGE (by invitation). [From the Department of Physiology, Cornell Medical College, Ithaca, N. Y.] The data were obtained from over 800 girls in the public schools of Ithaca, N. Y., whose ages ranged from 9 to 16 years and - from about an equal number of young women in Cornell Univer- sity, from 16 to 26 years old. 248 SCIENTIFIC PROCEEDINGS (121). Blood pressures were taken by Korotkow’s auscultatory ' method, using in all cases a Princo mercury sphygmomanometer. The reading for the diastolic pressure was made at the beginning of the fourth phase. All girls were examined in the sitting posture. There is a rapid rise in the systolic pressure from 104 mm. at 9 years to approximately 124 mm. at 14 years. This remains at the same level through the next year. Then there is a rapid fall of over 10 mm. to 18 years. From that age on the pressure remains fairly constant around IIo mm. up to 26 years. The diastolic pressure rises evenly from 63 mm. at 9 years to about 76 mm. at 14 years. It maintains about this level throughout the remaining years. The pulse rate drops rapidly from 98 at 9 years to 80 at 18 years and then continues with little change. Since, at 14 years of age, practically all girls in this climate have begun to menstruate, these curves would seem to indicate, that allowing 3 or 4 years for recovery from metabolic disturbances incident to the onset of puberty, the blood pressures and pulse rate vary little during the following 8 years. In the height curves, the systolic pressure rises gradually from 104 mm. at 50 inches to 113 mm. at 69 inches. The diastolic pressure rises slightly more rapidly—from 64 mm. to 74 mm. The pulse rate drops rather evenly from 106 to 82 per minute. The weights are arranged in classes of 10 lbs. each. The systolic pressure rises rapidly from 104 mm. for the 51-60 Ib. class up to 118 mm. for the 91-100 lb. class. Then it runs along without much change until it reaches the 151-160 Ib. group. Here the rise is abrupt up to about 130 mm. for the 200 Ib. class. The diastolic pressure shows a gradual rise from 58 mm. in the 41-50 Ib. class to about 90 mm. at 200 lbs. The pulse rate shows a decline from 102 at 51-60 lbs. to 78 at 200 lbs. The conclusion from a consideration of these data is that in determining the normal blood pressures and pulse rate for girls between 9 and 26 years of age, it is necessary to consider, not age alone, but weight and height as well. SEROLOGICAL STUDIES OF DIPHTHERIA GROUP. 249 109 (1856) Serological studies of the diphtheria group. By CARL O. LATHROP and CHARLES A. BENTZ. [From the Bacteriological Laboratories of the Medical Department, University of Buffalo, and Buffalo Bureau of Laboratories, Buffalo, N.Y.] A systematic qualitative and quantitative study has been made on the immunity developed in a group of young adults immunized against diphtheria with toxin-antitoxin. As a preliminary, a successful attempt was made to corroborate Havens’ contention that there are two serological groups of diph- theria bacilli with specific agglutinogenic properties and no evidence of cross agglutination. For our minor group antigen we used two cultures recovered from cases of diphtheria developed in persons immunized with toxin-antitoxin and yielding negative Schick tests. This specificity has been further substantiated by antibody absorption, which confirms the other findings completely. Incidentally, we used rabbit blood agar plates exclusively for isolation and study of the organisms, and noted that hemolysis, a sometime mooted point, is not characteristically allied with virulence, nor does it only occur in freshly isolated cultures, but may crop up as late as the 56th generation. We have also found, as we believe Park stated, that certain true diphtheria bacilli possess a factor of virulence not neutralized by antitoxin, concerning whose identity we are making a further study. Likewise we have confirmed Park and Havens in finding that there is some group antitoxin present in the antitoxin com- monly in use for the toxin of the minor group. Using a modified R6mer method, we titrated the antitoxin content of a group of young adults immunized against diphtheria with toxin-antitoxin. We found 20 out of 26 had developed anti- toxin in quantities varying from 1/30 unit up to 1/5, while three developed only 1/50, and three failed to develop any immunity. We then ran Schick tests with regular and minor group toxins. 250 SCIENTIFIC PROCEEDINGS (121). Without exception, all gave a strongly positive reaction to the minor group toxin, though 20 were protected completely against the regular toxin, and three more partially so. To further verify this stage of the work, we tried titrating any possible group antitoxin against the minor group by using a ripened minor group toxin, whose standardization, for obvious reasons, could not be attempted with standard antitoxin. Although we worked at the limit of sensitivity, approximately 1/500 M.L.D., we were unable to demonstrate the presence of any protective power in the blood of any of this group against the minor toxin. Seemingly, then, immunization with monovalent diphtheria toxin- antitoxin does not protect against infections of the minor group, as, of course, is manifest in the two cases of that type originally quoted and with whose cultures we started our work. Two of the most important laboratories in the country have recently told us that they were likewise studying the toxins of certain diphtheria organisms which are not neutralized by the antitoxin in current use, so we hope after their publication that steps will be taken to include the minor group in the preparation of antitoxin, toxin-antitoxin, and Schick test toxin. 110 (1857) An intramuscular method of digitalis assay. By M. S. DOOLEY and C. D. HIGLEY (by invitation). [From the Pharmacological Laboratory, Syracuse University, Syracuse, N. Y.| Many observers have criticized the one-hour method of stand- ardization on account of the failure of complete absorption. During the course of work on the elimination of digitalis sub- stances, one of us demonstrated the feasibility of making intra- venous injections in the frog by the insertion of a fine hypodermic needle into the abdominal vein. It was suggested by Dr. Hat- cher that an intravenous method of assay might be evolved upon the frog. Efforts to do so have not been successful but in testing the possibility the idea occurred to us to experiment with an intramuscular method. INTRAMUSCULAR METHOD OF DIGITALIS ASSAY. 251 Our method is to inject one half the total dose into the thickest part of each thigh, the needle being directed diagonally. The finest needle must be used and hemorrhage avoided. Otherwise the procedures are the same as for the Pharmacopeial method. Judging by our results, the intramuscular method very largely and, we believe, satisfactorily, solves the difficulty of poor absorp- tion. TABLE I. CRYSTALLINE STROPHANTHIN (OUABAIN). : Per Cent. Frogs. S. S. S. Dose in Mg./Gm. Difference. Lymph Sac. Intramuscular. BEE os vis sh as .00045 .00037 17.8 UIE ae ies ee es .00051 .00044 13.8 We Ee oie ai Wie aCe .00032 .00027 554 SoM ee ay hea ae .00039 .0003I 20.6 Oe es ee .00059 .00050 15-3 he SE Ma ee -00057 -00051 10.6 DIGITALIS-TINCTURES,. ST ee ee -70 -57 18.6 Peta wien ets 5 -59 -49 17.0 DIGITALIS-FLUIDEXTR “CTS. Se eee .90 Neg. -60 -70 22.3 50 DIGITALIN-MERCK. FORMERLY KNOWN AS ‘‘GERMAN.”’ eR ee ae | 025 | -022! | 12.0 Table I summarizes the results. In all cases the intramuscular S.S.S. (systolic standstill) dose has been compared with the lymph sac S.S.S. dose. In no instance has the intramuscular dose been found as large as that by the lymph-sac method. Tinctures, fluidextracts, digitalin and ouabain have been studied. The last column of the table shows the percentage differences between the lymph sac and the intramuscular S.S.S. doses in corresponding lots of frogs. The effective intramuscular dose of digitalin is 10 per cent. less than by the lymph sac while, 252 SCIENTIFIC PROCEEDINGS (121). with tinctures and fluidextracts, the difference is an average of 18.6 per cent. The intramuscular method gives more constant end points and, therefore, requires less time and material for an assay. Lot I, fluidextracts, illustrates this point. With this preparation we were unable to determine the effective dose on account of poor absorption even with injections of doses 20.6 per cent. above the intramuscular dose. Even ouabain, generally considered as satisfactorily absorbed from the lymph sac, has required an average dosage 15.6 per cent. less by the intramuscular method. Earlier experiments have been repeated at a different season on different lots of animals with similar results. | It is believed that division of the dose, better blood supply in muscle than in skin and massage from movements of the animal, account for the more constant results and for the smaller intra- muscular dosage required. ABSTRACTS OF THE COMMUNICATIONS, MINNESOTA BRANCH. Minneapolis, Minnesota, February 8, 1922. Third Meeting III (1858) Evidences of a structure in gelatin gels. By ROSS AIKEN GORTNER and W. F. HOFFMAN. [From the Division of Agricultural Biochemistry, University of Minnesota, St. Paul, Minn.| Bancroft! recently reported some conclusions drawn from unpublished data of a Mr. Cartledge who dried gelatin gels of different concentrations down to a 96 per cent. gelatin content and then allowed these dried sheets to again imbibe water. It was found that ‘‘each swelled rapidly to the original concentration and then took up water-slowly.”’ We have conducted experiments similar to those of Cartledge and have secured comparable results. Thus a 1o per cent. gelatin 1 Bancroft, W. D., ‘‘ Applied Colloid Chemistry,”’ 1921, p. 251. THE CONTROL OF RESPIRATION. 253 gel dried down to less than 3 per cent. moisture content had imbibed at the end of 72 hours 6.45 grams of water per gram dry gelatin as contrasted with 4.30 grams water for a 40 per cent. gel similarly treated. Comparable differences were observed when the dried sheets were ground and uniform sized particles sieved out and tested for hydration rate and maximum hydration capac- ity. Our experiments indicate that gelatin gels have a structure and that this structure is fixed at the time of gelation and is not appreciably altered by drying the gel at room temperature. A crystal structure in which the gelation temperature is actually the melting point of the crystals would explain the peculiarities ob- served. II2 (1859) The control of respiration. By C. C. GAULT and F. H. SCOTT. [From the Department of Physiology, University of Minnesota, Minneapolis, Minn.| The movements of respiration are carried out by voluntary muscles and ought to obey the laws of voluntary movement. One of the chief points in muscular action is the dependence of motor response on sensory impulses. Without the guidance of sensory impulses movements are ataxic. Just how ataxic or abnormal movements become depends on the extent of loss of sensory im- pulses and on the ability of the mechanism to guide itself by sen- sory impulses from other sources. These statements hold true in regard to the movements of respiration. It has long been known that a modified respiration results from cutting off the sensory impulses from the lungs by section of both vagi. Many investigators have, however, kept animals with divided vagi so that one cannot maintain that the vagi are essential to respiration. However, it was pointed out by one of us! a number of years ago that animals with vagi divided are not nearly as efficient in times _ of respiratory stress as is a normal animal. It was shown by 1 Scott, F. H., Jour. of Physiology, 1908, xxxvii, 301. 254 SCIENTIFIC PROCEEDINGS (121). Alcock, Einthoven and others that expansion of the lungs sets up electrical variations in the vagus, thus showing these sensory impulses actually exist. A number of years ago we! called atten- tion to the alteration in respiration after section of the cord or division of the posterior thoracic roots. That impulses are set up in the joints of the thoracic cage every time the thorax expands may be shown by connecting the peripheral end of a cut inter- costal nerve to a string galvanometer. There is an electrical variation each time the thorax is expanded. The respiratory center is thus informed of the position of the thorax as well as the position of the lungs. We will discuss in detail at a later time the effects of these impulses on the respiratory center. One of the means of testing ataxic muscle is to have it do cer- tain movements more strenuous than normal. The respiratory mechanism may be tested in this manner with either increased CO, or decreased O2 in the air. We finally adopted the method of rebreathing. By this method we found animals with divided vagi or with divided cord are much less efficient than normal animals. In a number of cases we found in animals with divided cord that rebreathing from the spirometer caused a marked slowing of respiration. The respirations always increased in depth but the animals were unable to make a deep and at the same time a rapid respiration. Inno case after division of the cord did we find an increase in rate nearly proportional to that in the intact animal. As examples two experiments may be quoted: Total c.c. per Min. Cat MORIN «ose dp ds 3m Mad eo 4 a bw bee 2464 After i min, rebreathing «(000% os tna eee wes 3780 = + 53% Cord cut 7th cervical 17 min. after operation... 2030 After 1 min. vebreathing ; - |. 4. :dsseny tenn tae 1628 = — 19% Cat, normal | 360% ep cha ero. 1965 3628 = + 84% 1134 1489 = + 30% 840 1050 = + 25% After z min. sebresthiing «so: :'s000s sae ek 30 minutes after division of vagi.............. After 1 minute rebreathing..............0000: 15 minutes after division of cord at 7th cervical. . After 1 minute rebreathing.............20008. 1 Gault, C. C., and Scott, F. H., Am. J. of Physiology, 1918, xlv, 555. EXPERIMENTAL ACTINOMYCOSIS. — 255 113, (1860) Potassium iodide does not influence the course of an experimental actinomycosis. By A. T. HENRICI and G. S. REYNOLDS. [From the Department of Bacteriology and Immunology, the Univer- sity of Minnesota, Minneapolis, Minn.| It is generally believed that iodides are almost specific in their favorable influence upon the course of various mycoses, notably sporotrichosis, aspergillosis, ‘‘blastomycosis,’”’ and actinomycosis. There can be no question about the value of iodides in sporotri- chosis, but concerning the other mycoses reports are not so uniform. It is generally believed, however, that the iodides may cure actinomycosis, particularly in cattle.! Some of the fungi are not virulent for lower animals, and most of the others rapidly lose their virulence when cultivated, so that little experimental work has been done. Renon? found that Aspergillus fumigatus grew in culture media containing as much as 10 per cent. of potassium iodide; but that inoculated rabbits treated by subcutaneous injections of the salt did not die until 26 and 32 days after infection, whereas the control died in 4 days. Davis’ found that in experimental sporotrichosis the injection of iodides previous to or simultaneous with inoculation had no inhibiting effect on the course of the disease; but when adminis- tered after the infection is under way, the lesions heal. He also found that Sporotrichum would grow in media containing consider- able quantities of iodide. | Henrici and Gardner‘ have isolated from a case of pulmonary infection a variety of Actinomyces very similar to but not quite identical with A. asteroides Eppinger, which they named A. gypsoides. ‘This fungus is very virulent for guinea pigs, and has maintained its virulence quite unaltered for several years. It is 1 Salmon, D. E., Eighth and Ninth Annual Reports of the Bureau of Anima! Industry, Washington, 1893. 2? Renon, L., “Etude sur l’Aspergillose chez les Animaux et chez l’Homme,”’ 1897. 3 Davis, D. J., Jour. of Inf. Dis., 1919, xxviii, 124. ‘ Henrici, A. T., and Gardner, E. L., Jour. of Inf. Dis., 1921, xxviii, 232. 256 SCIENTIFIC PROCEEDINGS (121). an acid-fast variety quite different from A. bovis, and there are no clinical reports of the use of iodides in this type of actinomycosis. Nevertheless, because of its constant virulence it is admirably suited for chemotherapeutic experiments, and it was thought desirable to see what influence iodides would have on the course of the infection in guinea pigs. The potassium iodide was administered by mouth in aqueous solution. The resultsare shown in the table below. The first guinea pig was treated with iodides alone; animals 2 and 4 received iodide both previous and subsequent to inoculation; animals 3 and 5 are untreated controls to 2 and 4 respectively. | Guinea Guinea Guinea Guinea Guinea Pig I. Pig 2. Pig 3. Pig 4. Pig 5. Date | S| A | 5 | HM Wt. | KI, | Wt KI, | Wt. KI, Wt 4S Wt KT, | gm. | gm. | gm gm gm. gm gm gm. gm gm JAR a? 2 743 | 0.12 | 838 0.25 654 | 0.01 pS ee 695 | 0.25 | 790 0.25 622 0.25 A) ee Uae 675 | 0.25 | 743 0.25 | I0I2 fe) 620 0.25 (inoculated) ! (inoculated) Jan. 11....| 650 | 0.25; 625 | 0.25 965 | o 595 | 0.25 jani.33% 5: died died Jan. 13....| 608 | 0.25 565 0.25 Jan. 16....| 580 | 0.25 543 0.25 Jan. 18; .2.}°S72 | 0.25 539 | 0.25 | — oO (inoculated) | (inoculated) Jan. 20....| 580 | 0.25 555 0.25 560 fe) Jan. 22. ..| -562°1:0.25 500 | 0.25 | — oO Jan. 23....| — 438 0.25 485 fe) Jan. 24....} 600 | 0.25 died died It will be seen from the above that the treated animals suc- cumbed simultaneously with the controls; and that the iodide itself, while given in relatively large doses, was not sufficient to hasten death. In vitro it was found that both A. gypsoides and A. asteroides, while retarded, still grew in broth containing Io per cent. of potas- sium iodide. It is clear, then, that potassium iodide has no specific action on this type of Actinomyces, and if it has any clinical value, it is due, as in sporotrichosis, not to an action on the parasite itself, but because of its action in stimulating the formation of granula- tion tissue. With A. gypsoides the course of the disease is too rapid to demonstrate this latter point in the guinea pig. EVIDENCE OF A STRUCTURE IN GELATIN GELS. 257 114 (1861) Evidence of a structure in gelatin gels.! By ROSS AIKEN GORTNER and W. F. HOFFMAN. [From the Division of Agricultural Biochemistry, University of Minnesota, St. Paul, Minn.| The question as to whether or not a colloid gel has a definite structure is. extremely important to the biologist because of its bearing on the structure of protoplasms. Bancroft? recently reported some unpublished results of a Mr. Cartledge in which gelatin gels of different concentration were dried down to 96 per cent. gelatin and then these dried sheets were allowed to again imbibe water. It was found that “each swelled rapidly to the original concentration and then took up water slowly,” or in other words, a film of dried gelatin made from 8 per cent. gelatin gel was still potentially an 8 per cent. gel and very different from a dried film from a 16 per cent. gel. Arisz* had previously reported data showing a marked difference in imbibition capacity of gelatin gels of different concentrations and also of discs of uniform concentration but of different thick- ness. Inasmuch as the experiments cited by Bancroft have such a profound bearing on all colloid-chemical studies in which gelatin has been used, it appeared worth while to attempt to duplicate Cartledge’s results. Accordingly a series of gels was prepared as follows: Weighed quantities (10, 15, 20, 25, 35 and 40 grams) of ‘‘ Bacto”’ gelatin? were added to 100 c.c. of distilled water in clean pyrex flasks. After soaking for 15 to 30 minutse the flasks were placed in a hot water bath and allowed to remain until all of the gelatin had dissolved to form a homogeneous solution. A measured quantity (25 c.c.) of this solution was then poured into petri dishes 89 mm. in diam., thus ensuring the same thickness of the 1 Published with the approval of the Director as Paper No. 311, Journal Series, Minnesota Agricultural Experiment Station. 2 Bancroft, W. D., ‘“‘Applied Colloid Chemistry,” 1921, p. 251. 8 Arisz, L., Koll. Beih., 1915, vii, 51-6. 4 Air-dry gelatin as received from manufacturer. 258 SCIENTIFIC PROCEEDINGS (121). gelatin gelineachinstance. After standing for 12 hours duplicate rectangles 5 x 2.5 cm. in surface area were cut and placed on watch glasses to dry in a current of warm air (30°—40° C.) to a moisture content which did not exceed 3.5 per cent. The rate of moisture loss was followed by frequent weighings of the gelatin plates during the drying process, but no marked differences in rate of moisture loss were observed. The dried sections were then placed 8 0 Original gel e=/Ogram « Os es ve Os o5 * ” GRAMS WATER /MB&/BED PER GRAM DRY GELATIN / 2 3 4 5 6 7 8 HOUVAS Fic. 1. Showing imbibition curves (in water) for sections of dried gelatin prepared from gels of different initial concentrations. EVIDENCE OF A STRUCTURE IN GELATIN GELS. 259 in distilled water and allowed to reimbibe moisture. The rate of swelling was followed by weighing the discs at frequent intervals after removing surface water by blotting with neutral filter paper. The data calculated in grams water tmbibed per gram dry gelatin are shown in Fig. 1. GRAMS WATER IMBIBED PEFR GRAM ORV GELATIN e¢/OGram gel OF 7) .« 020 ty > Q25 Me @35 4 040 26 32 36 40 44 + «46 HOURS Fic. 2. Showing imbibition curves (in water) for dried sections of a 10 per cent. gel and a 40 per cent. gel. Dried sections of equal surface area and of equal thickness. While these curves appear to indicate marked differences in the dried plates, depending on the original concentration of the gel, such a conclusion is open to the criticism that the dried plates, while of equal surface area, were necessarily of unequal thickness. 260 SCIENTIFIC PROCEEDINGS (121). We therefore prepared another series of Io per cent. and 40 per cent. gelatin gels (10 grams air-dry gelatin in 90 grams water and 40 grams gelatin in 60 grams water) and poured the Io per cent. gel 4 times as deep in the petri dish as was the 4o per cent. gel. These on drying should give gelatin plates of equal area and thick- ness. After drying these plates they were allowed to imbibe water asin the above experiment. Figure 2 shows the imbibition curves obtained. Here again there is a marked effect due to the initial concentration of the gel. Similar discs were placed in N/25 lactic acid (Py 2.49). It is well known that acid solutions greatly influence rate of imbibition and amount of swelling of GRAMS WATER (MBIBED PER GRAM ORV GELATIN /2 24 36 468 60 72 HOUVAS Fic. 3. Showing imbibition curves (in N/25 lactic acid) for dried sections of a 10 per cent. gel and a 40 per cent. gel. Dried sections of equal surface area and equal thickness. EVIDENCE OF A STRUCTURE IN GELATIN GELS. 261 proteins on the acid side of the isoelectric point. Fig. 3 shows the very marked differences which were obtained. In order to be perfectly sure that the shape or thickness of the gelatin plate was not responsible for the differences in swelling, portions of dried plates from I0-gram, 25-gram and 35-gram gels, as well as portions of the ‘‘original’’ Bacto gelatin, were ground and sieved, those particles passing through a 2-mm. sieve and remaining on a I-mm. sieve being retained for experimental work. A weighed quantity of these ‘‘granules”’ were placed in a Gooch crucible and allowed to imbibe water. At frequent intervals the crucibles were removed from the water, centrifuged at low speed for 2 minutes in order to remove excess moisture and weighed. Fig. 4 shows the form of the imbibition curves. In order to ascertain whether the lower imbibing capacity of the gels containing the most gelatin might be due to the dehydrat- ing effect of electrolytes in the gelatin ‘‘ash,’’ which of course would be present in increasing amount as the concentration of the gelatin increased, a series of experiments was conducted in which the swelling plates were allowed to imbibe in (1) the same portion (100 c.c. of distilled water), and (2) in frequent changes of distilled water. If electrolytes were present they should have been dialyzed out in series (2). No differences in excess of experi- mental error were observed; in fact, a slightly greater imbibition was noted in series (1). The effect of hydrogen-ion concentration upon the physical state of gelatin has been pointed out by many investigators, but it is not clear that we are dealing with any appreciable changes in hydrogen-ion concentration. It is impossible to secure by direct measurement the hydrogen-ion concentration of the dried gelatin plates which are immersed in the distilled water, and we have no means of being sure that it is identical with that of the gel from which the plates were prepared. Electrometric measurements on a 5 per cent. gel gave a Py of 5.19 and the same value was ob- tained for the 1o per cent. gel. The higher concentrations of gelatin were so viscous that electrometric determinations were not attempted. Inasmuch as there was no change between the 5 per cent. and Io per cent. gel we believe that it is safe to assume an initial Py of approximately 5.2 for all gels before drying. This 262 SCIENTIFIC PROCEEDINGS (121). is slightly on the alkaline side of the isoelectric point of gelatin.' The hydrogen-ion concentration of the water in which the plates were immersed was probably between Py 5.0 and 6.0. The water as it came from the still was a fair grade of conduc- tivity water and was free from carbon dioxide and ammonia. Naturally it afterward absorbed some carbon dioxide from the air. Kendall? has recently presented data showing the rapidity with which this takes place. According to his work distilled water is about Py 5.7 when in equilibrium within the carbon dioxide of laboratory air. We have tried a number of times to determine the Py of distilled water, as used in these experiments, but have failed to secure sharp readings on the potentiometer because of the slight conductance of the water, our values ranging between © Py 5.0 and 6.0.2 Colorimetric measurements of hydrogen-ion concentration could not give accurate readings due to the fact that the indicators are all acids or bases which have greater ionizing power than has the water which is being measured. If we assume, therefore, that the hydrogen-ion concentration of the dried gelatin plate is identical with that of the original gel from which it was prepared, 7.e., Py 5.2, and that the distilled water was in equilibrium with the carbon dioxide of laboratory air, 7.¢., Py 5.7, we should still have no appreciable effect of hy- drogen-ion concentration on our experimental results for the same water and gelatin were used in all experiments. We doubt whether the above assumptions as to Py values are justified, but we do feel that the differences in the swelling of dried gelatin plates which we have described are not due primarily to differences in hydrogen- ion concentration. | Conclusions.—The above data appear to indicate that gelatin gels have a structure and that this structure is more or less fixed at the time that gelation takes place. It would appear that the gelatin aggregates or micelles are more and more interlaced at increasing concentrations of gelatin. This structure is apparently not appreciably altered by drying at a temperature below the 1 Loeb, J., J. Gen. Physiol., 1918, i, 39. 2 Kendall, J., J. Amer. Chem. Soc., 1916, xxxviii, 2460. * As a matter of fact the Ra of the distilled water is probably not an important factor since the water is so feebly buffered that a mere trace of acid or alkali wil] change the hydrogen-ion concentration through a wide range. EVIDENCE OF A STRUCTURE IN GELATIN GELS. 263 ‘melting’? point of the gel. The fact that drying does not markedly influence the gel structure appears to argue against the formation of micelles by the adventitious coming in contact of dispersed particles of gelatin, for certainly many gelatin particles must touch each other in the dried sheet, but apparently they do not cohere to each other with any appreciable force, certainly not with a force at all comparable with the force of coherence between particles or micelles originating at the time of gelation. This GRAMS WATER IMBIBED PER GRAM ORY GELATIN 0 10% Ge/ ©F07. ° HOURS Fic. 4. Showing imbibition curves (in water) for granules of equal average size of dried gelatin prepared from gels of different initial concentration. 264 SCIENTIFIC PROCEEDINGS (121). might be explained by a crystal structure where the crystals melt or soften at the gelation temperature. The micelles would then be formed by the solidification of crystals and later when micelle touched micelle the hardened surface of the crystal would prevent cohesion. We have recently shown! that a crystal gel may be very dilute and yet possess considerable rigidity so that a crystal structure is not incompatible with the properties of gelatin gels. These experiments likewise show the marked influence that hysteresis may have on experiments where gelatin is involved. This is particularly noticeable in Fig. 4 where uniform-sized par- ticles prepared from gels of different concentration were used. Inasmuch as a dried sheet of gelatin apparently reflects the struc- ture of the original gel from which it was prepared, the production of different samples of dry gelatin possessing uniform physico- chemical properties would appear to be extremely difficult. It is possible that these experiments may account for the differences between the experimental data of various workers. It would be interesting to know whether alcohol precipitation from sols of differing concentration produces gelatin particles of uniform physico-chemical properties. It is our intention to in- vestigate this problem in the near future. ADDENDUM, MARCH I, 1922. Since the above MS. was prepared, Sheppard and Elliott? have published on the same question. They do not find it neces- sary to assume a gel structure and believe that the different swell- ing rates are due to ‘‘casehardening’”’ or surface drying effects. We believe that our experiment where uniform sized gelatin particles were employed excludes such an explanation and con- sequently prefer to adhere to the structure theory outlined above. 1 Gortner, R. A., and Hoffman, W. F., J. Amer. Chem. Soc., 1921, xliii, 2199. * Sheppard, S. E., and Elliott, F. A., J. Amer. Chem. Soc., 1921, xliv, 373. SCIENTIFIC PROCEEDINGS. ABSTRACTS OF COMMUNICATIONS. One hundred twenty-second meeting. Presbyterian Hospital, March 15, 1922. President Wallace in the chair. 115 (1862) The feeding of non-ketogenic odd-carbon fats to diabetic patients. By MAX KAHN. [From the Department of Laboratories, Beth Israel Hospital, New York City.| In certain states of the disturbances of the metabolism of fats and carbohydrates a condition of acidosis is established characterized by the fact that the blood is rich in ketonic acids of a certain type. To this condition the special name of ‘‘ketosis’’ has been applied. It is induced by starvation, by the toxic effect of lipin solvent anesthetics, and especially by that disturbance of carbohydrate metabolism known as diabetes. Under normal conditions, that is, in the presence of proper carbohydrate oxidation, there is a rapid breakdown of the fatty acid fraction of the fats to the four carbon acid, 7.e., butyric acid, which is then rapidly catabolized to carbon dioxide and water. This process is, however, markedly disturbed in states of deficient carbohydrate oxidation. In the latter circumstance the fats are primarily broken down to butyric acid, as in the normal condition, but in the absence of the heat of carbohydrate consump- tion, the further decomposition of the butyric acid proceeds very gradually. The butyric acid under these conditions is oxidized first to beta-oxybutyric acid, and then to acetoacetic acid, which _is decarboxylated to acetone. 265 266 SocIETY PROCEEDINGS (122). It is prohibitive to feed diabetic patients, who have a very low carbohydrate tolerance, even a moderate amount of natural fat, because of the danger of inducing a severe ketosis which may prove fatal. It was thought advisable, therefore, to prepare a synthetic fat to contain fatty acids of odd-carbon number, which, if they are absorbed and if the theory of intermediate fat meta- bolism described above, holds, should catabolize in the body without the production of the acetoacetic acid, etc. In collaboration with Dr. H. O. Nolan such synthetic fat was made and fed to typical diabetic and ketotic patients. It was found that the fat was absorbed, that large quantities of it could be fed to these patients without inducing any acidosis, and that the nutrition of such individuals was improved. We are now studying the intermediate metabolism of this fat, and its feeding effect on all types of diabetic and normal individuals. 116 (1863) Hydrogen-ion concentration studies of solutions used for intra- venous medication and clinical investigation. By JOHN R. WILLIAMS and MADELEINE SWETT (by invitation). [From the Highland Hospital, Rochester, New York.| We advance the hypothesis that there is a relationship between the hydrogen-ion concentration of fluids injected intravenously and some of the reactions which follow their use. We base this belief on clinical observations and on extended chemical analyses of fluids commonly used for therapeusis and clinical investigation. We wish to briefly report here some of our more important studies. It should be understood that the hydrogen-ion concentration of normal acid is P, 2; normal alkali P, 14; pure water P, 7; and human blood P, 7.4, and that this latter figure is fixed and cannot be varied without causing serious disaster or death. If a fluid with a much higher or lower P, than that of the blood is intro- duced into the circulation at a rate or in an amount greater than the blood can neutralize or buffer, reactions, as chills, fever and HyYDROGEN-ION CONCENTRATION. 267 prostration ensue. This is particularly true in individuals whose blood supply is impaired in either quantity or quality. Since distilled water is the solvent used in most intravenous medication we have studied it with care under the conditions which it is commonly produced in hospitals: and laboratories. Rochester tap water has a P, of 8.29. Boiling was found to increase this slightly. Various samples of stock distilled water varied from P, 6.89 to Py 5.05. Distilled water becomes acid even when the distillation is in progress from the absorption of carbonic acid of the atmosphere when the ordinary type of metal or glass still is used. Tap water, P, 8.27; first distillate (first Io c.c.) Py 7.30; after 200 c.c. had been distilled P, 6.90. When stored in container, whether sealed with a cotton plug, cork or glass stopper, dis- tilled water becomes quite acid. Example: freshly distilled water has a P, 6.8, after 48 hours, Py 5.23. Proximity to bottles containing fuming HCl or NH,OH made no appreciable difference. The stock glucose solutions of the hospital prepared from stock distilled water and so-called chemically pure glucose were found to have a P, of less than 5. Three well-known brands of glucose were tested. Boiling, autoclaving and storing for 24 hours all cause glucose to rapidly become acid. Fresh unheated, Io per cent. glucose, P,; 6.20; after boiling 10 minutes, Py 5.47; after boiling 20 minutes, P, 5.17; after boiling 30 minutes P, 5. A fresh unheated solution ranged in P, from 6.2 to 4.15 after stand- ing 48 hours. Glucose solutions were buffered with the salts of mono- and di-potassium phosphates so as to give a concentration approximat- ing that of the blood. They have been used repeatedly in the hospital without producing reactions. The method of buffering was devised and materials provided by Dr. E. A. Slagle and S. F. Acree. Buffering must be done after sterilization since the latter precipitates out the buffer substances. The normal salt solutions of the hospital were found to be very acid ranging from P, 6.4 to P, 4.95. This was due in part to the use of stock distilled water. One well-advertised brand of normal salt solution put up in ampoule form had a Py 4.95. This high concentration in normal salt solution was found to be due in 268 SOCIETY PROCEEDINGS (122). part to the salt but chiefly to the acid distilled water. A salt solution to be normal physiologically, should contain that amount of salt which will make it isotonic with the blood. It should also have the same hydrogen-ion concentration. One specimen of sodium citrate solution used in a transfusion where a violent reaction followed, had a Py of 10.25. We have had no reactions in seven transfusions with the citrate method since we have buffered the citrate solutions. Several specimens of phenolsulphonephthalein, the intravenous and intramuscular use of which was followed by reactions, had a P, 5.0. We have had no reactions since with phthalein solutions properly buffered. We have observed also in a series of twelve cases tested with a dye having a concentration of P, 5.0 and later with a dye with a P, 6.95 that from 10 to 40 per cent. more of the properly buf- fered dye is eliminated in two hours than is the acid type in the same time. We believe that the hydrogen-ion concentration of chemicals or drugs is an important feature in their absorption and elimination both when injected or applied to mucous membranes of the human body. Several preparations of salvarsan, when prepared according to the directions of the manufacturers, ranged in P, from 11.53 to 11.99. Silber-salvarsan, P, 10.71; Neo-salvarsan, Py 8.15 to 9.22. A long list of sera, vaccines, and antitoxins prepared by the New York State Department of Health approximated the P, of the blood except tetanus antitoxin which had a P, 8.60 and anti- pneumococcus serum, type I, P, 8.10. Triple pneumococcus vaccine, U. S. Army Medical School, Py 8.95. These might. be sufficiently alkaline to produce local reactions. All of the common drugs used for hypodermatic medication, both in tablet and ampoule form, were tested. Liquid digitalis and strophanthus preparations were found to have a high con- centration, as were preparations made from the pituitary body and suprarenal bodies. Novocaine becomes acid on standing. We found much of the medication sold in ampoule form much more acid than the body tissues or fluids. We believe this to be the main cause of sore arms after the subcutaneous injections of such drugs. ANTIGEN REMOVED FROM CIRCULATION. 269 117 (1864) On the mechanism by which antigen is removed from the circulation. By GEORGE M. MACKENZIE and EMILY L. FRUHBAUER. [From the Department of Medicine, College of Physicians and Surgeons, Columbia University and the Presbyterian Hospital, New York City.] In studying the rate of disappearance of horse serum and the curves for circulating precipitin in a group of serum-treated patients it was noted that individuals who have severe serum disease are good precipitin formers and that at the time the precipitin in the circulation reaches the crest of its curve or soon thereafter the precipitinogen rapidly disappears from the blood stream. On the other hand in those individuals who after a first administration of foreign serum, show very mild or no symptoms of serum disease little or no precipitin is demonstrable in the patient’s serum and the precipitinogen persists in the circula- tion for a long period. Intermediate types were also encountered. From these results it seemed at least plausible to assume that an important factor in determining the rate of disappearance of the foreign serum from the circulation was an intravascular union of antibody and antigen. That such an assumption is erroneous seems probable from the following experiments: In one series of experiments 12 previously immunized rabbits were injected intravenously with amounts of horse serum (3.00 c.c. Or 6.00 c.c.) comparable to the amounts used therapeutically in the group of patients studied. The animals were then bled every second or every third day and the precipitin and precipi- tinogen in the serum titrated. Six of the animals had a high titer (1:20,000 or higher) of precipitin in the circulation at the time of reinjection, 2 had a moderately high titer, 1 had only traces of precipitin, and 3 had nocirculating precipitin. Two of the 3 rabbits with no circulating precipitin had been immunized Io months previously and at that time had developed a high titer of precipitin which had entirely disappeared before the reinjection. Presumably such previously immunized rabbits which had shown 270 SOCIETY PROCEEDINGS (122). themselves to be good precipitin formers would form antibody in excess earlier than upon first immunization and would therefore dispose of injected antigen earlier than upon first immunization and earlier than fresh rabbits injected with the same amounts of antigen. A control series of 11 normal rabbits was injected with the amounts of horse serum used in the previously immunized rabbits; precipitin and precipitinogen determinations were made similarly on the controls every second or third day. Of the rabbits re- ceiving 3.00 c.c. of horse serum the time of disappearance in those previously immunized varied from 6 to 17 days with an average of 11.2 days and in the unimmunized controls the variation was 14 to 17 days with an average of 15.5 days. In the rabbits re- ceiving 6.00 c.c. of horse serum the figures were: Immunized, variation I to 37 days, average 13.7 days. Unimmunized, varia- tion 6 to 21 days; average 16.3 days. The difference, while in favor of the immunized animals, certainly falls short of theoretical expectations. Following these preliminary and somewhat inconclusive ob- servations an attempt was made to determine the réle of intra- vascular union of antigen and antibody by a different experimental procedure. Normal rabbits were injected intravenously with 6.00 c.c. of horse serum and then twice every day given large amounts of high titer anti-horse rabbit serum intravenously. Precipitin and precipitinogen determinations were made daily during the period of the experiment. No conclusive evidence of accelerated disappearance of antigen was observed. One rabbit which received 97 c.c. of potent (1:20,000 to I: 500,000) anti-serum during the 48 hours following the injection of 6.00 c.c. of horse serum had the foreign serum in the circulation for 7 days. Another rabbit receiving 126 c.c. of potent (1:20,000 to 1: 500,000) anti-serum during the 55 hours following the injection of 6.00 c.c. of horse serum continued to have the foreign serum in the circulation for 9 days after the antigen injection. While the rate of disappearance of antigen in these two rabbits is below the average for normal rabbits injected only with horse serum the rate is within the limits of variation of the control rabbits. If such a flooding of the circulation with specific anti- _——— ———— a a | a ANTIGEN REMOVED FROM CIRCULATION. 271 serum produces little or no acceleration of the disappearance of antigen it seems justifiable to assume that intravascular union plays an unimportant réle in the mechanism for removal of foreign serum from the circulation, and therefore that the cellular phase is of predominating significance. It is also evident from the results on the whole group of 28 rabbits studied that individual variation extends over a wide range both in ability to form antibody and in the rate at which foreign serum is removed from the circulation. 118 (1865) The effect of various proteins on streptolysin production. By FRANKLIN A. STEVENS and CLIFFORD LAMAR. [From the Department of Medicine, College of Physicians and Sur- geons, Columbia University, and the Presbyterian Hospital, New York City.] Variations in streptolysin production in horse and rabbit serum media were noted in a previous publication. In horse serum hemolysin was produced in titratable quantities later in the growth of cultures than in rabbit serum broth but the maximum concentration reached was greater. Unless glucose were present the curve of lysin production corresponded closely to that of growth since hemolysin was found in greater concentration during the period in which the bacteria were multiplying most rapidly. On account of these differences which were characteristic of these sera an attempt was made to discover the responsible factors. The albumen and globulin ratio was modified so that the horse serum contained the same proportions of horse-serum albumen and globulin as were found in rabbit serum, and rabbit serum the same percentages as were present in horse serum. Flasks pre- pared with 20 per cent. of these modified sera in plain infusion with 0.7 per cent. NaCl, were seeded with equal quantities of a 16-hour culture of hemolytic streptococcus in 20 per cent. horse serum broth. Hemolysin titrations were then made at intervals of an hour with a suspension of horse corpuscles in physiological salt. The percentages of albumen and globulin in normal rabbit 272 SOCIETY PROCEEDINGS (122). and horse sera were determined by fractioning the diluted serum with ammonium sulfate. The analyses in parts per hundred were as follows: Albumen. Globulin. ERR scene Nit mae eis eee BAS. Ps i iss ee, 29 se ee ee 3-5 Mabiaty oor 7h kaa ee SAE Fo sin Goss hoe ee Pe 2.27 Sterile albumen and globulin were obtained respectively from horse and rabbit sera under sterile conditions. The albumen was precipitated with ammonium sulfate. The globulin was prepared by saturating diluted rabbit serum with CQO, and so did not - represent the true globulin fraction of the serum. The horse serum was diluted until it contained 2.27 grams of globulin per hundred c.c. and sufficient horse-serum albumen added to bring the percentage of albumen to 3.41 grams per cent. Rabbit serum was diluted and rabbit-serum globulin added so that the percentages of albumen and globulin were the same as those of horse serum. Flasks of 20 per cent. media were prepared with the modified sera. When growth and streptolysin estima- tion were made with growing cultures in these media it was found that the reversed ratio of albumen to globulin had no effect on the type of curve obtained. The curves in the modified sera were similar to those in normal serum media. Hence it appears that albumen and globulin do not enter inte any of the peculi- arities of growth exhibited by hemolytic streptococcus. This fact was further substantiated by a study of the proteolytic enzyme of these bacteria, because the enzyme acted similar to erepsin and did not digest serum albumen and globulin. Further- more the addition of large amounts of albumen and globulin did not modify the curves. Growth and hemolysin were next studied in peptone. Two preparations of peptone were used. These were similar except one was partially hydrolyzed and the other broken until further digestion with active trypsin gave no increase in amino nitrogen. The peptone was boiled, filtered through a Io-pound filter and added to beef infusion. Analyses of the media prepared with these peptones after autoclaving were as follows: Total N. Amino N. Grams per 100. Per Cent. of Total N. PERT OORT FS OSS slits fo POS s a ab baw wee 14.9 STREPTOLYSIN PRODUCTION. 273 When streptococci which had not been animal-passed were grown in flasks of these media so that comparisons of growth and hemolysin could be made, the growth was more luxuriant, the lag shorter and the hemolysin stronger in the peptone which had been only partially hydrolyzed. This suggests that causes for variations in growth are to be sought in the partially split proteins of the blood serum. 119 (1866) The bicarbonate and chloride content of the blood in certain cases of persistent vomiting. By H. A. MURRAY, JR. (by invitation). [From the Presbyterian Hospital, New York City.] These researches were instigated by an interest in the phenom- enon of tetany, particularly in that form known as gastric tetany. Very little work has been done in this field on human subjects, none recently; and it seems that no blood analyses have been published. Whatever work was done was not convincing and the old hypotheses such as the dehydration and mechanical theories once offered as a result of clinical studies to explain the condition must be discarded as untenable. There has been some successful experimental work on dogs, however, in which tetany was produced by obstructing the pylorus and in which various disturbances in the salts of the blood were recorded. To summarize: McCann! (1918) was the one to discover that after pyloric closure there was a rise in the combined carbon di- oxide. This was confirmed by MacCallum? (1920) and in the surgical laboratories of this college (Hastings* and Murray 1921). MacCallum and ourselves also found a markedly diminished chloride content with normal values for calcium. In our lab- oratory, contrary to expectations, it was found that the H-ion 1McCann, W.S. J. Biol. Chem., 1918, XXXV, 553. * MacCallum, W. G., Lintz, J., Vermilye, H. N., Leggett, T. H.,.and Boas, E. Bull. Johns Hopkins Hosp., 1920, XxXxi, I. ’ Hastings, A. B., Murray, C. D.,and Murray, H. A. J. Biol. Chem., 1921, xIvi, 223. 274 | SOCIETY PROCEEDINGS (122). concentration was only slightly and what we considered insignifi- cantly raised. Finally, Dr. Greenwald, who was good enough to analyze specimens from three of our dogs, showed that there was no consistent change in the percentage of sodium. The analyses to be reported, indicate that the changes noted in dogs may occur in humans as well. Subjects with a carbon-dioxide tension of 70 vols. per cent. or over were considered abnormal and included in the group. In the chloride estimations anything below 5.5 grams per liter for _ plasma or 4.3 grams per liter for whole blood was deemed patho- logical. In all, we studied seven cases; three of them had obstructions at or near the pylorus from ulcer, and two from cancer; one man had subacute gastric dilatation following appendectomy, and the seventh member of the group had an annular carcinoma of the lower jejunum. All cases were associated with inordinate vomit- ing, four days to two months in duration. In a few cases gastric lavage was practised, a procedure which probably aggravated the condition. The abnormal values varied from 70 to 107 vols. per cent. In the three cases which showed tetany, the highest figures were respectively 103, 104 and 107 vols. per cent. The chloride values varied from 4.5 to 2.2 for whole blood and 5.1 to 3.7 for plasma. In several cases there was an accompanying rise of urea. In two cases, dilute solutions of hydrochloric acid were given intravenously with a resultant decrease in the COz tension. In the last case, 500 c.c. of 0.1 N HCl was combined with 500 c.c. of physiological salt solution as an infusion without apparent deleterious effects. These findings indicate then that persistent vomiting, in all our cases the result of high obstruction of the alimentary canal, will result in an increased bicarbonate and decreased chloride content of the blood. This is undoubtedly due to the loss of hydrochloric acid from the stomach. The cause of the tetany is not known definitely. Dr. Van Slyke‘ in a recent article stated that uncompensated alkalosis was a cause of tetany. Some of our findings suggest that this is ‘Van Slyke, D.D. J. Biol. Chem., 1921, xlviii, 153. STREPTOLYSIN PRODUCTION. 275 correct. For instance, it was observed that tetany occurred in the three cases in which the COz tension was highest. However, our results with dogs do not confirm this hypothesis. Dr. Hastings made the P, determinations by the gas chain method, using all the accepted refinements. He found that it was normal until just before death, when the usual sudden ante-mortem drop occurred, proving that in this condition, death at least, is not due to an intoxication by hydroxylions. Moreover, as far as I know, there are no fundamental experiments to prove that nerve tissue is hyperirritable in the more alkaline solutions. Thus, since we have no direct proof we are not prepared, yet, to say that alkalosis per se, is the cause. There are other ions to be considered besides the hydrogen ion, which may affect nerve irritability. Dr. Loeb® has pointed out the importance of the monovalent/divalent kation ratio or more specifically the Na/Ca ratio, and has shown differences in the anions as well. In this case, the sodium- calcium ratio seems to be normal but there is a marked disturbance in the anions which may be the important factor. We are investigating further into the problem, and hope to be able to determine by estimating the hydrogen-ion concentration and the carbon dioxide content in the same sample of blood whether we are dealing with a condition of compensated or un- compensated alkalosis. 120 (1867) Hen-feathering induced in the male fowl by feeding thyroid. By HARRY BEAL TORREY and BENJAMIN HORNING. [From the Depariment of Zoélogy, University of Oregon, Eugene, Oregon.| It is already well known to the members of this Society that the males of certain breed of fowls—notably Sebright bantams and Campines—are feathered so like the females as to be in this respect practically indistinguishable from them; and that such hen-feathered males, following castration (especially in early life), develop plumage of the usual male type. 5Loeb, J., and Ewald, W. F. J. Biol. Chem., 1916, XXv, 377. 276 SOCIETY PROCEEDINGS (122). This appearance of cock-feathering in normally hen-feathered males after castration is now familiar to us. It is the purpose of this paper to call attention to experiments that have led to a similar transformation but in just the opposite direction, namely, to the appearance of hen-feathering in cock-feathered males of ordinary breeds. We had begun a series of preliminary experiments upon the physiological correlations of certain of the ductless glands. Eighty Rhode Island Red chicks were under observation. They were of the same hatch. They had been divided into four lots of twenty each. All the birds in two of the lots, both males and females, had been castrated between two and four weeks after hatching. Those in the other two lots were unaltered. Four weeks after hatching, one lot of castrated birds and one lot of normal birds had begun to receive daily doses of dried thyroid (Armour and Company, containing 0.2 per cent. I) by mouth. The initial individual dose was 50 mg. It was increased from time totime. At the end of fifteen weeks it had become 330 mg.— a dose the birds were able to take without any disturbance of their normal health. There is a striking difference between the sexes of this breed of fowls with regard to the time at which the tail coverts make their appearance. These feathers began to show themselves in some of our birds six weeks after hatching. Five weeks later, they were well developed on the normal females, in both thyroid and control lots. But they had not yet appeared on the control males, either normal or castrated. This absence of tail coverts provided a ready recognition mark of the male. Less conspicuous at this time, but nevertheless unmistakably present on the neck, were the hackles characteristic of the male bird only. In sharp contrast with these hackled and tailless males were the thyroid-fed normal males. Though the latter were upstanding birds with well-developed comb and wattles, and unquestionably male in carriage and instincts, they were as unquestionably female in plumage, owing to the absence of hackles and the presence of well-developed tail coverts. All doubt as to the sex of these birds was removed when they were killed four weeks later. At HEN-FEATHERING IN MALE FOWL. 277 this time the control males were typically marked by long hackles, saddle feathers and definite sickles. The thyroid-fed males re- sembled females in each of these respects. In behavior, comb, wattles and spurs, they were male, in feathering female. It was clear that the addition of thyroid to the diet of unaltered males was somehow responsible for this condition. Turning to the castrated birds that had been fed thyroid, it was equally clear that the gonad was a necessary factor in the result. For not a single castrated bird of either sex showed the least evidence of thyroid feeding in any of the characters previously enumerated. The thyroid-fed castrated females like their castrated controls, approximated males in feathering. The thyroid-fed castrated males exhibited the ultra male plumage characteristic of capons. The facts thus far considered relating to the secondary sex characters are arranged for convenience in the accompanying table. Comb and Plumage. Wattles. Norsin! male occci eis ks. Male Male Wormnal females. oc. ue 5k Male Female Normal and thyroid male... . FEMALE Male Normal and thyroid female...Female Female Cpetrated male... lk Male Female Castrated female............ Male Female Castrated and thyroid male. . Male Female Castrated and thyroid femaleMale Female Males with hen-feathering induced by thyroid feeding are indicated by capitals. All normal males thus fed were hen- feathered without exception. In two instances, however, sickle feathers began to appear among the tail coverts toward the end of the thyroid-feeding period, suggesting a certain degree of escape from the influence of the thyroid. This was possibly connected with the fact that the birds were then getting less than the max- imum dosage of thyroid compatible with their health. Search for the portion of the gonad through which thyroid feeding achieved its effect led at once to the lutear cells originally demonstrated by Boring and Pearl!in the ovary of the hen and later in the testis of the hen-feathered Sebright, by Boring and Mor- gan.” The latter stated their belief that the secretion of these cells “1 Anat. Rec., 1917, xiii, 253. 2 Jour. Gen. Phystol., 1919, i, 127. 278 SOCIETY PROCEEDINGS (122). suppresses in the hen and in the Sebright male the character- istic cock-feathering. This view seems highly probable, although not, to our knowl- edge, completely demonstrated as yet. However, if it could be shown that as a consequence of thyroid feeding the lutear inter- stitial tissue had increased in our hen-feathered male birds, the fact would contribute strong support from a new direction. Our preparations, however, indicate no such hypertrophy. There seems to be little doubt of the complete or nearly complete absence of lutear cells from the normal testis of R. I. R. cockerels at the age of ten to fifteen weeks. The same appears to be true of the thyroid-fed birds, but we are not so confident of the facts in this case because of poor fixation of the tissues. New preparations from experiments now in progress will probably clarify the situation. The following series, however, adds some significance to the lutear tissue in the present connection. 1. In R. I. R. cockerels, ten to fifteen weeks of age, lutear cells appear to be entirely lacking, and these birds fail entirely to to develop tail coverts of the female type. 2. In White Leghorn cockerels of the same age, ieee cells are present and tail coverts of the female type are present also. In W. L. adult males, however, these cells are lacking, as in adult R. I. R. and correlated with their absence, the full male plumage is present. 3. In Sebright males, hen-feathering and lutear cells are present together in young cockerel and in adult as well. R. I. R. males do not pass through a juvenal plumage, at least so far as tail coverts are concerned. This is a rather ex- ceptional fact, contrasting sharply with what we have found to be true of the White Leghorn cockerel, and the Sebright, both cockerel and adult. Juvenal plumage would thus appear to be determined, not by age, but by cells whose presence, regardless of age or of sex, leads always to plumage of one type. According to this view, juvenal plumage is female plumage, and female, juvenal—or neither, as in the Sebright adult male. This problem has large implications, consideration of which would carry us outside the proper limits of this paper. RELATION OF SUPRARENALS TO OVULATION. 279 If the appearance of hen-feathering in normal males as a con- sequence of thyroid feeding is dependent on the presence of lutear cells, it remains to consider the nature of the relation between thyroid and lutear tissue. The former might lead to an augmenta- tion either of the mass or the activity of the latter, or both. There is no evidence as yet of an increase in the mass or number of lutear cells in the testis of thyroid-fed males. The second alternative, for which direct evidence is far less easy to obtain, may yet prove to be the correct one. I2I (1868) The relation of the diffusion constant to mountain sickness. By GEORGE HARROP (by invitation). [From the Presbyterian Hospital, New York City.] In connection with the recent physiological expedition to Peru, a series of determinations were made of the gas diffusion constant by the CO method described by Dr. Marie Krogh.! A series of preliminary determinations made at sea level agreed in the limits of error (5 per cent.) with those made at Cerro de Pasco, at 14,300 feet. It was found that the severity of the symptoms of mountain sickness, or seroche, exhibited by the eight members of the party were in direct proportion to the value found for their diffusion constants, those having values of over 40 suffering very little, or practically not at all. TABLE A. DETERMINATION OF DIFFUSION CONSTANT—CO METHOD. Cerro de Pasco, Dec. 27—Jan. 10, '22—14,300 Feet. Subject. K. Doz. | Clinical Phenomena. ye eee 5.50 25-4 Severe seroche, 3 days, symptoms chiefly nervous. Se? s, see 6.03 31.8 Severe seroche, 6 days, symptoms gastric and nervous. ee 8.44 36.0 Moderately severe seroche, 2 days in bed. Bass 9.3 38.3 Moderately severe seroche, 2 days in bed. BS vein vhs 5.91 41.5 Practically symptom free throughout stay. rs 6.85 42.9 Some headache; otherwise symptom free. aes ee 6.01 43.8 Practically symptom free. M. a ea 9.80 45.6 No effects from seroche. 1 Jour. of Physiol., 1915, xlix, 271. 280 SOCIETY PROCEEDINGS (122). TABLE B. DETERMINATIONS ON AMERICAN RESIDENTS AT CERRO—14,300 FEET. Subject. | K. | Dor. Pa vane 7.86 43-4 McL..... 9.76 44.9 All of these persons have lived two or more years os Bee Ape 7.85 44.7 at high altitudes carrying on their work, free Cay wk 11.37 4I.5 from symptoms. | ee aye 12.22 65.3 A further series on five acclimatized persons, none of whom had ever suffered from seroche, all gave values for the diffusion constant for oxygen (Doz) above 40. 122 (1869) An undescribed relation of the suprarenals to ovulation. By OSCAR RIDDLE. [From the Carnegie Station for Experimental Evolution, Cold Spring Harbor, N. Y.] The observations described here make it extremely probable that the suprarenal glands regularly and greatly enlarge in close relation to the time of liberation of ova from the ovary. The maximum size seems to be attained in the 44-hour interval between the ovulation of the first and second ova—which together con- stitute a definite period of ovarian activity in the pigeon. The early stages of the suprarenal enlargement are coincident with the 4 to 5 days of extremely rapid growth! which these ova undergo immediately before their expulsion from the ovary. Knowledge of the exact time (within an hour) of ovulation in the pigeon has made this result possible. An enlargement of the oviduct also occurs quite parallel with that of the suprarenals (both facts shown by curves and tables). That an hypertrophy of the suprarenals occurs in some sort of relation to the menstrua- tion, pregnancy and lactation of mammals has of course been described; so far as we are aware the nexus with ovulation has been overlooked. 1 Riddle, O., Amer. Jour. Physiol., 1916, xli, 387. a a RELATION OF SUPRARENALS TO OVULATION. 281 Before undertaking the present study we had learned that birds dead of tuberculosis, or from the presence of round-worms (Ascaridia), usually show eniarged suprarenals. The normal size of each suprarenal had been found to lie between 0.006- 0.009 gram; those dead of tuberculosis weighed as much as 0.051 g. and the Ascaridia-infested were almost equally enlarged. En- largement of these glands doubtless occurs under many other infections. Forty-three females with fully known reproductive history were taken for this study; they were killed at several intervals with reference to ovulation; they were placed in one of two groups according to whether they showed or failed to show round-worms or tuberculosis. The data obtained from the healthy birds show that in nearly (not quite) all cases the supra- CuRVE SHOWING DATA FOR HEALTHY PIGEONS. The 44-hour ovulation period (shown between the two vertical lines) occupies for convenience only one half the space it should occupy. All other time intervals are properly spaced. The ordinates represent weight. The number of birds con- cerned at each point in the curve—from left to right—follows: 3, 4, 6, 3, I, 2, 3, 1 DATA FOR SIZE OF SUPRARENALS IN RELATION TO THE PERIOD OF OVULATION. SS SS FS RS EE Ave. Weight (Grams). Period with Reference No. of to Ovulation. Birds. Body. | Oviduct. | Adrenals. Healthy Common Pigeons. ee Ee Soo, ea hs cee (3) 312 0.883 -0076 eee sie Hee Ss Saree wee CaR eeS (4) 351 4.957 .0148 a Rae ROR SS Ss Le eiaitin e'ccais ous as (6) 360 10.356 .0207 SL I ee oR SO ee (3) 355 8.781 0219 Pe ES wives Gs a es wae dere e (1) 373 9.483 .O197 meee reer oe ek ook es ee ie. (2) 344 7.886 -0136 a ae ER a ac uc avisp a's oe ae me (3) 343 2.987 .0164! OG Bee ae ens eee (1) 338 0.883 -O108 Common Pigeons bearing Ascaridia or Tuberculosis. Be ee eI Ote Sues bese Te ew eek (2) 314 1.499 .0097 Ie TS SIE 0S pie cain, opin brid eo eo (3) 348 3.725 .0166 WARE PRIS Cree os a ek oos Bee SS (3) 331 6.823 .0096 A a ee (1) 323 8.382 -OIIQ MDC MEIONE SS CUS. es hos pica ws a eee ee (7) 336 8.907 .0122 Poa ME eS 2 eee Soe et. ese Sh (2) 366 8.649 .OI4I OR RPS ee a (1) 340 7.828 .0138 UES ARIE Se ogre cals od eae 5 eta (1) 340 4.708 .0137 Fs Ma ROT oss ois is Re AR ee a be ce sen (2) 314 1.499 -0097 1 For two of the three the average is 0.0095. 282 SOCIETY PROCEEDINGS (122). renals were enlarged, and most enlarged in the middle of the ovulation period. ‘The diseased birds fail to show this relation; and, though these glands are larger than the normals in the periods most removed from ovulation, they do not show a comparable en- largement during ovulation. The appearance of the hypertrophied glands in the normal birds is otherwise wholly normal. Their appearance certainly suggests an increased activity of the glands during this period. Sections of the gland have not been made and we do not know - whether both parts share in the hypertrophy. If increased secre- tion occurs, it doubtless has an important bearing upon the gen- eral physiology of the gland; and upon the effects which “repro- ductive overwork”’ (Whitman, Riddle) has been found to produce on the egg size and offspring of pigeons; for, a bird made to produce as many as 30 or 40 pairs of eggs per year would thus be almost continuously subjected to a hypersecretion of the suprarenals. 123 (1870) Studies in the physiology of vitamins. III. A comparison of the effects of feeding extracts of muscle and yeast respectively. By GEORGE R. COWGILL. [From the Sheffield Laboratory of Physiological Chemistry in Yale University, New Haven, Conn.] In a previous communication! experiments were reported wherein it was shown that the feeding of extracts of rice polishings, wheat embryo, and navy bean to dogs which had been fed on a diet lacking vitamin-B resulted in a recovery of appetite which lasted for varying periods. Vitamin-B was suggested as the appetite-promoting factor in the preparations used. The present report concerns control experiments in which an extract lacking this factor was tested. Commercial Liebig’s extract of beef muscle and the extract of yeast vitamin as prepared by the Harris Laboratories were used for these experiments. Tests were made using the Liebig 1 Cowgill, PRocEEDINGS Soc. Exp. BIOL. AND MED., 1921, xviii, 290-291. oe a ee STUDIES IN THE PHYSIOLOGY OF VITAMINS. 283 extract administered either with single large doses or with large doses repeated daily for as many as fourteen (14) days, the material being introduced by stomach sound in order to eliminate the taste factor. In another series of experiments the meat extract was mixed with the food which had been refused. The meat extract did not restore the appetite to such animals. On the other hand relatively small amounts of the yeast extract given by stomach produced a prompt recovery of appetite which lasted for from four (4) to nineteen (19) days depending on the amount ad- ministered. Liebig’s extract in doses such as were employed in these ex- periments promotes the flow of gastric juice in normal animals.? This fact, together with our own observation that products con- taining vitamin-B do not promote the flow of saliva, pancreatic juice, or bile,’ suggests that the recovery of the desire to eat in our animals is not to be ascribed to an increased flow of gastric juice. 124 (1871) Studies in the physiology of vitamins. IV. Parenteral administra- tion of products containing vitamin-B— mammalian experiments. By GEORGE R. COWGILL. [From the Sheffield Laboratory of Physiological Chemisiry in Yale Uniwwersity, New Haven, Conn.| That relief from the symptoms due to a lack of vitamin-B in pigeons may be brought about in a very short time by intra- muscular injection of products containing this vitamin has been shown by many investigators. Studies of parenteral administra- tions of vitamin-B to mammals do not appear to have been made. In the course of our studies into the physiology of this vitamin, using dogs as experimental animals, the protein-free concentrate of vitamin-B from yeast as prepared by the Harris Laboratories 2 Pawlow, ‘‘ The Digestive Glands,’ 1902, 96. . § Cowgill, PRocEEpINGS Soc. Exp. BIOL. AND MED., 1921, xviii, 148-149; ibid., 290. 284 SOCIETY PROCEEDINGS (122). was used, sufficient amounts of this product being generously furnished by Dr. I. Harris. Intravenous injections of a neutral aqueous solution of this product into dogs showing severe nervous symptoms (clonic spasms, etc.) due to a lack of vitamin-B resulted in a complete cessation of such symptoms, in one instance within a half hour, and in another case within three hours after the injection. Ina third instance the relief from symptoms occurred after a longer period. An intravenous injection of the vitamin-containing product was also made into an animal which had lost its appetite after subsisting on the vitamin-free food for a period of days but which showed none of the nervous symptoms characteristic of vitamin-B deficiency. The injection was followed by a complete recovery of appetite which lasted six (6) days. Intraperitoneal injections were also made. Such an injection, while bringing about relief from nervous symptoms in all cases, did not prevent death from supervening from ten (10) to twelve (12) hours later in those instances where large amounts of material were injected. A control injection into a normal animal resulted in phenomena indicating that the fatal outcome in the instances cited was probably due to the too sudden introduction of large amounts of material and the effect of this procedure on the tissues within the peritoneal cavity. The effect of subcutaneous injections into such animals will be studied shortly. 125 (1872) The conductance of unicellular organisms. By S. C. BROOKS.! [From the Division of Pharmacology, Hygienic Laboratory, U. S. Public Health Service, Washington, D. C.] The electrical conductance of bacteria (B. coli and B. butyricus) unicellular alge (Chlorella sp.) yeasts (Saccharomyces sp.) and mammalian red-blood cells has been studied by a method which yields figures for the gross conductance dependable within 1/1o per ! Approved for publication by the Surgeon General. CONDUCTANCE OF UNICELLULAR ORGANISMS. 285 cent. Polarization capacity was compensated for by variable condensers in parallel with the variable resistance. While most of the studies here reported were made on the alga, Chlorella, there is apparently no fundamental difference between this and the other organisms. The difference between the conductance of the suspending fluid and that of the suspension of cells is expressed in per cent. of the former and called the net conductance. This will always be negative in sign, because living cells are poor conductors of electricity. Rather large variations in the net conductance are produced by factors, such as irregular distribution of cells in the suspension, which it is impracticable to eliminate and which may affect the net conductance of any one sample by several per cent. For this reason enough samples were used in each experiment to make the error of the mean less than three per cent. It is possible to calculate the limits between which the ob- served net conductance should fall, the relative conductance of the cells themselves and their volume concentration being known. If r is the resistance of the suspending fluid and er that of the cells and the concentration of the cells in volume per cent., then the total resistance should lie between ob Vn(k —1)+ 10 (Io Vn — n)(kR — 1) + 100 I and n(k — 1) ). r(r+— Vn(1 — k) + 100k These formulas give curves convex to the axis of concentrations when they are plotted against resistances as ordinates. But when the cells form less than about 65 per cent. of the total volume the observed curves are nearly linear, possibly because of varia- tions in the relative extent to which the current lines are able to evade the more highly resistant cell material. It is fortunate that this is so, since in case the volume of cells varies during the course of an experiment (e.g., because of osmotic changes) a linear correction may be introduced. Different organisms and even different lots of the same organism differ considerably in their net conductance at any given concentration. 286 SOCIETY PROCEEDINGS (122). The relation between the conductance of the suspending fluid and the net conductance of the suspension was somewhat un- expected. Perrier’s artificial sea-water' was diluted to 2, 4 and 8 volumes with distilled water. Chlorella which had been grown in a solution somewhat more dilute, was transferred successively to increasing concentrations of this artificial sea-water and the net conductance determined several times in each case until upon further renewal of the solution between each determination no further change occurred. For example, in changing from 1/8 to 1/4 strength sea-water, the net conductance changed from 28.9 to 27.2 per cent. Corrected for the decrease in the volume of cells, the figures would be 28.9 and 28.3 per cent. The change in net | conductance is within the limits of error of the method. This shows that if by any chance the conductivity of the surrounding medium is changed during the course of an experiment only the net conductivity in per cent. will normally remain constant. Ex- pressed in any other way the results might easily be misleading. In the above case the net resistance expressed in ohms instead of remaining constant fell from 80 to 45 ohms approximately. It is also of interest to note that the net conductance of dead cells is relatively greater in those organisms which normally encounter a fluctuating environment. ‘The ratio of the net con- ductances of dead and living cells is about as follows: red blood cells .45, yeast .65, bacteria .70, Chlorella .80-.90. This fact suggests that the cell walls (or at least some non-living structure), help to protect such cells from extreme fluctuations in electrolyte concentrations, insofar as the changes inside the cell walls would be retarded and therefore less abrupt. Multicellular forms such as Laminaria need such protection only at the outer surfaces of the superficial cells, and it is therefore not surprising that such tissues have when dead only about one tenth the electrical re- sistance that they have when living. 1 Perrier, E., Comptes Rendus, 1890, cx, 1076. HyDROGEN-ION CONCENTRATIONS. 287 126 (1873) The hydrogen-ion concentrations of joint exudates in acute arthritis. By RALPH H. BOOTS and GLENN E. CULLEN. [From the Hospital of the Rockefeller Institute for Medical Research, New York City.| The hydrogen-ion concentrations of exudates aspirated from joints of patients ill with acute rheumatic fever and other forms of arthritis were determined. This was done: (1) to compare the reactions of the exudates of the various forms of arthritis; (2) to determine if an acidity existed in inflamed joints of acute rheumatic fever patients sufficient to permit the liberation of free salicylic acid following salicylate therapy. Salicylic acid can not exist as such in alkaline solutions; and its salts have not been shown to have bactericidal power in low con- centrations. Although the acid can not exist in normal blood and tissues, their reactions being slightly alkaline, it has been suggested for a number of years that its liberation might occur in the inflamed tissues of patients with acute rheumatic fever; these tissues were supposed to have been under considerably in- creased COs tension. The bactericidal action of this liberated salicylic acid could explain the seemingly specific action of the salicylates on the arthritis of acute rheumatic fever. Hanzlik! examined exudates from inflamed joints of acute rheumatic fever patients directly for the presence of salicylic acid. The results showed none to be present; but the author offers the criticism that no precaution was taken to prevent the escape of COs. In our work, the hydrogen-ion concentrations of all of the exudates were determined colorimetrically at room temperature and corrected to 38° C. by a method recently described by Cullen.? If sufficient fluid was obtained from a joint, the determination was also made electrometrically. With both methods, in order 1 Jour. Pharm. and Exp. Therapeutics, 1917, ix, 217. 2 Jour. Biol. Chem., 1922, 1, 17. 288 SOCIETY PROCEEDINGS (122). to prevent the escape of COs, the fluid was not allowed to come into contact with the air. Results —The reactions of 16 joint exudates from patients with acute rheumatic fever were all slightly alkaline; their hydrogen- ion concentration varied from P, 7.2 to 7.38. Seven exudates from patients with chronic arthritis varied in P, from 7.27 to 7.4. An exudate aspirated from a knee infected with Staphylococcus aureus had a P, of 6.63 and that from a knee infected with Strepio- coccus hemolyticus was also acid, having a Py, of 6.14. Since a definitely acid medium is necessary for the liberation of salicylic acid, and since all of the joint exudates from acute rheumatic fever patients were slightly alkaline, no free salicylic acid can exist in these joint fluids following the administration of salicylates. 127 (1874) The selective bactericidal effect of acid fuchsin and sodium chloride. By JOHN W. CHURCHMAN. [From the Department of Hygiene, Cornell University Medical School, New York City.] In 1912 it was found that if bacteria be stained with gentian violet and planted on plain agar a sharp selective activity of the dye could be readily demonstrated. All the commoner gram- negative organisms survived even long exposure to the stain, while all the commoner gram-positive spore-bearing aérobes were ‘“‘killed,’’ even by a relatively short exposure. Even the spores— though not deeply, if at all, stained by gentian violet—were “killed”” by exposure to the dye. What was true of gentian violet was found to be true also of other basic dyes of the tri- phenyl-methane group. It is now found that a cleavage in exactly the opposite sense occurs if organisms be exposed to acid fuchsin—an acid dye of the tri-phenyl-methane series. Whereas gentian violet kills the gram-positive spore-bearing aérobes and spares the commoner gram-negative bacteria, acid fuchsin spares the former and kills SELECTIVE BACTERICIDAL EFFECT. 289 the latter. The experiments on which this statement is based were done with B. subtilis, B. megathertum, B. anthracis, B. typhosus, B. coli communis, B. prodigiosus, B. pyocyaneus, B. proteus vulgaris. In the case of gentian violet the reaction is evident if the stain is applied to the organisms at room tempera- ture; but in the case of acid fuchsin, while long exposure to the dye at room temperature produces the reaction, a slight increase in temperature (to 45° C.) makes it much sharper and speedier. In the case of gentian violet it was shown, by a study of the whole bacterial field, that the gentian-violet reaction and the gram reaction ran parallel in a striking way. Not only did the cleavage hold between gram-negative organisms and the commoner spore-bearing aérobes; it also held between gram-negative bacteria and the great majority of non-spore-bearing gram-positive organisms. That a reverse parallelism holds for acid fuchsin has not been proven. Of the greater susceptibility to this dye of the gram- negative organisms (as compared with the gram-positive spore bearers) there is no doubt; it is not established that a similar cleavage holds between gram-negatives and gram-positive non- spore bearers. An interesting practical point is the susceptibility of the gram-negative B. pyocyaneus to acid fuchsin; this organism is often a great nuisance in open wounds and resists ordinary antiseptics. What has been said of acid fuchsin is true also of another and wholly unrelated acid dye—ponceau P.R. Itisalso true of sodium chloride. When the commoner gram-negative organisms are exposed to saturated solutions of any of these three substances at 45° C., they are killed; while gram-positive spore bearers are unaffected under similar conditions. This is the exact reverse of the behavior of these two classes of bacteria toward basic gentian violet. In the case of gentian violet, the selective effect of the dye may be demonstrated either by staining the organisms and planting them on plain agar or by planting them unstained on agar con- taining the dye. The gram negatives grow well in the presence of the dye, while the gram positives will not grow at all, even when .the stain is present in the media in very small amounts. For this and other reasons the activity of the dye has been referred to 290 SOCIETY PROCEEDINGS (122). as bacteriostatic, since it seems to consist essentially in an in- hibition of growth. It is probable that this parallelism between the bacteriostatic and the bactericidal activity of the dye, so clear cut in the case of gentian violet, does not hold in the case of acid fuchsin and sodium chloride. The evidence thus far gathered would indicate that the mechanism by which growth is inhibited may be entirely different from the mechanism by which organisms are killed. 128 (1875) On the method of macronuclear disintegration during endomixis | in Paramecium aurelia. By LORANDE LOSS WOODRUFF and HOPE SPENCER. [From the Osborn Zoélogical Laboratory, Yale University, New Haven, Conn.] . In the original studies! of the nuclear phenomena involved in endomixis, it was found that macronuclear disintegration, both in Paramecium aurelia and Paramecium caudatum, was effected by the elimination of spherical chromatin-bodies from the macro- nucleus, instead of by the transformation of most of the macro- nucleus into long tangled chromatin-ribbons such as occurs during conjugation in these species. Regarding Paramecium aurelia, it was stated that the ‘‘differences between the macronuclear changes during conjugation and during the process (endomixis) are only morphological; on the one hand, the macronucleus forms ‘wurstformige Schlingen,’ while on the other, the macronucleus eliminates its chromatin by extruding it in the form of spherical bodies.” ? This contrast proved valid not only in Woodruff’s pedigree race’ (1) of Paramecium aurelia in which endomixis was discovered, but also in animals from such diverse sources as Germany and Ohio. Only one cell (Paramecium aurelia, 1), 4087th generation, Decem- 1L. L. Woodruff and Rhoda Erdmann, Journ. Exper. Zodlogy, 1914, xvii, 425- 517. Erdmann and Woodruff, Journ. Exper. Zodlogy, 1916, xx, 59-97. 2? Woodruff and Erdmann, loc. cit., pp. 438-39, and 444; Plate 1, Figs. 9 and 11; Plate 2, Fig. 14; Plate 3, Fig. 32. ® Woodruff, Biological Bulletin, 1917, xxx, 51-56. tattle ite ee er MACRONUCLEAR DISSOLUTION. 291 ber 6, 1913) was observed during endomixis which in any way even suggested ribbon formation and a figure was given of this animal with the legend ‘‘An atypical form of macronuclear disintegration, slightly resembling the ribbon-like formation characteristic of conjugation.” 4 It is therefore interesting to record that the study of animals from this same pedigree culture (I) of Paramecium aurelia at about the 89o00th generation (November, 1921) showed some cells successfully undergoing endomixis with macronuclear disintegra- tion by ribbon formation and others by chromatin-body formation. Thus it is clear that although all the data thus far at hand indicate that chromatin-body formation is the typical method of destroying the macronucleus in endomixis, nevertheless under certain unknown conditions the formation of chromatin-ribbons, until now regarded as diagnostic of conjugation, occurs in endo- mixis. This fact, of course, in nowise narrows the significant and crucial difference between endomixis and conjugation—the absence of synkaryon formation in the former and its presence in the latter. 129 (1876) Nutritive factors in plant tissues. V. Further observations on the occurrence of vitamin-B. By THOMAS B. OSBORNE and LAFAYETTE B. MENDEL. [From the Laboratory of the Connecticut Agricultural Experiment Station, and the Sheffield Laboratory of Physiological Chem- istry in Yale University, New Haven, Conn.] In the course of our studies of the distribution of vitamins in plant products we have collected data regarding a number of important edible foods for which no information in this respect seems to be available at present, with the possible exception of indirect suggestions obtained by other than animal feeding trials. Our experiments, made with rats, supplement numerous earlier ones! conducted by the same technique and indicate that asparagus, 4 Woodruff and Erdmann, loc. cit., Plate 4, Fig. 37. 1 Osborne, T. B., and Mendel, L. B., Jour. Biol. Chem., 1919, xxxvii, 187; xxxix, 29; 1920, xli, 549; xlii, 465. 292 SOCIETY PROCEEDINGS (122). celery, dandelion, lettuce, and parsley all contain noteworthy amounts of vitamin-B. The details of the investigation will be published elsewhere. 130 (1877) An experiment on the absorption of glucose given by rectum. By ROGER S. HUBBARD and DAVID C. WILSON. [From the Clifton Springs Sanitarium, New York.] For many years it has been accepted by clinicians that glucose given by rectum is absorbed immediately into the blood, and sur- geons constantly use such a procedure as a part of their post- operative therapy. Because of the difficulties attending the pro- cedure there are comparatively few controlled experiments on the effect of glucose so administered in the literature. The effect of carbohydrate feeding on acetonuria has been clearly demonstrated by many experiments during the past few years, and it was deter- mined to test the rectal absorption of glucose by studies of the effect produced upon experimental acetonuria. The subject of the experiment (one of the authors, D. C. W.) was a man 5 ft. 9¢ in. tall who weighed 165 pounds, and whose basal metabolism, as measured by the portable Benedict calorim- eter was 1700 calories. He received the diet recently discussed by Hubbard and Wright! for four days. This diet furnished 2,142 calories—twenty per cent. more than the basal requirement—and consisted of 54 grams of protein, 54 grams of carbohydrate, and 190 grams of fat. Ten per cent. of the calories in this diet are furnished by protein, ten per cent. by carbohydrate, and eighty by fat?*. The total food intake was probably not sufficient for the needs of the subject. Acetonuria developed gradually as shown in Table I, and on the fourth day of the experiment, before the acetone excretion had reached its highest level, an enema consisting of 300 c.c. of a 5 per cent. glucose solution was given. Table II shows figures for the morning of the day before the enema was given, for the speci- 1 Hubbard, R. S. and Wright, F. R., J. Biol. Chem., 1922, I, 361. 2 Zeller, H., Arch. Physiol., 1914, p. 213. ® Shaffer, P. A., J. Biol. Chem., 1921, xlvii, 449. ABSORPTION OF GLUCOSE GIVEN BY RECTUM. 293 men collected just before the enema was given, and for the speci- men which corresponded to the period during which the enema was retained. Both tables show that there was a decrease in the- concentration of acetone plus acetoacetic acid caused by the glucose, while the 8-hydroxybutyric acid did not show the increase which is usually caused by such diets.1. The decrease was not as great as that caused by similar amounts of glucose when taken by mouth. Shaffer? has recently advocated the theory that the failure of acetoacetic acid to burn completely in the absence of glucose is responsible for the appearance of increased amounts of the acetone bodies in the urine. The observation that the aceto- TABLE I. Urine. Date, Vol. Acetone. 8-hydroxy. Alv,CO:z 1921 ec. mm mg mg. 100 ¢c.c aie 100 c.c ae | o-oo |: gIo I.I 0.010 1.7 0.015 40 16-17 780 3.6 0.029 2:4 0.021 36 17-18 770 7.8 0.060 3-4 0.026 i i ae 852 25.2 0.215 II.9 0.IOI 32 19-20 628 18.7 0.118 15.2 0.096 300 c.c. 5 per cent. glucose at 9:30 A.M. on December 19. Results of the determinations of the acetone bodies are expressed in terms of acetone. Under acetone the results of the determination of acetone from acetone plus acetoacetic acid are given. TABLE II. Urine. Date, Time, Vol. Acetone. B -hydroxybutyric 1921. A.M. c.c. Acid. pas gms. ms gms. 100 c.c. 100 c.c. tafeGes<'. 8:30 to II 102 19.2 0.020 13.3 0.014 Bato... 8:30 to 9:30 40 13.0 0.005 5.5 0.002 12/19... . 9:30 to 12 108 7:8 0.008 4.6 0.005 300 c.c. of 5 per cent. glucose given by rectum at 9:30 A.M. on December Io. 1 Hubbard and Wright, loc. cit. 2 Shaffer, loc. cit., p. 433- 3 Day before the diet was taken. 294 SOCIETY PROCEEDINGS (122). acetic acid shows a decrease in these experiments, while the B- hydroxybutyric acid only fails to show the ‘“‘expected increase” seems to be in accordance with this view. The experiment shows that glucose is absorbed rapidly by rectum, and is taken into the blood in sufficient quantities to decrease the excretion of acetone produced by a diet high in fat. 131 (1878) Effect of dilution on the precipitation reaction for syphilis proposed by author. By R. L. KAHN. [From Bureau of Laboratories, Michigan Department of Health, Lansing, Michigan.| One of the important differences between the precipitation reactions of Meinicke, Sachs and Georgi, Dryer and Ward and that proposed by the author! is that in the last reaction, the amount of normal salt solution is reduced to a minimum. It was early observed, when adding given amounts of serum and antigen to a series of tubes and subsequently adding increasing amounts of normal salt solution to that series, that the degree of TABLE SHOWING DELAYING EFFECT OF PHYSIOLOGICAL SALT SOLUTION ON PRECIPI- TATION REACTION FOR SYPHILIS PROPOSED BY AUTHOR. | Tube No. | I | 2 3 4 5 6 Syphilitic serum, | CEio aia coe ae ae A 1>: Ho 235 179 93 132 res > Np ie Se ML 8 263 206 82 IOI Fee Ee ta ee ae Sere 290 247 90 97 1a giewad Marat ee 88 104 180 “ Te Es Dia es hacia aaa 186 161 90 TABLE II. EFFECT OF INGESTION OF MALTOSE AND OF FRUCTOSE. Blood Sugar. Mg. per 100 c.c. Whole Blood. Time. Tap aa, Exp; 233 Pep. 25, ' Exp. 26; 75 gm. 100 gm. 75 gm. go gm. Maltose. Maltose. Fructose. Fructose. Belore i271. oe eo ee 100 95. 84 95 Sm, alters: S72. ae — — 101 125 BE S51 Gig Tae 144 157 108 126 "5 ea OS gas cet Sores — — — 128 30-,°"! ray ig Sak eee Oe 89 165 105 III OS coat Mt ocak ot were QI 118 — 101 a0 Te a Me thas Se Ls — — 92 — FOr ey pre arate re Le. — —. — 104 Sy--"" Sig alk. ee he QI — — 140°°*T Te en ee ee 104 101 — — It is well known that fructose is much more rapidly oxidized in the body than glucose! *»* and there is some evidence that it is a better glycogen former.* Taking our blood samples at short and frequent intervals we could detect a rather small but unmis- takable rise of blood sugar in two fructose tests. In studying the effects of repeated doses of glucose we have obtained results quite different from those of McLean and de ! Johansson, J. E., Skand. Arch. Physiol., 1908, xxi, I. 2 Lusk, G., Jour. Biol. Chem., 1915, xx, 555. * Biirger, M., Biochem. Zeits., 1921, cxxiv, I. ‘Weinland, E., Zeits. f. Biol., 1899, xxxviii, 16 and 607. a eee —_ —_ eS aa a... 4 s BLoop SUGAR STUDIES. A4II Wesselow. They gave fifty grams of glucose which caused the blood sugar to rise and fall sharply. Then, when the curve had returned to normal, they gave a second dose of fifty grams which produced a response almost identical with the first. We find that the second dose may cause little or no response, depending upon the time at which it is taken (Table III). If the second dose is TABLE III. EFFECT OF REPEATED DOSES OF GLUCOSE. Two doses of 100 gm. each at interval as noted by asterisk. Blood Sugar. Mg. per 100 c.c. Whole Blood. Time. Exp. 21. Exp. 24. exp. 24. E 6h ea ines enemas N. Y. Health Department BARBER, W. HOWARD.........- iPad ease teket.s 5 cable New York University Banpovur, HEMRy G..., «04d bavede cena vs cane vet ae McGill University BARDEEN, CHARLES R.. .o5 6 s.3 bs Bude we eee cha utes ts. wie University of Wisconsin BaRNuetT, Georce D.. . ¢ s's:\Jdwelon Ue oe en abe eas Leland Stanford University DARE DAVE Py 5k k's nas cndciae ee Cornell University Medical College, N. Y. City Es Ae ih a Ee eS On RA Sly 7 Montefiore Home, N. Y. City BAUMANN, LOUIS. soit oe.co tb oe boat ee eae eas Presbyterian Hospital, N. Y. City 464 ROLL OF MEMBERSHIP. 465 NE MN Tes. cay buns 3 ow RAIA e Dees Ree Johns Hopkins University eG a aT Sg ts eS oa: ae ae University of California RRM te oes wk Woah ah a2 us ae cneuale Guan eae University of Minnesota ee Ee ee a ag Cornell University Medical College, N. Y. City eee, Wataser Ne ik. one won oo on Bureau of Animal Industry, Washington, D. C. Pe REI So a wk ws I a oa ated asin aes Seca Jefferson Medical College ERE SRR IRS AS ee i a ee University of Pennsylvania ES OS RE Se a er ee Hospital of the Rockefeller Institute BLAKESLEY, ALBERT F.....Station for Exp. Evolution, Cold Spring Harbor, N. Y. BLATHERWICK, NORMAN R..............-; Potter Metabolic Clinic, Santa Barbara ET ee ee ee eee University of California SM MEE EMME ie good as aoa Seek ges bee week ots n ls University of Wyoming SOMME ERE Seong Do aiicln ask chk bee ce ows Rockefeller Institute, N. Y. City PETIT. oo Ska sc sk a we ne ele anew enw es Harvard Medical School oo 3 SG SERS ce ea University of Alabama PCN a SG ge vk haa had kanes New York University Brome wet ieak soll nd Hygienic Laboratory, Washington, D. C. ee ES RR oe geile. kk ca Rca bea b Ube bbe d be University of Minnesota Re TS IO WARD oi iS. esa e ecw Rockefeller Institute, Princeton, N. J. Ren OY aie Aas tet SRS he Sis o ce ew eae Rockefeller Institute, N. Y. City REN NE SIM ee PEL a to6 Soe Sig le. cng reba tore hes College of the City of New York PUMA ON SII tone a ON cep ic baka eb otha be har Johns Hopkins University IDES ta ee Ee ie eee ce tues a da xls University of Wisconsin ree Un ed OREN Mong oS a. aces Ue ake ct sade edaes University of California PIPING es oop ann nde ec akie eee « Washington University Medical School NE REA RIE es Pierce nsan bode wore gabe hep esas Columbia University SIRES EMER ad foe ns ae cin Ge akicww Weta y Ae wee bees Columbia University oe en ltthhy OS A eer iene ere Harvard Medical School IIR OE ts on ra aoe Gece Sowa crorne wiligta's « Sarees ads aaa é University of Chicago Co EES Ss oq Cala okisie mice acs eee ta we wee Rockefeller Institute, N. Y. City Da PtP teary 6s ae Wy tive ajw yaa dim ends Susre si panhoin 0 University of Toronto, Canada a NE em street e's 5 ae Bellevue Hospital, N. Y. City eee Par rege ieee ee ke hee es N. Y. Post-Graduate Medical School CHAMBERS, ROBERT.............. Cornell University Medical College, N. Y. City Ce er Sate irate mr So SS, hin oe ne University of West Virginia Ree eRe eto 8) ns fo NG se ce sae oo os he note oe eres Yale University CHURCHMAN, JOHN W............. Cornell University Medical College, N. Y. City er ee OP aE Pr a NG ne he a Cedar biavesnieiese University of California erage Se” Braet WO tet Oop 5 Pe ke ode a's wees University of Wisconsin Cray ha, Fas Pied ks es SPE ee ee eee ee. Indianapolis, Indiana EE pA ES Ee RD ee ee eRe Cornell University Medical College, N. Y. City Cae a 9 Ce er era U. S. Hygienic Laboratory, Washington, D. C. Seat SUA DUR ere L . Sn w nm oo vin ole Mei Sat N. Y. Post-Graduate Medical School SEE: SEs er ree ee ane cee Rockefeller Institute, N. Y. City ee es Soe ne bos OE LK See a Th a ed es ae University of Wisconsin SECIS Wak ions). sa vo sk os she Swe eena as Rockefeller Institute, N. Y. City Ce A Tis oh eck se coo bs wd see eee Lake Forest College, Lake Forest, Ill. Cee PAS VE AMRUI SS cw on hs beh cee Or ckabeeaAeewninn to tenes New York University COLLINS, KATHARINE R.......... Division of Laboratories, Buffalo City Hospitals 466 SCIENTIFIC PROCEEDINGS (124). Commit, Bi G5 555 S82 hae ona Re ge dae eee ee Princeton University Contes Frcs si 2b headed ee Washington University Medical School Consaue, Tima (ie oo oasis cb a ele aoe Columbia University Comer, Geet Vii. eens Son es Eee eee ae Johns Hopkins Medical School Cowate SORE. .a6e ee «as oe tee Stanford University Hospital, San Francisco Crampron, (> WARS 0. o-2u\s sess os oe Department of Education, New York City Cais Somes Wis hides licen eee Western Reserve University Chine; Dargis, Be eo. aces oe hee ee ak bee Mt. Sinai Hospital, N. Y. City Gur We Toe sc ee ie iw bg DO Ware ivtecnte Fi hee ate Rutgers College Cowes, Gomes Fee ie ee Panktace fh ocacu she neh oes Coleen Yale University CURES, LM Ecos ak os oe ee ae ee ee eee University of Pennsylvania CAIRNE EE” Pheer crs, Bis uo act ec ha eee wack Johns Hopkins University COMERS, MLRYNTE MG cams. ood De cael anit eees Pea ceukawe agree Columbia University CUS SAVY Wien anv en pana ae ee ee ae nee aa: Harvard Medical School Pha, TE ai Ss es Se on 8 sad ace ek ee apn eee Ossining, N. Y. DAVENPORT, C. B......... Station for Exp. Evolution, Cold Spring Harbor, N. Y. LS Sas Fis as 2 ea ee eee Caer Rockefeller Institute, N. Y. City Pcmmen BG... kein ky i eile Sh sake Stanford University Medical School DOE AN, Regia ned tna SS ope f ice pie bit Presbyterian Hospital, N. Y. City Diptera TAs The, ited Se hates er ee es ee Wistar Institute, Philadelphia oot eV, Bs Saw eet os as 55 3S eee eee Syracuse University nies, Grint Wass. isn oc awe aces ee eee ae Columbia University Daren. Jom Weide os aivga ccdchs vo deex arts an Me ata New York City Daemenace. DO o 5. 55 est ine eae 3. eae ae ks Albany Medical College Toons, HAMA Ts oo cc cast Sate ak es ee N. Y. City DuoBow, E. Fess ies) 2 ee ts Cornell University Medical College, N. Y. City DuUGGAR, Bis. 8 cette ede toe eee Missouri Botanical Garden Dorcume, EE ADAMS. is aaa is we Sade ee ohn asab en eeee Pennsylvania State College PR CC. Wes 506g div see Ginn d Ue cleien ai enn eke sks ome Se Tulane University Bocwies6, Ciikl... 36 steels ou bs dante ts need ee University of Minnesota Shey Wa tan Te cis Gs ce aw ha hen ss ogieatat clad eaae Columbia University Ronse, CoWs 2155 25S St oe ears soak eens University of Michigan Eowénns, Ds Fic cs Saicods:. seated: Cornell University Medical College, N. Y. City EGGLESTON, CARY......... Tn? Soe Cornell University Medical College, N. Y. City BGGBIEEN, AMOI | 66S So ine ee Sls Pd a hee Columbia University Museenety, is" sc nic 2. 25 eae oa ea 2k ae Western Reserve University Misamac, CARE AAG io sii acc okay t0 8 nds Carters Mt. Sinai Hospital, N. Y. City Misuns Wi Je oak an ere Cornell University Medical College, N. Y. City MPUSBIN, Bis Ad si odds ceded Fahl apd in eed ae Mt. Sinai Hospital, N. Y. City ERDMANN, RHODA............. alse bak eb es SV aSy on XS enw ere Berlin, Germany ERLANGie, JOOP 00:5: 9a ws eae ee Washington University Medical School Evans, Eimear Mas vite Se oe ite etn ied Cabs Che Poa University of California wine, B. Ma. 30s 2 ewe Sot el es ee ole adunwedee Napoleonville, Miss. SEWING, JAMES 2. is bs ee ek a kowe dak Cornell University Medical College, N. Y. City BIRERR Gs Fo OE Mos ah 6 Fs HACKS ih ee ci case eevawceween University of Wisconsin VaEER, HAROLD Bok fois cece Ue taretes Stanford Medical School, San Francisco AREER COGIER «0 sre nw 8 kp datdikie tata tena University of Minnesota ROLL OF MEMBERSHIP. 467 RS IIE os a Ln 5's vow ts movie piven RD Roosevelt Hospital, N. Y. City SO rae ern rrr St. Luke’s Hospital, N. Y. City Me ge So. 1g Sang tn n'y Rehan s sak OP a RE Se hae in oe N.. ¥. City NR SEES at cE Ee Pee eT gen pres Snr re ts a EE Battle Creek, Mich. RMN MUP EIEIET oe as gg chi Stapds 6 dela ww a OE Rockefeller Institute, N. Y. City Rermre MON PRO lg go a gS wi no Mo bons Sle ew bok General Hospital, Cincinnati RRS Ss noes Sic. d ors cid cee vis! bi dais o Saied © x SUG pilin © ae Cornell University Ra ied aie Sar a te ie. asd'e x eau e'u Sole University of Minnesota, St. Paul BE TON IOUS al IS aa eg a University of Toronto I EE let ty kx cas oo | wage ecala Oia. ee oon dn we St. Louis University em TE Bene onal icta 2 w asic ase vo es University of California, San Francisco em IE ag 2 bikie. once iv Dae wre eas a Rockefeller Institute, N. Y. City PROUT T IOMAS So ae oa ss ndale wmce ae House of Mercy Hospital, Pittsfield, Mass. ne MOEN MEE hg oo aco feb a wlai ed ow eehla 8g eek wera, Se University of California ROmmnI EEe B aa es a oa td onc nd we eS New York Hospital, N. Y. City Rie A TASES EADIE TON . 5 oc 25 Soa kc eet cos da vee viens « New York University UEC ae ek wana alors y a an ee aa eee 8 NS Columbia University RIMM eS Arak SL Shs Sn Si cal aa GIG bib are tem'acc's'w Fs wsceslataad ces N.Y. City Ca es OS *y geuledl US SS SR RB Re A a Rao Brooklyn Botanical Gardens og I SS 3p IRS od iar Se PRE Rockefeller Institute, N. Y. City CN ie ee el gh ies aa de ae eR ae University of California en RIDMRE Sonesta Sek OES 4 te cee Gratwick Laboratory, Buffalo CeO RMIT Phe ye Cg ee a SO RN TAI EAS we University of California Oe gly gee Reh ns RE oy eR NOSE Be ad Soe paige Pe New York University og el neat ie 0 i Ea a AE i psy i Iowa State University OEM Tor rt! yee eS Neo eS rons aN Columbia University ce ice Mig eytns gdh SA ag an ap Se el As Shpall a Rl Ra Bee Philadelphia, Pa. GIVENS, MAurRICE H......... She bn bh Ne Western Pennsylvania Hospital, Pittsburgh Co RES a Et i oe Amherst College reer i MT ate ed shee vino bw ee eee Long Island College Hospital, N. Y. City MS NE Sg SID ERE ORR CEES Nice ea 8 gm ES ae ae Cornell University 2 LS OS CSET eae College of the City of New York eee CN AMER IETS Sc, nua kdis ba wies bls ease ee bee University of Pennsylvania Res PR RR a 5S ic ian) Sie & cee cae aw we University of Minnesota, St. Paul Ren NU NE hs PERO So oc cu ce ep ga woe Rhee S ose Roosevelt Hospital, N. Y. City ea Ea ee tS ies sik pina avecm © oases’ OhRs bars Barnard College ERR E NR S CLS areal. oP ices sated Sakis Vos We ge ee guiplaw aes University of Nebraska eR AA acca is Mae rho) Ses 22. cada “aon din Sib eee oa *. Xs University of Pittsburgh RR WN CORRE feo Sa OW os. 2 es ine. Seal ew Stow wlee Bm al Harvard Medical School I Mi aE ol ss ahh ce a a uel ou tare Mirena sia.e wa University of California MM, SERCMERMERDIIN 3. ww Hato, Ww eee cabal Rie ere N. Y. Post-Graduate Medical School Pe We os 5 Sie a ste ne Meee ee wees ves Johns Hopkins University PAM Ds oi ike no! o, o auth » ae © WE aie aw ere Stanford University, San Francisco sR i a a a's Sma ns oe ER le OW AE Clee at hae Tuckahoe, N. Y. HARRIS, J. ARTHUR........ Station for Exp. Evolution, Cold Spring Harbor, N. Y. PEIN OLS Cee ain lien, Bo eats Lie ined Sena Ane wk wea Yale University EPARROP GEORGES PR's 5% ood fo EVE iin eed Oe Sats Presbyterian Hospital, N. Y. City BART WELL; JOR AS. viii one ins ¥ vee Cornell University Medical College, N. Y. City 468 SCIENTIFIC PROCEEDINGS (124). HARVEY; TH. IMR WEON <8 och ost ape h ete e ook tae ee Princeton University HASTINGS, A. BADER 2S Moos ica ae eee Rockefeller Institute, N. Y. City SIREAS, SHUI oo ease nlacs a's Ca ee ee end Tohoku Imperial University — EAAVCHGS, SO ictes oo aero eae Cornell University Medical College, N. Y. City TIAWE. Boe casa icc he as oe oe Oe a ee ee eee Jefferson Medical College POA TRGS hw F wicca s ol niet ee ee eee University of Minnesota, St. Paul TEEMDEESON, LAWEENCE Foe oi ok ek es ook cee eae Harvard Medical School TISROMEE Th. Me steer vodke Can cub oid ck eae University of Pennsylvania RA MSER SCE, PoWT PEI Ts eo re ee an) ance University of Minnesota Paes, ALrewy Ho ee See ee oie i a ame etalon New York University SAWETS, Bu Wie 5 cee ee nl Soa eo eis Stanford University Medical School ESOT Sree, ARR. oes os oes ee eee ees University of Minnesota HOFFMAN, GEORGE L............. Western Pennsylvania Hospital, Pittsburgh, Pa. PERMEATED ¢ Siac « ving Deda d ae ee Se Re Ate eae ee Ee eee Stanford University Tae See wk aes es ear ee Ee ee Ge ae University of California SOREN, TENVENE ORE as ok Pee eo emehalnataae University of Pittsburgh FIOUPER AOMARLES WSS Fe ee oie klnowinin ae o ee eee Brooklyn, N. Y. FROCK ING, F--GReO ee eS wee So ae eee VS ee aes Columbia University RA OM UI hn Aoi Fs nile «Pn WAG o tah) oO nee Ohio State University er, 84: FS os 5 3i il a es See Oe Kristiania, Norway Mowe: Pace By ei. ek cata Ue eoe a reese Rockefeller Institute, N. Y. City PROWVELL,. WWSIELIAM Bless ata t ok oh ee dee eee eee Johns Hopkins University MOWGAND, JOB. sete esc on bee eee ewes Johns Hopkins Hospital, Baltimore EIVEBARD, ROGER Bice. cee ee es aeas Clifton Springs Sanitarium, N. Y. HMunis, (:- CAR... €cccasis.« Sear eee ees University of Michigan © RENT, | BR eG boc be dod Me naa lee de eee Harvard Medical School PION TER, ANDREW oo 548 cele sR EES Ca ie es Ce EN University of Toronto HuRwirt; SAU 6 icc cK wane aes baer University of California, San Francisco PACRON, Co RR ow ee ts es ee University of Minnesota JACason, D.Don 7s be wee th ree tenth tae tian University of Cincinnati JACESON, Horpttes (oo i5 5 tae cece OR eo ae ee New York University JACONS, WALTER A ino ede cee eevee stone Rockefeller Institute, N. Y. City JARMRY, NOE GON WH oes ees 55 Sere cet aie eet eae Los Angeles, Cal. Jind, Ti. 'S....'5.35 chutes Pe een rn Crete eee Johns Hopkins University JOSLING, J. Wiss oe cians vas Cole heath coe a ree Columbia University SOMES, FREDRICK B53. oi ea Rockefeller Institute, Princeton, N. J. JORDAN, Th Bie sao. 6b. ee hae le atl ak eal tl a aan University of Virginia juste, TOM Boi bos. iced hoes Beet eae St. Louis University Medical School AMM, BEAR o.oo Sook ro EE ok ti eee ane Beth Israel Hospital, N. Y. City AM, MONI Tl... fesoece Peps eee fo Beth Israel Hospital, N. Y. City BRAM, Mes dais 3c cs das ve cdanermse ect encacks hoheetaeie nts Roosevelt Hospital, N. Y. City EYNGE EAR Juche nen bik sea Oe ee Rockefeller Institute, N. Y. City LYON Be Bioiaa V koi ks eaten een Gre eae see University of Minnesota RE ACALS AIM A Tee og sk sccs aha She eee eee McGill University, Montreal BEACDONR AE TOTS sen oan Gee ee tee Desert Laboratory, Tucson, Arizona MacDowELtL, E. CARLTON. .Station for Exp. Evolution, Cold Spring Harbor, N. Y. Macninrizn, Groner Moo. sla: fl ickaeeeee Presbyterian Hospital, N. Y. City MACE ROD, J, JsiRisves ¥ ovewatae’ «ve Mob REs priest University of Toronto BEACNSAL, WORRD Bios Gi eer ne 2G wane eee N. Y. Post-Graduate Medical Schooi MAcNipge, WIELIAMIDED:. 6 os f.5 cons oa execs emer University of North Carolina RAAC AWN SILESIA RO 2 Lis oc tn uaa re pacar ea eee Johns Hopkins University DARCINS, LINE Ss cls So 1 aloe sige ace sky Oe OE Johns Hopkins University PRC ERMC, 3. FRANCIS, 6.0% «coat u's oes Poke Oo University of Minnesota PEGA -OLLUME, Bs Vee Freeh 6 coe edd Ate Ea aOR Johns Hopkins University BeCUCRUDDEM, omAwers Mino 55% aes So bee U. S. Public Health Hsopital, Boston MCLEAN, HRANELIN Nooo 5 <5 sacks eacsldeae Peking Union Medical College, China BERBERA RO 75 Wcdiikos cin coe sak Oe ules pce e Hes University of Pittsburgh MALTANER; PRANK <%..0.0°-. 4.5% co canw ane N. Y. State Department of Health, Albany Missa,“ ARTIS Mess ooo do ee ed hk De ee New York University Pe Anwie, Fema As cree vs oy cae ce ke be be oa eee New York University DEAN, Wine K es A205 Ne ware hk we od ee Ce eens University of Minnesota Deine ERS i OLS Ae Peet aW OR a sh ee nea Mt. Sinai Hospital, N. Y. City DM awwininig Wi Beet >. . Sa noah Se res a he Leland Stanford University Manure Davies io i dc baucn dee Montefiore Home and Hospital, N. Y. City MaAwsratn,, Ft. Teh JR iis ce 9a ka eS ee ak Johns Hopkins University PORTE TE Sees he co's ht ces ove en OA eee Leland Stanford University Marr, See Ais a ad. ee Dc reco we ah eek University of Rochester DAA Wwats., Bi BMY ye oe net OSes ae Len eae University of California MAYES A. Gisied t linded sole babias ees Carnegie Institute, Washington, D. C. DA awn La Ris io ig hot aa ths Ea SEK hated Cee RoR ee Cornell University MeASTENS, HIENEY Gis ciety ie Stanford University Hospital, San Francisco PamtGe, Th Bie. ni se ee ta diigo ca ee kee Experiment Station, Beltville, Md. PAMELON: TALPR I oii ba tena ae bale weak Highland Hospital, Rochester, N. Y. Dmwat., LiAFATMICE Bic Fes o's Sine oe bee ew RKO SN Ee Cee Yale University METz, CHARLES W........ Station for Exp. Evolution, Cold Spring Harbor, N. Y. DEM AIOE fo SS ia Foo ocd 1d Johns Hopkins Hospital, Baltimore Dis iR A Tasch hl i Ra MS nae DSA Johns Hopkins University Meyun, 6. Bs ae ik wie oes caw eens Rockefeller Institute, New York City PSV ie, Me. Baked base ad Bas a Scie ow bone, Gb ee an ee University of California DOOR, i Bex. eis patel Reems SE arto SERENE ee ee 5 A Rutgers College MOORE, WSLEAAM 5 icc dé sine cs Veda U.S. Bureau Entomology, Riverton, N. J. DEQOAGAM, 1S 0 Ths itech n:teh 4G ean 0 Rae hid Ace ee Columbia University DEGREE, WEIR 6 ic dhb ho CUR aOR ie Ga eke hae University of West Virginia MosrnTHar, H. Quy Ji vc ve Sake tw taeanvees N. Y. Post-Graduate Medical School DEM Mek OMG oleic pf acclae si x onniwad Tee anaes tas Columbia University DAVLAMR, TAURMAM Te ok cerane ve cds hia rds veces e tie University of Texas muaian, Jone BA iio is oct caret rere tesa venekere University of Rochester PEURIES, 30 Baa oO sis nd GAPE Laan hota ete t Rockefeller Institute, N. Y. City ROLL OF MEMBERSHIP. 471 ME TN le ocr ong n'a n'a a ‘stv ao oie G's ele cele Wels University of Pennsylvania ee Cs Rei eet e sa deeds cn canst N. Y. Post-Graduate Medical School aa) Oe MERTEN Ogre we wis vara bob RC Ed cS HOES wie Bik a Ohe b Pl suw ees Rutgers College PS WTAE TER Mass oOo sis Cornell University Medical College, N. Y. City PE ORM rt Oe eS awa Up eieaa, ey oa eRe eae New York University MIM SA ptt Ses air OS 6 Gas aw wap ee, oa Rockefeller Institute, N. Y. City pene ee UR En ick sb oie sini d vio w:0,s 0,0. orn we s.e'e Chief Medical Examiner, N. Y. City eR POI baa dw is ais 2 alewviets/t ales say ve eens Rockefeller Institute, N. Y. City Dea Me es oc cc wb fn Sc ene Sawedsicneeveess University of Michigan NINE ND a oe te oo bre Dc in Deas wae aes. 8 Rms McGill University, Montreal I ae ce ain «9h. ding 44 4. 0b Re wisi 6 we Rockefeller Institute, N. Y. City SO RIAN orc herviak Fg. 0a <0 0.0 sis is we «ya wren Dare Stanford University Medical School mee WUE TM a oo a oe has sida o te yo Soe 2 Stanford University Medical School RM SOCIO Na ae ic ea wate ie elude Washington University Medical School Oe ERE RE BS PCA Rak ao wa «2 hiloba Wack Wis wate «Sle na WES ote as Columbia University WSEBORNE, PHOMAS Be. cin ceca ees Agricultural Exp. Station, New Haven, Conn. SMR EM ADEN os ie oie wines tide ave who Wk ace iae * hws ae Harvard Medical School ie ONIN ee Rare ga oe ra a ae UM US gw lhe Mt. Sinai Hospital, N. Y. City Pn NN ae on. dW) ia oie we Nite en gp alwelb mals University of Minnesota, St. Paul Ri ss a oo alu one's sb we eee Presbyterian Hospital, New York City RNS EN EN a 5 bets) ay debe GO Se OH hoe LS, DE a ee Columbia University NE NER he 1 hed oak aa scat hems a «Xe d vk ae pe Ree GEES Yale University PAM LIAM Pe yc oie ote ees bbe ole ves es department of Health, N. Y. City UR CG RM PB a Ox! a ie a nese o pe va welaee Mewes. Harvard University ReMi oe Gh dum, eto olaly's adn e Kine wewile a wanda ooh Columbia University PEABODY, FRANCIS W........ PARR EE Nee Peter Bent Brigham Hospital, Boston RMP MIEN oe a as gh aie’ a's Ga ce abe oks Sis oe oh ae Rockefeller Institute, N. Y. City EMM UUM oy a hctonigic d Gino a cic’ eid bdwie be sale tebe es Johns Hopkins University Dp OU AET, Cos se a Ss we ae ek N. Y. Post-Graduate Medical School Let [LEDS [QR ae tis aaa ap eR oe New York University PMMBeRTOW IAEPH. ols a. kas ween a bes os Presbyterian Hospital, Philadelphia a OS aS Ta a ee ee RE Sa University of Pennsylvania 8 0 eR RR) ae ary Toe ot a Yale University Pee ee CAR ay rere Ce ints hen Gee Sake Soa en la eG een’ University of Illinois Pee Re So Wee we a heh on ne pur cathe ioe ieee cs University of Minnesota SUR DEA ee pai oe ti as Pe oie a a Oe el eae ecewere Harvard University pee FOR i Ey Sire a aes Sn el cle a ek To oe old “ory caw os Baltimore, Maryland Remreee arabs Reet lk Rowe Tate Un Mtge Le Columbia University een MRE eS RS De Lak Tea Sine ead bin EK, Gre SO ee ww Oe Ose NH NS Paris, France POUMEMAMS ALIGUSIUIS [ose x 3.4 sos Cae Sak he ae ek Eee Wen St. Louis University pee PM Ue TEE MAME Pee Sk sy athe ew te Oe tee Shoe Kew gow ke hot Harvard University See, WEPREPER Cle Se es, See le ROI EE PE See peo e dbase Harvard University PementM EROUEUS WU. Fo 5S 4 Sc foien anew ss eke ok eae cues New York University SPR De Se ois Ao Se od es Coes wd AOS bbe De wed oS Wethersfield, Conn. RAIZISS,"GRORGE Wie ss see 3k 6 Dermatological Research Institute, Philadelphia Reet eee Mere ee Ske ea dias de ae oat edie Me witic University of Minnesota Sey REEL AUR MS Sean arene! a ble slaltie su wacew ek wae leh wi University of Missouri 472 SCIENTIFIC PROCEEDINGS (124). REIMAMN, STANLET Py 5 a5 bs sp aen's ne bees sake eee eee University of Pennsylvania REPS ER Fossa s cece ea $8 a Wha A Re eee Yale University Rwwanns; ALFRED Noe... 2.5 baa eas Vs eee University of Pennsylvania Ricwanys, TeeBEayT Moo SoS ai ee ase te eich oat Columbia University Rimpes, OSCak So. oo sb s Station for Exp. Evolution, Cold Spring Harbor, N. Y. RINGER, AST. 55 oa ee eee te ee eeean eee te Ge eee ene New York City RimGer, MICHARD 60.00.00 esa. es Cornell University Medical College, N. Y. City ROBERTSON, EI. Bass sn Pe ka AS REE CORES University of Minnesota ROSERETSON, *T.Bee Se iA sh 5 te Bexd University of Adelaide, South Australia ROBINSON, G.CANBT 8 ee aie it el ee wale i ate Johns Hopkins University Rogers Freep Tos ooo tn cee otk e eet eee Baylor University, Dallas, Texas ROGUE I. Bee oo ck cae oes Sap lalate he eee Western Reserve Medical School PRG, ANON Dhis eas sais 85.42 0 das bs ee ae ee N. Y. Post-Graduate Medical School Boge MARY BWARTE os << o 6 ee oie als a BRR SRR SRR Columbia University Se, VRLLAAM TG. ok ok ss Sala at ok ake Mee eka R Oe Ue Eee University of Texas BOGENAU SE. Fac. 6Se RoE Sain wt lre hime Punt eos Harvard Medical School ROSENBLOOM, JACDB 62 oii 0s’ x sports was aa a Se ead eee ee Pittsburgh, Pa. Rors; (e08GE.B.s0 re fase eae U. S. Department of Health, Washington, D. C. RorHsct 3, S05. ook. Saas tee saa ew oe Mt. Sinai Hospital, N. Y. City ROS -PAVION.. 225 627. hes dene era aE Rees Rockefeller Institute, N. Y. City Rvan,- Aisi te os wha eh Me bes Soh see ee eee eee Waterbury, Conn. SALANT | WHAIAM. 655 54se ook Biden sO oe ee ee ee ‘University of Georgia SSRN, Wc dle Shi oe ut eke ee ee on uke Potter Metabolic Clinic, Santa Barbara SCAMMOM, Ths Bos. o: ces wh 's Fg aa pol po c:\0 eu eee Si ae wae University of Minnesota Scwt0Gk, USCA Mies nso sc cnb Seek ens eeaese een ee Children’s Hospital, Boston SCHBROTS: Fo Wi ot hing h 2 eRe oc Owns BAe wae eke halos heme University of Minnesota Some? CAR Cs: Pace 5 hn Sane eb Ree eed a ein ty ee ee University of California SCHNBUER, TL WAR Soak 4 Eas bss nicks hin awa sa eee youn Wesleyan University SCHOO. 3. Fe os dak oak CAO A ee oe babe eee University of Minnesota . SICMLURE, WV «Eke as 5p os Ap ae Mae ees a hee tora George Washington University SCHW WER, FREES 5c 5 oh kei sb hd ee oe teen Kastanienbaum, Switzerland SOONIE iy Elo eres iss 5 a, 'w wg Vina aa See me en ee Lee Columbia University SCOTT, Fo Tia ckendsaucnckan ce titans ie eee te University of Minnesota BOOTT, Ge. ABis nd creterd oc ak aKa ee ae si College of the City of New York SCOTT, Ra Woes sce co wteeets Pa oy a Re ee SS” Western Reserve Medical College See, FARCLD Wen oiscdwd tink wh eb thee os ee ia eee New York University SHAVFER, PULAP Pion on cada bee et suns nkeae Washington University Medical School SAREE; Ay Qi. 6. one's KU nha ae 2 6 Hak hae eee aa eed Rit St. Louis University SORBRMAM, Bh. Cis aed 6a oan eRe bk ko eer we Lee aeade che Columbia University SHERMAN, JAMES M...........ecee0e> U. S. Dept. Agriculture, Washington, D. C. Semis, CAnt. 3, . 608s who vn write th 2d io i lle Vedas eee eae Fordham University SOLE, Pave Goes soos cwde ialetnnkea secu teeta Johns Hopkins University = At | SSR ie tk < on" N. J. State Agr. Exp. Station, New Brunswick, N. J. Scns, ALPRER 'T. ..6.0 én vinnie dA Te a bas Leak ce Yale University CIR, 35 Boe snc adacesadtaveete Office of the Surgeon General, Washington, D. C, SxherGont DOTSRRLAMD . oy 2.0.5.0 K Ai hae vi bac tbe s 03 ob od cade on Cornell University CRE TMCIMER, Bh, Boe so tak sVakaent awit ruse d ex atEhinaaene Columbia University POET. MABTAUR Fis 6c s 4 6 vas's 0 ob a mlodeaa ava e abs cxuteaeuin Yale University SNE, CAEP Whe o's SW ben ce nde y es ikakiaeek ets ee aes University of California ROLL OF MEMBERSHIP 473 Sere MORAL 258 Pe ook bes aint Rockefeller Institute, Princeton, N. J. ee NI OTE Fe NNN cis uh pias chien ps ak oak es ah ek po ao aNek amen nTL wn Pee Western Reserve University STEN SE 2 oi ww nes eS Eee Lier eos Wee be ble Johns Hopkins University Pepe Baas Sp eines eo Lee ra paver cabs ke sak University of Minnesota PAE, DEAR cece seve. N. Y. Homeopathic Medical College, N. Y. City SREVMNG, PP RANEDIN Ase 6. Oli Sea cous Presbyterian Hospital, N. Y. City ern NN AERA PW Rn tlt se see bd che vn boots aw « 6 Western Reserve Medical School eer rd IMT tein PE hes Je. wie Fu x4) sc-n gs dherre eivaa re lo ta rwned dives Harvard Medical School MUR PE EER eat DEES ots oe rs eke ese i8 8S 8 New York Hospital, N. Y. City STOCKARD, CHARLES R............ Cornell University Medical College, N. Y. City ee, LTA ok warn tin edie dinocdevuwst University of Southern California SeMENRNEM et ROMER Fis te ooo cc ky Nnle ice wsa seas College of the City of New York eee ee IAIN RIO Ae Sa een See ewe whee eea ee Harvard Medical School RPE MNMIMN Es od Goda ob ike nebo ned bw uw owen ea mone Northwestern University MRR INOS OMT Me MN RCS is ick scissors foram So vd a (nce vos Naw Ge naar Columbia University RENE LENE hh PES Ma vd One eels > dd eee asa sess Leland Stanford University PR | ee a ee a University of Pennsylvania PEM R eee Rts, SPN sop bd vewy eee daas Rockefeller Institute, N. Y. City Sep Nema St FON NP A gg oo is ay salss Ua Any Sa 7G wv 0 /OMEjahewa bare a Seveasto mee New York University men MRD EON Bay Sees Sera ie ho ee Osea d weidle 4 Harvard Medical School aI ae ae ale oases aes qunin chav ena phadad alelandtiwne se University of Cincinnati RE II READIN TE a 2.5 Geshe k 5 (a> aha lah aon v oreies wo ak Re wate nin oes Soe Stanford University ete RES 2) Ses Es bho PRCT oul aN be aN Maceo, Kentucky MMR IMNEMEE aes S er aw kart ail ns old. 6au'm Gan a yrs Mok ata lao Columbia University ran Per, OE Py oe oe ae nota ee Peking Union Medical College, China IMME sey seer Lo alk y bialdlohe km bok die den’ 6 Vanderbilt School of Medicine UWMATCHER, OBERT W.. .-.-. 550s )ecad $4 Sao N. Y. Agric. Exp. Station, Geneva, N. Y. ee ee EOMICONN ati cl ig wricr as big tg ba ee kip Sg els He awe a hee Columbia University RT ey et ie et each» wwe ac St. Louis University Se UMMM NN RE TEE Pie os ag se wisls wa 8.6 ne es es Clifton Springs Sanitarium, N. Y. eetMes WVIEEGAME Coo 2S Peek oe be Cornell University Medical College, N. Y. City MERION Rar Soe sie Lig Sk SG pew awe Goare ea wow o alg es University of Oregon TOsREN, OHM Coos oe bes ee es Cornell University Medical College, N. Y. City fe EIN RID se ods a acu kf 4 oe ERR A bee ww ws Lane Hospital, San Francisco LT by RS COO ep a OS rt | Sa ee Harvard Medical School Dime, BOAR 6. oa e sb ae ob cee Ree es Rockefeller Institute, N. Y. City eee ei em te ay Nip rg Wh wna wee wc oven ace Yale University A Cie, SIOMATIE IDS) eienc g Ojemk kobe ka ee de Rockefeller Institute, N. Y. City Pe eeeter ae MRE Mins ag Ate an Aig ck Gk iowa + adie Oe een e N. Y. Agric. Exp. Station, Geneva TeECOMa, MUMBERR INE TES Sitti SS ae nS, ly Gin eR Na ened ie a as Columbia University © Wapswortu, Aucustus B.....N. Y. State Department of Health, Albany, N. Y. WAKSMAN, S.A....... N. J. State Agr. Experiment Station, New Brunswick, N. J. ann Ma Teles hirano Soule sip au.s che Dein oo University of California, San Francisco ie nee RTI Eyer er Ss i Pare Sere ee eS oe New York University a Se INE SS cS nae Dawa seek eke Ghee ow ee ware University of Michigan WR ea ee oo he ena sake akan clebee ne kd University of Toronto, Canada Pe Re SU Ss ae aa ee se ele Re RUN gS oe Dg RAS ig RO Tokyo, Japan 474 SCIENTIFIC PROCEEDINGS (124). Wraiss; CaARENS jis Sots dose Dermatological Research Institute, Philadelphia, Pa. Waueme. Ws Beco) 05. eho) ok ean ele ee he nee cena University of Illinois Wiettrn, (CARE Wik cok Soo co. ent eile pa eee eee University of Michigan Wars Gide tie era ec dncencoe National Research Council, Washington, D. C. Watwrte, 17 BamGe Fi 662. 2 ek ie ee ae ees University of Rochester Warre, BENJAMIN. 0..46 6.60508 2s scene Antitoxin and Vaccine Laboratory, Boston Ware, 0.-Bioe 2586350 eee econ Brooklyn Botanical Garden, Brooklyn, N. Y. Waeeemis, (Ans Fioteiss iw sie eacercs ener eee Western Reserve University WEELAWAN,. Ji Fav antes. fe ake ood eet ore University of Minnesota, St. Paul ‘Waszams, Anna Wc oad Jae xs cS eS Department of Health, N. Y. City . WHAIAMS, BORAT Gait Bi Pirie hve ne cao a ek eee Columbia University Wirt wan: 88. A, > etd Oo Sica i ak Kees he ie eee University of Buffalo WSO ae. PURI ois Se He ee Bs Sete See Johns Hopkins University WS ENT Oe Ei acid cciscsavp S pe puis le a eee ie Columbia University PRMD, CAs Pass ok ere 5 oy BS a Toe hos aie a bis ki bale al Yale University MBAS as. ES ola Gare baw vod o's Cais eRe eee ee ee Harvard University ie, Co Sods heed ke eae Addenbrooke’s Hospital, Cambridge, England Wiesrit, MABTEAS fic Sisko i ee re Rockefeller Institute, N. Y. City Weary Pre ae oe oa oo ais Sa as Raed ke Bod eadae en eee Columbia University WOOnmUEE, 1s. Lae. sgn te ink doc Reale ee ee eae eee ee Yale University Years; NMG i 2586 23 re eee eee ek eee Keo University, Tokyo, Japan Vewens, Rosner. 5.3 )o or a Ran oe has ede ee ee Washington, D. C. ZAMGEH Eh, - AURATAM «5c weak woes eed bik an ee Department of Health, N. Y. City ZIRE, HAWS oo 56505 ks 5 Sek k ARERR lene is She lea Columbia University FUCKER, ‘THHOUOGE A 24255 ans Slats es we wees weak ap eeenen Columbia University Total number of members at the close of the academic year, 1921-22 :—492. i Treasurer Secretary Sec’y-Treas Additional members of Council Sec’y-Treas Additional members of Council ! IQ2I-'22 Wallace Jobling F 8, 2 6 66 & 6 2 2) Bie Uo cece ee eevee Myers OFFICERS. 1903-1922. 1904-05 1905-’06 Meltzer Wilson Ewing Dunham Lusk Lusk Calkins Calkins Gies Gies I9QIO—"IO IQII—I2 Morgan Morgan Gies Levene Lusk Lusk Opie Wallace I91I6—"17 1917-18 J. Loeb Gies Gies Auer Jackson Jackson ‘Auer DuBois DuBois Wallace 1922-—'23 Wallace Jobling 1906-07 Flexner Dunham Calkins Gies I9I2—13 Ewing Levene Norris Wallace 1918-19 Gies Auer Jackson Wallace Sherman 1907-08 Flexner Morgan Calkins Gies 1913-14 Ewing Field Norris Jackson IQIQ—’20 Calkins Wallace Jackson Sherman Jobling Pacific Coast Branch—Ophiils Minnesota Branch—Hirschfelder Western New York Branch—Murlin Jackson Jackson Myers DuBois 1 The Past Presidents are also members. 475 1908-09 Lee Morgan Lusk Gies I9I4-15 Lusk Gies Murlin Jackson 1920-21 Calkins Wallace Jackson Jobling Hess CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Honorary. William T. Councilman, Harvard University. Edward T. Reichert, University of Pennsylvania. William H. Welch, Johns Hopkins University. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. Columbia University—C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, George Draper, Walter H. Eddy, Andrew Eggstein, G. A. Friedman, William J. Gies, Louise H. Gregory, George Harrop, Jr., J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, Thomas H. Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M. Richard s, Mary S. Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. Cornell University Medical College—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert Chambers, A. F. Coca, John W. Churchman, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, Robert A. Hatcher, Graham Lusk, Walter L. Niles, Michael Ringer, Charles R. Stockard, W. C. Thro, John C. Torrey. Fordham University School of Medicine.—Car1 P. Sherwin. Hospitals. Babies—Martha Wollstein. Beth Israel—Max Kahn, Morris H, Kahn. Long Island College-—Emil Goetsch. Montefiore Home.—Emil J. Baumann, George Fahr, B. S. Kline, Michael Levine, David Marine. Mt. Sinai.—George Baehr, Burrill B. Crohn, Charles A. Elsberg, Albert A. Epstein, Hubert Mann, R. Ottenberg, Harry Plotz, M. A. Rothschild. New York.—Nellis B. Foster, Ralph G. Stillman. Presbylerian.—Louis Baumann, A. R. Dochez, George M. Mackenzie, Walter W. Palmer, Franklin A. Stevens. Roosevelt—K. G. Falk, I. Greenwald, W.G. Lyle. St. Lukes —L.W.Famulener, Karl M. Vogel. New York City Departments. Health—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Park, Anna W. Williams, A. Zingher. Chief Medical Ex- aminer.—Charles Norris. New York Homoeopathic Medical College.—I. S. Kleiner, Mary B. Stark. New York Post-Graduate Medical School.—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. 476 CLASSIFIED List OF MEMBERS. 477 New York University —W. H. Barber, Harlow Brooks, Warren Coleman, Florence Hulton Frankel, A. O. Gettler, Alfred F. Hess, Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Emil J. Pellini, Pro. V. Prewitt, H. D. Senior, Douglas Symmers, George B. Wallace. Psychiatric Institute.-—Nicholas Kopeloff. Rockefeller Institute for Medical Research.Harold L. Amoss, O. T. Avery, Carl A. L. Binger, Ralph H. Boots, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, Paul DeKruif, Albert Fischer, Simon Flexner, F. L. Gates, A. Baird Hastings, Walter A. Jacobs, I. J. Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Christian Lundsgaard, Clara J. Lynch, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke. Industrial Laboratories (New York City). Research Laboratory of H. A. Metz.— Harry E. Dubin, Casimir Funk. Research Laboratory of H. A. Meiz (Brooklyn).— Charles W. Hooper. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N.Y. City.—E. E. Butterfield. 17 E. 38th St., N. Y. City.—-J. W. Draper. 126 E. 64th St., N. Y. City —Cyrus W. Field. 141 W. 78th St., N. Y. City.—A. I. Ringer. Non-Resident. Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick).—J. W. Shive, S. A. Waksman. New York (Geneva).—Rudolph J. Anderson, R. W. Thatcher, L. L. Van Slyke. Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, Albert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, Charles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D.T. MacDougal. (Wash- ington, D. C.).—Alfred G. Mayer. State Boards of Health. Michigan (Lansing).—R.L. Kahn. New York (Albany). —Mary B. Kirkbride, Frank Maltaner, A. B. Wadsworth. Hospitals. Addenbrooke's (Cambridge, England).—C. G. L. Wolf. Buffalo City. —Katharine R. Collins. The Children’s (Boston, Mass.).—Oscar M. Schloss. General (Cincinnati, Ohio).—Martin H. Fischer. Highland (Rochester, N. Y.).— Ralph R. Mellon. Mercy (Pittsfield, Mass.)—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Philadelphia General.— E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.).—Ralph Pemberton. Riks- hospitalet (Kristiania, Norway).—H.F.Hést. U.S. Public Health (Boston).—F. M. McCrudden. Western Pennsylvania (Pittsburgh)—Maurice H. Givens, George L. Hoffman. Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton Springs).—Roger S. Hubbard, Walter S. Thomas. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. Gratwick Laboratory (Buffalo)—H. R. Gaylord. Juvenile Psychopathic (Chicago).— Herman M. Adler. Potter Metabolic Clinic (Santa Barbara, Calif.).—H. R. Blather- wick, W. D. Sansum. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. Phipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—J. Howard Brown, Paul E. Howe, F. S. Jones, Theobald Smith. Trudeau Sanitarium (Saranac Lake, N. Y.).—R. A. Kocher. Wistar (Philadelphia).—H. H. Donaldson. 478 SCIENTIFIC PROCEEDINGS (124). U.S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs. Bureau of Entomology (Riverton, N. J.).—William Moore. Hygienic Laboratory (Washington, D. C.).—C. S. Brooks, Barnett Cohen, J. P. Leake, George B. Roth. Surgeon General's Office (Washington, D. C.).— J. F. Siler. National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Australia).—T. Brailsford Robertson. Amherst Otto Glaser, Alabama.—Clyde Brooks, Baylor.—J. H. Black, Fred T. Rogers Buffalo.—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwith, W. R. Bloor, T. C. Burnett, Guy W. Clark, H. M. Evans, E. C. Fleischner, G. L. Foster, F. P. Gay, Robert A. Gesell, Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles A. Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Cornell. —S. A. Goldberg, L. A. Maynard, Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. George Washington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Worth Hale, Lawrence J. Henderson, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Percy G. Stiles, Richard P. Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. Illinois —H. B. Lewis, W. F. Peterson, William H. Welker. Iowa State-—R. B. Gibson. Jefferson.—O. Bergeim, P. B. Hawk. Johns Hopkins. —John J. Abel, S. Bayne-Jones, C. G. Bull, George W. Corner, R. S. Cunningham, W.S. Halsted, William H. Howell, John Howland, H. S. Jennings, Benjamin Kramer, Paul D. Lamson, W. T. Longcope, W. G. MacCallum, David I. Macht, E. K. Marshall, Jr., Wm. S. McCann, E. V. McCollum, Adolph Meyer, Arthur L. Meyer, Raymond Pearl, G. C. Robinson, Paul G. Shipley, Reynold Albrecht Spaeth, D. Wright Wilson. Kansas.—Bennett M. Allen. Keo (Japan).—Naohide Yatsu. Lake Forest (Ill.).—William H. Cole. Leland Stanford.—Thomas Addis, Carl “L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. J. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophiils, R. E. Swain, A. E. Taylor, E. B. Towne. Liver- pool.—J. G. Adami. McGill (Montreal) —Henry Gray Barbour, Horst Oertel, A. B. Macallum. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Brown, C. H. Eckles, George Fahr, C. P. Fitch, R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, C. M. Jackson, E. C. Kendall, F. E. Kingsbury, W. P. Larson, K. S. Lashley, E. J. Lund, E. P. Lyon, Frank C. Mann, J. F. McClendon, Leroy S. Palmer, C. J. V. Pettibone, A. T. Rasmussen, H. E. Robertson, R. E. Scammon, J. P. Schneider, F. W. Schultz, F. H. Scott, E. C. Stakman, J. J. Willaman. Missouri —Maz ‘ck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.—W. deB. MacNider. Northwestern.—Solomon Strouse. Ohio State-—R. G. Hoskins. Oregon.—D. E. Lancefield, C. F. Hodge, Harry Beal Torrey. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, D. H. Bergey, Glenn E. Cullen, Samuel Goldschmidt, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., O. H. Perry Pepper, Stanley P. Reimann, Alfred N. Richards, J. Edwin Sweet. Pennsylvania State.— R. Adams Dutcher. Pittsburgh.—C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.—Edwin G. Conklin, E. Newton Harvey. Rochester. —Henry A. Mattill, John R. Murlin, G. H. Whipple. Rutgers.—John F. Anderson, W. J. Crozier, A. R. Moore, Thurlow C. Nelson. Southern California (Los Angeles). CLASSIFIED List OF MEMBERS. 479 —Lyman B. Stookey. Springfield (Mass.).—William B. Kirkham. St. Louis.— John Auer, Moyer S. Fleisher, Don R. Joseph, Ralph A. Kinsella, Albert Kuntz, Augustus G. Pohlman, A. O. Shaklee, J. E. Thomas. Texas.—Herman J. Muller, William C. Rose. Tohoka Imperial (Japan).—Shinkishi Hatai, C. K. Watanabe. Toronto.—A. H. Caulfeild, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Waste- neys. Tulane.—Charles W. Duval. Union (Albany Medical College).—Melvin Dresbach, Arthur Knudson. Vanderbilt (Nashville) —B. T. Terry. Virginia.— H. E. Jordan. Washington (St. Louis)—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Eugene L. Opie, Philip A. Shaffer. Wesleyan.—E. C. Schneider. Western Reserve (Cleveland) .—George W. Crile, A. B. Eisenbrey, H. T. Karsner, J. M. Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia. —F. E. Chidester, Withrow Morse. Wisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Wyoming. —William C. Boeck. Yale.——George A. Baitsell, R. H. Chittenden, George R. Cow- gill, Ross G. Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo F. Rettger, Alfred T. Shohl, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Industrial Laboratories: Battle Creek, Mich., Postum Cereal Co., Glenolden, Pa. —M. S. Fine. Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. New Bruns- wick, N. J., E. R. Squibb and Son.—P. A. Kober. Peekskill, N. Y., Fleischman Company.—A. K. Balls. Waterbury, Conn., Scovill Mfg. Co—A. H. Ryan. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer. Los Angeles, Calif., 533 Lucerne Blud.—N. W. Janney. Missouri Botanical Garden, St. Louis, Mo.— B. M. Duggar. WNapoleonville, La.—Ephraim M. Ewing. Ossining, N. Y., R. F. D. 2.—H.D. Dakin. Pittsburgh, Pa., Jenkins Arcade.—Jacob Rosenbloom. Tuckahoe, N. Y.—Isaac F. Harris. Washington, D. C., 1701 Mass. Ave-—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St.—Alexander L. Prince. Paris, France.—Harry Plotz. Berlin, Germany.—Reinhard Beutner, Rhoda Erdmann. Kastanienbaum, Switzerland.—Fritz Schwyzer. INDEX OF THE SCIENTIFIC PROCEEDINGS. (THE NUMERALS IN THE INDEX CORRESPOND WITH THE NUMERALS IN PARENTHESIS ABOVE THE TITLES OF THE ABSTRACTS. PAGES ARE NOT INDICATED.) Absorption, effect of purgatives on, of | Antiscorbutic potency of strawberries, drugs, 1904. Acetonuria produced by fat, 1795. Acid base equilibrium after muscular exercise, 1828. Acid fuchsin, selective bactericidal effect of, 1874. Acidity, effect of, upon bacteria, 1888. Actinomycetes, immunologic study of, IQIS. Actinomycosis, influence of potassium iodide on, 1860. Adrenalin, effect of, upon glycogen in tissues, 1796. Agglutinability, relation of, to antagonis- tic cation action, 1854. Agglutination reaction in tuberculosis, 1921; acid, differentiation of microbes by, 1765; acid, factors influencing microbes, 1766; phenomena with diphtheria antitoxin, 1847. Agglutinins, accumulation of globulins and of, 1903. Alcohol, effect of, on fertility of rats, 1783. Alcoholism, testicular changes in, 1812. Alkaloid test for synapse function, 1900. Almond, source of vitamine-A, 1939. Amino acid, sulphur containing, from casein, 1822. Ameba, rhythms in reproduction of, 1942. Ammonium chloride, in gastric tetany, 1940. Anerobe, putrefactive, tyrosine by, 1769. Anesthetic value of isopropyl alcohol, 1791. Anaphylactic reaction from x-rays, 1760. Anaphylactoid phenomena, 1883. Anemia, effect of germanium dioxide on, 1944. Antigen and complement fixation tests, 1810; removal of, from circulation, 1864. Antigenic properties of red-cell golublin, 1913. Antiketogenesis, 1797. production of 1759; vitamine in cabbage, effect of cooking upon, 1821; vitamine in milk, effect of copper upon, 1804. Appetite, relation of splenectomy to, 1809. Arthritis, hydrogen-ion concentration of exudates in, 1873. Asphyxia, effect of, on vagus stimulation of stomach, 1846. Atropine, relation of, and homatropine, 1832. Auricular fibrillation, effect of quinidine upon, 1794. B. acidophilus, therapeutic application of, 1785; effect of, 1927. B. botulinus, determination of the thermal death time of, 1798; different strains of, 1758; toxin, nature of, 1748. B. enteriditis, effect of, on mice, 1784. B. pertussis, different types of, 1840. Bact. abortus Bang, 1951. Bacteria, effect of acidity and salt con- centration on, 1888; effect of salts upon, 1889; halophilic, 1895; vitamine requirement of, 18309. Bacterial content of stomach, influenced by saliva, 1800; cultivation, collodion sacs for, 1806; growth, influence of surface tension on, 1780. Bactericidal action, effect of ethoxy group upon, 1782; effect of acid fuchsin and sodium chloride, 1874. Bacteriostatic activity, effect of sulfanilic acid on, 1892. Benzoate sodium, effect upon glomerular circulation, 1916. Benzoyltaurin, synthesis of, 1948. Bicarbonate content of blood in vomiting, 1866. Blastophoria, relation of testicular changes to, 1812. Blepharisma undulans, regeneration in, 1938; variations in, 1908. Bile in pancreatitis, 1917. 480 INDEX. Blood, changes in, by sunlight, in rickets, 1763; determination of urea in, 1838; effect of sodium citrate on, I9I2; re- action of, in intestinal intoxication, 1897. Blood cells and plasma, inorganic phos- phates in, 1825. Blood coagulation, effect of pancreatic rennet on, 1789. Blood phosphate, seasonal tide of, in infants, 1932. Blood pressure, effect on, by partial spinal cord removal, 1829; in girls, 1855; waves of, produced by breathing, 1777: Blood sugar and ovulation, 1931; studies of, 1949. Blood volume, relation of, to tissues, 1773; 1776. Body surface, relation of metabolism to, at different ages, 1887; in infants, 1960. Botulinus toxin, aggregation of particles of, 1881. Botulism, use of morphine in, 1880. Breathing, effect of, upon waves of blood pressure, 1777. Bronchus, effect of purin derivatives on, 1848. Cabbage, effect of cooking upon antiscor- butic vitamine in, 1821. Calcium depositing substances, test for, 1807. Calcium salts, effect of heat on, 1816. Casein, sulphur containing amino acid from, 1822. Cations, antagonistic action of, in relation to agglutinability, 1854. Chloride content of blood in vomiting, 1866. Citrate, sodium, effect on peristalsis, 1898; effect on central nervous system, 1899; effect of, on blood, 1912. Cocaine, effect of, on growth of lupine alba, 1849. Cod-liver oil and rickets, 1824. Collodion sacs for bacterial cultivation, 1806. Colloid, hydrophilic, estimation of, in plant fluids, 1919. Colon, anti-, streptococcus serum, 1852; as preoperative measure, 1853. Colorimeter, modified Hellige, 1788. Complement fixation tests, amount of antigen in, 1810. Conductance of organisms, 1872. Conduction, effect of quinidine on, in heart, 1827. Copper, effect of, upon antiscorbutic vitamines in milk, 1804. 481 Corpora striata, lesions in, by radium, 1933- Cottonseed meal, feeding, 1957. Cretin sheep, electrocardiogram of, 1955. Cystic growths in ovary and uterus, 1946; 1947. Cystin, method for preparation of, 1770. tissue changes from Demarcation current, velocity of develop- ment of, 1817. Depancreatinization, ligation of thyroid arteries after, 1843. Depancreatinized dog, effect of thyroidec- tomy on, 1936. Detoxication in the fowl, 1893. Development of central nervous system, attracting stimulus in, I9IOo. Diabetes insipidus, polyuria of, 1886. Diabetes, odd-carbon fats in, 1862. Diffusion constant in mountain sickness, 1868. Digitalis assay, intramuscular method of, 1857. Digitalis bodies, emetic action of, 1751. Diphtheria antitoxin, agglutination phe- nomena with, 1847. Diphtheria group, serological studies of, 1856. Diphtheria toxin, 1; study of, 1761. Disinfection theory, 1787. Dying tissue, reaction of, 1762. Electrocardiogram of cretin sheep, 1955. Electrolysis, separation of hexone bases by, I9QI4. Emesis, action of, by tartar emetic, 1935. Emetic action of digitalis bodies, 1751. Encystment in Spathidium, 1909. Endomixus, in Spathidium, 1909; macro- nuclear dissolution during, 1875. Ethoxy group, effect of, upon bactericidal action, 1782. Exercise, acid base equilibrium after, 1828. Expiration, appliance for recording vol- ume of, 1896. Exudates, hydrogen-ion concentration of, in arthritis, 1873. Fat, acetonuria produced by, 1795. Fats, odd-carbon, in diabetes, 1862. Fetuses, weight increments of premature and full-term infants compared with, 1814. Fibrillation, normal rhythm restored in, 1934- Fibroblasts, a ten-year-old strain of, 1890. Food, influence of, during lactation period, 1786. 482 Gall bladder, physiology of, 1803. Gastric analysis, comparison of urine with, 1958. Gastric contents, variations in, 1820. Gelatin gels, structure of, 1861. SCIENTIFIC PROCEEDINGS (124). Intelligence, effect of thyroidectomy on, in sheep, 1954; estimation of, in sheep, Igri. Intestinal intoxication with phospho- tungstate reagent, 1897. Germanium dioxide, effect of, in anemia, | | Intratracheal immunization, 1907. I9g44. Globulin from pecan nut, 1771. | Immunization, intratracheal, against pneu- mococcus, 1907. Globulins, accumulation of, and agglutin- | Implantation, inheritance of susceptibility ins, 1903; antigenic properties of red to, of splenic tissue, 1823. cell, 1913. | Ischemia, influence of, in infection, 1945. Glomerular circulation, effect of drugs Isopropyl alcohol, anaesthetic use of, 1791. upon, 1916; effect on, by phenol- ! sulphonephthalein, 1952. | Kala-Azar, diagnosis of, by blood culture, Glucose, absorption of, by rectum, 187 77- | 31882. Glycogen, effect of adrenalin upon, in | Kidney, tissues, 1796. Growth, bacterial, effected by substances | of plant tissue, 1801; distentive | Lactation period, influence of food upon, agencies in cell, 1799; relation of| 1786. splenectomy to, 1809; of spore-bearing | Lactose, with B. acidophilus milk, 1927. bacillus, 1813. | Lasiopogon, spermatocytes of, 1928. Gum saline, effect of, upon stimulation of | Lecithin, collodion membrane, diffusion of dog muscle, 1775; effect of, upon | sodium chloride through, 1802. volume flow, 1774- | Light, effect of, on toxicity, 1943. Gonorrheal infections, precipitin reaction | | Light rays, prevention of rickets by, in, 1841. | 1851. Lipoid, stainable material in pathological stainable lipoid material, in pathological, 1845. Heart rate, in girls, 1855. Heat, effect of, on calcium salts rennet, 1816. Hecht-Weinberg-Gradwohl reaction in syphilis, 1924. DuBois, 1772. Hemolytic properties of pneumococcus, 1926. Hemorrhage, effect of, upon stimulation of dog muscle, 1775; upon volume flow, 1774. Hen-feathering feeding, 1867. Hexone bases, trolysis, 1914. H-ion concentration, effect of sodium cirtate on, 1912; of exudates in arthritis, 1873; in medication, 1863. Hippurate, sodium, effect on glomerular circulation, 1916. Histamine, showing anaphylactoid- phe- nomena, 1883. Homatropine, relation of, and atropine, 1832. Hypophysis cerebri, volumetric study of, 1953. Infants, basal metabolism of, 1960; pre- mature and full-term, weight incre- ments of, compared with fetuses, 1814. Infection, influence of, on ischemia, 1945. Inheritance of susceptibility to implanted spleen, 1823. in male after thyroid separation of, by elec- mr pe renal epithelium, 1845. upine alba, effect of cocaine on growth of, 1849. | Macronucleur dissolution during endo- Height-weight formula, modification of Merogeny experiments, 1894. mixus, 1875. Metabolism, relation to body surface at different ages, 1887; of infants, 1960. Microbes D and G, differentiation of, by acid agglutination, 1765; virulence of, 1767. Micro-injection, apparatus for, 1792. Micro-manipulation, apparatus for, 1792. Moisture, method for determination of, in plant fluids, 1918. Morphine, use of, in botulism, 1880. Mountain sickness, diffusion constant in, 1868. Muscle extract, effect of feeding, 1870. Muscle of dog, effect of hemorrhage and gum saline, upon stimulation of, 1775. Muscular movements, effect of prostatec- tomy on, 1833. Nerve cord, stimulation of, 1905. Nervous system, central, attracting stimulus in development of, I910; effect of sodium citrate on, 1899. Oranges, vitamine-A in, 1834. Organisms, unicellular, conductance of, 1872. Orientation, heliotropic, 1902. INDEX. Ova, twins in pigeons from varying size, 1753- Ovary, cystic growths in, 1946; 1947. Ovulation, blood sugar and 1931; relation of suprarenals to, 1869. Oxalic acid poisoning, 1842. Oxygen, quantitative determination of, dissolved, 1781. Pancreatic diabetes, relation of thyroid and parathyroid to, 1844. Pancreatitis, bile factor in, 1917. Paralysis, posterior, in swine, 1956. Paralysis agitans, therapy of, 1757. Paramecium aurelia, endomixus in, 1875. Parathyroid in pancreatic diabetes, 1844; tetany, I8II. Pecan nut, globulin from, 1771. Peptones, buffering value of, 1755. Peristalsis, effect of sodium citrate on, 1898. Phenolsuphonephthalein, effect of, on glomerular circulation, 1952. Phosphate, inorganic, in plasma and blood cells, 1825; of blood in rickets, 1826. Phosphates, metabolism of, in rickets, 1920; 1950. Phosphotungstate reagent, intoxication from, 1897. Pigeons, twins in, from varying sized ova, 1753- Pituitary extract, effect of, on tempera- ture of poikilothermous animals, 1808. Plant fluids, method for determination of moisture in, 1918; estimation of hydro- philic colloid content of, 1919. Plant tissue, substances of, influencing bacterial growths, 1801. Plasma and blood cells, inorganic phos- phate in, 1825. Plethora, experimental, 1835. Plumbism, experimental, 1837. Pneumococcus, intratracheal immuniza- tion against, 1907; grouping, 1925; hemolytic properties of, 1926. Poison ivy, undetermined principle from, 1815. Potassium iodide, influence of, on actino- mycosis, 1860. Potato tubers, 1922. Precipitin reaction in gonorrheal infec- tions, 1841. Precipitation reaction for syphilis, 1878; relation between serum and antigen for syphilis in, 1879. Prostatectomy, effect of, on muscular movements, 1833. Protoplasm, movement of, by changes in consistency, 1793. Purgatives, saline, effect on absorption of drugs, 1904. 483 Purin derivatives, effect of, on bronchus, 1848. Quinidine, effect of, in auricular fibrilla- tion, 1794; light on toxicity of, on con- duction and refractory period in heart, 1827. Radiological studies, 1891. Radium, lesions caused by, in corpora striata, 1933. Rats, alcohol affecting fertility of, 1783. Rectum, absorption of glucose given by, 1877. Refractory period, effect of quinidine on, in heart, 1827. Regeneration, in Spathidium, and Ble- pharisma, 1938. Rennet, pancreatic, 1750; pancreatic, effect of, on blood coagulation, 1789; effect of heat on, 1816. Reproduction, rhythms in the rate of, 1942. Resistance, 1836. Respiration, control of, 1859. Rhythm in rate of reproduction, 1942; normal, restored in fibrillation, 1934. Rickets, cod-liver oil and, 1924; experi- mental, 1778; infantile, sunlight caus- ing changes in blood of, 1763; inorganic phosphate in blood in, 1826; more than one kind of, 1819; phosphate metab- oJism in, 1920; 1950; prevention of, by light rays, 1851; prevention of, by mercury vapor quartz lamp, 1805; prevention of, by sunlight, 1752; 1768; spontaneous cure of, 1850. Salicyl, urinary excretion of, 1884. Saligenin, pharmacological action of some ethers and esters of, 1818. Saliva, effect of, upon bacterial content of stomach, 1800. Salt concentration, effect of, on bacteria, 1888. Salts, effect upon bacteria by, 1889. Salvarsan, toxicity of, 1885. Seasons, effect of, on blood pressure, 1932. Septicemia, rabbit, mutation in pure-line strains of, 1764. Shock, peripheral resistance during, 1836. Sodium chloride, diffusion of, through collodion membrane, 1802; selective bactericidal effect of, 1874. Spathidium, endomixus and encystment in, 1909; regeneration in, 1938. Spermatocytes, nucleus and chromosomes of, 1928. Spinal cord, effect on blood pressure by partial removal of, 1829. peripheral, during shock, 484 Splenectomy, relation of, to growth and appetite, 1809. Splenic tissue, susceptibility to implants of, 1823. Spore-bearing bacillus, study of form and growth of, 1813. Stomach, vagus stimulation of, 1846; bacterial content of, influenced by saliva, 1800. Strawberries, antiscorbutic potency of, 1759- Streptococcus, preparation of anti-colon serum, 1852. Streptolysin production, effect of various proteins on, 1865. Strophanthus, assaying of, 1937. Sugar, excretion of, in urine, 1790. Sulfanilic acid, bacteriostatic activity of, 18902. Sunlight, causing changes in blood of infantile rickets, 1763; prevention of rickets by, 1752; prevention of rickets by, 1768. Suprarenals, relation of, to ovulation, 1869. Surface tension, determination of, 1779; influence of, on bacterial growth, 1780. Synapse function, alkaloid test for, 1900. Syphilis, precipitation reaction for, 1830; 1878; 1879; micro-procedure, 1831; Hecht-Weinberg-Gradwohl reaction in, 1924. Swine, posterior paralysis in, 1956. Tartar emetic, emetic action of, 1935. Temperature, effect of pituitary extract on, of poikilothermous animals, 1808. Testicular changes in alcoholism, 1812. Tetany, use of ammonium chloride in, 1940. Thalamus, optic, effect of pituitary ex- tract on animals with destroyed, 1808. Thyroid arteries, ligation of, in depan- creatized dogs, 1843; hen-feathering in male after feeding of, 1867; relation to pancreatic diabetes, 1844. Thyroidectomized sheep, estimation of intelligence in, 1911. SCIENTIFIC PROCEEDINGS (124). Thyroidectomy, effect of, 1936; effect of: on sheep’s intelligence, 1954. Tissue changes, from feeding cottonseed meal, 1957. Tissues, relation of blood volume to, 1773; 1776. Tuberculosis, agglutination reaction in, 1921. Tyrosine, produced by putrefactive anzrobe, 1769. Urea, determination of, in blood, 1838. Urine, alkaline tide of, in relation to gastric analysis, 1958. Uterus, cystic growths in, 1946; 1947. Vagus, effect of, on stomach of dog, 1846. Vitamine-A, in oranges, 1834; in almonds, 1939; antiscorbutic, in milk, effect of copper on, 1804; antiscorbutic, effect of cooking on, in cabbage, 1821; -B in the wheat kernel, 1941; -B obtained from yeast, 1923; occurrence of, 1876; parenteral administration of, 1871; re- quirements of yeast, and bacteria, 1839. Vitamines-B and -C, plant sources of, 1749. Vitamines of yeast, 1754. Volume flow, effects of hemorrhage and gum saline upon, 1774. Vomiting, bicarbonate and chloride con- tent of blood in, 1866. Wassermann reaction, mathematical rela- tion in, 1756. Weight increments of premature and full- term infants and fetuses, 1814. Wheat kernels, vitamine-B in, 1941. Xerophthalmia, earliest changes in experi- mental, 1929; 1930; lesions in lacrimal glands in, 1930. X-rays, anaphylactic reaction from, 1760. Yeast, as source of vitamine-B, 1923; effect of feeding, 1870; vitamine re- quirement of, 1839; vitamines of, 1754. Addenda. To be bound at page 183. The paragraph on top of page 183 should end with the follow- ing sentence: These should be cholesterinized to 0.4 per cent. To be bound at page 210. P. 210. Fig. 1 should be Fig. 2, Dog 108. se “és ‘ ce Pett. 2 * $23. P 212 sé 3 cc é sé I sé 120 a . +] . P.213. ‘“ 4 From normal dog. To be bound at page 258. P. 258. Graph should be Fig. 4, which legend is on p. 263. é cé é ‘é I, ‘sé ‘é P. 259. en! oe ae P. 260. éé “é é ins 2. é sé sé éé p. 259. P. 263. sé éé “ec ‘é a sé sé ‘é éé p. 260. 485 PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED SEVENTEENTH MEETING CORNELL UNIVERSITY MEDICAL COLLEGE NEW YORK CITY OCTOBER Io, 1921 AND THIRTIETH MEETING PACIFIC COAST BRANCH BERKELEY, CALIFORNIA OCTOBER 15, 1921 AND FIRST MEETING MINNESOTA BRANCH MINNEAPOLIS, MINNESOTA OCTOBER 12, 1921 VOLUME XIX No. 1 NEW YORK 1921 CONTENTS. J. BRONFENBRENNER AND M. J. SCHLESINGER: Further studies on the nature of botulinus toxin. 1 (1748). Francisco O. SANTOS (by invitation): Some plant sources of vitamins B and C. 2 (1749). ALBERT A. EPSTEIN: Observations on pancreatic rennet. 3 (1750). ROBERT A. HATCHER AND SOMA WEISS: Further observations on the seat of the emetic action of the digitalis 4 (1751). ALFRED F. HEss, L. J. UNGER AND A. W. PAPPENHEIMER: The prevention of rickets by exposure to sunlight. 5 (1752) OscaR RIDDLE: Identical twins in pigeons arise from ova of markedly aberrant size. 6 (1753). CASIMIR FUNK AND Harry E. DUBIN: The vitamines of yeast and their réle in animal nutrition. 7 (1754). J. BRONFENBRENNER, G. G. DE BorD AND P. F. Orr: Comparative buffering value of American peptones. 8 (1755). STERNE Morse (by invitation): Some mathematical relations in the Wassermann reaction. 9 (1756). M. H. WEINBERG AND T. SCHUBB (by invitation): Experiment in new method of therapy of paralysis agitans. (1757)- J. BRONFENBRENNER, M. J. SCHLESINGER AND S. C. CALAZANS: Typing of different strains of bacillus botulinus by im munologic methods. 11 (1758). CLARENCE A. SMITH, OLAF BERGEIM AND PuHiLip B. HAwK: The antiscorbutic potency of strawberries. 12 (1759) - R. G. Hussey (by invitation): A modified anaphylactic reaction induced by x-rays. 13 (1760). P. J. MOLONEY AND L. HANNA (by invitation): Contribution to study of diphtheria toxin I. 14 (1761). WINTHROW MorRSE AND H. C. VAN DER HEYDE: The change in reaction of dying tissue. 15 (17762). ALFRED F. Hess AND P. GUTMAN: The cure of infantile rickets by sunlight as demonstrated by a chemical alteration o the blood. 16 (1763). Pau H. De Kruir: Dissociation of microbic species. II. Mutation in pure-line strains of the bacillus of rabbit sep- ticemia. 17 (1764). : Pau H. De KrutF: III. Differentiation of microbes D and G by acid agglutination. 18 (1765). PauLt H. De Krutr: IV. Factors influencing the acid agglutination optimum of types Dand G. 19 (1766). Paut H. De KrutF: V. Further considerations in regard to the virulence of microbes Dand G. 20 (1767). : P. G. SurpLey, E. A. Park, G. F. Powers, E. V. MCCOLLUM AND NINA Srmmonps: The prevention of the development of rickets in rats by sunlight. 21 (1768). . IvAN C. HALL AND FLORENCE FINNERUD: The production of tyrosine by a putrefactive anaérobe. 22 (1769). Cart L. A. ScuHmipt: A method for the preparation of cystin. 23 (1770). F. A. Cajori (by invitation): A globulin as the principal protein of the pecan nut. 24 (17712). HAROLD K. FABER AND MARGARET S. MELCHER: A modification of the DuBois height-weight formula for surface | areas of newborn infants. 25 (1772). ROBERT GESELL, CHARLES S. CAPP AND FREDERICK S. Foote: On the relation of blood-volume to the nutrition of the tissues. I. 26 (1773). RosBERT GESELL: II. The effects of hemorrhage and subsequent injections of gum-saline upon the volume flow through striated muscle of the dog. 27 (1774). ROBERT GESELL: III. The effects of hemorrhage and subsequent injection of gum-saline upon the response of sartorius muscle of dog to rapid electrical stimulation. 28 (1775). ROBERT GESELL, CHARLES S. CAPP AND FREDERICK S. Foote: IV. On the relation of blood-volume to the nutrition of the tissues. 29 (1776). ROBERT TROTTER, PHILIP EDSON AND ROBERT GESELL: A comparison of the waves of blood pressure produced by slow and by rapid breathing. 30 (1777). ‘ J. F. McCLenNpON AND Harry BavuGueEss (by invitation): Experimental rickets. 31 (1778). Rosert G. GREEN (by invitation): Rapid determination of surface tension. 32 (1779). W. P. Larson: The influence of the surface tension of the culture medium on bacterial growth. 33 (1780). E. J. Lunp: A micro-Winkler method for the quantitative determination of dissolved oxygen. 34 (1781). ARTHUR D. HIRSCHFELDER AND L. J. PANKOW: Does the introduction of an ethoxy group into aromatic compounds in- crease their bactericidal action upon the pneumococcus and the gonococcus? 35 (1782). - The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after each meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of October to @ May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume is three dollars each, postage pre- § paid. The price of copies of the proceedings of any meeting is fifty cents each, postage prepaid. Subscriptions are payable in advance. ‘ PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. VICE-PRESIDENT—]J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, Post-Graduate Medical School, Alfred F. Hess, University and © Bellevue Hospital Medical College, and ex-Presidents. MANAGING Ep1ToR—The Secretary-Treasurer, 338 East 26th St., New York City. CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Honorary. William T. Councilman, Harvard University. Edward T. Reichert, University of Pennsylvania. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. Columbia University —C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, seorge Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, Louise H. Gregory, A. Baird Hastings, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M. Richards, Mary Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. | Cornell University Medical College—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert Chambers, A. F. Coca, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, Samuel Goldschmidt, A. Hartwell, Robert A. Hatcher, Graham Lusk, William S. McCann, Walter L. Niles, Charles R. Stockard, W. C. fhro, John C. Torrey. Fordham University School of Medicine.—Carl P. Sherwin. Hospitals, Beth Israel—Max Kahn, Morris H. Kahn. Montefiore Home.—Emil J. Baumann, George Fahr, B. S. fline, Michael Levine, David Marine. Mt. Sinai.—George Baehr, Burrill B. Crohn, Charles A. Elsberg, Albert A. -pstein, R. Ottenberg, Harry Plotz, M. A. Rothschild. New York.—Nellis B. Foster, Ralph G. Stillman. Presbyte- an.—Louis Baumann, A. R. Dochez, George M. Mackenzie, Walter W. Palmer, Franklin A. Stevens. Roosevelt.— G. Falk, I. Greenwald, W. G. Lyle. St. Lukes —L.W. Famulener, Karl M. Vogel. New York City Departments. Health—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Park, ona W. Williams, A. Zingher. Chief Medical Examiner.—Charles Norris. New York Homoeopathic Medical College.—I. S. Kleiner, Mary B. Stark. New York Post-Graduate Medical School_—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, nton R. Rose, R. M. Taylor. | New York University—W. H. Barber, Harlow Brooks, Warren Coleman, E. K. Dunham, Florence Hulton Frankel, O. Gettler, Alfred F. Hess,Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Emil J. ellini, H. D. Senior, Douglas Symmers, George B. Wallace. Rockefeller Institute for Medical Research.—Harold L. Amoss, J. H. Austin, O. T. Avery, Carl A. L. Binger, Wade a. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, Glenn E. Cullen, Paul DeKruif, Simon Flexner, F. L. Gates, Walter Jacobs, I. J. Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Clara J. Lynch, Gustave M. Meyer, James B. Mur- hy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. _ Industrial Laboratories (New York City). Research Laboratory of H. A. Metz.—Harry E. Dubin, Casimir Funk. | Research Laboratory of H. A. Metz (Brooklyn).—Charles W. Hooper. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 17 E. 38th Si., N. Y. City—J. W. Draper. 126 E. 64th St. N. Y. City— Cyrus W. Field. 141 W. 78th St., N. Y. City.— . I. Ringer. Non-Resident. Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick) .— . W. Shive, S. A. Waksman. Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, | 4 pert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, } Charles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal, H. A. Spoehr. (Washington, | oD . C.).—Allfred G. Mayer. 1 State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank faltaner, A. B. Wadsworth. | \ Hospitals.. Addenbrooke's (Cambridge, England).—C. G. L. Wolf. Buffalo City—Katharine R. Collins. The J dees ).—Thomas Flournoy. Peter Bent Brigham (Bosion).—Harvey Cushing, Francis W. Peabody. Philadelphia al—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.) —Ralph Pemberton. Rikshospitalet (Kristiania, Nor- way).—H. F. Hést. U.S. Pubic Health (Boston).—F. M.McCrudden. Western Pennsylvania (Pitisburgh).—Maurice . P Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton Springs).—Roger S. Hubbard. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. PS atwick Laboratory (Buffalo)—H. R. Gaylord. Juvenile Psychopathic (Chicago)—Herman M. Adler. Potter Meta- a Phipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F.S. Jones, Theobald Smith. Trudeau ir ildren’s (Boston, Mass.).—Oscar M. Schloss. General (Cincinnati, Ohio).—Martin H. Fischer. Mercy (Pittsfield, . Givens. olic Clinic (Santa Barbara, Calif.)—H.R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. Sanatorium (Saranac Lake, N. Y.).—R. A. Kocher. Wistar (Philadelphia).—H. H. Donaldson. U.S. Departmenis. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, J Mohler. Hygienic Laberatory (Washington, D. C.).—Wm. C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, Georg B. Roth. Surgeon General's Office (Washington, D. C.).—J. F. Siler. ‘ National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Ausiralia).—T. Brailsford Robertson. Amherst.—Otto Glaser. Baylor.—J. Hi Black. Buffalo.—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwith, W. R. Bloor, T. C. Burnett™ H. M. Evans, E. C. Fleischner, F. P. Gay, Ivan C. Hall, Robert A. Gesell, S. J. Holmes, Samuel H. Hurwitz, Charles A@ Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker’ Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati. —D. E. Jackson, Shiro Tashiro. Copen™ hagen.—Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. Georg Washington. — W. H. Schultz. Harvard. — Jacob Bronfenbrenner, Walter B. Cannon, Otto Folin, Worth Hale, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Percy G. Stiles Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach. Illinois.—H. B. Lewis, William H. Welker. Jefferson.—O. Bergeim®, P. B. Hawk. Johns Hopkins.—John J. Abel, C.G. Bull, George W. Corner, W. W. Cort, R. S. Cunningham, W. W Ford, W. S. Halsted, R. W. Hegener, William H. Howell, John Howland, H. S. Jennings, Benjamin Kramer, Paul D) Lamson, W. G. MacCallum, David I. Macht, E. K. Marshall, Jr.. Wm. S. McCann, E. V. McCollum, Adolph Meyer Arthur L. Meyer, Raymond Pearl, G. C. Robinson, Paul G. Shipley, Reynold Albrecht Spaeth, W. H. Taliaferro, Wil! liam H. Welch, D. Wright Wilson. Kansas.—Bennett M. Allen. Keo (Japan)—Noahidé Yatsu.—Leland Stanford. Thomas Addis, Carl L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. J. Hanzlik, A® W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophiils, R. E Swain, E. B. Towne. Liverpool.—J. G. Adami. Maryland.—Charles E. Simon. McGill (Montreal). —Henry Gra Barbour, Horst Oertel, A. B. Macallum, John L. Todd. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lom bard, Frederick G. Novy, Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Brown, C. H. Eckles C. P. Fitch, R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, E. C. Kendall, F. E. Kingsbury, W. P Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, William Moore, Leroy S. Palmer, H. E. Robertson, R. E. Scammon, F. . W. Schultz, F. H. Scott, E. C. Stakman, J. J. Willaman. Missouri—Mazyck P. Ravenel. Nebraska.—A. E. Guen- ther. North Carolina.—W. deB. MacNider. Northwestern.—Solomon Strouse. Ohio State-—R. G. Hoskins. Oregon. D. E. Lancefield, C. F. Hodge. Peking Union Medical—Franklin C. McLean. Pennsylvania.—Alexander C. Ab bott, D. H. Bergey, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., O. H. Perry Pepper, Alfred N Richards, J. Edwin Sweet, A. E. Taylor. Pennsylvania State-—R. Adams Dutcher. Philippine.—R. B. Gibson. Pittsburgh.—C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.—Edwin G. Conklin, E. Newton Harvey. Rochester —John R. Murlin, G. H. Whipple. Rutgers—John F. Anderson, W. J. Crozier, A. R. Moore, Thurlow C. Nelson. Sheffield—J. B. Leathes. Southern California (Los Angeles).—Lyman B. Stookey. St Louis.—John Auer, Don R. Joseph, Ralph A. Kinsella. Texas.—Herman J. Muller, William C. Rose. Tohoka In perial (Japan).—Shinkishi Hatai, C. K. Watanabe. Toronto. — A. H. Caulfeild, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane.—Charles W. Duval. Union University (Albany Medical College).—Melvin Dres-@ bach, Arthur Knudson. Vanderbilt (Nashville).—B. T. Terry. Virginia.—H.E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Eugene L. Opie, Philip A. Shaffer. Wesleyan.—E. C. Schnei- der. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenbrey, H. T. Karsner, J. M. Rogoff, R. W. Scott, Torald) Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia. — F. E. Chidester, Withrow Morse. Wéisconsin.—Charles R.@ Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Yale——George A. Baitsell, R. H. Chittenden, J. W. Churchman, Ross G. Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo F. Rettger, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Industrial Laboratories: Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. Newark, N. J., Calo Chemical Co.—M. S. Fine. New Brunswick, N. J., E. R. Squibb and Son.—P. A. Koher. Peekskill, N. Y., Fleischman Company.—A. K. Balls. Philadelphia, Pa., 1524 Chestnut St., H. K. Mulford Co.—T. S. Githens. Water- bury, Conn., Scovill Mfr. Co.—A. H. Ryan. . Asheville, N. C., 43 Austin Ave.—Ephraim M. Ewing. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer. Chicago, Illinois, 31 East Elm St.—W.F. Petersen. Los Angeles, Calif., 533 Lucerne Blud.—N. W. Janney. Misso Botanical Garden, St. Louis, Mo.—B. M. Duggar. Ossining, N. Y., R. F. D. 2.—H.D. Dakin. Pittsburgh, Pa., Jenki Arcade.—Jacob Rosenbloom. —Tuckahoe, N. Y. — Isaac F. Harris. Washington, D. C., 1701 Mass. Ave. — Robert M.§ Yerkes. Wethersfield, Conn., 4 Wilcox St.—Alexander L. Prince. Paris, France.— Harry Plotz. Berlin, Germany.— Reinhard Beutner, Rhoda Erdmann. Kastanienbaum, Switzerland.—Fritz Schwyzer. | Members present at the one hundred seventeenth meeting: 4 Bailey, C. V., Binger, Bronfenbrenner, Calkins, Cecil, Chambers, Cohn, Curtis, Dochez, Dubin, Eddy, Edwards, § Eggleston, Epstein, Fine, Funk, Greenwald, Hatcher, Hess, Jackson, H. C., Joblin, Kahn, Max, Kahn, M. oT Myers, Ottenberg, Pappenheimer, Park, E. A., Riddle, Ringer, Sherwin, Stark, Stevens, Swift, Teague, Thomas, Th Torrey, Wallace. aif Members elected at the one hundred seventeenth meeting: Tadachika Minoura, Fritz B. Talbot. Dates of the next two meetings: November 16, 1921—December 21, 1921. . PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED EIGHTEENTH MEETING NEW YORK POST-GRADUATE MEDICAL SCHOOL NEW YORK CITY NOVEMBER 16, 1921 VoLUME XIX No. 2 NEW YORK 192! CONTENTS. E. CARLTON MACDOwWELL: Alcohol and white rats; a study of fertility. 36 (1783). LESLIE T. WEBSTER (by invitation): Experiments with B. enteriditis (murium) on normal and immune mice. 37 (1784 Lro F. RETTGER AND HARRY A. CHEPLIN: The therapeutic application of bacillus acidophilus. 38 (1785). H. C. SHERMAN AND MARIE MUHLFELD: Growth and reproduction upon simplified food supply, II. Influence of foo upon mother and young during the lactation period. 39 (1786). BARNETT COHEN: Some phases of the disinfection theory. 40 (1787). Victor C. Myers: A modified Hellige colorimeter for the comparison of solutions containing two colors. 41 (1788). ALBERT A. EPSTEIN AND NATHAN ROSENTHAL: The effect of pancreatic rennet on blood coagulation. 42 (1789). Lupwic Kast AND Hitpa M. Crott: Observations on the excretion of sugar in the urine in health and disease. 43 (1790 Davip I. Macut: Isopropyl alcohol; a convenient laboratory anesthetic for cats. 44 (1791). ROBERT CHAMBERS: Apparatus for micro-manipulation and micro-injection. 45 (1792). ROBERT CHAMBERS: Effect of experimentally induced changes in consistency of protoplasmic movement. 46 (1793 Rosert L. Levy: Alterations in the cardiac mechanisms after administration of quinidine to patients with auricul: fibrillation. 47 (1794). ROGER S. HUBBARD AND FLOYD R. WRIGHT: Studies on the acetonuria produced by diets high in fat. 48 (1795). A. I. Rincer, H. DUBIN AND F. HULTON FRANKEL: I. The glycogen content of the tissues of diabetic animals and tk influence of adrenalin thereon. 49 (1796). A. I. RINGER: II. Concerning antiketogenesis. 50 (1797). + 3 Ernest C. DICKSON AND GEORGINA S. BURKE: Botulism: a method for determining the thermal death time of spores of Bacillus botulinus. 51 (1798). The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after e cl meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of October May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume is three dollars each, postage f paid. The price of copies of the proceedings of any meeting is fifty cents each, postage prepaid. Subscriptions a payable in advance. PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College, VicE-PRESIDENT—J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, New York, Post-Graduate Medical School, Alfred F. University and Bellevue Hospital Medical College, and ex-Presidents. MANAGING Epitor—The Secretary-Treasurer, 338 East 26th St., New York City. _ CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL . BIOLOGY AND MEDICINE. Honorary. William T. Councilman, Harvard University. Edward T. Reichert, University of Pennsylvania. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. _ Columbia University —C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, Louise H. Gregory, A. Baird Hastings, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M. Richards, Mary Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. ii Cornell University Medical College—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert Chambers, A. F. Coca, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, bert A. Hatcher, Graham Lusk, Walter L. Niles, Charles R. Stockard, W. C. Thro, John C. Torrey. _ Fordham University School of Medicine.—Carl P. Sherwin. _ Hospitals, Beth Israel—Max Kahn, Morris H. Kahn. Montefiore Home.—Emil J. Baumann, George Fahr, B. S. Kline, Michael Levine, David Marine. Mt. Sinai.—George Baehr, Burrill B. Crohn, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Rothschild. New York.—Nellis B. Foster, Ralph G. Stillman. Presbyte- 4an.—Louis Baumann, A. R. Dochez, George M. Mackenzie, Walter W. Palmer, Franklin A. Stevens. Roosevelt.— K. G. Falk, I. Greenwald, W. G. Lyle. St. Lukes.—L. W. Famulener, Karl M. Vogel. | New York City Depariments. Health—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Park, Anna W. Williams, A. Zingher. Chief Medical Examiner.—Charles Norris. _ New York Homoeopathic Medical College.—I. S. Kleiner, Mary B. Stark. _ New York Post-Graduate Medical School.—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, mton R. Rose, R. M. Taylor. _ New York University —W. H. Barber, Harlow Brooks, Warren Coleman, E. K. Dunham, Florence Hulton Frankel, 4. O. Gettler, Alfred F. Hess,Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Emil J. Pellini, H. D. Senior, Douglas Symmers, George B. Wallace. _ Rockefeller Institute for Medical Research.—Harold L. Amoss, J. H. Austin, O. T. Avery, Carl A. L. Binger, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, Glenn E. Cullen, Paul DeKruif, Simon Flexner, F. L. Gates, Walter 4. Jacobs, I. J. Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Clara J. Lynch, Gustave M. Meyer, James B. Mur- ohy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. _ Industrial Laboratories (New York City). Research Laboratory of H. A. Metz.—Harry E. Dubin, Casimir Funk. Research Laboratory of H. A. Metz (Brooklyn).—Charles W. Hooper. _ Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City —E. E. Butterfield. 17 E. 8th St., N. Y. City—J. W. Draper. 126 E. 64th St. N. Y. City.—Cyrus W. Field. 141 W. 78th St., N. Y. City.— A. I. Ringer. Non-Resident. Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick).— ', W. Shive, S. A. Waksman. § Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, MAlbert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, ®-harles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal, H. A. Spoehr. (Washington, #2. C.).— Alfred G. Mayer. _ State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank faltaner, A. B. Wadsworth. _ Hospitals. Addenbrooke’s (Cambridge, England).—C. G. L. Wolf. Buffalo City.—Katharine R. Collins. The children’s (Boston, Mass.).—Oscar M. Schloss. General (Cincinnati, Ohio)—Martin H. Fischer. Mercy (Pittsfield, Yass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Philadelphia reneral—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.).—Ralph Pemberton. Rikshospitalet (Kristiania, Nor- : ied F. Host. U.S. Pubic Health (Boston).—F.M.McCrudden. Western Pennsylvania (Pitisburgh).—Maurice . Givens. _ Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton iprings).—Roger S. Hubbard. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. watwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago).—Herman M. Adler. Potter Meta- olic Clinic (Santa Barbara, Calif.).—H. R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. 7h ipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F.S. Jones, Theobald Smith. Trudeau ‘- ‘ Sanatorium (Saranac Lake, N. Y.).—R. A. Kocher, Wistar (Philadelphia).—H. H. Donaldson. . U. S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, J. 1 Mohler. Hygienic: Laboratory (Washington, D. C.).—Wm. C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, Georgia B. Roth. Surgeon General's Office (Washington, D. C.).—J. F. Siler. National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Austrelia).—T. Brailsford Robertson. Amherst.—Otto Glaser. Baylor.—J. Black. Buffalo—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwith, W. R. Bloor, T. C. Burnet’ H. M. Evans, E. C. Fleischner, F. P. Gay, Robert A. Gesell, Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles ; Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walke Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Copenhagen.- Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. George Was ington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Otto Folin, Worth Hale, Reid Hun W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Percy G. Stiles, Richard F 1 Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. JIllinois.—H. B. Lewis, William H. Welker. Jefferson. d Bergeim, P. B. Hawk. Johns Hopkins.—John J. Abel, C. G. Bull, George W. Corner, W. W. Cort, R. S. Cunningham W. W. Ford, W. S. Halsted, R. W. Hegener, William H. Howell, John Howland, H. S. Jennings, Benjamin Krame? Paul D. Lamson, W. G. MacCallum, David I. Macht, William S. McCann, E. K. Marshall, Jr., Wm. S. McCann, E. | McCollum, Adolph Meyer, Arthur L. Meyer, Raymond Pearl, G. C. Robinson, Paul G. Shipley, Reynold Albrecht Spaeth W. H. Taliaferro, William H. Welch, D. Wright Wilson. Kansas.—Bennett M. Allen. Keo (Japan).—Noahidé Yatsv Leland Stanford.—Thomas Addis, Carl L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophi H R. E. Swain, A. E. Taylor, E. B. Towne. Liverpool.—J.G. Adami. Maryland.—Charles E. Simon. McGill (Montr eal, —Henry Gray Barbour, Horst Oertel, A. B. Macallum, John L. Todd. Michigan.—C. W. Edmunds, G. Carl Hube Warren P. Lombard, Frederick G. Novy, Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Browr® C. H. Eckles, C. P. Fitch, R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, E. C. Kendall, F.B Kingsbury, W. P..Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, William Moore, Leroy S. Palmer, H. E. Robertson, R E. Scammon, F. W. Schultz, F. H. Scott, E. C. Stakman, J. J. Willaman. MissouriimMazyck P. Ravenel. Nebrask —A. E. Guenther. North Carolina.—W. deB. MacNider. Northwestern.—Solomon Strouse. Ohio State-——R.G. Hos Oregon.—D. E. Lancefield, C. F. Hodge. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander © Abbott, D. H. Bergey, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., O. H. Perry Pepper, Alfred } Richards, J. Edwin Sweet. Pennsylvania State—R. Adams Dutcher. Philippine-——R. B. Gibson. Pittsburgh. C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton—Edwin G. Conklin, E. Newton Harve Rochester —John R. Murlin, G. H. Whipple. Rutgers.——John F. Anderson, W. J. Crozier, A. R. Moore, Thurlow C Nelson. Sheffield —J. B. Leathes. Southern California (Los Angeles).—Lyman B. Stookey. St. Louis.—John Auer Don R. Joseph, Ralph A. Kinsella. Texas.—Herman J. Muller, William C. Rose. Tohoka Imperial (Japan).—Shink sh Hatai, C. K. Watanabe. Toronto.—A. H. Caulfeild, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys Ss Tulane.—Charles W. Duval. Union University (Albany Medical College).—Melvin Dresbach, Arthur Knudson. Vander bilt (Nashville). —B. T. Terry. Virginia.—H.E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Josep Erlanger, Leo Loeb, Eugene L. Opie, Philip A. Shaffer. Wesleyan.—E. C. Schneider. Western Reserve (Cleveland).—@ George W. Crile, A. B. Eisenbrey, H. T. Karsner, J. M. Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C._ J | Wiggers. West Virginia.—F. E. Chidester, Withrow Morse. Wéisconsin.—Charles R. Bardeen, C. H. Bunting, J. bg Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Yale.—George A. Baitsell, R. H. Chittenden, J. W. 5 i: man, Ross-G. Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., ! Rettger, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. = | Industrial Laboratories: Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. Newark, N. J., Calco Chemica: Co.—M.S. Fine. New Brunswick, N. J., E. R. Squibb and Son.—P. A. Koher. Peekskill, N. Y., Fleischman Company.— A. K. Balls. Philadelphia, Pa., 1524 Chestnut St., H. K. Mulford Co.—T. S. Githens. Waterbury, Conn., Scovill Mfr Co.—A. H. Ryan. Asheville, N. C., 43 Austin Ave-—Ephraim M. Ewing. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer Chicago, Illinois, 31 East Elm St.—W. F. Petersen. Los Angeles, Calif., 533 Lucerne Blud.—N. W. Janney. Misso Botanical Garden, St. Louis, Mo.—B. M. Duggar. Ossining, N. Y., R. F. D. 2.—H. D. Dakin. Pitisburgh, Pa., Jenk Arcade.—Jacob Rosenbloom. Tuckahoe, N. Y.—Isaac F. Harris. Washington, D. C., 1701 Mass. Ave.—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St.—Alexander L. Prince. Paris, France.—Harry Plotz. Berlin, Germanys Reinhard Beutner, Rhoda Erdmann. Kastanienbaum, Switzerland.—Fritz Schwyzer. Members present at the one hundred eighteenth meeting: Baumann, E. J., Bauman, L. Barnett, Benedict, Cohen, Barnett, Coleman, Crohn, Dubin, Edwards, Frankel, Jat son, H. C., Kleiner, Lynch, Myers, Ottenberg, Riddle, Ringer, Torrey, Wallace. Members elected at the one hundred eighteenth meeting: Albert Fischer, Emil Goetsch, Clarence M. Jackson, W. B. Kirkham, Nicholas Kopeloff, Jesse Francis McClend¢ Proviso V. Prewitt, Stanley P. Reimann, John P. Schneider, Harry Beal Torrey. Dates of the next two meetings: December 21, 1921I-January 18, 1922. PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED NINETEENTH MEETING ROCKEFELLER INSTITUTE FOR MEDICAL RESEARCH NEW YORK CITY DECEMBER 21, 1921, AND SECOND MEETING MINNESOTA BRANCH MINNEAPOLIS, MINNESOTA DECEMBER 14, 1921 VoLUME XIX No. 3 NEW YORK 1921 eer a ee AL CONTENTS D. T. MacDouGAL: The distentive agencies in the growth of the cell. 52 (1799). NICHOLAS KOPELOFF: The bacterial content of the stomach as influenced by saliva. 53 (1800). | O. T. AVERY AND HuGH J. MorRGAN: The effect of the accessory substances of plant tissue upon etaweh of bacteria: 54 (1801). Haro_p A. ABRAMSON AND SAMUEL H. Gray (by invitation): The diffusion of sodium chloride through a “lecithin” - collodion membrane. 55 (1802). Cc LIONEL S. AUSTER AND BURRILL B. CROHN: Notes on studies in the tno of the gall bladder. 56 (1803). ALFRED F. HESS AND LESTER J. UNGER: The destruction of the antiscorbutic vitamin in milk by the catalytic action of minute amounts of copper. 57 (1804). E. A. Park, G. F. Powers, P. G. SHIPLEY, E. V. MCCOLLUM AND NINA SIMMONDS: The prevention of rickets in the rat by means of radiation with the mercury vapor quartz lamp. 58 (1805). FREDERICK L. GATES: Collodion sacs for aérobic and anaérobic bacterial cultivation. 59 (1806). E. V. McCottum, NINA Srmmmonps, P. G. SHIPLEY AND E. A. PARK: A delicate biological test for calcium depositing — substances. 60 (1807). FRED T. ROGERS (by invitation): The effects of pituitary extract on the body temperature of animals rendered poikilo-_ thermous by destruction of the optic thalamus. 61 (1808). ARTHUR H. SMITH AND LEAH ASCHAM: Relation of splenectomy to growth and appetite in the rat. 62 (1809). ; R. L. KAHN AND S. R. JoHNSON: Determination of optimum amount of antigen in complement fixation tests. 63 (1810). Arno B. LUCKHARDT AND PHILIP J. ROSENBLOOM: The prevention and control of parathyroid tetany. 64 (1811). CARL VERNON WELLER: Testicular changes in acute alcoholism in man and their relationship to blastophthoria._ 65 (1812). ) ARTHUR T. Hewnrici: A statistical study of the form and growth of a spore-bearing bacillus. 66 (1813). 3 Z RICHARD E. SCAMMON: On the weight increments of premature infants as compared with those of fetuses of the same gestation age and those of full-term children. 67 (1814). ( E. D. Brown: An undetermined principle obtained from poison ivy. 68 (1815). Leroy S. PALMER: The effect of heat on the calcium salts and rennet. 69 (1816). GEORGE EDMESTON FAHR: The velocity of development of the demarcation current in the frog’s sartorius. 70 (819) ArTHUR D. HIRSCHFELDER AND HERMAN H. JENSEN: The pharmacological action of some ethers and esters of saligenin. — 71 (1818). Hd iki : I The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after each y I meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of October to. h May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume is three dollars each, postage pre-_ paid. The price of copies of the proceedings of any meeting is fifty cents each, postage prepaid. Subscriptions are © payable in advance. ; PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. ViICE-PRESIDENT—J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, New York, Post-Graduate Medical School, Alfred F. Hess, University and Bellevue Hospital Medical College, and ex-Presidents. MANAGING EpiITOR—The Secretary-Treasurer, 338 East 26th St., New York City. CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL | Z BIOLOGY AND MEDICINE. 4) id Honorary. aa William T. Councilman, Harvard University. - Edward T. Reichert, University of Pennsylvania. i 4 | Resident (Greater New York). a College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. a Columbia University.—C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, | George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, Louise H. Gregory, A. Baird Hastings, J. Gardner "Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M.. Richards, Mary S$. Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. y Cornell University Medical College—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert Chambers, A. F. Coca, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, Robert _A. Hatcher, Graham Lusk, Walter L. Niles, Charles R. Stockard, W. C. Thro, John C. Torvey. Fordham University School of Medicine.-—Carl P. Sherwin. Hospitals, Beth Israel Babies —Martha Wollstein, Max Kahn, Morris H. Kahn. Long Island College —Emil Goetsch. ontefiore Home.—Emil J. Baumann, B.S. Kline, Michael Levine, David Marine. Mt. Sinai.—George Baehr, Burrill -B. Crohn, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Rothschild. New York.—Nellis B. ; ‘Foster, Ralph G. Stillman. Presbyterian.—Louis Baumann, A. R. Dochez, George M. Mackenzie, Walter W. Palmer, | Franklin A. Stevens. Roosevelt.—K. G. Falk, I. Greenwald, W. G. Lyle. St. Lukes.—L. W. Famulener, Karl M. Vogel. a e New York City Departments. Health—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Parks Anna W. Williams, A. Zingher. Chief Medical Examiner.—Charles Norris. New York Homoeopathic Medical College.—I. S. Kleiner, Mary B. Stark. _ New York Post-Graduate Medical School.—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, ‘Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. _ New York University —W.H. Barber, Harlow Brooks, Warren Coleman, E. K. Dunham, Florence Hulton Frankel, £ AO. Gettler, Alfred F. Hess, Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Emil J. ® Pellini, H. D. Senior, Douglas Symmers, George B. Wallace. Psychiatric Institute-—Nicholas Kopeloff. Rockefeller Institute for Medical Research.—Harold L. Amoss, O. T. Avery, Carl A. L. Binger, Wade H. Brown, exis Carrel, A. E. Cohn, Rufus Cole, Paul DeKruif, Albert Fischer, Simon Flexner, F. L, Gates, Walter A. Jacobs, I. . Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Clara J. Lynch, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Industrial Laboratories (New York City). Research Laboratory of H. A. Metz—Harry E. Dubin, Casimir Funk. Research Laboratory of H. A. Metz (Brooklyn).—Charles W. Hooper. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 177 E. 38th St., N. Y. City—J. W. Draper. 126 E. 64th St., N. Y. City—Cyrus W. Field. 141 W. 78th St., N. Y. City.— A. I. Ringer. . ; f ss J m ie Non-Resident. _ Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick) .— J. W. Shive, S. A. Waksman. Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, é Ibert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, Charles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal, H. A. Spoehr. (Washington, 2. C.).— Alfred G. Mayer. _ State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank Maltaner, A. B. Wadsworth. _ Hospitals. Addenbrooke's (Cambridge, England).—C. G. L. Wolf. Buffalo City.—Katharine R. Collins. The Children’s (Boston, Mass.).—Oscar M. Schloss. General (Cincinnati, Ohio) —Martin H. Fischer. Mercy (Pittsfield, ‘Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Philadelphia General—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.).—Ralph Pemberton. Rikshospitalet (Kristiania, Nor- 2. F. Host. U.S. Public Health (Boston).—F.M.McCrudden. Western Pennsylvania (Piitsburgh).—Maurice H. Givens. _ Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton Springs).—Roger S. Hubbard. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. Gratwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago).—Herman M. Adler. Potter Meta- 5 lic Clinic (Santa Barbara, Calif.).—H.R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. "hipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F.S. Jones, Theobald Smith. Trudeau Sanotarium (Saranac Lake, N.Y.).—R. A. Kocher. Wistar (Philadelphia).—H. H. Donaldson. U. S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, J. R.| Mohler. Hygienic Laboratory (Washington, D. C.).—Wm. C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, George B. Roth. Surgeon General’s Office (Washington, D. C.).—J. ¥. Siler. National Research Council, Washington, D. C.—V.L. Kellogg, Clarence J. West. Universities. Adelaide (South Austiralia)—T. Brailsford Robertson. Ambherst.—Otto Glaser. Baylor.—J. H: Black. Buffalo—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwith, W. R. Bloor, T. C. Burnett: ® H. M. Evans, E. C. Fleischner, F. P. Gay, Robert A. Gesell, Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles A-| Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker. | Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnatim—D. E. Jackson, Shiro Tashiro. Copen- hagen.—Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.——Richard V. Lamar, William Salant. George Washington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Otto Folin, Worth Hale, Law- rence J. Henderson, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J.| Rosenau, Percy G. Stiles, Richard P. Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. Iilinois.—H. B. Lewis, W. F. Petersen, William H. Welker. Iowa State-—R.B. Gibson. Jefferson.—O. Bergeim, P. B. Hawk. Johns Hopkins. | —John J. Abel, C. G. Bull, George W. Corner, W. W. Cort, R. S. Cunningham, W. S. Halsted, R. W. Hegener, Wiliiam } H. Howell, John Howland, H. S. Jennings, Benjamin Kramer, Paul D. Lamson, W. G. MacCallum, David I. Macht, E. K. Marshall, Jr., Wm. S. McCann, E. V. McCollum, Adolph Meyer, Arthur L. Meyer, Raymond Pearl, G. C. Robin-/ son, Paul G. Shipley, Reynold Albrecht Spaeth, W. H. Taliaferro, William H. Welch, D. Wright Wilson. Kansas.—/ Bennett M. Allen. Keo (Japna).—Naohidé Yatsu.—Leland Stanford.—Thomas Addis, Carl L. Alsberg, George D.) Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. J. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, & Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophiils, R. E. Swain, A. E. Taylor, E.B. Towne. Liverpool.—J. G. Adami. Maryland.—Charles E. Simon. McGill (Montreal).—Henry Gray Barbour, Horst Oertel, A. B. Macallum, John L. Todd. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, Alfred S. Warthin, Carl) Vernon Weller. Minnesota. —E. T. Bell, E. D. Brown, C. H. Eckles, George Fahr, C. P. Fitch, R. A. Gortner, H.) K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, C. M. Jackson, E. C. Kendall, F. E. Kingsbury, W.P. Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, J. F. McClendon, William Moore, Leroy S. Palmer, H. E. Robertson, R. E. Scammon, F.W. Schultz, F. H. Scott, E. C. Stakman, J. J. Willaman. Missouri—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.—W. deB. MacNider. Northwestern.—Solomon Strouse. Ohio State—R.G. Hoskins. Oregon.—D. E. Lancefield, C. F. Hodge, Harry B. Torrey. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, D. H. Bergey, Glen E. Allen, Samuel Goldschmidt, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., O. H. Perry Pepper, Stanley P. Reimann, Alfred N. Richards, J. Edwin Sweet. Pennsylvania State. —R. Adams Dutcher. Pitisburgh.—C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.— Edwin C. Conklin, E. Newton Harvey. Rochestey—John R. Murlin, G. H. Whipple. Rutgers——John F. Anderson, W. J. Crozier, A. R. Moore, Thurlow C. Nelson. Shefield—J. B. Leathes. Southern California (Los Angeles).—) Lyman B. Stookey. Springfield, Mass.—William B. Kirkham. St. Louis ——John Auer, Don R. Joseph, Ralph A. Kinsella. Texas.—Herman J. Muller, William C.Rose. Tohoka Imperial (Japan).—Shinkishi Hatai, C. K. Watanabe. T oronto. —A.H. Caulfeild, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane.—Charles W. Duval. Union University (Albany Medical College) Melvin Dresbach, Arthur Knudson. Vanderbilt (Nashville)—B. T. Terry. Virginia.—H. E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Eugene L. | Opie, Philip A. Shaffer. Wesleyan.—E.C. Schneider. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenbrey, H. | T. Karsner, J. M. Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.—F. E. Chidester, Withrow Morse. Wéisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Yale.—George A. Baitsell, R. H. Chittenden, J. W. Churchman, Ross G. Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo F. Rettger, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Industrial Laboratories: Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. Newark, N. J., Calco Chemical) Co.—M. S. Fine. New Brunswick, N. J., E. R. Squibb and Son.—P. A. Koher. Peekskill, N. Y., Fleischman Company. @ —A. K. Balls. Philadelphia, Pa., 1524 Chestnut St., H. K. Mulford Co.—T. S. Githens. Waterbury, Conn., Scovill Mfg. Co.—A. H. Ryan. Asheville, N. C., 43 Austin Ave-—Ephraim M. Ewing. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer: Los Angeles, Calif., 533 Lucerne Blud.—N. W. Janney. Missouri Botanical Gardens, St. Louis, Mo.—B. M. Duggar. Ossining, N. Y., R. F. D. 2.—H.D. Dakin. Pittsburgh, Pa., Jenkins Arcade.—Jacob Rosenbloom. Tuckahoe, N. Y.— Isaac F. Harris. Washington, D. C., 1701 Mass. Ave.—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St. Alexander L. Prince. Paris, France-—Harry Plotz. Berlin, Germany.—Reinhard Beutner, Rhoda Erdmann. K tanienbaum, Switzerland.—Fritz Schwyzer. : Members present at the one hundred nineteenth meeting: r Austin, Avery, Bailey, C. V., Burton-Opitz, Cohen, M., Eddy, Funk, Gates, Greenwald, Hastings, Hess, Jackson, H. C., Kopeloff, Levy, MacNeal, Myers, Pellini, Park, E. A., Salant, Sittenfield, Uhlenhuth, Van Slyke, Wallace, Zucker, | Members elected at the one hundred nineteenth meeting: S. Bayne-Jones, Guy W. Clark, George R. Cogwill, G. L. Foster, G. A. Friedman, Herbert Mann, Michael F Alfred Shohl. ; Dates of the next two meetings: January 18, 1922—February I5, 1922. PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED TWENTIETH MEETING COLLEGE OF PHYSICIANS AND SURGEONS : NEW YORK CITY JANUARY 18, 1922, VoLUME XIX No. 4 NEW YORK 1922 CONTENTS E. A. Park, P. G. SHIPLEY, E. V. McCoLtum AND Nina Srumonps: Is there more than one kind of rickets? 72 (1819) NICHOLAS KOPELOFF: Variations in aliquot fractions of gastric contents. 73 (1820). s WALTER H. Eppy, E. SHELOW AND R. A. PEASE: The effect of cooking upon the antiscorbutic vitamin in cabbage 74 (1821). J. HowarD MUELLER: New sulphur-containing amino acid isolated from casein. 75 (1822). C. C. LittLe AND B. W. JoHNSON: The inheritance of susceptibility to implants of splenic tissue in mice. 76 (18 ) T. F. ZucKErR, A. M. PAPPENHEIMER AND MARION BARNETT: Observations on cod liver oil and rickets. 77 (1824). — Be T. F. ZUCKER AND MARGARET B. GUTMAN: The distribution of inorganic phosphate of the blood between plasma anc# ii cells. 78 (1825). = M. B. GUTMAN AND V. KNEELAND FRANZ (by invitation): Observations on the inorganic phosphate of blood in « mental rickets of rats. 79 (1826). ; : ALFRED E. COHN AND ROBERT L. Levy: Experiments with quinidine on conduction and on the refiactory period in 1 dog’s heart. 80 (7827). Davip P. Barr: The acid base equilibrium ot the blood following vigorous musculai exercise. 81 (1828). R. J. BowEN, HELEN C. Coomss AND F. H. PIKE: The effect on blood pressure of removal of portions of the spinal in the thoracic region. 82 (1829). R. L. Kaun: A simple quantitative precipitation reaction for syphilis. 83 (1830). R. L. Kaun: A simple quantitative precipitation reaction for syphilis—Micro procedure. 84 (1831). 2 Davip I. Macut: Contribution to the chemico-pharmacodynamic relationship of atropine and homatropine. 85 (183 2) Davip I. MAcuT AND J. L. ULricH: Effect of prestatectomy on integration of muscular movements of the white ra 86 (1833). THoMasS B. OSBORNE AND LAFAYETTE B. MENDEL: Vitamin A in oranges. 87 (1834). E. B. KRUMBHAAR AND A. CHANUTIN: Studies in experimental plethora in dogs and rabbits. 88 (1835). : DONALD D. FORWARD AND Louis J. PERME (by invitation): Changes in total peripheral resistance during experimenta shock. 89 (1836). ci P. J. HANZLIK, Mary McINTYRE AND ELIZABETH PRESHO: Experimental plumbism: therapeutic efficiency of some ¢ and comparative toxicity of other metals. 90 (1837). The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after e meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of Octobe May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. ; The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume, three dollars each, postage p paid. The price of copies of the proceedings of any meeting is fifty cents each, postage prepaid. Subscriptions payable in advance. , PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. VICE-PRESIDENT—J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, New York, Post-Graduate Medical School, Alfred F. ¥ University and Bellevue Hospital Medical College, and ex-Presidents. : MANAGING Epitor—The Secretary-Treasurer, 338 East 26th St., New York City. : CLASSIFIED LIST_OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL ie BIOLOGY AND MEDICINE. ; Honorary. 1, William T. Councilman, Harvard University. Edward T. Reichert, University of Pennsylvania. : Resident (Greater New York). 4 College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. Columbia University—C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, _ George Draper, Walter H. Eddy, Andrew Eggstein, G. A. Friedman, William J. Gies, Louise H. Gregory, A. Baird Hast- ie ings, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. k “Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M. _ Richards, Mary S. Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, i ‘Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. ; Cornell University Medical College-—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert Chambers, A. F. Coca, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, _ Robert A. Hatcher, Graham Lusk, Walter L. Niles, Michael Ringer, Charles R. Stockard, W. C. Thro, John C. Torrey, le Fordham University School of Medicine.—Carl P. Sherwin. a Hospitals. Babies —Martha Wolstein. Beth Israel—Max Kahn, Morris H. Kahn. Long Island College -——Emil Goetsch. Montefiore Home.—Emil J. Baumann, George Fahr, B. S. Kline, Michael Levine, David Marine. Me. Sinai. —George Baehr, Burrill B. Crohn, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Rothschild. _ New York.—Nellis B. Foster, Ralph G. Stillman. Presbyterian.—Louis Baumann, A. R. Dochez, George M. Mackenzie, _ Walter W. Palmer, Franklin A. Stevens. Rooseveli—K. G. Faik, I. Greenwald, W. G. Lyle. St. Lukes.—L. W. . _ Famulener, Karl M. Vogel. 5} New York City Depariments. Healih—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Park, | Anna W. Williams, A. Zingher. Chief Medical Examiner.—Charles Norris. New York Homoeopathic Medical College—I. S. Kleiner, Mary B. Stark. New York Post-Graduate Medical School—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, "Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, : ton R. Rose, R. M. Taylor. ¢, ‘ New York University —W. H. Barber, Harlow Brooks, Warren Coleman, E. K. Dunham, Florence Hulton Frankel, "A. O. Gettler, Alfred F. Hess, Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Pro. V. Prewitt, Emil J. Pellini, H. D. Senior, Douglas Symmers, George B.Wallace. Psychiatric Institute —Nicholas Kopeloff. Rockefeller Institute for Medical Research.—Harold L. Amoss, O. T. Avery, Carl A. L. Binger, Wade H. Brown, “Alexis Carrel, A. E. Cohn, Rufus Cole, Paul DeKruif, Albert Fischer, Simon Flexner, F. L. Gates, Walter A. Jacobs, ia. J. Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Clara J. Lynch, Gustave M. Meyer, James B. Murphy, “3 ideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke. 4 Industrial Laboratories (New York City). Research Laboratory of H. A. Meiz.—Harry E. Dubin, Casimir Funk A earch Laboratory of H. A. Metz (Brooklyn).—Charles W. Hooper. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 17. Fe a St., N. Y. City—J. W. Draper. 126 E. 64th Si., N. Y. City—Cyrus W. Field. 141 W. 78th St., N. Y. City.— I. Ringer. ; 3 Non-Resident. _ Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick).— J. W. Shive, S. A. Waksman. t _ Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, pert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, arles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal, H. A. Spoehr. (Washington, D C. ).—Alfred G. Mayer. ei State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank Maltaner, A. B. Wadsworth. Hospitals. Addenbrooke's (Cambridge, England).—C. G. L. Wolf. Buffalo City.—Katharine R. Collins. The Cc ildren’s (Boston, Mass.).—Oscar M. Schloss. General (Cincinnati, Ohio)—Martin H. Fischer. Mercy (Pittsfield, Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Philadelphia G re eral.—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.).—Ralph Pemberton. Rikshospitalet (Kristiania, Nor- we Sie F. Hést. U.S. Public Health (Boston).—F.M.McCrudden. Western Pennsylvania (Pittsburgh).—Maurice H. Givens. ye _ Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton Springs).—Roger S. Hubbard. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. Gratwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago)—Herman M. Adler. Potter Meta- bolic Clinic (Santa Barbara, Calif.).—H. R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. Phipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F.S. Jones, Theobald Smith. Trudeau Sanotarium (Saranac Lake, N.Y.).—R. A. Kocher. Wistar (Philadelphia).—H. H. Donaldson. U.S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, J. R. Mohler. Hygienic Laboratory (Washington, D. C.).—Wm. C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, George B. Roth. Surgeon General's Office (Washington, D. C.).—J. F. Siler. National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Australia) —T. Brailsford Robertson. Amherst.—Otto Glaser. Baylor.—J. H. Black. Buffalo—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwith, W. R. Bloor, T. C. Burnett, H. M. Evans, E. C. Fleischner, G. L. Foster, F. P. Gay, Robert A. Gesell, Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles A. Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip @ E. Smith, E. L. Walker. Chicago—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati—D. E. Jackson, @ Shiro Tashiro. Copenhagen.—Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. George Washington—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Otte # Folin, Worth Hale, Lawrence J. Henderson, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph . | H. Pratt, M. J. Rosenau, Percy G. Stiles, Richard P. Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. IJllinoi —H. B. Lewis, W. F. Peterson, William H. Welker. Jowa State-——R. B. Gibson. Jefferson.—O. Bergeim, P. B. Haw Johns Hopkins.—John J. Abel, S. Bayne-Jones, C. G. Bull, George W. Corner, W. W. Cort, R. S. Cunningham, W. Ss. Halsted, R. W. Hegener, William H. Howell, John Howland, H. S. Jennings, Benjamin Kramer, Paul D. Lamson, W. G. MacCallum, David I. Macht, William S. McCann, E. K. Marshall, Jr., Wm. S. McCann, E. V. McCollum, Adolpk Meyer, Arthur L. Meyer, Raymond Pearl, G. C. Robinson, Paul G. Shipley, Reynold Albrecht Spaeth, W. H. Taliaferr id William H. Welch, D. Wright Wilson. Kansas.—Bennett M. Allen. Keo (Japan).—Noahidé Yatsu. Leland Stanford. —Thomas Addis, Carl L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. J. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophiils, R. E. Swain, A. E. Taylor, E. B. Towne. Liverpool.—J. G. Adami. Maryland.—Charles E. Simon. McGill (Monitreal).— Henry Gray Barbour, Horst Oertel, A. B. Macallum, John L. Todd. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Bro C. H. Eckles, George Fahr, C. P. Fitch, R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, C. M. Jackson, E. C. Kendall, F. E. Kingsbury, W. P. Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, J. F. McClendon, William | Moore, Leroy S. Palmer, H. E. Robertson, R. E. Scammon, F. W. Schultz, J. P. Schneider, F. H. Scott, E. C. Stakme ne J. J. Willaman. Missouri.—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.—W. deB. MacNider. § Northwestern.—Solomon Strouse. Ohio State-—R. G. Hoskins. Ovegon.—D. E. Lancefield, C. F. Hodge, Harry B fe | Torrey. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, Pe | Bergey, Glen E. Cullen, Samuel Goldschmidt, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., I | Perry Pepper, Stanley P. Reimann, Alfred N. Richards, J. Edwin Sweet. Pennsylvania State-—R. Adams he her. Pittsburgh.—C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.—Edwin G. Conklin, E. Nev v= | ton Harvey. Rochester—John R. Murlin, G. H. Whipple. Rutgers——John F. Anderson, W. J. Crozier, A. R. Moore, Thurlow C. Nelson. Sheffield.—J. B. Leathes. Southern California (Los Angeles)—Lyman B. Stookey. Spring (Mass.).—William B. Kirkham. St. Louis——John Auer, Don R. Joseph, Ralph A. Kinsella. Texas.—Herman J. M William C. Rose. Tohoka Imperial (Japan).—Shinkishi Hatai, C. K. Watanabe. Toronto.—A. H. Caulfield, J Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane.—Charles W. Duval. Union University (Albanj Medical College).—Melvin Dresbach, Arthur Knudson. Vanderbilt (Nashville).—B.T. Terry. Virginia.—H. E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Eugene L. Opie, Philip A. Shaffer Wesleyan.—E: C. Schneider. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenbrey, H. T. Karsner, J. W Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.—F. E. Chidester, Withrow Morse Wisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F, Clark, J. A. E. Eyster, Arthur S. Loevenhart Yale.—George A. Baitsell, R. H. Chittenden, J. W. Churchman, George R. Cowgill, Ross G. Harrison, R. A. Lamber Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo. F. Rettger, Alfred T. Shohl, Arthur HB Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. = Industrial Laboratories: Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. Newark, N. J., Calco Chemie Co.—M.S. Fine. New Brunswick, N. J., E. R. Squibb and Son.—P. A. Koher. Peekskill, N. Y., Fleischman Company, —A. K. Balls. Philadelphia, Pa., 1524 Chestnut St., H. K. Mulford Co.—T. S. Githens. Waterbury, Conn., Scovi Mfg. Co.—A. H. Ryan. ; Asheville, N. C., 43 Austin Ave-—Ephraim M. Ewing. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeifi Los Angeles, Calif., 533 Lucerne Blud—N. W. Janney. Missouri Botanical Garden, St. Louis, Mo.—B. M. Dugg Ossining, N. Y., R. F. D. 2.—H. D. Dakin. Pittsburgh, Pa., Jenkins Arcade.—Jacob Rosenbloom. Tuckahoe, N. ¥- Isaac F. Harris. Washington, D. C., 1701 Mass. Ave-—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St.—Alexan L. Prince. Paris, France.—Harry Plotz. Berlin, Germany.—Reinhard Beutner, Rhoda Erdmann. Kastanienbau Switzerland.—Fritz Schwyzer. i ¥ a SS a v4 7 ie a er cee ws Members present at the one hundred twentieth meeting: Bailey, C. V., Barr, Burton-Opitz, Cohn, A. E., Eddy, Edwards, Glaser, Greenwald, Friedman, Halsey, Hastin Hess, Jobling, Kopeloff, Levy, Lieb, Little, Lynch, Mueller, Myers, Pappenheimer, Prewitt, Ringer, M., Scott, E. | Scott, G. G., Sherwin, Stark, Stevens, Teague, Wallace, Williams, H. B., Zucker. a Members elected at the one hundred twentieth meeting: Rudolph J. Anderson, Clyde Brooks, J. Howard Brown, Harry D. Clough, William H. Cole, Karl S. Lashey, He A. Mattill, Ralph R. Mellon, Chauncey John V. Pettibone, Andrew Theodore Rasmussen, Lucius L. Van Slyke. — Dates of the next two meetings: February 15, 1922—March 15, 1922. PROCEEDINGS * OF THE i ; SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE = 25 ST ee Oh TY ae — en ee ul wae 4 ; ONE HUNDRED TWENTY-FIRST MEETING UNIVERSITY AND BELLEVUE HOSPITAL MEDICAL COLLEGE NEW YORK CITY FEBRUARY 15, 1922 AND THIRD MEETING | MINNESOTA BRANCH, , MINNEAPOLIS, MINNESOTA, FEBRUARY 8, 1922 VoLUME XIX No. 5 NEW YORK 1922 CONTENTS ISRAEL S. KLEINER: A method for the rapid determination of urea in minute amounts of blood. 91 (1838). Louris FREEDMAN and CASIMIR FUNK: The vitamine requirements of certain yeasts and bacteria. 92 (1839). CHARLES KRUMWIEDE and Lucy MISHULOW (by invitation): The existence of different immunological types of B. pertussis. 93 (1840). MARGARET S. KELLEY (by invitation): The applicability of the precipitin reaction in determining the infectivity discharge from gonorrheal infections. 94 (1841). SAMUEL A. BROWN and ALEXANDER O. GETTLER: A study of oxalic-acid poisoning. 95 (1842). : G. A. FRIEDMAN and J. GOTTESMAN: Further studies on ligation of the thyroid arteries in depancreatized dogit 96 (1843). _ G. A. FRIEDMAN and J. GOTTESMAN: The relation of the thyroid and parathyroids to pancreatic diabetes in dogs. 97 (1844). | WILLIAM DEB. MACNIDER: Concerning the amount and distribution of stainable lipoid material in renal epithelium in normal and acutely nephropathic animals, with observations on the functional response of the kidney. 98 (1845). Z. Bercovitz (by invitation): Preliminary report on the effects of vagus stimulation on the stomach of a dog and the influence of asphyxia of these effects. 99 (1846). | P. J. MoLoney and L. O. Hanna (by invitation): Agglutination phenomena with diphtheria antitoxin. 100 (1847). Davin I. MacuT and Gui CHING TinG: Action of some purin derivatives on the isolated bronchus. 101 (1848). Davip I. MAcHT and MARGUERITE LIVINGSTON: Effect of cocaine on the growth of Lupine Alba: a contribution to phyto-| pharmacology. 102 (1849). ALFRED F. Hess, LESTER J. UNGER and A. M. PAPPENHEIMER: Spontaneous cure of rickets in rats. 103 (1850). ¢ ALFRED F. Hess, LESTER J. UNGER and A. M. PAPPENHEIMER: A further report on the prevention of rickets in rats by light rays. 104 (1851). 7 JouN F. ANDERSON: A note on the preparation of anti-colon streptococcus serum. 105 (1852). JoHN WILLIAM DRAPER: The use of a colon-streptococcus anti-serum as a preoperation measure. 106 (1853). RALPH E. MELLON: Spontaneous agglutinability of bacteria in relation to the antagonistic action of certain cations. 107 (1854). STANLEY Ross BurRLAGE (by invitation): Blood pressures and heart rate in girls during adolescence. A preliminary study of 1,700 cases. 108 (1855). Cari O. LATHROP and CHARLES A. BENTZ (by invitation): Serological nsiding of the diphtheria group. 109 (1856). M. S. Doorey and C. B. HIGLey (by invitation): An intramuscular method of digitalis assay. 110 (1857). Ross AIKEN GORTNER and W. F. HorFMAN: Evidences of structure in gelatin gels. 111 (1858). C. C. GAULT and F. H. Scott: The control of respiration. 112 (1859). . A. T. Henrici and G. S. REYNOLDS: Potassium iodide does not influence the course of an experimental actinomycosis, bi 113 (1860). cm | R. A. GorTNER and W. F. HorrMan: Evidence of a structure in gelatin gels. 114 (1861). | | The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after eac meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of October May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume, three dollars each, postage pre paid. The price of copies of the es of any meeting is fifty cents each, postage prepaid. Subscriptions are payable in advance. PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. ViICE-PRESIDENT—J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, New York, Post-Graduate Medical School, Alfred F. E University and Bellevue Hospital Medical College, and ex-Presidents. MANAGING Epitor—The Secretary-Treasurer, 338 East 26th St., New York City. CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Honorary. William T. Councilman, Harvard University. Edward T. Reichert, University of Pennsylvania. William H. Welch. Resident (Greater New York). . College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, T. A. Storey. \ Columbia University.—C. H. Bailey, Russell Burton-Opitz, Gary N. Calkins, Helen C. Coombs, Maynie R. Curtis, _ George Draper, Walter H. Eddy, Andrew Eggstein, G. A. Friedman, William J. Gies, Louise H. Gregory, _ J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. - Morgan, J. Howard Mueller, B. S. Oppenheimer, Alwin M. Pappenheimer, Julia T. Parker, F. H. Pike, Herbert M. _ Richards, Mary S. Rose, E. L. Scott, Henry C. Sherman, M. J. Sittenfield, A. H. Sturtevant, H. F. Swift, Oscar Teague, _ Arthur W. Thomas, H. B. Williams, Edmund B. Wilson, Francis C. Wood, Hans Zinsser, Theodore F. Zucker. is Cornell University Medical College.-—H. L. Alexander, Harold Bailey, David P. Barr, Stanley R. Benedict, Robert _ Chambers, A. F. Coca, E. F. DuBois, D. J. Edwards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, _ Robert A. Hatcher, Graham Lusk, Walter L. Niles, Michael Ringer, Charles R. Stockard, W. C. Thro, John C. Torrey. F Fordham University School of Medicine.—Carl P. Sherwin. fe Hospitals. Babies—Martha Wollstein. Beth Israel—Max Kahn, Morris H. Kahn. Long Island College-—Eme _ Goetsch. Montefiore Home.—Emil J. Baumann, B. S. Kline, Michael Levine, David Marine. Mt. Sinai.—Georgd } Baehr, Burrill B. Crohn, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Rothschile, I _ New York.—Nellis B. Foster, Ralph G. Stillman. Presbyterian.—Louis Baumann, A. R. Dochez, George M. Mackenzie, _ Walter W. Palmer, Franklin A. Stevens. Roosevelt.—K. G. Falk, I. Greenwald, W. G. Lyle. St. Lukes.—L. W. _ Famulener, Karl M. Vogel. NY New York City Depariments. Healih—James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, William H. Park, ks Anna W. Williams, A. Zingher. Chief Medical Examiner.—Charles Norris. i New York Homoeopathic Medical College—I. S. Kleiner, Mary B. Stark. 5 New York Post-Graduate Medical School.—C. V. Bailey, Arthur F. Chace, Martin Cohen, C. Ward Crampton, _ Robert H. Halsey, Ludwig Kast, John A. Killian, W. J. MacNeal, H. O. Mosenthal, V. C. Myers, Marshall C. Pease, _ Anton R. Rose, R. M. Taylor. % New York University.—W. H. Barber, Harlow Brooks, Warren Coleman, E. K. Dunham, Florence Hulton Frankel, _A. O. Gettler, Alfred F. Hess, Holmes C. Jackson, Isaac Levin, Arthur R. Mandel, John A. Mandel, W. C. Noble, Emil J. Pellini, P. V. Prewitt, H. D. Senior, Douglas Symmers, George B. Wallace. Psychiatric Institute-—Nicholas Kopeloff. Rockefeller Institute for Medical Research.—Harold L. Amoss, O. T. Avery, Carl A. L. Binger, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, Paul DeKruif, Albert Fischer, Simon Flexner, F. L. Gates, A. Baird Hastings, Walter A. Jacobs, I. J. Kligler, P. A. Levene, Robert L. Levy, Jacques Loeb, Christian Lundsgaard, Clara J. Lynch, ‘(Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, ‘Eduard Uhlenhuth, Donald D. Van Slyke. Industrial Laboratories (New York City). Research Laboratory of H. A. Metz—Harry E. Dubin, Casimir Funk, Research Laboratory of H. A. Metz (Brooklyn).—Charles W. Hooper. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 17 E. 38th St., N. Y. City —J. W. Draper. 126 E. 64th St., N. Y. City—Cyrus W. Field. 141 W. 78th St., N. Y. City.— A. I. Ringer. 3 ny — ee Non-Resident. Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick) .— J. W. Shive, S. A. Waksman. New York (Geneva).—Rudolph J. Anderson, L. L. Van Slyke. Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, Albert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, Charles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal, H. A. Spoehr. (Washington, D. C.).— Alfred G. Mayer. State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank Maltaner, A. B. Wadsworth. ‘ Hospitals. Addenbrooke’s (Cambridge, England).—C. G. L. Wolf. Buffalo City.—Katharine R. Collins. The Children’s (Boston, Mass.)\—Oscar M. Schloss. General (Cincinnati, Ohio).—Martin H. Fischer. Highland (Rochester, N. Y.).—Ralph R. Mellon. Mercy (Pittsfield, Mass.).—Thomas F lournoy. Peter Bent Brigham (Boston.)—Harvey Cushing, Francis W. Peabody. Philadelphia General.—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.).—Ralph Pemberton. Rikshospitalet (Kristiania, Norway).—H. F. Host. U. S. Public Health (Boston).—F. M. McCrudden. Western Pennsylvania (Pittsburgh).—Maurice H. Givens. Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton éngs).—Roger S. Hubbard. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. r atwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago).—Herman M. Adler. Potter Meta- oléc Clinic (Santa Barbara, Calif.).—H. R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. Phipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—J. Howard Brown, Paul E. Howe, F. S. Jones, Theobald Smith. Wistar (Philadelphia).—H. H. Donaldson. U. S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, J. R Mohler. Bureau of Entomology (Riverton, N. J.)—William Moore. Hygienic Laboratory (Washington, D. C.).—Wm C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, George B. Roth. Surgeon General's Office (Washington, D. C.). J. F. Siler. National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Australia).—T. Brailsford Robertson. Alabama.—Clyde Brooks. Amherst.—O Glaser. Baylor.—J. H. Black. Buffalo—Herbert U. Williams. California.—Walter C. Alvarez, T. D. Beckwitk W. R. Bloor, T. C. Burnett, Guy W. Clark, H. M. Evans, E. C. Fleischner, G. L. Foster, F. P. Gay, Robert A. Gesel Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles A. Kofoid, Lovell Langstroth, W. P. Lucas, S. S. Me 1 K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D.E. Jackson, Shiro Tashiro. Cornell.—Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. George WashingtonW. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Otto Folin, Worth Hale, Lawrence J. Henderson, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Percy G. Stiles, Richard P. Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. Jilinois. —H. B. Lewis, W. F. Peterson, William H. Welker. Jowa State-——R. B. Gibson. Jefferson.—O. Bergeim, P. B. Hawk. § Johns Hopkins.—John J. Abel, S. Bayne-Jones, C. G. Bull, George W. Corner, W. W. Cort, R. S. Cunningham, W. S. Halsted, R. W. Hegener, William H. Howell, John Howland, H. S. Jennings, Benjamin Kramer, Paul D. Lamson, W. G. MacCallum, David I. Macht, E. K. Marshall, Jr., William S. McCann, E. V. McCollum, Adolph Meyer, Arthu L. Meyer, Raymond Pearl, G. C. Robinson, Paul G. Shipley, Reynold Albrecht Spaeth, W. H. Taliaferro, D. Wright: Wilson. Kansas.—Bennett M. Allen. Keo (Japan).—Noahidé Yatsu. Leland Stanford.—Thomas Addis, Carl L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, P. J. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. Ophiils, R. E. Swain, A. E. Taylor, E. B. Towne. § Liverpool.—J. G. Adami. Maryland.—Charles E. Simon. McGill (Montreal).—Henry Gray Barbour, Horst Oertel, § A. B. Macallum, John L. Todd. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, § Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Brown, C. H. Eckles, George Fahr, C. P. Fitch, § R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, C. M. Jackson, E. C. Kendall, F. E. Kingsbury K. S. Lashley, W. P. Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, J. F. McClendon, William Moore, Leroy Palmer, C. J. V. Pettibone, H. E. Robertson, R. E. Scammon, J. P. Schneider, F. W. Schultz, F. H. Scott, E. Co) Stakman, J. J. Willaman. Missouri—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.—W. deB MacNider. Northwestern.—Solomon Strouse. Ohio State-—R. G. Hoskins. Oregon.—D. E. Lancefield, C. F. Hod Harry Beal Torrey. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. Austin, D. H. Bergey, Glen E. Cullen, Samuel Goldschmidt, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Mus Jr., O. H. Perry Pepper, Stanley P. Reimann, Alfred N. Richards, J. Edwin Sweet. Pennsylvania State-—R. Ad: Dutcher. Pittsburgh—C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.—Edwin G Conklin, E. Newton Harvey. Rochester—John R. Murlin, G. H. Whipple. Rutgers.—John F. Anderson, W. J. Crozie: A. R. Moore, Thurlow C. Nelson. Sheffield—J. B. Leathes. Southern California (Los Angeles).—Lyman A. O. She Texas.—Herman J. Muller, William C. Rose. Tohoku Imperial (Japan).—Shinkishi Hatai, C. K. Watanabe. Tora —A. H. Caulfield, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane——Charles W. Duval. Unto University (Albany Medical College) —Melvin Dresbach, Arthur Knudson. Vanderbilt (Nashville.—B. T. Terry Virginia.—H. E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Euger L. Opie, Philip A. Shaffer. Wesleyan.—E. C. Schneider. Western Reserve (Cleveland).—George W. Crile, A. B. Eiset brey, H. T. Karsner, J. M. Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.—F. Chidester, Withrow Morse. Wéisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyste Arthur S. Loevenhart. Yale—George A. Baitsell, R. H. Chittenden, J. W. Churchman, George R. Cowgill, Ross € Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo. F. Rettger, Alfre T. Shohl, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Industrial Laboratories: Indianapolis, Ind., Eli Lilly and Co.—G. H. A. Clowes. Newark, N. J., Calco Che Co.—M. S. Fine. New Brunswick, N. J., E. R. Squibb and Son.—P. A. Koher. Peekskill, N. Y., Fleischman Comt —A. K. Balls. Philadelphia, Pa., 1524 Chestnut St., H. K. Mulford Co.—T. S. Githens. Waterbury, Conn., Mfg. Co.—A. H. Ryan. Asheville, N. C., 43 Austin Ave-—Ephraim M. Ewing. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiff Los Angeles, Calif., 533 Lucerne Blud.—N. W. Janney. Missouri Botanical Garden, St. Louis, Mo.—B. M. Dugg Ossining, N. Y., R. F. D. 2.—H. D. Dakin. Pittsburgh, Pa., Jenkins Arcade.—Jacob Rosenbloom. Tuckahoe, N. ¥. Isaac F. Harris. Washington, D. C., 1801 Mass. Ave.—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St.—Ale: L. Prince. Paris, France.—Harry Plotz. Berlin, Germany.—Reinhard Beutner, Rhoda Erdmann. Kastanie: Switzerland, Fritz Schwyzer. Members present at the one hundred twenty-first meeting: Anderson, Draper, Eddy, Gettler, Jackson, H. C., Kopeloff, Levin, I., Maltaner, Mueller, Myers, MacNeal, F Rose, A. R., Teague, Torrey, Wallace. Members elected at the one hundred twenty-first meeting: S. A. Goldberg, George L. Hoffman, L. A. Maynard, Fred T. Rogers, R. W. Thatcher, W. S. Thomas me PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED TWENTY-SECOND MEETING PRESBYTERIAN HOSPITAL NEW YORK CITY MARCH 15, 1922 AND THIRTY-SECOND MEETING PACIFIC COAST BRANCH, SAN FRANCISCO, CALIFORNIA FEBRUARY 15, 1922 VOLUME XIX No. 6 NEW YORK 1922 CONTENTS Max Kaun: The feeding of non-ketogenic odd-carbon fats to diabetic patients. 115 (1862). JOHN R. WriiiaMs and MADELEINE SweETT (by invitation): Hydrogen ion concentration studies of solutions used for intravenous medication and clinical investigation. 116 (1863). GEORGE M. MACKENZIE and EMILY FRUHBAUER: On the mechanism by which antigen is removed from the circulation. . 117 (1864). FRANKLIN A. STEVENS and CLiFForD Lamar: The effect of various proteins on streptolysin production. 118 (1865). Henry A. Murray (by invitation): The bicarbonate and chloride content of the blood in certain cases of permistents ¥ vomiting. 119 (1866). bh . Harry BEAL TORREY and BENJAMIN HORNING: Hen feathering induced in the male fowl by feeding thyroid. 120 (1867). GrorGE Harrop (by invitation) : The relation of the diffusion constant to mountain sickness. 121 (1868). Oscar Riwpe: An undescribed relation of the suprarenals to ovulation. 122 (1869). : GEORGE R. CowcGILt: A comparison of the effects of feeding extracts of muscle and yeast respectively. 123 (1870). GEORGE R. CowGILL: Parenteral administration of products containing vitamin-B—Mammalian experiments. 124 (18712). S. C. Brooks: The conductance of unicellular organisms. 125 (1872). RALPH H. Boots and GLENN E. CULLEN: The hydrogen ion concentrations of joint exudates in acute arthritis. 126 (1873). a & L. L. WoopruFF and Hope SPENCER: On the method of macronuclear dissolution during endomixis in Paramecium aurelia. 128 (1875). THOMAS B. OSBORNE and LAFAYETTE B. MENDEL: Further observations on the occurrence of vitamin B. 129 (1876). Joun W. CHURCHMAN: The selective bactericidal effect of acid fuchsin and sodium chloride. 127 (1874). Rocer S. HUBBARD and Davin C. WiLson: An experiment on the absorption of glucose given by rectum. 130 (2877). R. L. Kaun: Effect of dilution on the precipitation reaction for syphilis proposed by author. 131 (1878). t R. L. Kaun: Relation between serum and antigen in precipitation reaction for syphilis proposed by author. 132 (1879). J. BRONFENBRENNER and H. Weiss: The use of morphine in connection with serumtherapy of botulism. 133 (18808 on Zz | CHARLES W. YounG and HELEN M. Van SANT (by invitation) : The diagnosis of Kala-Azar by blood culture. 135 (1882). | J. BRONFENBRENNER and M. J. SCHLESINGER: The state of aggregation of particles of botulinus toxin. 134 (1881). 2 t i PauL J. HaNzLikK and Howarp T. KaArsNeErR: Further observations on anaphylactoid phenomena from different agen including histamin. 136 (1883). P, J. HANzLIK, FLoyp DE Eps and ELIZABETH PRESHO: Urinary excretion of salicyl after the administration of sali and salicyl esters. 137 (1884). eI JEAN OLIVER and So SaBro YAMaDA: The effect of the administration of salvarsan in combination with various colloid 4 bo substances on its toxicity. 138 (1885). a: E. B. Towne: The so-called permanent polyuria of experimental diabetes insipidus. 139 (886). The Proceedings of the Society for Experimental Biology and Medicine are published as soon as possible after ez meeting. Regular meetings of the Society are held in New York on the third Wednesday of the months of Octobe May inclusive. A volume of the Proceedings consists of the numbers issued during an academic year. The price of Vols. I, II and III is five dollars each, of Vol. IV to current volume, three dollars each, postage p paid. The price of copies of the proceedings of any meeting is fifty cents each, postage prepaid. Subscriptions ¢ payable in advance. - 2] . PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. VICE-PRESIDENT—J. W. Jobling, Columbia University. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—V. C. Myers, New York, Post-Graduate Medical School, Alfred Fi B University and Bellevue Hospital Medical College, and ex-Presidents. =% . af AS ae Non-Resident. Agricultural Experiment Stations. Connecticut (New Haven).—Thomas B. Osborne. New Jersey (New Brunswick).— J. W. Shive, S. A. Waksman. New York (Geneva).—Rudolph J. Anderson, R. W. Thatcher, L. L. Van Slyke. Carnegie Institution of Washington. (Station for Experimental Evolution, Cold Spring Harbor, N. Y.).—A. M. Banta, Albert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little, E. Carleton MacDowell, Charles W. Metz, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D.T. MacDougal. (Washington, D. C.).—Alfred G. Mayer. State Boards of Health. Michigan (Lansing).—R. L. Kahn. New York (Albany).—Mary B. Kirkbride, Frank -Maltaner, A. B. Wadsworth. Hospitals. Addenbrooke's (Cambridge, England).—C. G. L. Wolf. Buffalo City.—Katharine R. Collins. The Children’s (Boston, Mass).—Oscar M. Schloss. General (Cincinnati, Ohio).—Martin H. Fischer. Mercy (Pittsfield, Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Philadelphia General.—E. B. Krumbhaar. Presbyterian (Philadelphia, Pa.)—Ralph Pemberton. Rikshospitalet (Kristiania, Nor- ) way).—H. F. Host. U.S. Public Health (Boston).—F. M. McCrudden. Western Pennsylvania (Pitisburgh).—Maurice H. Givens, George L. Hoffman. Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Clifton Springs Sanitarium (Clifton ) Springs).—Roger S. Hubbard, Walter S. Thomas. Dermatological Research Institute (Philadelphia).—George W. Raiziss, Charles Weiss. Gratwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago).—Herman M. Adler. § ) Potter Metabolic Clinic (Santa Barbara, Calif.)—H. R. Blatherwick. Nela Research Laboratory (Cleveland, Ohio).—Ralph S. Lillie. Phipps (Philadelphia).—Paul A. Lewis. Rockefeller (Princeton).—J. Howard Brown, Paul E. Howe, F. S. prs. ior Smith. Trudeau Sanatorium (Saranac Lake, N. Y.).—R. A. Kocher. Wistar (Philadelphia).— H. H. Donaldson. =. U.S. Departments. Bureau of Animal Industry (Washington, D. C.).—William N. Berg, Edward B. Meigs, James — M. Sherman. Bureau of Entomology (Riverton, N. J.).—William Moore. Hygienic Laboratory (Washington, D. C.).—_ Wm. C. Boeck, C. S. Brooks, Barnett Cohen, J. P. Leake, George B. Roth. Surgeon General's Office (Washington, D. C.). —J. F. Siler. National Research Council, Washington, D. C.—V. L. Kellogg, Clarence J. West. Universities. Adelaide (South Australia).—T. Rrailsford Robertson. Alabama.—Clyde Brooks. Amherst.—Otto | Glaser. Baylor.—J. H. Black, Fred T. Rogers. Buffalo.—Herbert U. Williams. California.—Walter C. Alvarez, T. ] D. Beckwith, W. R. Bloor, T. C. Burnett, Guy W. Clark, H. M. Evans, E. C. Fleischner, G. L. Foster, F. P. Gay, Robert | A. Gesell, Ivan C. Hall, S. J. Holmes, Samuel H. Hurwitz, Charles A. Kofoid, Lovell Langstroth, W, P. Lucas, S.S. | Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker. Chicago.—A. J. Carlson, Frank R. Lillie, } Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Copenhagen.—Christen Lundsgaard. Cornell— P. A. Fish, S. A. Goldberg, L. A. Maynard, Sutherland Simpson. Georgia.—Richard V. Lamar, William Salant. George | Washington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B. Cannon, Worth Hale, Lawrence J. Hender- son, Reid Hunt, W. J. V. Osterhout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Percy G. Stiles, Richard P. Strong, Fritz B. Talbot, E. E. Tyzzer, S. Burt Wolbach. Iilinois.—H. B. Lewis, W. F. Peterson, William H. Welker. Jowa Siate-—R.B. Gibson. Jefferson.—O. Bergeim, P. B. Hawk. Johns Hopkins.—John J. Abel, S. Bayne- } Jones, C. G. Bull, George W. Corner, R. S. Cunningham, W. S. Halsted, William H. Howell, John Howland, H. S. ball, | nings, Benjamin.Kramer, Paul D. Lamson, W. G. MacCallum, David I. Macht, William S. McCann, E. K. Marshall, _ Jr., Wm. S. McCann, E. V. McCollum, Adolph Meyer, Arthur L. Meyer, Raymond Pearl, G. C. Robinson, Paul Gr Shipley, Reynold Albrecht Spaeth, D. Wright Wilson. Kansas.—Bennett M. Allen. Keo ‘Japan).—Noahidé Yatsu. — Leland Stanford.—Thomas Addis, Carl L. Alsberg, George D. Barnett, J. F. Cowan, E. C. Dickson, Harold K. Faber, at . P. J. Hanzlik, A. W. Hewlett, W. L. Holman, W. H. Manwaring, Ernest G. Martin, H. G. Mehrtens, Jean Oliver, W. — Ophiils, R. E. Swain, A. E. Taylor, E. B. Towne. Liverpool.—J. G. Adami. McGill (Montreal).—Henry Gray Barbour, — Horst Oertel, A. B. Macailum. Michigan.—C. W. Edmunds, G. Carl Huber, Warren P. Lombard, Frederick G. Novgaae Alfred S. Warthin, Carl Vernon Weller. Minnesota.—E. T. Bell, E. D. Brown, C. H. Eckles, George Fahr, C. P. Fitch, |} R. A. Gortner, H. K. Hayes, Arthur T. Henrici, A. D. Hirschfelder, C. M. Jackson, E. C. Kendall, F. E. Kingsbury, : oe S. Lashley, W. P. Larson, E. J. Lund, E. P. Lyon, Frank C. Mann, J. F. McClendon, Leroy S. Palmer, C. J. V. Petti- mm |. bone, A. T. Rasmussen, H. E. Robertson, R. E. Scammon, F. W. Schultz, J. P. Schneider, F. H. Scott, E. C. Stakma J. J Willaman. Missouri.i—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.—W. deB. MacNiders| Northwestern—Solomon Strouse. Ohio State-—R. G. Hoskins. Oregon.—D. E. Lancefield, C. F. Hodge, Harry Bea Torrey. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, D. H. Bergey, Glen E. Cullen, Samuel Goldschmidt, B. M. Hendrix, J. A. Kolmer, Baldwin Lucké, J. H. Musser, Jr., O. H. Pe Pepper, Stanley P. Reimann, Alfred N. Richards, J. Edwin Sweet. Pennsylvania State—R. Adams Dutcher. Pittsburgh. C. C. Guthrie, Davenport Hooker, Oskar Klotz, J. W. McMeans. Princeton.—Edwin G. Conklin, E. Newton Harve Rochester.—Henry A. Mattill, John R. Murlin, G.H. Whipple. Rutgers.—John F. Anderson, W. J. Crozier, A. R. Moc Thurlow C. Nelson. Southern California (Los Angeles).—Lyman B. Stookey. Springfield (Mass.).—William B. ham. St. Louis.—John Auer, Don R. Joseph, Ralph A. Kinsella, A. O. Shaklee. Texas.—Herman J. Muller, Willi C. Rose. Syracuse.-—M.S. Dooley, Frank P. Knowlton. Tohoka Imperial (Japan).—Shinkishi Hatai, C. K. Watana Toronto.—A. H. Caulfield, J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane.—Charles W. Duv Union University (Albany Medical College).—Melvin Dresbach, Arthur Knudson. Vanderbilt (Nashville).—B. T. Te Virginia.—H. E. Jordan. Washington (St. Louis).—M. T. Burrows, J. V. Cooke, Joseph Erlanger, Leo Loeb, Eugene L Opie, Philip A. Shaffer. Wesleyan.—E. C. Schneider. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenb: H. T. Karsner, J. M. Rogoff, R. W. Scott, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.—F. E. Ch dester, Withrow Morse. Wisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthu S. Loevenhart. Yale.—George A. Baitsell, R. H. Chittenden, J. W. Churchman, George R. Cowgill, Ross G. Ha on R. A. Lambert, Henry Laurens, Lafayette B. Mendel, E. A. Park, John P. Peters, Jr., Leo F. Rettger, Alfred T. She Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Industrial Laboratories:—Glenolden, Pa.; H. K. Mulford Co.—T. S. Githens. Indianapolis, Ind., Eli Lilly and Co —G. H. A. Clowes. Newark, N. J., Calco Chemical Co.—M. S. Fine. New Brunswick, N. J., E. R. Squibb and Son. P. A. Koher. Peekskill, N. Y., Fleischman Company—A. K. Balls. Waterbury, Conn., Scovill Mfg. Co.—A. H. Ryan. Baltimore, Md., 1421 Edmonson Ave.—J. A. F. Pfeiffer. Los Angeles, Calif., 533 Lucerne Blud.—N. W. Jann Missouri Botanical Garden, St. Louis, Mo.—B. M. Duggar. Napoleonville, La —Ephraim M. Ewing. Ossining, Y., R. F. D. 2.—H.D. Dakin. Pittsburgh, Pa., Jenkins Arcade.—Jacob Rosenbloom. Tuckahoe, N. Y.—Isaac F. He Washington, D. C., 1701 Mass. Ave.—Robert M. Yerkes. Wethersfield, Conn., 4 Wilcox St.—Alexander L. Prince. Pé France.—Harry Plotz. Berlin, Germany.—Reinhard Beutner, Rhoda Erdmann. Kastanienbaum, Switzerland.—¥ Schwyzer. A A A Se AN Members present at the one hundred twenty-third meeting: Bailey, C. V., Baumann, E. J., Browne, W. W., Chambers, Churchman, DuBois, Eddy, Edwards, Famulener, F : man, Hess, Hooper, Jackson, H. C., Jobling, MacNeal, Mann, H., Marine, Myers, Noble, Prewitt, Ringer, Rose, A. B } Ryan, Scott, G. A., Sherwin, Thro, Torrey, J. C., Wallace, Winslow. |) Members elected at the one hundred twenty-third meeting: re Ralph Boots, Moyer S. Fleisher, George Harrup, Jr., Albert Kuntz, August G. Pohlman, William D. Sansum, J. omas. PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED TWENTY-FOURTH MEETING COLUMBIA UNIVERSITY NEW YORK CITY MAY 17, 1922 AND FIFTH MEETING MINNESOTA BRANCH MINNEAPOLIS, MINNESOTA MAY 10, 1922 AND SECOND MEETING WESTERN NEW YORK BRANCH ITHACA, NEW YORK MAY 20, 1922 VoLUME XIX No. 8 NEW YORK 1922 CONTENTS L. W. FAMULENER and JuLia A. W. Hewitt: The Hecht-Weinberg-Gradwohl reaction in syphilis. 177 (1924). Z { L. W. FAMULENER and LucILE ROBEY: Pneumococcus grouping on 1000 cases. 178 (1925). es iN Jutia A. W. Hewitt and L. W. FAMULENER: The hemolytic properties of the pneumococcus. 179 (1926). = os | NiIcHOLAS Kope.orF and C. O. CHENEY: Studies on the therapeutic effect of B. acidophilus milk and lactose. 180 (1927). | CHARLES W. Metz and Jos& F. NONIDEz: Observations on the behavior of the nucleus and chromosomes in spermato of Lasiopogon (diptera). 181 (1928). A. M. YupDKIN and R. A. LAMBERT: Location of the earliest changes in experimental xerophthalmia of rats. 182 (1929). A. M. YupKIN and R. A. LAMBERT: Lesions in the lacrimal glands of rats in experimental xerophthalmia. 183 (1930). | HANNAH ELIZABETH HONEYWELL and OscAR RIDDLE: Increased blood sugar coincident with ovulation in pigease a 184 (1931). ALFRED F. Hess and MARION A. LUNDAGEN: Seasonal tide of blood phosphate in infants. 185 (1932). 3: a D. J. Epwarps and H. J. Bacc: Localized lesions in the corpora striata produced by buried radium emanation. 186 = | : (1933). a HUBERT MANN: An hypothesis of the mechanism by which normal rhythm is restored in atrial fibrillation. 187 (1938). 2 M Soma Weiss and Ropert A. HATCHER: The emetic action of antimony and potassium (tartar emetic). 188 (1935). 7k G. A. FRIEDMAN and J. GOTTESMAN: The effect of thyroidectomy in two sittings upon depancreatized, non-glycosurie, “