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Veg pen re bhi hee aii ew jane ae hae idea setae a ’ O° ge “4 F ve phpew cute Se mn mit int gigs Pee Tr ie rye wake ei 4 ; j ta» 8 OO cy CHEE Rite apr indy pear garag ow bbe byte Pe ete baectae Re Sp acd, Aagh whet) 6d . yatane ‘ Peer Mer rire brs rere ea ae - ‘ > .4.7 3 He we dy at sates says Suet stot Tdvautene uh ae me Rees. Keehaeetts a ee he »- To's a < yaa rr? vee DONOR gepertd ere eg ae ath pe laet a . . gta ‘ ' ei etubigagears {debe bs va arate: ng 4 ran ere. ae hy ayy q ‘ . pee} ir * et OU ee eh) P e's FY «eae Bad A hae jaan o er, SU ee a | FOR THE PEOPLE FOR EDVCATION FOR SCIENCE LIBRARY OF THE AMERICAN MUSEUM OF NATURAL HISTORY . Pa h ot ae aN With leva : i i Wap ea ih ra i hi A t tH Wy 7 ie if) Wa an A ais s/i Mii a) PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE VOLUME XVII 1919-1920 EDITED BY THE SECRETARY NEW YORK 1920 SE reoek ae \3 ? ee ¥ «2 = s — cU ~ ae ot | CONTENTS. PAGE SCIENTIFIC PROCEEDINGS (10Ist-108th meetings): Communications of the one hundred first meeting . I Communications of the one hundred second meeting 29 Communications of the one hundred third meeting . 49 Communications of the one hundred fourth meeting 65 Communications of the one hundred fifth meeting . 87 Communications of the one hundred sixth meeting . II5 Communications of the one hundred seventh meeting 143 Communications of the one hundred eighth meeting ae api RECAPITULATION OF THE NAMES OF THE AUTHORS AND OF THE ia ies OF THE COMMUNICATIONS 222 EXECUTIVE PROCEEDINGS Gan pote Beran) ; “B35 REGISTER OF NAMES AND ADDRESSES OF THE MEMBERS : 238 LIST OF OFFICERS ; . 247 CLASSIFIED LIST OF MEMBERS . 248 INDEX OF THE SCIENTIFIC PROCEEDINGS iii ot ie A < soe prem SCIENTIFIC PROCEEDINGS. One hundred first meeting. ABSTRACTS OF COMMUNICATIONS. Cornell University Medical College, October 15, I9I9. President Calkins in the chatr. I (1461) The developmental stages at which mutations occur in the germ tract. By CaLvIN B. BRIDGEs (by invitation). [From the Zoélogical Laboratory, Columbia University.] In the fruit-fly Drosophila melanogaster about 300 primary mutations have been found, most of which arose in cultures car- ried on in the laboratory. A study of the critical cases among these mutations has shown that a large majority of them origi- nated at or very near the maturation stage; that a few occurred in the gonial cells some time prior to maturation; and that a few occurred early in the segmentation stage. The conclusion that most mutations occur at the maturation stage is based largely on the proportion of sex-linked recessives and of dominants that have been first found as a single individuai. Approximately half of the sex-linked recessives have been dis- covered as a single male. This is a surprisingly large proportion and clearly means that in these cases the actual mutation occurred in the mother, and at, or not more than a very few cell generations before, the maturation of the egg. Those sex-linked recessives that did not first appear as a single male have in the main appeared as half the sons of a female already heterozygous for the gene. In these cases the actual mutation had occurred at some inde- terminate stage one or more generations previous to the appearance I 2 SCIENTIFIC PROCEEDINGS (101). of the character. There are now about 30 known dominants in Drosophila melanogaster, of which fully two-thirds were first found as single heterozygous individuals. This very large proportion of dominants appearing as single individuals means that the actual mutation has occurred very close to the final stage in the forma- tion of the gamete—probably little if any prior to maturation. That mutations may occur in the oégonial cells prior to matura- tion is proved by a few cases in which a single female has given rise to more than a single individual of a new sex-linked recessive character. In the first of these cases the mutant called ‘‘cut”’ occurred as six males among the 131 sons of a particular female. The mutation “tiny-wing’’ occurred as two sons among about 150; ‘‘sable-duplication”’ occurred as three males among 133; and “ivory” as about Io percent. of the sons. There are one or two other such cases known. In all of these cases the facts are in accord with the hypothesis that the actual mutation occurred in the odgonial cells of particular females, and from one to a few cell-generations prior to maturation. ‘There has been one case in which a female homozygous for an autosomal recessive was outcrossed to a wild- type male, and produced among 61 offspring 6 flies that were heterozygous for a new allelomorph of the recessive. The muta- tion responsible for the new allelomorph occurred in the wild- type male in the spermatogonial stage far enough previous to spermatogenesis so that approximately a sixth of the sperm carried the gene. The proportion of gametes that carry the mutant character corresponds to the time previous to maturation at which the particular mutation occurred. That mutation may occur in the zygote immediately after fertilization is proved by the discovery of nearly a dozen mosaic individuals in which a new mutant has appeared as part of a fly. If the new mutant is a dominant it may appear as part of a female, and if a sex-linked recessive as part of a male. Such mosaics arise by mutation in the zygote, and the parts descended from the mutant cell show the new character while the remainder of the animal shows the original type. HypROGEN ION CONCENTRATION OF ACID SOLUTIONS. 3 2 (1462) Contrasting effects of chlorides and sulphates on the hydrogen ion concentration of acid solutions. By ARTHUR W. THOMAS and MABEL E. BALDWIN. [From the Laboratory of Food Chemistry, Columbia University.] In the course of an investigation upon the effect of neutral salts upon hydrogen ion concentration we have noted a striking contrast in the effect of chlorides, sulphates and magnesium salts on the apparent hydrogen ion concentration of solutions of hydro- chloric and of sulphuric acids. The results of our measurements are given below. In all cases a solution of acid was mixed with a solution of a given salt and diluted to 100 c.c. The concentration of this solution of the mixture was always 0.I normal with respect to the acid and that particular concentration of the salt whose effect upon the hydrogen ion concentration was to be studied. The hydrogen ion concentra- tions were measured two days after the solutions were made up. The effect of chlorides in increasing the hydrogen ion con- centration of hydrochloric acid solutions is not new. This has previously been shown by Harned! and by Fales and Nelson?. Arrhenius* found that the hydrogen ion activities of acetic and of hydrochloric acids were increased by neutral salts which he thought was due to the salts increasing the dissociation of the acids. It is possible that recent work on the hydration of ions in aqueous solution will offer a more plausible explanation. When the chlorides are arranged in order of their ability to increase the hydrogen ion activity, we get the following series: MgCl, > BaCl, > LiCl > NaCl > NH,Cl = KCl which, with the exception of magnesium and barium is in inverse order to a series of these cations arranged in order of their equivalent con- ductivities. In the above series the salts are arranged in order of the in- creasing hydration (i.e., number of molecules of water combined 1J. Am. Chem. Soc., 37, 2460 (1915). 2J. Am. Chem. Soc., 37, 2769 (1915). 8 Z. physik. Chem., 31, 197 (1899). 4 SCIENTIFIC PROCEEDINGS (IOI). with the ions at infinite dilution) of their cations—Kt, 9.6; NH,*, 10.7; Nat, 16.9; Lit, 24.0;! Bat+, relatively higher;? Mgtt, higher than Bat+.3 The contrast in the effect of sodium and ammonium sul- phates in decreasing the hydrogen ion concentration as compared with the increasing action of the chlorides is noteworthy. The peculiar effect of magnesium salts is interesting, especially in that the concentrations for bend in the curves is in each case between 0.5 and 1 molar. We have tried to find an explanation for these peculiarities in Jones’ ‘‘ Hydrates in Aqueous Solutions.’ Jones states, ‘‘ Magnesium sulphate, like all the other sulphates studied, gives abnormal results. It appears to form no hydrates in aqueous solution, notwithstanding the fact that it crystallized with seven molecules of water of crystallization. It is almost TABLE I. EFFECT OF SALTS UPON THE CONCENTRATION OF HYDROGEN ION OF 0.1 NORMAL SULPHURIC ACID. Conc, Salt. Log. Cyy+. | Conc, Salt, Log. Cy+. Sodium Chloride Ammonium Chloride. Doe i*, hoy Pees — 1.21 OF tase? AS aie — 1.21 I yo. ee eS — 0.99 eee | a ee — 1.08 2 PORE okra: — 0.78 2 ig ee ey — 0.98 3 “ETS Oy Oe — 0.57 3 ork ich aac eas — 0.84 4 Pat bes tatot 2s — 0.36 4 pee S aaB eS — 0.72 Magnesium Chloride. Sodium Sulphate. Do) (33 Aisne — 1.21 9495 ah Aes — 1.21 0.25 Molar........ = 0.84 0.25 Molar...< ..2;5 — 1.60 0.5 ere tA Fe — 0.56 0.5 sie Pee yf ee — 1.72 I sare a Eee — 0.56 I Daa citancaeel — 1.80 2 eae te Ses + 0.03 2 ee tt — 1.88 s bath a 644 ah + 0.60 3 eR de Pie — 1.92 Ammonium Sulphate. Magnesium Sulphate. We AR Trea — 1.21 ee he a. — 1.21 Os DMolar «oct. — 1.61 O85 Molar... — 1.46 0.5 er eet — 1.76 0.5 gee ghee — 1.52 I a Ga nt haa — 1.90 I eG ta a eS — 1.53 2 Pehle aide ned ahs — 2.04 2 eh eS Se — 1.39 3 ee ee ee — 2.15 3 Deb bee — 1.15 4 76s & aetna — 2.24 1Smith, J. Am. Chem. Soc., 37, 722 (1915). 2 Washburn, Tech. Quarterly, 21, 360 (1908). * Jones, Carnegie Institution of Washington, Publication No. 60 (1907). HYDROGEN ION CONCENTRATION OF ACID SOLUTIONS. 5 TABLE II. EFFECT OF SALTS UPON THE CONCENTRATION OF HYDROGEN ION OF 0.1 NORMAL HYDROCHLORIC ACID. Conc. Salt, Log. Cy+. Conc. Salt. : Log. Cy+. - Sodium Chloride. Potassium Chloride. lt ake yy eee — 1.038 Gr G25 deh das xs — 1.038 Oe Wiilak. eo kc — 0.88 I Molar. os if%- — 0.95 2 tt es ea — 0.72 2 TM te ty — 0.85 3 aire setae — 0.52 3 SCE Ripe — 0.75 4 Rat as 8 — 0.36 4 2 exdiise ss Be — 0.63 Ammonium Chloride. Lithium Chloride. > SE Real Bae Se a — 1.038 aad eh eee ae — 1.038 I I ese x. i — 0.94 I 1) 2 a — 0.81 2 3 os ae eee — 0.87 2 gk EWE A — 0.60 3 OMe aS, epee — 0.75 3 PSAs ace: Bak be — 0.35 4 See Ee Soe — 0.65 4 Dee 2 ae bt — 0.12 Barium Chloride. Magnesium Chloride. ee Oe Ee — 1.038 eis ape hl Si ey sy kas — 1.038 @25 Molar. .).5..%: — 0.96 O25 Molaro 22. = 0.73 0.5 Bee nian Sra, — 0.88 0.5 sate Cees — 0.55 0.75 ee ae — 0.80 I a oe ea ee — 0.55 I Beebe iy a aes — 0.71 2 fe as hte + 0.005 3 eee bee + 0.68 4 aya = OC ae + 1.37 Sodium Sulphate. Ammonium Sulphate. OG baad we — 5.035 i RS kate tr ene — 1.038 ea Molar... >... — 1.51 29 Molar... 2... — 1.52 0.5 PRA re — 1.65 0.5 are Latane Seer — 1.72 I sane es SN — 1.79 I waw Raia eee — 1.90 2 os aa ke bea ae ee — 1.86 2 se were Oa — 2.05 3 Se ae — 1.89 3 eM icciitwhia's os — 2.14 4 Peete = nec — 2.18 Magnesium Sulphate A Be eee — 1.038 RES ee Ree fae ee —~ Az G25 Biotar... 6 2u.. — 1.36 2 Pe Oaks F — 1.34 0.5 eho ig Re araee — 1.45 3 Oe gts LR ee — 1.12 certain that this substance has considerable hydrating power, but this is masked in our results by the large amount of poly- merization which the sulphates undergo.” The curve of the freezing point depression of magnesium sul- phate plotted against concentration shows a depression to about 0.5 molar, from which point the depression decreases for higher concentrations. In this respect there is a slight similarity to its 6 SCIENTIFIC PROCEEDINGS (I0I). effect on hydrogen ion concentration, i.e., it increases hydrogen ion concentration to 0.5 to I molar beyond which it decreases it. Magnesium chloride, however, gave a similar effect on freezing point depression, although not so pronounced as magnesium sul- phate. We could not find anything in the hydrate theory as developed at present to account for the peculiar bend in the magnesium chloride hydrogen ion curve. We do not believe that the figures for hydrogen ion concen- trations in the presence of salts given in this paper, are the true values. They should be termed apparent concentrations of hydrogen ion as determined by the method in general use at this date. For the determination of the hydrogen ion concentrations, a Wolff 15,000 bridge with a galvanometer to determine the null point was used. As hydrogen electrode, a No. 16 Browne and Sharp gauge platinum wire (platinized) fused in a glass tube in- serted in the Clark! cell was used. The calomel element con- tained 3.5 molar potassium chloride solution saturated with calome] and was the same design as that described by Fales and Vosburgh.?, The E.M.F. was determined by means of a Weston cell that had been checked by the Bureau of Standards. The hydrogen contained in a tank under pressure, was carefully washed through saturated mercuric chloride solution, alkaline permanga- nate, alkaline pyrogallol and a tower of cotton fibers. The measurements were made at room temperatures which varied between 22° and 26° C., the proper corrections being made. No correction for barometric pressure was made since it is so small. (See Harned, loc. cit.) No attempt was made to calculate and correct for the solution contact potential because we know of no satisfactory method of doing so, especially where solutions con- taining divalent ions are concerned. It is emphasized however, that the differences in effects reported in this paper cannot be attributed to solution contact potential. This point was demon- strated by Harned, and Fales and Vosburgh proved there is no contact potential at 25° C., between a saturated solution of po- 1 Clark, J. Biol. Chem., 23, 475 (1915). 2 Fales and Vosburgh, J. Am. Chem. Soc., 40, 129 (1918). UTILIZATION OF SALEP MANNAN. 7 tassium chloride (4.1 M) and hydrochloric acid solutions ranging in concentrations between 0.1 molar and 1 molar. The salt bridge between the hydrogen and calomel elements used in our measurements was a saturated solution of potassium chloride. | 3 (1463) On the utilization of salep mannan. By Mary SWARTZ ROSE. [From the Department of Nutrition, Teachers College, Columbia University.] Some experiments on the utilization of salep mannan were reported in 1911.1 It was found that this anhydride of mannose was not hydrolyzed by the enzymes of saliva, pancreatic and intestinal juice, nor by malt diastase, but disappeared almost completely from the human alimentary tract when eaten, the coefficient of digestibility in three out of four experiments being 100 per cent. and 94 per cent. in the fourth. Studies of the effect of fecal bacteria indicated that some of them could produce appre- ciable amounts of sugar from this polysaccharide, and stimulated further research as to its precise fate in the animal organism. Investigations were interrupted in 1914, when the war cut off the supply of salep, and what has been accomplished along several lines is now reported as it is doubtful when these studies can be resumed. Four more determinations of the coefficient of digestibility were made, two on healthy young women and two on diabetics. The young women, consuming identical and uniform diets, free from cellulose, throughout a fore, mid, and after period, took in the mid period of three days 75 grams of salep mannan, equivalent to 61 grams of glucose. The coefficient of digestibility was 97 per cent. in one case and 95 in the other. A diabetic man given in one day 45 grams of salep mannan, with no other food but broth, coffee and whiskey, had a coefficient of 98 per cent. A diabetic boy fifteen years old, took in three days 33, 65 and 70 grams of 1 Trans. Conn. Acad. Arts and Sciences, XVI, pp. 247-382, IQII. 8 SCIENTIFIC PROCEEDINGS (IOI). mannan respectively, and the coefficient of digestibility was 96 per cent. This was the largest amount administered to any sub- ject. There was in no case any discomfort from gas formation, or other evidence of fermentation. A detailed study of the nitrogen intake and output in the experiments on the two young women showed an increase in the fecal nitrogen accompanied by a marked increase in the volume of dry feces. There was a slight fall in the urinary nitrogen in the salep period and an increase of 16 and 17 per cent. re- spectively in the after period. This may have been due to de- crease in the urine volume in the salep period and a subsequent “‘flushing out.” There was no evidence of sugar formation in the diabetic organism. In one case 10 grams were given in a day with no other food but broth, coffee and whiskey, and in another 45 grams, the urine in both cases remaining sugar free. For a five- day period a fifteen-year-old diabetic boy was kept on a controlled diet, and salep averaging 56 grams per day given for three days. Salep did not stop the production of $-oxybutyric acid, which rose from 3.4 grams on the day before the salep feeding to 15 grams on the third day. | Glycogen storage in the livers of rabbits could not be demon- strated, though mannose has been shown to form glycogen reaidly. The animals were starved five or six days, then fed salep by stomach sound for from one to three days. They were killed twelve to fifteen hours after the last feeding, but only traces of glycogen were found after administration of as much as 30 grams of salepinaday. The largest amount was 35 milligrams, whereas a rabbit fed 15 grams starch as a control had 209 milligrams. In two rabbits 16 and 60 per cent. respectfully of the mannan was recovered from the alimentary tract and identified. It was thought that since creatine elimination induced by starvation may be made to disappear by administration of calori- cally insufficient carbohydrate, carefully controlled experiments with rabbits might afford evidence as to the utilization of this mannan in metabolism. A series of experiments in which rabbits were starved from three to six days, then salep fed by stomach sound for two or three days, showed nothing especially significant SIMPLIFIED Foop SUPPLY. 9 in the output of creatine, creatinine and total nitrogen, when these animals were compared with controls similarly treated, but fed soluble starch alone or combined with maltose or lactose in amounts equivalent to the salep administered. There was a faint tendency for creatine and total nitrogen to fall in the starch period, but the results cannot be considered conclusive. Salep resembles inulin in its ease of acid hydrolysis, its re- sistance to digestive enzymes, and its failure to form sugar in the diabetic organism. It is not so fermentable as inulin. 4 (1464) Growth and reproduction upon simplified food supply. By H. C. SHERMAN, M. E. Rowse, B. ALLEN and E. Woops. [From the Laboratory of Food Chemistry, Columbia University.] Rats were fed rations consisting of white bread (made without milk or butter) either alone or with only one other article of food. Later, ground whole wheat was substituted for white bread in several cases. In preliminary experiments with animals placed upon the experimental rations at the time of weaning, bread alone resulted in cessation of growth at once and death after about six weeks. With bread and meat there was some growth at first, but the survival period was only slightly longer than with bread alone, with bread and apple there was no growth, but the survival period was considerably longer; with bread and turnip there was con- tinuous slow growth; with bread and milk there was continuous growth at a normal rate. In this case the bread and milk ration consisted of equal weights of fresh bread and market milk, making a food mixture in which the white bread furnished four fifths and the milk one fifth of the total calories. On this ration young rats of both sexes (taken at weaning time from mothers which were receiving mixed food) made normal growth and the males were capable of normal reproduction but the females usually failed to breed and none of them raised any young. On a ration containing the same proportion of milk (about 10 SCIENTIFIC PROCEEDINGS (IOI). one fifth of the total calories) but with whole wheat instead of white read or patent flour, young were successfully suckled (though at the cost of considerable loss of weight on the part of the mother) and are growing at somewhat less than the average rate. When about two fifths of the total calories were supplied by milk and the rest by whole wheat, the mother has suckled the young without undue loss of weight and the young have made a fully normal rate of growth. When the market milk used has been replaced by dried milk, or when it has been incorporated into the bread in bread-making and, therefore, subjected to the heating involved in the baking of the bread, there has been no evidence of any serious destruction of either “fat-soluble A” or ‘‘water-soluble B.’’ Since the ex- periments were made upon rats, they would, of course, throw little if any light upon the destruction of the antiscorbutic vitamine. We plan to continue the study of the effects of heating upon the vitamines in some of the staple articles of food. 5 (1465) The rate of change of hereditary factors in Drosophila. By H. J. MULLER and E. ALTENBURG. [From the Rice Institute, Houston, Texas, and Columbia University, New York City.] A knowledge of the rate at which hereditary changes of various sorts occur is the necessary groundwork for an adequate under- standing of evolution. The wide recognition given to this fact is attested to by the vast amount of literature on the subject of “variation,” but, with our new exact knowledge of the Mendelian — and chromosomal method of inheritance of the so-called ‘“ varia- tions,”’ it is evident that this literature has very little bearing on the real question of how often changes in the hereditary factors, 7.e., mutations, actually occur: for the breeding procedures used in the experiments there considered were not of the type necessary for ferretting out the new mutant factors as they arise, and for distinguishing between them and the apparent variations RATE OF CHANGE OF HEREDITARY FACTORS. II caused by the sorting out of old mutant factors into new com- binations. There is, to be sure, enough work to show that the real mutations are ‘‘rare’’-—whatever that term may mean; but, so far as an approximate quantitative determination of the rate of factor change is concerned, it is not possible, from the published work, to determine even its general order of magnitude. Some special scheme of crossing is required for this purpose. In the present series of experiments with Drosophila, the X chromosome was chosen as the most convenient one for the detection of mutation, since every hereditary factor in either of the X chromosomes of the female fly stand revealed in the charac- ters of one half of her male offspring, no matter what their father was. Thus, if the female has a new mutated factor in one of her X chromosomes, even though she does not usually show that factor herself, and even though her mate does not contain it, nevertheless one half of her sons are bound to show it and the mutation will thus be recognized. There is reason to believe that by far the commonest type of mutation is that which gives rise to a lethal factor—which kills the organisms containing it—and such lethal factors, also, in the X chromosome of a female, would be revealed; in this case, by the fact that half the male offspring, receiving it, would die before hatching. There would thus be half as many sons hatched as daughters, giving a sex ratioof 2 9:10’, instead of I Q : I co’, the usual ratio. In the first set of experi- ments these lethal factors were looked for primarily, by making a count of the sex ratios. As a preliminary measure, about 90 females were bred, and the sex ratios of their progeny counted. ‘Those families which gave a lethal, 7.e., 2:1, sex ratio (there were three of these) were then discarded, since there was no way of knowing, in this pre- liminary cross, whether these lethals had just arisen by mutation or were of ancient origin. Females from the normal families (with I:1 sex ratio) were bred, however, since any lethal later dis- covered in the descendants of these flies must be due to a really new mutation, inasmuch as the ancestors had been certified as normal. It was necessary, moreover, in. selecting females for breeding the next generation, not to breed many females from the same family, but to choose them from as many separate families I2 SCIENTIFIC PROCEEDINGS (101). as possible, in order to be sure that any mutations that might be discovered later had arisen separately, and were not merely sister representatives of one original mutation. By continuing this method of breeding from separate families over five or more generations after the preliminary tests, the sex ratios in 385 families were counted. Thirteen of these were found to be 2:1 ratios; this is a proportion of one new lethal mutant among each thirty females that are bred. This figure is of a far higher mag- nitude than any which had been anticipated. It should be noted that at the same time as all these lethals arose, no mutations causing ordinary visible character variations were observed. The correctness of classification of most of the thirteen lethals was verified by further breeding tests, but there were a few doubt- ful cases, and it was realized that ratios intermediate between I: I and 2:1 aresometimes brought about in other ways. Although the possible error due to these cases was not enough to change the order of magnitude of the frequency found, a new set of experi- ments was undertaken in which a still more definite test of lethal factors than the sex ratio was used—namely, the test of linkage to known factors in the X chromosome. The breeding procedure —having preliminary tests, breeding from many separate families, etc.—was the same as before, but instead of using pure wild type flies for the work, the following cross was made in each genera- wv f WVF X chromosome makes itself known not only by the 2:1 sex ratio, but by the practically total absence of all males containing factors on both sides of the lethal. By noting whether any expected class of males was absent, it could thus be determined whether a lethal was present, and, if so, approximately where it was located tion: 2 Xw'vfi oo. In this case a lethal arising in either in the chromosome. 1,062 families were examined in this way, » after the preliminary tests, and twenty lethals were found—a ratio of one in fifty-three. Enough work has been done on them thus far to know that they occurred in at least ten different loci scat- tered along the X chromosome—but this is a bare minimum. Four of the lethals (perhaps five) are more strictly speaking ‘‘semi- lethals,’’ as they occasionally allow the male possessing them to live (and then produce some curious morphological effects in him) RATE OF CHANGE OF HEREDITARY FACTORS. 13 but lethal mutations are so much more frequent than the type of visible character variation ordinarily dealt with, that none of the latter were observed in the whole experiment. The above figure of 20 in 1,062 has a probable error due to chance of about + 3 in 1,062. There can, therefore, be no doubt about the correctness of the order of magnitude of the ratio 1: 53, so far as any error caused by random sampling is concerned. The ratio I: 53 is, however, a composite result, for the families were kept in two main lots, one at about 66° F., the other at about 80°. The 445 grown at the lower temperature produced five lethals, or one in ninety; the 517 at the higher temperature produced 13 lethals, or one in forty. The other two were new lethals which occurred in the 100 bottles kept at room temperature, in which the lethals found in the two main series were being tested out. In this connection, it should be pointed out that the high ratio of one in thirty observed in the earlier experiment was obtained in bottles kept at room temperature in the warm climate of southern Texas. Although the absolute numbers of lethals are small, the difference between the two series in the later experiment is probably statis- tically significant,—at least, it may be calculated that if the lots had really been similar, the chances would have been about twenty to one against a difference of this magnitude occurring between figures of the given size. Taking the figures at their face value, we should obtain Qo for mutation between 2 and 3, as is usual for chemical reactions. If we accept the one in fifty-three ratio as representing the average frequency for the X chromosome, and if, as there is reason to believe, mutation occurs at the same rate in the other chromosomes as in the X chromosome, then, since the X’s form about one fourth of the entire chromosome mass, we may figure that about one fly in every thirteen has a new lethal mutation in some chromosome or other. It is evident that, at this rate, with- out natural selection to weed out the “‘unfit,’’ the race would soon become filled with lethal factors. For the X chromosome alone, since each female has two X’s, and one female in fifty has a new lethal, we may figure that one X chromosome in every 100 con- tains a lethal factor just arisen in the present generation. Or, to put the matter differently, each X chromosome would, on the 14 SCIENTIFIC PROCEEDINGS (IOI). average, tend to contain one lethal factor after 100 generations— which means about four years in Drosophila. The rate of change for the X in Drosophila is thus about one detectable muta- tion in four years. This immediately shows us that Drosophila must have a different rate from some other organisms—man for example—for if the X chromosome of man mutated at anything like a similar rate, all the X chromosomes in a female would contain several lethal factors by the time she was ready to re- produce, and none of her sons would be viable. The rate of one mutation in four years is the rate for the whole chromosome. It is of greater interest to know the rate for the individual factors. There is good reason to believe that there are at least 500 factors in the X chromosome of Drosophila— probably many times that number. But, taking this undoubtedly much too low minimum figure, it is easy to see that, if 500 factors show only one mutation in four years, each individual factor must on the average show a change in its composition only once in 2,000 years. (Yet this is in the mutable Drosophila.) It will be interesting to observe the difference in mutation rate in different organisms and under different conditions. 6 (1466) The influence of lactic acid upon the metabolism of the dog.! By GRAHAM LUSK and H. V. ATKINSON. [From the Physiological Laboratory, Cornell University Medical College.| Lactic acid, when given to a dog, causes the same increase in metabolism that is noticed when a similar amount of alanin is administered. It was also noted that the metabolism was in- creased after giving 500 c.c. of water in which there was 2.5 c.c. of Liebig’s extract of beef, whereas the administration of 150 c.c. of water had no influence whatever. When a large quantity of water was given about 100 c.c. per hour were eliminated in the urine. This indicates that for the transport of a large volume of fluid through the circulation increased energy is needed. 1 A brief report of this work was also published in the Compt. rendus de l'academie des sciences, 1919, 168, No. 20, 1012. REJUVENESCENCE IN UROLEPTUS. 15 7 (1467) Rejuvenescence without encystment and without nuclear fusion in Uroleptus? By Gary N. CALKINS. [From the Department of Zoélogy, Columbia University.] Experiments made during the last two years on Uroleptus mobilis have shown that renewal of vitality follows conjugation, both parents, coming from the same protoplasm and all kept under identical conditions of food and environment. It was also shown that asexual reorganization occurring during encystment, like- wise results in rejuvenescence. It was argued that, since the only Mommon phenomenon in conjugation and encystment is the nuclear dissolution and absorp- tion in the protoplasm, the renewal of vitality after each is due to the chemical and physical changes in the protoplasm set up by the addition of relatively large quantities of nucleoproteins added to it. To test this working hypothesis the following experiments were undertaken last April and May. Individuals of the same ancestry (P series) in the 140th generation, were allowed to con- jugate; a normal ex-conjugant was isolated (V series), and this, with the parent P series, were maintained as controls, the latter to show the degree of vitality without conjugation, the former to show the effect of conjugation. Other pairs of conjugating individuals were isolated in a minute drop of culture medium, a pair at a time, and cut with a scalpel across the angle of the V made by the two individuals in conjugation. The detached angle of the V and one of the arms of the V were immediately fixed and stained to determine the stage of conjugation at the time of cutting. The other part was isolated in culture medium and set aside for observation. Seven pairs were successfully cut in this way, and in the desired plane; four of these died within 10 days, three continued to live forming the X series, the X6 series, and the X7 series. The period required for reorganiztion of the normal individual after conjugation and prior to the first cell division, varies from 16 SCIENTIFIC PROCEEDINGS (IOI). 3 to 6 days and the process may be watched in the living cell in which the changes of the nucleus are clearly visible. The in- dividual forming the V series required seven days for this reor- ganization. It was most unexpected and interesting to find that the cut individuals invariably went through the same nuclear reorganiza- tion states as those of a normal ex-conjugant. If they died, death was always at the last phase of the reorganization process. The individuals destined to live and form the X series, the X6 series, and the X7 series, required eight, nine, and eight days respectively before the first division of the young cells. From the start each series represented by five different lines, was carried on in cul- ture like any normal ex-conjugant series, with similar records of the daily division rate, the average rate for ten days, and the average rate for the first and second sixty day periods. The results for the four months following the operations are given in the following table. AVERAGE DIVISION RATES IN 10-DaAy PERIODS, OF CONTROLS AND EXPERIMENTAL SERIES. Series V Seriea|P, Series a7) cond ee gating. | gating. | jugant. | gant. | 17.6 | 18.2 ) 12.4 15.8 heya, 13.3 | 16.4 | 16.8 | 12.2 MR hg 15-8 | 18.6 | 18.2 | 16.8 pa TA -*. 14.0 | 15.8 | 14.8 | 13.2 Sa eatage 13-2 | 15.4 | 17.0 | 16.4 ma: ™ 14.0 | 16.8 | 16.6 | 14.4 Ist 60 “ 14.3 | 16.8 | 16.9 | 14.2 ad. Go. <= 10.3 | 12.0 | 12.9 7-7 Average 120 12.3 | 14.4 | 14.9 | 10.9 days. ) It has been shown in an earlier paper! that the extent of re- juvenescence as indicated by the division rate, varies with the age of the parent series at the time of conjugation. The greater the vitality of the parent series the less is the difference in vitality between parent and filial series. For example, if the filial series starts by a conjugation occurring during the first 60 days (ap- 1 Calkins, “‘Uroleptus mobilis. II. The Renewal of Vitality through Con- jugation.” Jour. Exper. Zodl., Vol. 29, No. I, 1919. REJUVENESCENCE IN UROLEPTUS. 17 proximately 100 generations) the average difference in vitality amounts to only 1.5 divisions in 10 days for the first 60-day period of the offspring. If the filial generation starts from a parent series which is 130 to 200 generations old, the difference in vitality between parent and offspring increases to between 3 and 4 divisions in 10 days, and if the filial generation starts from a parent series that is about 300 generations old, the difference in vitality rises to a maximum of 17 or more divisions in Io days. In the present experiments the parent P series was in. the 140th generation when the filial V series was started, and in the 160th generation when X6 and X7 were cut. The difference in vitality between the V series and the P series, 3.1 divisions in 10 days for the first 60 days, agrees with the results obtained with all ex-conjugants started at corresponding periods of the parent cycle, and shows the normal rejuvenating effects of conjugation. The cut conjugants show different results in regard to renewal of vitality. The X series shows no rejuvenescence at all but agrees with the non-conjugant P series throughout. The X6 series agrees with the non-conjugant P series in the first 60-day period but has a higher average division rate for the second 60- day period, while its average for four months is intermediate be- tween the non-conjugant P series and the ex-conjugant V series. The X7 series, on the other hand, shows for the entire period the same vitality as the ex-conjugant V series and gives the same evidence of rejuvenescence. These three divergent cases are difficult to interpret. Each recovered perfectly after the operation; each underwent reor- ganization processes apparently identical with those of normal ex-conjugants; each gave rise to a population similar to that of a normal ex-conjugant except that in none of them have I seen a case of encystment; epidemics of conjugation have been fre- quent in each of these populations. A possible explanation may be found in the fact that the con- jugating pairs which were operated upon to give the X, X6, and X7 series were cut at different phases of the process of conjugation. In none of them had nuclear interchange taken place. One pair was cut at the very outset of conjugation; this gave rise to the X series which showed no rejuvenescence. Another pair was cut 18 SCIENTIFIC PROCEEDINGS (IOI). at the stage of the first maturation spindles; this gave rise to the X6 series with evidence of partial rejuvenescence. The third pair was cut at the stage of the second maturation division and this gave rise to the X7 series which showed the same rejuvenes- cence as the normal ex-conjugant from the same source. Many more experiments of the same nature are now under way and must be carried out before conclusions can be drawn. These three cases indicate, however, that the absorption of nucleo-proteins in the cytoplasm and which occurred in all cases, is not, by itself at least, the secret of renewed vitality. 8 (1468) The total carbonate content of the arterial and venous plasma in normal individuals. By R. W. Scott (by invitation). [From the Department of Medicine, School of Medicine, Western Reserve University, Cleveland.] In the course of some observations on the respiratory dis- turbances seen in certain diseased conditions it became necessary for the sake of comparison to determine the total carbon dioxide of the arterial and venous plasma in individuals with normal cardio-respiratory mechanisms. In all the bloods of nineteen normal individuals at rest have been examined. In each case samples of arterial and venous blood were obtained within a few minutes of each other. | Method.—The arterial blood was obtained by direct punctur of the radial artery using a technique similar to that employed by Stadie.t The venous blood was collected without stasis from one of the large veins at the bend of the elbow. To avoid con- tact with air both samples were carefully delivered under albolene into paraffin coated centrifuge tubes and immediately centrifuged at high speed. One c.c. of the separated plasma was delivered under carbonate free ammonia water contained in a receiving cup 1 Stadie, W. C., J. Exp. Med., 1919, XXX, 215. CARBONATE CONTENT OF PLASMA. 19 of the Van Slyke apparatus and the total Coz content determined by the method devised by Van Slyke.!. The method adopted was considered to be more advantageous for my purpose than the widely used method of first exposing plasma to 5.5 per cent. of CO, and ascertaining the CO, combining power. Results: The results presented in the accompanying table in- dicate that the total CO, content of the arterial plasma is a fairly constant figure, averaging fifty-six volumes per cent. The venous plasma is always a little higher than the arterial in in- dividual cases, the discrepancy being from three to eight volumes per cent. This discrepancy has been found to increase if the individual is allowed to take some light exercise, such as walking, just before the blood samples are taken. Under these conditions the arterial figures remain about normal while the venous are from twelve to fifteen volumes higher. THE TOTAL CARBONATE CONTENT OF THE ARTERIAL AND VENOUS PLASMA OF NoR- MAL INDIVIDUALS AT REST. CO: reduced to 0°—760 mm. in 100 c.c. plasma. Arterial Venous Arterial Venous c.c. c.c. c.c. c.c. 62.0 66.0 55-0 61.0 57.8 64.4 58.0 61.4 54.0 61.0 57.5 62.8 59.1 67.2 62.8 65.4 54.9 62.2 53-4 60.0 58.1 64.2 59.0 65-7 Be 59.9 60.3 67.4 So5 61.5 54.9 62.2 55-7 61.0 60.5 68.9 51.5 59-9 9 (1469) The total carbonate content of the arterial and venous plasma in patients with chronic heart disease. By R. W. Scott (by invitation). [From the Department of Medicine, School of Medicine, Western Reserve University, Cleveland.]| Twenty-eight determinations of the total carbon dioxide content of the arterial and venous plasma have been made on ten 1 Van Slyke, D. D., J. Biol. Chem., 1917, XXX, 347. 20 SCIENTIFIC PROCEEDINGS (101). individuals with chronic heart disease. All patients in this group have been carefully selected because they were suffering primarily from a failure of the heart to maintain an adequate circulation. They have been free as far as could be determined from any vascular or renal disease. For the most part they were young patients with chronic rheumatic myocarditis and valvulitis. In some a normal cardiac mechanism was present, as deter- mined by the electrocardiograph. In others auricular fibrillation usually associated with mitral stenosis was found. Moribund patients and patients with a marked degree of venous stasis and edema have not been included. Samples of the arterial and venous blood were obtained at the same operation; immediately centrifuged, and the total CO, content of the separated plasma determined directly. In all cases the blood was obtained while the patient was at rest in bed. ‘The results are presented in the accompanying table. THE TOTAL CARBONATE CONTENT OF THE ARTERIAL AND VENOUS PLASMA IN PATIENTS WITH CHRONIC HEART DISEASE. CO: reduced to 0°—-760 mm. in 100 c.c. plasma. Arterial Venous Arterial Venous c.c. c.c, c.c. c.Cc. 42.8 50.6 32.0 44.5 33-7 37-3 41.6 48.4 38.2 50.3 48.5 54.2 47.6 52.5 39-7 44.6 46.9 50.6 51.0 63.0 It is seen that both the arterial and venous plasma have a total CO, lower than that found in normal individuals. The arterial values for CO, show wider variation and the discrepancy between the CO, of the arterial and venous plasma is more marked in heart cases than in normal individuals. ‘These differences are. attributed to the varying degrees of cardiac efficiency in the heart patients. In the type of cases studied there has been a certain relation between the integrity of the circulation and the level of CO, in the plasma. The more dyspneic the patient the lower has been the CO, in the arterial plasma. The following case will serve as an example: A patient walked into the hospital complaining of CARBONATE CONTENT OF PLASMA. 21 shortness of breath. On examination a moderate cardiac en- largement with mitral stenosis and auricular fibrillation was found. The CO, of the arterial plasma was thirty-eight volumes per cent. Five days later when he was much improved the CO, of the arterial plasma had increased to forty-nine volumes per cent. The results of this study seem to indicate that when the minute volume of air respired at rest is definitely above normal (10 to 12 liters) the plasma CO, is low. With improvement in the circulation and the accompanying fall in the minute volume the CO, of the arterial plasma shows a definite increase toward the normal. 10 (1470) The total carbonate content of the arterial and venous plasma in patients with chronic pulmonary emphysema. By R. W. Scott (by invitation). [From the Depariment of Medicine, School of Medicine, Western Reserve University, Cleveland.] This study represents six determinations of the total carbon dioxide content of the arterial and venous plasma over a period of six months on three patients with chronic pulmonary emphy- sema of the so-called ‘‘large lunged”’ type. The patients were males between forty-five and fifty years of age with definite enlargements in all diameters of the thorax, particularly the anterior-posterior diameter; thus presenting the typical “barrel shaped”’ chest. They were singularly free from cardio-renal disease so that all their symptoms and signs were attributed to the disturbance in the respiration resulting from the degenera- tive process in the lung. From observations to be reported in detail elsewhere, it has been found that this type of patient will tolerate an unusually high percentage of CQ, in the inspired air with little increase in the minute volume over that at room air and without any subjective symptoms of distress. As a rule such patients breath eight to ten per cent. CO, for from ten to fifteen minutes with no apparent discomfort. That is, one man who has been under observation for the past nine months had a 22 SCIENTIFIC PROCEEDINGS (IOI). respiratory rate of twenty-two and a minute volume of ten liters while breathing room air. During an experiment in which air con- taining 11.4 per cent. CO, was inspired for a period of six minutes, the respiratory rate remained at twenty-two and the minute volume increased to only fourteen liters. At this high concentra- tion of inspired CO, the patient complained of a little dizziness and nausea which disappeared when he was allowed to breath room air for a few minutes. In an attempt to explain the unusual tolerance to CO, in the inspired air shown by the type of case described, the CO, content of the arterial and venous plasma of three patients have been de- termined and found to be consistently above normal as indicated by the data in the accompanying table. 7 TABLE. THE TOTAL CARBONATE CONTENT OF THE ARTERIAL AND VENOUS PLASMA IN THREE PATIENTS WITH CHRONIC PULMONARY EMPHYSEMA. COz reduced to 0°-760 mm. in 100 c.c. plasma. Arterial Venous Arterial Venous Tite ETA RP Wapigs a) ed eee, . ae aia c.c. 76.1 82.7 80.2 88.4 72.4 78.4 70.2 76.4 71.0 76.0 74.5 80.2 II (1471) Concerning the influence of antipyretics on the acuity of hearing. By D. I. Macut, J. P. GREENBERG and S. ISAACS. [From the Johns Hopkins University, Baltimore, Md.] In connection with an extensive pharmacological and psycho- — logical study of antipyretic drugs, the effect of same was tried on the acuity of hearing. Experiments were performed on normal human subjects, and in a few cases on persons suffering with mild deafness. The tests were made, by means of a watch, in a quiet room, with the subject in a sitting position; and all necessary controls were carried out for the elimination of error. In every experiment the normal limit of hearing was first determined; the drug to be studied was then administered by mouth; and the - * “oe. 2 ee dingy 4S. INFLUENCE OF ANTIPYRETICS ON HEARING. 23 acuity was afterwards tested at definite intervals of time. Only therapeutic doses of the drugs were administered. The following substances were studied: acetanilid, acetphenetidin, antipyrin, pyramidon, lactophenin, salol, aspirin, quinin, sodium salicylate, and ‘‘melubrin.’’ After studying the effects of individual drugs, certain combinations were administered. The following were among the combinations studied: acetanilid plus sodium bicar- bonate, acetphenetidin plus salol, acetanilid plus salol, acetanilid plus acetphenetidin, antipyrin plus aspirin, and antipyrin plus salol. The results obtained were both interesting and unexpected. It was found that some drugs decrease the acuity of hearing while others increase it. Furthermore, it was found that certain com- binations of antipyretics produce synergistic effects not explain- able by the simple arithmetical sum of the effects produced by the components individually. Among the agents found to decrease the acuity of hearing were acetanilid, salol, and aspirin. Among those found to increase the hearing were antipyrin, pyramidon, and small doses of quinin. Among the most remarkable com- binations studied were those of acetanilid plus sodium bicarbonate and acetanilid plus salol. It was established that whereas aceta- nilid given alone decreases the acuity of hearing, and ordinary doses of sodium bicarbonate given alone produce no change; a combination of the two produced a definite improvement in the acuity of auditory perception. Again, whereas acetanilid and salol when administered separately, each by itself tends to im- pair the hearing, a combination of the two actually increases the acuity of perception. The peculiar synergism of acetanilid with sodium bicarbonate recalls the experiments of Hale who called attention to the fact that such a combination is less toxic for animals than the same dose of acetanilid given alone. Experi- ments are in progress with a view to attempt an explanation of this peculiar synergism, and complete data of the research will be published in due time. 24 SCIENTIFIC PROCEEDINGS (101). I2 (1472) Serologic method for detecting infection in foods. By J. BRONFENBRENNER and M. J. SCHLESINGER. [From the Department of Preventive Medicine and Hygiene, Harvard Medical School.] The isolation and identification of the infecting organism in con- taminated foods is usually beset with difficulties and success- ful results are not the usual outcome. Negative findings often are due to the fact that it is impossible to examine the whole sample of the incriminated food bacteriologically. Since the pathogenic bacteria are usually few in number, and are not distributed throughout the food, it is more or less a matter of luck if one suc- ceeds in isolating an organism which might justify the suspicions. If one attempts to increase the number of the specific bacteria by enrichment through incubation, he at the same time increases the number of saphrophytes, and thus adds to his difficulties. We find that the entire sample of suspected food can be advan- tageously analyzed for the presence of any suspected organism, or their split products (in addition to an attempt to isolate in- dividual bacteria), by the following procedure. The whole of the sample of food is chopped up and an extract made from it. This extract is concentrated so that all the speci- fic bacterial protein is collected in a very small volume of liquid." This concentrated solution is then tested against a set of specific immune sera. We have been able to detect by this method the presence of B. botulinus protein in 20 gram samples of artificially inoculated food, where the concentration of toxin was so small that it would have required giving at least 7 grams by mouth or 1.3 . grams by injection into a mouse of 15-20 grams to obtain a positive result. This method enables one to determine the presence of a sus- pected organism in contaminated food within 24 hours after receiving the specimen. It is, of course, necessary to have on hand a collection of specific sera of high titer. 1 J. Bronfenbrenner and M. J. Schlesinger, Jour. Med. Res. (in press). ——————eEeEOEee—E—eEe—EE——E—E———— IDENTIFICATION OF COLON-TyYPHOID BACTERIA. 25 (This work is a part of the investigation of food poisoning, conducted under the direction of Dr. M. J. Rosenau, professor of preventive medicine and hygiene, Harvard Medical School. The investigations are done under the auspices of the Advisory Com- mittee of the National Research Council on the Toxicity of Preserved Foods, and under a grant to Harvard University from the National Canners’ Association.) 13 (1473) On methods of isolation and identification of the members of the colon-typhoid group of bacteria. Further studies on C. R. indicator. By J. BRONFENBRENNER, D. SOLETSKY and M. J. SCHLESINGER. [From the Department of Preventive Medicine and Hygiene, Harvard Medical School.| Some time ago we described an indicator for the direct measure- ment of the hydrogen ion concentration in growing bacterial cultures.1. This indicator consists of the mixture of China blue and rosolic acid, and covers the range of changes in the hydrogen ion concentration between C, = 1 X 107° and Cy = 5 X 1075. The choice of the dyes was made on the basis of their possessing suitable turning points coupled with the fact that their respective phases of highest color lie on the opposite sides of the point of neutrality. China blue is colorless at the concentration of hydro- gen ions below I X 107’, and gives graded intensity of blue with the increase in the hydrogen ion concentration up to the point of about C, = 5 X 10-5, when it reaches its maximum color. Rosolic acid, on the other hand, gives graded intensities of pink beginning with the hydrogen ion concentration close to I X 107”, and increasing gradually with the increase of the hydroxyl ions concentration reaching its maximum color at the Co, = I X 1075 (or C, = I X 107%). With the increase in concentration of hydro- gen ions above I X 10~’ the rosolic acid has a pale yellow (straw) color, which is masked by the color of ordinary culture media. 1J. Bronfenbrenner, Jour. Med. Res., XXXIX, I, 25, 1918. 26 SCIENTIFIC PROCEEDINGS (IOI). The combination of the two dyes thus offers an indicator which has a faint gray tinge at the neutral point with pure blue and pink on the acid and alkaline sides respectively. Due to the high tinctorial power of the dyes composing it, this indicator is incorporated into the media in very minute quantities. The actual concentrations of China blue and rosolic acid in the medium . are respectively 0.0025 per cent. and 0.005 per cent. by weight. This indicator, however, can be used only for the study of the Gram negative organisms, because the rosolic acid it contains exerts selective bactericidal action against the Gram positive organisms.!_ While this property of the rosolic acid is useful in certain circumstances (as for instance, for the purpose of sup- pressing Gram positive bacteria while isolating the Gram negative from the mixtures of both)?, it limits the usefulness of the mixture as an indicator. In order to permit the use of our indicator in the study of Gram positive as well as Gram negative bacteria, we suggest the substitution of corallin (Harmer) for the rosolic acid (Merk) in the above mixture. The turning point of corallin is the same as that of rosolic acid, and its tinctorial power, as well as the color, are much the same: Corallin (Harmer), however: has no bactericidal action upon Gram positive organisms. As the supply of either China blue, rosolic acid or corallin of foreign manufacture is very low at present, and as the domestic dyes sold under the same names are manifestly different from the foreign dyes used by us, it was necessary to try a number of re- lated dyes before a suitable choice could be made. While this work is still in progress, we have already found a few preparations which seem to answer the requirements. | (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, Harvard Medical School. The investigations are done under the auspices of the Advisory Committee of the National Research Council on the Toxicity of Preserved Foods, and under a grant to Harvard University from the National Canners’ Association.) 1 J. Bronfenbrenner, M. J. Schlesinger and D. Soletsky, Jour. Bact. (in press). 2 J. Bronfenbrenner, M. J. Schlesinger and D. Soletsky, Jour. Med. Res. (in press). BLoop SERUM OF NEPHRITIC PATIENTS. 27 14 (1474) The protein and lipin content of blood serum of nephritic patients. By Max KAgBn. [From the Department of Laboratories, Beth Israel Hospital, New York City.] It has been reported by Erben,! in 1902-3, that in the serum of patients suffering from chronic parenchymatous nephritis, the albumin-globulin ratio (which, normally is about 1.5-2.0: I) is entirely disturbed, the figure being 0.2593 of albumin to 7.0352 of globulin. This deviation in the ratio was confirmed by Epstein,’ in 1912. The latter author (basing his conclusion on his analyses of nine cases of chronic parenchymatous nephritis, in whom, besides the globulin increase, he also found a huge amount of cholesterol in the serum) propounded the following theory of the etiology of certain cases of chronic parenchymatous nephritis. According to him, there is a group among the cases of chronic parenchymatous nephritis which is due to a constitutional dis- order—of a metabolic or endocrinic nature—in which the renal manifestations are concomitant or secondary in point of develop- ment and importance. He does not, however, describe cases of chronic parenchymatous nephritis in whom the protein or lipin fractions should vary from the so-called “‘metabolic”’ type, and which he should point out as “‘non-metabolic”’ in nature. In the series of cases investigated, the writer did not find one case of the so-called ‘metabolic or endocrinic”’ type of chronic parenchymatous nephritis, which would seem to indicate that the type described by Epstein is very rarely met with. The al- bumin-globulin ratio does not seem to be markedly disturbed by various diseases. Feeding of patients suffering with chronic parenchymatous nephritis on a protein-rich, fat-poor diet is a rather risky undertaking. 1 Erben, Zeit. f. klin. Med., 1903 XLVIII, 450. 2 Epstein, Am. J. Med. Sc., 1917, CLIV, 638. 3 Epstein does not state what percentage of the cases of chronic parenchy- matous nephritis are of his so-called ‘‘metabolic or endocrinic’’ group. . 28 SCIENTIFIC PROCEEDINGS (IOI). 15 (1475) Method and results of a study of the distribution of iodine be- tween cells and colloid of thyroid glands. By ARTHUR L. TATUM (by invitation). [From the Laboratory of Physiological Chemistry and Pharmacology, University of Chicago.] Method.—Use was made of the fact that from frozen sections, as prepared for histological study of fresh unfixed tissues, the colloid of the thyroid gland completely disappears out of the acini when the sections are floated in physiological salt solution. The sections were picked out by means of a needle, washed in another solution of salt solution, dried at 105° C., weighed and analysed for iodine by the technique of Kendall. The iodine in the colloid portion was sometimes determined by evaporation of the salt solution containing the colloid, and at other times differentially by analysis of the whole dried gland. Results —In a series of about thirty experiments on dogs’ thyroids the thyroid iodine was found in the majority of in- stances to be wholly in the colloid as the cell portion was free from iodine. In a smaller percentage of cases the cell mass gave only qualitative tests for iodine, which at the present stage of this investigation might be considered due to unopened small acini. The physiological significance of these findings is being in- vestigated. 16 (1476) On the certain dietary factors to be considered in the treatment of cases of hyperthyroidism. By JacoB ROSENBLOOM. [From the Laboratory of Dr. Jacob Rosenbloom, Pittsburgh, Pa.] The writer has obtained clinical evidence that in formulating a diet for patients suffering from hyperthyroidism, two important factors must be considered; first, the diet should contain the minimum amount of protein and second, foods low in iodine content should be selected. SCIENTIFIC PROCEEDINGS ABSTRACTS OF COMMUNICATIONS. One hundred second meeting. New York Post Graduate Medical School, New York City, November 19, 1919. President Calkins in the chair. 17 (1477) Blood sugar curves with glucose, lactose, maltose, mannite, and cane sugar. By Cyrus W. FIELD. These curves were obtained by feeding normal males colored, with 100 grams of the pure sugar, on a fasting stomach. The dose was as a rule given after the first sample of blood had been taken; this was asarule at8 A.M. The second, third and fourth samples were taken one, two and three hours after the ingestion of the sugar, which had been dissolved in a large glass of water. The urines were tested for glucose up to three hours after the last sample of blood had been taken, and in none of the cases did a specimen ever show the slightest trace of a reducing substance with Benedict’s qualitative solution. The glucose curve was that with which all are familiar, that is rising to its highest point one hour after the ingestion, and then dropping to the normal at the end of the next hour or two. Maltose gave the same curve as the glucose. Mannite gave the same time curve as the glucose, and maltose, but did not rise to the same height as the other two; the average of five cases gave an increase of only 10 milligrams per 100 c.c. of blood, while that for glucose was 40 milligrams, and for maltose 34 milligrams. Cane sugar showed a curve that reached its height at the end of the second hour after its ingestion, and had dropped to normal at the third hour. Its average rise for 10 cases was 20 milligrams 29 30 SCIENTIFIC PROCEEDINGS (102). per 100 c.c. of blood. Lactose shows only a very slight rise, 4 milligrams, and that at the end of the second hour, as was the case with the other disachrade, cane sugar. I have been unable to consult the literature and so will offer the figures for what they are worth. 18 (1478) The bacteriology of infectious gaseous gangrene. By MARSHALL C. PEASE. [From the New York Post-Graduate Medical School.] Infectious gaseous gangrene can no longer be conceived of as being necessarily a monomicrobic disease. On the contrary it is frequently the result of an association of bacteria, not all of which are by themselves pathogenic or even under the most favorable condition of animal inoculation capable of causing a pathological lesion. The causative agents of infectious gaseous gangrene are found in a certain group of anaérobes, all of which are capable of — elaborating a powerful toxin which has not only a local but also a systemic action. Death in gaseous gangrene is not the direct result of the local lesion but of the absorption of toxin into the general circulation with a consequent general toxemia. The spread of the local lesion is dependent upon local tissue necrosis. The tissue necrosis in turn is dependent upon the elaboration of bacterial toxins, which are distributed along the line of the muscle sheaths and facia, and through the lymph spaces. There is no evidence that the toxin producing the local tissue necrosis differs from the toxin which is the cause of the general toxemia. If for any reason toxins are not elaborated within the wound or are not absorbed from the wound a gaseous gangrene does not develop despite the fact that there may be within the wound a large number of potentially pathogenic anaérobes. All the aérobes can be dismissed as a cause of infectious gaseous gangrene. Any effects which they produce are in the nature of a complication. At the most their réle in this disease process is confined to the absorption of oxygen, the turning upon BACTERIOLOGY OF INFECTIOUS GASEOUS GANGRENE. 31 themselves of the processes which tend to produce an immunity and in causing the death of tissue thus preparing a favorable media for the multiplication of the anaérobes. They have never been isolated in pure culture from a case of infectious gaseous gangrene and have never produced typical lesions when inoculated into animals. There are a number of anaérobes which cause characteristic lesion in animal inoculations and one or more of which are always isolated from cases of gaseous gangrene. In the following arrange- ment the organisms that are found in cases of gaseous gangrene are grouped with regard to their importance as causal agents of infectious gaseous gangrene; and there is in addition an indication of their main action toward the carbohydrates and proteids. Grovp I. Essential Causal A gents of Gaseous Gangrene. Saccharolytic: 1. Vibrion septique. 2. B. edematiens (B. Novyii). 3. B. welchit. | Proteolytic: B. hastolyticus. Group II. Accessory Agents of Gaseous Gangrene. Comprises organisms capable of causing gaseous phlegmons (even of a severe type). Probably rarely or never an essential agent of gaseous gangrene. Important in association with the essential causal agents and generally unimportant when occurring alone. I. Anaérobes: A. Proteolytic group: B. sporogenes. B. aérofetidus. B. Saccharolytic group: B. fallax. II. Aérobes: B. colt. 32 SCIENTIFIC PROCEEDINGS (102). B. proteus. (Both very doubtful as causal agents of true gaseous gangrene.) Group III. Organisms Merely Present in Gaseous Gangrene. Never essential causal agents, though they may be capable of causing complications; and may have an importance in symbiosis with other organisms; and may be a cause of a great modification of the clinical picture. A. Anaérobes: Proteolytic: B. putrificus. . B. bifermentans (also has a powerful saccharolytic action). | B. tetant. B. tertius. B. Aérobes. Cocci: Streptococci, staphylococci and diplococci. Bacilli. (Gram negative.) B. proteus, coli, pyocyaneous, etc. Bacilli. (Gram positive.) B. anthracoides group B. subtilis group B. mesentericus and myscoides group, etc. The pathogenicity of this entire series exhibits great variation. There are strains of B. welchit and B. edematiens that have almost no virulence. The B. welchii is notable in this respect, a few strains showing great pathogenicity, while many others have little or none; and the average is not sufficiently high to make it an easy matter to produce an antitoxin of high titer. Under exceptional circumstances the B. sporogenes, fallax and aérofetidus are capable of producing a lesion alone, though as a rule these lesions are of a benign character of a type of a gaseous phlegmon. In infectious gaseous gangrene these organisms are commonly associated with organisms of greater pathogenicity such as the B. welchii or edematiens, so that their r6le approaches that of accessory micro- HYDRATION EFFECTS OF AMINO-COMPOUNDS. 33 érganisms. In a bacteriological analysis of 308 cases of gaseous gangrene, of which 91 were derived from Weinberg’s series and 217 from wounded American soldiers, the percentage of incidence of the various pathogenic anaérobes is as follows: Per Cent. PERS f era OCR po ea oe co ee arte 85 EO Bp ae ns eis eee ae rer eee ee 35-4 DOTS Tate oe ke POOL pt wet eae dined a dee oo Be 8 wee 12.6 ens OMEN. 2 Chas co ten ye a awe ev ach assent sedans 17.2 Te ac thes 1a ta aide as Ae ats ée bie dice WiolnthieisieW © 6.4 This is not the true incidence of the B. edematiens, fallax, histoly- ticus or aérofetidus as the difficulties in the isolation of these organisms in pure culture is great, but it does serve to emphasize the fact that infectious gaseous gangrene is usually a mixed infection. In this group of 308 cases of infectious gaseous gangrene only 79 were infected with a single pathogenic anaérobe, the remaining 229 having from two to six anaérobes in the local lesion, and nearly always at least two pathogenic anaérobes. 19 (1479) Hydration effects of amino-compounds. By D. T. MAacDouGaL and H. A. SPOEHR. [From Desert Laboratory, Tucson, Arizona.| The chief interest in the results presented in this brief paper depends upon the following facts and conditions: A. The amino-compounds furnish the only known solutions in which agar and other pentosans or mucilages undergo a greater hydration than in distilled water. Tentative conclusions to this effect have been confirmed by all of the results obtained during the past year.! ‘B. The pentosans, or anhydrides of the 5-carbon sugars are universally and abundantly present in plant cells, originating by transformations of wall-material, starch, etc., in any part of 1 MacDougal, D. T. and H. A. Spoehr, ‘‘The effect of organic acids and their amino-compounds on the hydration of agar and on a biocolloid,’’ Proc. Soc. FOR EXPER. BIOL. AND MED., 1918, xvi: 33-35. 34 SCIENTIFIC PROCEEDINGS (102). the protoplast, and presumably intimately interwoven into its colloidal mesh. In animals the pentosans seem to be confined to the nucleo-proteins, and the manner of their origin is not so clear in this case. C. The mucilages, gums and slimes in which form these sub- stances appear in definite masses in syneretic cavities and in layers in the plant cell have a hydration capacity enormously greater than that of the sugars from which they are derived, and show a wide range of solubility and other qualities. D. The pentosans are subject to digestion in animals to an extent variously assigned by different authors. These sub- stances undergo metabolic changes in the plant but slowly. Wherever they occur they must show changes in volume and form according to the colloidal structure in which they occur and to the nature of the solutions penetrating them. Our own experimentation has been made principally with agar and some of the common plant gums, separately and in mixtures with albumins. The revised generalizations which we are pre- pared to support may be briefly stated in the following summary: 1. The pentosans are weak acids which dissociate so slightly that I per cent. solutions of agar, acacia and cherry gum showed pH values of 5.1 as determined by the indicator method. The mucilage of Opuntia showed a value of 5.8. The swelling of these substances in the amino-acids which dissociate strongly, as aspartic acid which shows a pH of 3 at 0.01 M is less than in water. 2. Such acids and other amino compounds as asparagin, pH = 6.2, alanin pH = 7.0, phenyl-alanin pH = 4.8, glycocoll pH = 6.2 ato.o1 WW as tested by the indicator method, ammonium hydroxide and ethylamine facilitated the hydration of agar so that it showed swellings greater than in distilled water. 3. Other factors than the hydrogen ion concentration are - determinative in this action as illustrated by the fact that swellings in phenyl-alanin and glycocoll are fairly equal. The total swelling of agar as compared with that in water as unity was 1.26 in aspar- agin, 1.52 in alanine, 1.65 in glycocoll and in phenyl-alanin, in ammonium hydroxide and in ethylamine, all in 0.01 M solutions. Propionamide the only amide tested did not exert any marked effect on swelling except to retard it slightly at 0.01 M and in stronger solutions. HyDRATION EFFECTS OF AMINO-COMPOUNDS. 35 4. Glycocoll was used in extending these experiments, and it was found to exert an accelerating effect on agar when mixed with soy bean albumin, gelatine and similar increases were also shown when the agar was partly replaced by such mucilages as that from Opuntia, acacia and cherry gum. 5. Living and dried sections of tomato fruits, growing cell- masses of stems of Phytolacca, and joints of Opuntia, the cell-sap of which has varying acidity, showed the greatest swellings in alanin, phenyl-alanin, glycocoll and ammonium hydroxide as compared with results in water and organic acids, with but few exceptions. Taking water as unity living sections of Opuntia discata showed swellings of I. in alanin, 1.5 in phenyl-alanin, and an equal increase in glycocoll. Dried sections of the same material showed swellings of 1.3 in alanin, 1 in phenyl-alanin, and 1.4 in glycocoll solutions. Living sections of Opuntia leptocaulis swelled 1.4 in alanin, 1.2 in phenyl-alanin, 1.55 in ethylamine, while dried sections of this species gave increases of 2 in alanin and 1.5 in ammonium hydroxide and 2.2 in phenyl-alanin as compared with water. 6. The swellings or increases due to hydration were determined by the use of the auxograph. It has been found that the hydration of dried sections of such pentosans as agar causes changes which are the reverse of those which ensue during desiccation. Plates of this substance poured from a warm 2.5 per cent. solution, which were fastened at the margin in such manner as to prevent shrinkage in area and to allow decrease in thickness only, when swelled showed increases of not more than 4 and generally as little as 2 per cent. laterally, while swelling 4,000 per cent. in thickness. 7. Sections of gelatine from plates cast in the above manner may show a lateral expansion of 8 to 40 per cent. while swelling 500 to 2,000 per cent. in thickness, in water and in acid solutions with a pH value of 2. _ 8. The swelling of gelatine which showed a pH value of 5.2 in an 8 per cent. solution in nitric and hydrochloric acid at a pH value of 2, in succinic acid (0.01N) at a pH value of 3.05 and in amino-succinic acid (aspartic acid) at a pH value of 3 was much greater than water, but the swelling in alanin, phenyl-alanin and glycocoll was less than that in water. 36 SCIENTIFIC PROCEEDINGS (102). g. The pentosans on the one hand and the albuminous com- pounds on the other are intimately intermixed or interwoven in the protoplast of the plant and in the nucleus of the animal cell. The conditions which accelerate the hydration of the first may not affect the volume of the other except to cause a slight shrinkage. The effect of the hydrogen ion is to increase the hydration capacity of albumin and its derivatives, while lessening the hydration of the carbohydrates. 10. In addition to these differential effects of the solutions upon the principal components of the plasmatic colloids, the changes in volume are not isotropic but may be determined by the manner in which the mesh, masses of colloidal material or organs of the protoplast are laid down or fall into place. These two classes of variables may well be considered as prolific sources of differentiation in the procedure of the cell. ; 20 (1480) Profound effects of digitalis on the vagus producing severe detrimental subjective symptoms, as shown by simultaneous electro-cardiograms and pneumograms. By ROBERT H. HALSEy (by invitation). [From the New York Post-Graduate Medical School.| In a case of syphilis, chronic interstitial nephritis, dilatation and hypertrophy of the heart, auricular fibrillation, arterio- sclerosis, periodic breathing, Cheyne-Stokes type, in which because the auricule was fibrillating, digitalis had been given for the pur- pose of showing the ventricular rate. Counts of the pulse showed the ventricle to be contracting at a rate approximating 100 during apnea and slowing to about half this rate as soon as hyperpnea . began. That is, the heart rate was highest while there was no lung ventilation (apnea) and the heart beat less frequently during the period the lung ventilation was greatest (hyperpnea). As the digitalis was continued the man complained of increasing distress, most severe during his hyperpnea. The ventricle rate did not become slower and evidently the patient was much more uncomfortable. The thought occurred that releasing the heart EFFECTS OF DIGITALIS ON VAGUS. 37 from vagus control might give relief, therefore, atropine gr. 1/30 was given by hypodermic injection. Record taken 27 minutes later shows the ventricle beating irregularly at the rate of 150 per minute but uninfluenced by breathing. The patient became comfortable and the cyanosis was less. Discussion.—As the slowing of the ventricular rate followed the first inspiration after apnea and was maintained until the last part of the hyperpnea stage, it was considered to be due to the inhibition of the vagus which was stimulated by the reflexes from the lung generated by the respiratory efforts of hyperpnea. The average hyperpnea periods were of 37 sec. duration. The reflex from the lung became much less effective as a stimulant to the vagus during the descending phase of the hyperpnea; for, the ventricular rate became accelerated several inspirations before apnea began. This loss of vagus effect upon the heart rate was due, probably, to the fact that reflex stimuli from the lung tissue when it is con- tracting are not so strong as when the pulmonary tissue is being distended, but not to true vagus escape of the heart. That the vagus controlled the ventricular rate was shown by absence of alterations of the ventricular rate while the atropine was effective. During atropine effect the hyperpnea persisted for 47 seconds, a ten second increase over the digitalis period. The distress of the patient was due to the diminished inter- change of O, and CO, between the lungs and blood. This inter- change was least during apnea, when the pulmonary ventilation was lowest though the heart rate was highest, but during hyper- pnea, when the ventilation of the lung was greatest, the heart rate was slowest. The sensitiveness of the vagus to the pul- monary reflex was increased by digitalis to such a degree that the number of ventricular contractions during hyperpnea were very much less than when digitalis was not given. _ To state this in another way; it appears that without digitalis the amount of interchange of O2 and CO, was sufficient, to keep the patient comfortable, notwithstanding the inverse periodic changes in rate of heart and breathing; but, when digitalis was given, the vagus became much more susceptible to the lung re- flexes slowed the circulation so as to diminish the interchange of 38 SCIENTIFIC PROCEEDINGS (102). O, and CO, between the alveolar air and the blood sufficiently to cause real distress. Relief followed the discontinuance of digitalis and the administration of belladonna. Conclusion.—In this case of auricular fibrillation digitalis did not improve the circulation because it inrceased vagus affects which diminished the O, and CQ, interchange between alveolar air and blood. 21 (1481) The action of camphor on the central nervous system of the squid. By A. R. Moore. [From the Physiological Laboratory of Rutgers College, and the Marine Biological Laboratory, Woods Hole, Mass.| Newly hatched squid (Loligo pealiz) if put into a solution of camphor gum in sea water, 1/10 saturation, show characteristic mantle spasms, involving play of the chromatophores, after a latent period which is about 40 seconds at 24° C. This effect, it has been shown, is due to the action of camphor on the stellar ganglia.!. In appearance, camphor spasms are indistinguishable from those caused by nicotine.? Thedifference does not lie in the character of the response of the end organs, muscles and chromato- phores, but in the locus and the nature of neuronic excitation. The value of the temperature coefficient Qi» for the action of camphor, based on the lengths of the latent periods, is 2.4. The function connecting the velocity of the reaction with the concen- tration of the drug is, for camphor, expressed by v = RC? in I latent period’ whose value is approximately .75. The camphor spasms soon pass off and the animals lie inert with chromatophores relaxed. This is not due to paralysis of the end organs for the reason that they may again be thrown into activity by treatment with strychnine or nicotine. The absence of any interference between the action of nicotine and that of cam- phor may be demonstrated by the following experiment. Let which v = C = concentration, and & is a constant 1 Moore, A. R., Proc. Nat. Acad. Sc., 1917, III, 598. 2 Moore, A. R., J. Gen. Physiol., 1919, I, 505. STANDARDIZING BACTERIAL SUSPENSIONS. 39 the specimens of squid be immersed in nicotine solution for 1 minute. They are then put into the solution of camphor. Cam- phor spasms take place, after which the animals lie quiet for several minutes. At the end of the required latent period! nicotine spasms occur just as typically as in animals treated with nicotine alone. 22 (1482) A method of standardizing bacterial suspensions. By FREDERICK L. GATES. [From the Department of Pathology and Bacteriology of the Rocke- feller Institute for Medical Research, New York.| If a wire loop is thrust down into a suspension of bacteria in a test tube, and viewed by looking down into the mouth of the tube, the depth at which the loop disappears will be determined by the opacity of the supervening suspension. If, however, a second suspension of the same organism containing half as many bacteria per cubic centimeter is similarly examined, or if an equal amount of the diluent is added to the original suspension, and the ‘‘depth of disappearance’? again measured it will be found to be less than twice as great as in the original suspension. In other words, the observed depths of disappearance are not in proportion to the bacterial concentrations or the corresponding volumes. This discrepancy is due to the presence in each reading of a constant which is apparently related to the size and opacity of the individual organisms. It is found that this constant may be elimi- nated, thus bringing the opacity observations into inverse ratio with the corresponding bacterial concentrations, and a corrected reading (the observed reading minus the constant) for any sus- pension may be obtained by making two readings at different dilutions of the suspension, and substituting the observed values in the following equation: _ vola (6 — a). ~ vol 6 — vol a’ A 1Moore, A. R., J. Gen. Physiol., 1919, I, 505. 40 SCIENTIFIC PROCEEDINGS (102). in which A = the corrected reading for the first volume of the suspen- sion = (a — constant) a = first observed reading. vol a = first volume of the suspension on which reading a is made. b = second observed reading. vol b = second volume of the suspension (diluted), on which reading 0b is made. A concrete example will illustrate the method. In 4 c.c. (vol a) of a given suspension the loop disappears 1.2 cm. below the menis- cus (reading a). The suspension is diluted to 10 c.c. (vol 3). The loop now disappears 2.7 cm. below the meniscus (reading }). Then the corrected reading for the opacity of the suspension (A), jp (27 — = or 1.0 cm. Such a corrected reading may be directly compared with cor- rected readings on other suspensions of the same organism, since the corrected readings on such suspensions stand in inverse ratio to their bacterial concentrations. Thus, if two suspensions of the same organism are to be compared and the corrected reading for one is half that for the other, the first suspension contains twice as many bacteria as the second. If the actual bacterial count is required, a standard for the given organism must first be established by the correlation of several corrected readings with the corresponding counts, obtained by the usual methods. Thereafter the concentration of the organism per cubic centimeter - in suspensions under examination is obtained by inverse propor- tion: The required count The standard corrected opacity The standard count. The given corrected opacity The simplest possible instrument for making the determinations consists of a piece of 18 gauge nichrome, chromel, or black iron wire about 20 cm. long, bent into a small loop at right angles to one end, and with the other end thrust through a cork, near one side, so that a view may be obtained past the cork into an ordinary test tube, 1.6 X 16 cm. on whose lip it rests. The sterile test tube is partly filled with a measured quantity of the suspension, and the wire loop raised or lowered through the cork until the SURFACE LAYER IN THE LivING EGG CELL. 41 point is found at which the loop just disappears from view. The distance of the loop below the meniscus is then measured with a centimeter scale laid along the tube, giving the first reading. A second reading is obtained after the addition of a measured amount of the diluent, and the data required for substitution in the equation for the corrected reading is at hand. The readings and calculations require but two or three minutes. In the zone of most accurate measurement, with suspensions of such opacity that the loop disappears between I and 4 cm. below the surface, repeated readings may be made with a variation of about one milli- meter, an error of less than 10 per cent. The wire loop, which alone comes in contact with the bacteria, may be flamed after each determination, thus reducing the danger of contamination to a minimum. A more complete explanation of the method, with a description of the more convenient and accurate instrument shown at the meeting will appear in a forthcoming number of the Journal of Experimental Medicine. 23 (1483) Some studies on the surface layer in the living egg cell. By ROBERT CHAMBERS. [From Cornell University Medical College.| The results recorded here were obtained through the use of Barber’s mechanical pipette holder somewhat modified for micro- dissection purposes. The cells experimented upon were the egg cells of the starfish and of the sea urchin. The eggs, which are somewhat over 1/10 of a millimeter in diameter, were placed in a drop of sea water hanging from the roof of a moist chamber. The microscopically fine tips of the glass dissecting needles projected into the moist chamber and up into the hanging drop. By manipulation of the screws of the mechanical pipette holder the cells in the hanging drop could be dissected with considerable accuracy and an esti- mate ascertained of their physical consistency. Detailed accounts 42 SCIENTIFIC PROCEEDINGS (102). of Barber’s apparatus and its application to microdissection have already been published.! The egg cells studied consist of a decidedly fluid interior sur- rounded by a more solid surface layer of appreciable thickness. This surface layer is most solid on its external surface. Internally its consistency seems to merge insensibly into that of the fluid interior. The inner surface of this layer adheres to the touch. This is demonstrated by introducing a microdissection needle into an egg and pushing the needle through until its tip comes into contact with the inner boundary of the surface layer on the side of the egg opposite the puncture. On withdrawing the needle the layer adheres to the needle tip and strands are drawn into the interior of the egg. | If the surface layer be torn while the egg is kept under compres- sion the fluid interior will bulge out through the tear. The cytoplasm, on coming into contact with the surrounding water, tends to establish a definite surface film which prevents the cyto- plasm from mixing with the water. If the internal pressure be not too great this film persists and, in time, strengthens into a definite ectoplasmic layer. The bulge then slowly retracts until the orig- inal contour of the egg is reéstablished. If the neck of the pro- truding mass of cytoplasm be small it may pinch off a spherule of cytoplasm which to all appearances is normal. If the internal pressure be too great a succession of films may form as, one after the other, they succumb while the escaping cytoplasm disperses and disintegrates in the surrounding water and the film which finally holds out may enclose only a fraction of the original cell but what it encloses will be normal protoplasm.? Churning of the contents of a mature unfertilized sea-urchin egg causes the ectoplasmic layer to revert to the fluid condition of the interior. The surface film of such an egg is very thin and very easily tears upon which the entire egg disintegrates. On standing, however, the surface film steadily strengthens until the normal condition is reéstablished. That the distribution of substances throughout the egg cell is 1 Barber, Philippine Journal of Sc., Vol. X, Sec. B, Tropical Medicine, 1914; Chambers, Biol. Bull., Vol. 34, 1918. 2 Chambers, Amer. Journ. Physiol., Vol. 43, 1917. eer SURFACE LAYER IN THE LIVING EGG CELL. 43 not uniform can be demonstrated by the following experiment on the starfish egg: If the surface of a mature unfertilized egg be torn while the egg is kept under compression almost all of the internal cytoplasm may be made to flow out to form a spherule of cytoplasm which pinches off from the rest of the egg. What is left behind is a collapsed remnant consisting mainly of protoplasm which originally enveloped the egg. This remnant consisting largely of the more solid ectoplasm tends only slowly to round up. The extruded mass, which is very fluid, immediately assumes a shape approximating that of a sphere. This may be termed an endoplasmic sphere. The remnant containing the original ecto- plasmic substance of the egg is readily fertilizable and undergoes segmentation. The endoplasmic sphere is unfertilizable. If, on the other hand, the endoplasmic sphere remains for some time connected by means of a bridge of protoplasm with the remnant containing the original ectoplasmic substance it is fertilizable. The ability of the endoplasmic sphere to approximate normal conditions of segmentation is a function of the length of time that it remains in organic continuity with the original ectoplasmic mass. Possibly there exists a substance necessary for develop- ment which normally accumulates in the surface layer of an egg. This substance is diffusible and will distribute itself over new protoplasmic surfaces. If a bridge of protoplasm connects the ectoplasmic remnant with the endoplasmic sphere this substance will diffuse into the sphere thereby rendering it fertilizable. The nature of the surface film produced by cutting an egg cell differs in an unfertilized egg from one which has been fertilized. Before fertilization the needle may be pushed vertically into the side of the egg and moved through the egg from one side to the other without cutting the egg in two. The cytoplasm close, behind the needle thus obliterating the furrow. Shortly after fertilization, however, such a procedure cuts the egg cleanly in two. The sides of the furrow produced by the needle do not fuse although contiguous. The character of the surface film which forms over a cut is thus changed upon fertilization. This change prepares the egg for the ensuing segmentation process by causing the formation of a type of surface film which prevents contiguous blastomeres from fusing with one another. 44 SCIENTIFIC PROCEEDINGS (102). 24 (1484) Concerning the toxicity of acetanilid and bicarbonate combina- tions for muscle-nerve preparations. By Davip I. MAcurT. Pharmological Laboratory, Johns Hopkins University.] It is well known, through the work of Worth Hale,! that ace- tanilid when given in combination with small doses of sodium bicarbonate, is less toxic than when administered in the same doses alone. The studies of Macht, Greenberg and Isaacs on the influence of antipyretics on the acuity of hearing? have also shown an interesting difference between the effects of acetanilid when given alone and acetanilid when combined with sodium bicarbonate. The explanations given for the above peculiar synergism are most unsatisfactory. In connection with a pharm- acological study of various antipyretics, the author investigated the effect of acetanilid solutions on muscle-nerve preparations which may throw some light upon the above-mentioned phenomena. Two gastrocnemius muscle-nerve preparations of a frog are immersed simultaneously in physiological sodium chloride solution and the limits of both the muscle and nerve excitability to the electric shocks of an induction coil are determined. One of the preparations is then immersed in a solution of acetanilid in the same saline solution, and the other preparation is immersed in a solution of acetanilid of exactly the same strength but containing © some sodium bicarbonate (I-1000 or even weaker). On testing the excitability of the muscles and nerves at regular intervals after treatment with the drugs, it was found that the plain acetanilid solution tends to paralyze and finally kill the sciatic nerve more quickly than the solution of acetanilid plus bicarbonate. The following protocol of an experiment thus performed will serve as an illustration. The slower toxic action of the acetanilid and bicar- bonate combination, all other factors being equal would, it is fair to assume, render the drug less poisonous when injected into an 1U. S. Hygienic Lab. Bulletin No. 53 (1909). ? Proc. Soc. FOR Exper. BIOL. AND MED., I919, xvii, 22, 23. ee nn — -_ = ee ee SS CARBOHYDRATES OF THE ROOT OF THE CAT TAIL. 45 animal than acetanilid would be if injected alone, for in case of the more slowly acting combination the animal has a longer period of time for the excretion of the poison. The author does not at all presume to offer these experiments as a complete explanation of the above-described synergism, but it is thought that the present findings are at least a little tangible contribution towards the explanation of the peculiar phenomenon. EXPERIMENT, OCTOBER 28, IQIQ. Stimulation of Gastrocnemii Muscles of Rana Clamaia. Left Side. Right Side. Stimulation of Nerve. Position of Secondary Coil. Stimulation of Stimulation of Nerve. Position Muscle. Position of Secondary Coil. | of Secondary Coil. Time. Stimulation of Muscle. Position of Secondary Coil. Immersedin physiological NaCl Sol.| Immersed in physiological NaCl Sol, 2:45 p.m. 23.0 cm. 37.0 cm. 24.5 cm. 47.0 cm. 2:50 ae 23. re) ae S70 ae 24. 5 “é 43.0 sé 3:05 ae 22. fe) “é 39. fe) sc 24. 5 sé 44.0 “se cy Immersed in physiological NaCl | Immersed in physiological NaCl Sol. containing acetanilid 1:500 | Sol. containing acetanilid 1:5000 and sodium bicarbonate 1:1,000 ast." 20.1 cm. 27.0 cm. 22.5 cm. 23.5 cm Ee: a ee Pa NL) oll wanes Ss ane ante * oe 25.0‘ 22'a° °° oa. '*! Po rete roe“? yA ia 12:0)" | PA 8 (ai 2.46..." mero." 25.6: 20 Gi 23 20.0. * 4:02.“ rs ah soigiei ! Ta:0n7 | 18.0 “ cic a 1 A eg a0)" T0.0: ."* oe asag.5 c ree 29.0. -" Cee a5 3. 4:460-.“* nO. S700" ),* - Bt et: eae ui 1 50.52'" S20) ** ee ea 25 (1485) A note on the carbohydrates of the root of the cat-tail (Typha latifolia). By ZALIA JENCKS (by invitation). eC the Sheffield Laboratory of Physiological Chemistry, Yale University, New Haven, Conn.| In a recent communication! the root of the cat-tail has been recommended as a valuable food product for man. An analysis is recorded to indicate that the material contains 81 per cent. of carbohydrates. No evidence is presented however, as to the 1P, W. Claasen, The Scientific Monthly, 1919, Vol. 9, No. 2. 46 SCIENTIFIC PROCEEDINGS (102). precise identity of the latter. In view of the fact that various roots are known to contain carbohydrates, like inulin, which are by no means identical in physiological value with starch although they have various reactions in common with it, I have separated the most abundant carbohydrate of the cat-tail root for identifi- cation. It gives a blue color with iodine, forms a characteristic paste with hot water, is readily digested (in contrast with inulin) by saliva, and yields on hydrolysis a dextro-rotatory solution from which an osazone, identical with glucosazone, was prepared. The carbohydrate thus corresponds with starch. Our “flour’’ indicated a carbohydrate content of 56.8 per cent., estimated in the conventional way from the reducing sugar formed by hydrolysis with acid. To test the innocuousness of the cat-tail root as a food mice were fed for a week on otherwise adequate diets containing 30 per cent. of the ‘‘flour’’ without evident untoward results. The animals gained in weight upon the ration. 26 (1486) Do fruits contain water-soluble vitamine? By THOMAS B. OSBORNE and LAFAYETTE B. MENDEL. [From the Laboratory of the Connecticut Agricultural Experiment Station, and the Sheffield Laboratory of Physiological Chemistry in Yale University, New Haven, Conn.| Although fresh fruits have long been classed as valuable anti- scorbutic foods there are comparatively few recorded scientifically planned tests of their potency aside from the familiar studies of the juice of oranges and lemons. With respect to the possible presence, in fruit, of water-soluble vitamine (water-soluble B) comparable to this essential factor in yeast, scarcely anything has been published. We have begun experiments on rats in the otherwise adequate diet of which fruits and fruit juices furnish the the sole source of the water-soluble vitamine. When larger portions (more than 5 grams per day) of fresh apples and pears are fed the characteristic decline in weight observed where vita- BIOLOGICAL CHARACTERISTICS OF BACILLUS BOTULINUS. 47 mine-free diets devoid of water-soluble vitamine are used, is averted. The bulky character of such fruits has made it imprac- ticable to feed more than 10 grams per day without decreasing too greatly the intake of other essential nutrients. Ten c.c. of orange juice per day suffice to promote considerable growth. The inner peel of the orange (which Hess has found to be antiscor- butic) seems also to contain some of the other water-soluble vita- mine. It is already evident that the proportions of the latter in the fruits tested is not large in relation to the quantities edible. 27 (1487) Some observations on the biological characteristics of bacillus botulinus. By Pau F. Orr (by invitation). [From the Department of Preventive Medicine and Hygiene, Harvard University Medical School, Boston, Mass.| In a study of sixteen strains of Bacillus botulinus, which have been isolated in connection with outbreaks of food and forage poisoning occurring in different parts of the United States during the past five years, a number of interesting facts have been ob- served. While a complete report of the findings will be published elsewhere, it seems justifiable to record at this time some of the salient facts; namely: 1. Contrary to the general view that the optimum temperature for growth of B. botulinus is about 22° C., we have found that the body temperature 37° C., is most favorable for the growth and spore production of all of the sixteen strains of B. botulinus studied. Toxin is readily formed at this temperature. 2. At autopsy of guinea pigs which have been either fed or injected with cultures of B. botulinus it has been possible to recover this organism quite frequently from the liver and spleen and also occasionally from the heart’s blood, the kidneys and the pancreas. 3. Contrary to the results obtained by previous investigators, guinea pigs, which have been fed or injected with toxin-free spores of B. botulinus, Nevin strain, have died with symptoms resembling those of botulinus poisoning. At autopsy of these animals B. botulinus was recovered from the liver and spleen. ver a Agree oh ene han oy = ~~ mina <2 « a ae reo oy eitted ha ace SCIENTIFIC PROCEEDINGS ABSTRACTS OF COMMUNICATIONS. One hundred third meeting. Rockefeller Institute for Medical Research, New York City, December 17, 1919. President Calkins in the chair. 28 (1488) The role of fat-soluble vitamine in the dietary of infants. By ALFRED F. HEss and LESTER J. UNGER. [From the State Laboratory, Department of Health, New York City.] It has been shown that the fat-soluble vitamine is an essential constituent of the dietary of rats. There have also been clinical reports attributing marked malnutrition in infants and children to a lack of this dietary factor (Japan, Denmark). As a result of these experiences it has been accepted that this vitamine is highly important for man, and that the lack of it leads to nutritional disorder in children. This has been emphasized all the more as this vitamine is not nearly as widely distributed in nature as is the water-soluble vitamine. -In order to study this question five infants, varying in age from 5 to 12 months, were given a diet which was complete except for a very small amount of fat-soluble vitamine. It consisted of 180 g. daily of highly skimmed milk (Krystalak 0.2 per cent. fat), 30 g. of cane sugar, 15 to 30 g. of autolyzed yeast (to supply water-soluble vitamine), 15 c.c. of orange juice, 30 g. of cottonseed oil, and cereal for the older infants. _On this diet the children have done well for a period of eight to nine months. They have shown no anemia, no eye trouble, no bone changes, as seen by the X-ray, nor has their growth in length or in weight suffered. We believe, therefore, that either a very small amount of this vitamine suffices to supply the needs of human nutrition, or that this deficiency has to be maintained 49 50 SCIENTIFIC PROCEEDINGS (103). for a period of years in order to bring about any harmful result. Danger from a lack of this dietary factor need not be apprehended if the diet is otherwise complete. The development of rickets has been attributed by Mellanby, as a result of experiments on dogs, to a lack of fat-soluble vitamine, and Hopkins and Chick have termed this vitamine the “‘anti- rachitic factor.”’ It was found, however, that infants fed on this ‘fat-soluble vitamine minimal diet’’ did not develop the well- established signs of rickets—beading of the ribs, enlargement of the epiphyses, weakness of the muscles, etc. We cannot believe, therefore, that rickets is brought about by a deficiency of this principle; all the more so, as this disorder developed in infants receiving large quantities of milk containing ample fat-soluble vitamine. It may be added that neither cream nor the leafy vegetables, both of which are rich in this principle, were found to be comparable to cod liver oil as growth stimulants. 29 (1489) A method for the determination of calcium, magnesium, potas- Sium, sodium, chlorides and “acid-soluble”? sulfur and phosphorus in one sample (25 c.c.) of blood. By IsIDOR GREENWALD and JOSEPH GROSS. [From the Harriman Research Laboratory, The Roosevelt Hospital, New York.] The blood is laked with eight volumes of water and the protein is then precipitated by the addition of one volume of 1:1 nitric acid. Ina portion of the filtrate chlorides are determined gravi- metrically by precipitation with silver nitrate and filtering on a Gooch crucible. The excess of silver in the filtrate is removed with hydrochloric acid and the resultant filtrate and the remainder of the blood filtrate are combined and treated with copper nitrate and perchloric acid, evaporated to dryness and then to blackness. The residue is treated with concentrated hydrochloric acid, again evaporated to dryness and then dissolved in dilute hydrochloric 1 Redness must be avoided, or potassium will be lost. This is a modification of Benedict's method (Journal of Biological Chemistry, 6, 363, 1909). — — —— A BENIGN TUMOR IN DROSOPHILA. 51 acid. Sulfates are precipitated with barium chloride and the filtrate is made ammoniacal, then acid with acetic acid and phos- phates are precipitated with ferric chloride. The precipitate is dissolved in nitric acid and the phosphoric acid is precipitated with ammonium molybdate and the yellow precipitate is titrated. The filtrate from the ferric acetate precipitate is treated with sulphuric acid to remove the barium and with hydrogen sulfide to remove the copper and the filtrate is evaporated to small volume. Calcium is precipitated in a centrifuge tube as oxalate, washed and titrated. The supernatant liquid and washings are evaporated, heated to remove ammonium salts and the magnesium is precipitated in a centrifuge tube as magnesium ammonium arsenate. - This is washed and then dried, 7m vacuo, over sulfuric acid. It is dissolved in acid, transferred to a distillation flask and the ammonia determined by distillation and Nesslerization. The supernatant liquid and washings are treated with hydrochloric and hydrobromic acids, evaporated to dryness, freed of ammonium salts and arsenic, sulfuric acid is added and the potassium and sodium are weighed as mixed sulfates in a platinum dish. Potas- sium is then precipitated as the cobaltinitrite and weighed.! 30 (1490) A benign tumor in Drosophila. By Mary B. STARK. (by invitation). [From the Zoélogy Laboratory, Indiana University.] In a strain of flies with a lethal tumor, 7.e., a tumor occurring in one half of the males and causing their death, another tumor has appeared as a mutation. The new tumor differs from the lethal one in that it is not sex-linked, 7.e., it appears in females as well as males, and further in that it does not cause the death of the flies in which it occurs. Linkage experiments show that one at least of the genes essentially for tumor development is in the third chromosome closely linked to dichaete. 1 Garola and Braun, Annales de falsifications, 10, 572, 1917. 52 SCIENTIFIC PROCEEDINGS (103). The tumor may occur in any segment of the larva but seems to occur more often in the twelfth and thirteenth segments. When the tumor occurs in the thoracic region, there may be an ingrowth of tumor cells into the imaginal discs of the appendages checking the development of these parts. The cells of the tumor are rounded or polygonal in shape and show the presence of pigment. 31 (1491) The suitability of the ‘‘Bachman Test” for water-soluble B. By WALTER H. EDDY and HELEN C. STEVENSON. [From Teachers College, Columbia University, New York City.] Two recent publications by Drs. Bachman! and Williams? dealing with the vitamine requirements of yeasts suggest that the methods developed are adaptable to quantitative measurement of vitamine content (B variety). At the suggestion of the senior author Miss Stevenson has conducted experiments with both methods and in this report are presented some of the results with the Bachman test. Briefly this method consists in planting yeast cells in a culture medium (Nageli’s solution: 100 c.c. distilled water; 10 gms. dextrose; I gm. ammonium nitrate; 0.05 gms. calcium phosphate; 0.5 gms. potassium acid phosphate; 0.25 gms. magnesium sulfate) contained in a Durham or Smith fermentation tube and incubating the tubes at 28-32° C. to obtain gas formation. ‘To these tubes are added vitamine ‘‘B’’ extracts from various sources and Dr. Bachman’s results showed that in the absence of such extracts gas formation either fails to take place or at least very slowly. Our experiments aimed to confirm Dr. Bachman’s results, to determine whether the method gave promise of use quantitatively and whether it might be used to detect the ‘“B’’ vitamine quali- tatively. The results of our experiments follow: 1*The ‘‘ Vitamine Requirements of Certain Yeasts,"’ by Freda Bachman, Journ. Biol. Chem., XX XIX, 235 (1919). 2‘*A Simple Biological Test For Vitamines,’’ by R. W. Williams, Journ. Biol. Chem., XXXVIII, 465 (1919). —_—— -" THE ‘‘ BACHMAN TEST.” 53 EXPERIMENT I. To test the applicability of the method to ‘‘B’’ Vitamine extract as prepared by McCollum! from Navy Bean. Per Cent. of Gas-formation by Days. Contents of Tubes. 1. Control tube. Contained Nageli solution and one loopful of yeast cell suspension..| 0 | 0|!0]0j/]0!{]0!]0)0 oO 2. Control tube 2. Contained Nageli solu- tion, loopful yeast cell suspension and 1 c.c. of a dextrin solution made by dis- solving 20 gms. dextrine in 150 c.c. of dis- tilled water. The dextrin solution was sterilized in the Arnold machine at 100° C.|} 0 | 0 | 0}0/]0}]0]0/)]0 fe) 3. Control tube 3. Contents same as tube 2 but the dextrin solution was sterilized again, after Arnold treatment, for 20 minutes in the autoclave at 15 lbs. and minte TOM Coot s ees Pel bi aes ely o}/ol|o!;|o};o];/o|]ojo () 4. Tube contained Nageli, loop of yeast and I c.c. of a dextrine vitamine extract from Navy Bean (for preparation see explana- tion below) sterilized in the Arnold at Mr Oa.) Oe apenas eae erie eae es ....| 0 | 90] 87| 63] 18 | upset at this point 5. Same as tube 4 but sterilized in the auto- clave in addition to Arnold sterilization. .| 0 | 93} 85| 58] 56/ 40] 38 | 37 33 Explanation: The vitamine dextrine solutions were obtained by first extracting 86 gms. of Navy bean flour with ether in the Soxhlet for 18 hours. The residue was then extracted for two six hour periods with boiling alcohol (95 per cent.) in a reflux condenser. The alcohol extract was mixed with 20 gms. of dextrine and evaporated to dryness. The solution was made by dissolving this dextrin vitamine in 150 c.c. of distilled water. The control dextrin solution was made by dissolving 20 gms. of dextrine in 150 c.c. distilled water. All the solutions were subjected to two 30 minute periods in the Arnold sterilizer with a 24-hour incubation period intervening. The materials tested in tubes 3 and 5 were then given an additional 20 minutes in the autoclave at 15 lbs. and approximately 120° C. A pure culture of the Fleisch- man round yeast was used. The suspension was made by transferring to 5 c.c. of sterile water several loopfuls of the agar slant culture and the tubes were inoculated with one loopful of this suspension. Our tubes were incubated at 32° C. The CO: is very rapidly reabsorbed by the Nageli solution and sealing the tube with mineral oil was not effective. Consequently readings may be discontinued after the per cent. of gas reaches the maximum. 1“ A Study of the Dietary Essential, Water-soluble B.,’”’ by E. V. McCollum, Journ. of Biological Chemistry, XXXIII, 55. 54 SCIENTIFIC PROCEEDINGS (103). EXPERIMENT 2. To determine the effect of variation in the concentration of the dextrine vitamine solution. Tube Contents. ............ o.1/Cc. Dextrine V. o.2/Cc. Dextrine V. Sterilization Method..... Arnold. Arnold Plus Arnold. Arnold Plus Autoclave. Autoclave. GEREN 55d ecpnasenswemur sioner I. 2 3 I 2 3 I 2 | 3 I | 2 3 Days 1 IE EE A oe Ae ae et () ) fe) fe) ) oO fe) fe) te) fe) fe) oO Wanda wa rine heels aie fe) fe) fe) fe) fe) Oo | 16 2 I | 50 oO fe) eps Rape Vay AO Se oO oO re) re) fe) Oo 1/63°] 35°) 78 + Je 2S. 195 WE eintag EM hate ee ees fe) fe) oO fe) oO © | 29°1'25,1°090 | 35 7 30 ) a Se icata ai Bias oie iote MRS oO fe) II 72 ialol ace £ pista Soamietereta oO I 3 oO fe) 0:1 20°) Bo) 9o +o tae Bo Fe abe tele’e erates fe) fe) 5 52 iuieS Saad ke ewe dette oO fe) 4 48 Pe inct teste fe) fe) 3 45 Tube Contents............ o.5/Cc. Dextrine V. z.0/Cc. Dextrine V. Sterilization Method..... Arnold. Arnold Plus Arnold. Arnold Plus Autoclave. Autoclave. CB ick cove Metdadociinenes I 2 3. I 2 3 I 2 3 I 2 3 Days Mic de helen ts dhs ec fe) o| oO oO oO o| oO fe) o| oO oO fe) SB iiadabics arcane 95 | 95] 53 | 26 | 60} 23] 90] 99| 68] 93 | 99] 68 ae odie Sy ietatakene in tie thse OA QI | 100] 95 | 5I | 100/ 100) 87 | 100} 100! 85 | 100} 100 Boys eet eda os tele 85 | 99195 | 58 84| 96) 63 75| 82) 58 | 65| 80 © i als piety win ee as 68 25 18 56 TD eid ax pice aah andi a arene 10 | 10] I0 8 I2 8| 40 | 20 8/ u 30| 28 te ee eet ot 8 7 38 u Ds dblinmncs els ead 8 7 37 u S2..i Siva ae sees + | 6 33 u Explanation: The only variants were the amounts of vitamine solution used and the yeast suspensions. All the Vitamine solutions were sterilized as in experiment 1 and the second set in the Autoclave for additional 20 minutes. In series 2 and 3 a new yeast suspension was made. The Vitamine solution was the same as used in Experiment I. THE ‘‘ BACHMAN TEsT.”’ 55 EXPERIMENT 3. To determine the suitability of the test asa qualitative agent in determining the presence and relative amount of ‘‘B’’ Vitamine. N.B. All tubes contained the Nageli solution and a loopful of yeast suspension as in Experiments 1 and 2. In this experiment only Arnold sterilization was used (two 30-minute periods with 24 hours incubation period intervening). Per Cent. of Gas Formed by Tube Contents. ct Fol anpes Cm heey: I. I c.c. of a vitamine extract of a commerical farina. I10 gms. farina was used and the extract made up to DERE 9 oO wie a Wile a cin ee eee eS be ed wae 0| 60/100} 83 | 33| 28 2. Vitamine extract from 25 gms. alfalfa. Extract made Her to. F560 tes. MRIS G5 CG: 8 ion ageless & nisla winiesse's 3] 98 pr SY 2 ge ee eee Pan 24| 97 48 4. Protozoa food mixture after Calkins made by boiling 130 gms. flour with 100 gms. timothy hay and 100 c.c. distilled water, allowing to cool and decanting. een TE eS Ste Vian Si atcha ah ea A Nle WAS ace oie awn A niaha Of OL. 6} ol. oF .O}-.0 Pe TEA eh a ea Scag Ph Ce Wee tins Sroka eis oe © oO} oO} I5| 32] 12| 18 5. 35 c.c. of the Protein free milk obtained from I qt. MMR S et eters ee eae OLE ote ak g O| 20} 40] 53 6. 1 c.c. Walker Gordon whole milk................. 0 |I100/100/100| 100} 60 7. Duplicate of (6) with varying concentrations: MINE It ard a oa Sas tt eh Mere Ry ae 6 0} 68|/r100|100| 86 ened eave oS 4. Bee oe sy tw atat eee ek Ses 0} 52/100) 88} 50 em i eri ty oe sg uae e Beas ees bs O| 25} 60} 50] 32 8. Cow’s milk to which was added varying amounts of N(NaOH) solution before sterilizing. Using 1 c.c. of solution (25 c.c. milk cont. I c.c. Po sass Cie 5 oa ate eed andi Ga ae aes 0] 3! 70/100 Using 1 c.c. of solution (25 c.c. milk cont. 0.5 c.c. RRMA OP da ey Ske chowidc a dae ceRs cee ys 0} 4! 58! 95 Using 1 c.c. of solution (25 c.c. milk cont. 0.4 c.c. REAR ee ie poet tite be vias videnie ne Sues was 0} 55|100/100 Using 1 c.c. of solution (25 c.c. milk cont. 0.3 c.c. Oe 2 ER oe 5 ne 0} 55|100/100 Using 1 c.c. of solution (25 c.c. milk cont. 0.2 c.c. PY a py sare 2a wale he ena yD ae 's ES Ss 0| 60/100/100 Using 1 c.c. of solution (25 c.c. milk cont. 0.1 c.c. POUR eMON Sah ese ORL oe ee Sadat eka ew 0} 55/100|100 9. Breast milk from six different sources. Each specimen used sterilized two 30- minute periods in the Arnold Sterilizer before using. 1 c.c. used in each inocula- tion. 3 series of experiments. Series 1 used a different yeast suspension from that used for Series 2 and 3. Oo oO 0/10/30 24/65\90 3 |33/50/48 0 |28/78\90 0 |20/68/64 Series 1. Series 2. Sources. SPU TET lie oll 3 che I OUECES: Sources Fall 23} Sopigicd Ber bOu te Fe r.|2./3-| 4. |5 As Pec. 0 |12|/63|/80|100/100|100} I c.c. 0| 0|60}100 68} 0.5 c.c. Be ce 0 /21|59|70| 70) 65] 65] I c.c.}| 0'31/70| 72/62) 0.5 c.c Circe 0/17/47/68| 82| 68} 65) I c.c. o| 8/82} 60/65] 0.5 c.c D,.:1:¢.¢ 0 |21|72|72| 72] 68] 68] 1 c.c.| 0)65\90| 80!70] 0.5 c.c Ee ee 0} 1/23/56) 65] 45] 44] I c.c.| 0] 5/70 90165 0.5 c.c Bit ee O| 0/20/35] 59] 55] 54| Ic.c.| Oo} 3/78 72145 G.5 ce eooo0o0o°0 0 |48/68/50 56 SCIENTIFIC PROCEEDINGS (103). CONCLUSIONS. The results of the various experiments are obvious from the records. It need merely be pointed out that in none of the dupli- cates were we able to repeat the gas figures exactly and that for quantitative measurement the test needs further standardization to be efficient in comparing solutions. The great variability of the yeast loopfuls obtained in this method would easily give rise to considerable variation and experiments are being made along this line to be reported later. As a means of studying presence of ‘“‘B”’ Vitamine in large or small quantity and as an index more reliable than rat feeding experiments the test offers such marked advantages in sensitive- ness and in speed of observation that it seems well worth while to devote more time to its improvement. 32 (1492) Two sex-linked lethals of simultaneous appearance in Drosophila obscura. By D. E. LANCEFIELD (by invitation). [From the Zoélogical Laboratory, Columbia University, New York City.] A pair mating in Drosophila obscura (Fallén) produced a sex ratio of 106 females to 22 males. This was about a 1:4 ratio and indicated a case of two sex-linked lethals, or lethals at two loci on the X-chromosome. Both lethals (1; and 13) appeared simul- taneously in the same culture from a female whose mother did not carry a sex-linked lethal, as was shown by the normal sex ratio produced by her; and the father could not have carried such a lethal and lived. Three daughters inherited both lethals in the same chromosome with a sex-linked gene producing the character ‘‘short’’ wing veins. These three females produced a total of 352 females to 40 normal and 50 “‘short’’ males. Such a count suggested that the gene for ‘‘short’’ was between the two lethals, which were far enough away from it for each to segregate almost independently from it, ANALYSIS OF CYTOPLASMIC STRUCTURES. 57 that is, giving approximately 50 per cent. crossing over between each lethal and ‘‘short.’’” This shows more crossing-over than is known to occur in the X-chromosome of other species of Drosophila and may correspond to the greater length of the X-chromosome in obscura. Further breeding tests showed the presence of two separate sex-linked lethals and agreed with the original assumption as to their loci. One (le) was found to be very close to the locus for ‘‘beaded”’ and gave 10 cross-overs with itin 155 males. ‘‘Beaded”’ was already known to be far enough from short to show no appre- ciable linkage to it. The other lethal (l;) was independent of “‘beaded”’ but showed some linkage to ‘‘short’’ by a ratio of 21 “short” to 34 ‘‘not-short’’ males, while the number of ‘‘beaded”’ and ‘‘not-beaded”’ males was equal. That established its locus in the other end of the chromosome from (lz) with ‘“‘short’’ between. 33 (1493) New methods for the analysis of cytoplasmic structures. With demonstrations. By ROBERT H. BOWEN (by invitation). [From the Deparimenit of Zoédlogy, Columbia University.] The year 1898 marked the recognition of two fundamental cytoplasmic structures,—mitochondria and the Golgi apparatus. The mitochondria soon became of interest to every biologist through Meves’ theories concerning their importance in cellular differentiation and inheritance, but we are still very much in the dark regarding many features of their behavior. The Golgi apparatus, on the other hand, was almost forgotten and until recently even its status as an independent cytoplasmic element was in doubt. The last few years have witnessed in Europe a revival of interest in these and other cytoplasmic inclusions, an interest not yet fully reflected in this country. It would seem, then, an opportune time for calling the attention of biologists to these structures, so little reckoned within our physiological concept of the cell. 58 SCIENTIFIC PROCEEDINGS (103). I have been experimenting with a variety of methods while studying the formation of the insect sperm, more particularly of Hemiptera belonging to the Family Cimicide (Pentatomide)- Two new methods were found which have yielded interesting results. The first is a modification of the Kopsch method for Golgi bodies, which depends on the reduction of osmic acid by the Golgi apparatus more quickly than by other cell constituents. This modification is more rapid than Kopsch and often gives good fixation of the cell as a whole. The procedure is as follows: Fix in glass-stoppered bottles for 20 to 30 hours in Mann’s sublimate-osmic :— I per cent. egmic ached sommtion < 60: \/ h 7 x posite ack = 18 ta Pe ee v he hele ee x wo} g |toFinl |G] gimgieg) S$] @ | esleg) S| S/S ies) = se ~ Del hw] . Oo <4 Ra iu -oO Nay & +) ° — A} g | mre | 2) 2 lsdles| &] & | sd les| S| = | Sg lba) > [Ae a < {A < (A z (A : ~ = = 4 > > = | > << = a %- = _ A .|/tI- 6} 11.6 |476|77| 75} § |23.1 |22.2 |23.0 |0.8 |15.6 |14.7 |15.2 |0.5 | 7.8 |66.1 I- 6 8.3 78| 68173] 5 |28.7 |26.9 |26.3 |2.5 |21.7 |18.5 |19.0 |2.7| 7.3 |71.6 B .\10-30| 16.6 | 95 | 83| 81 | 14 |17.7 |16.1 |17.1 |1.0| 9.3] 8.2] 9.3 |z.1| 7.8 154.2 C .|10—-31 8.6 | 63] 76] 67] 14 |19.9 |15.5 |17.9 |2.4 |13.5 | 7-4 |10.3 |3.2| 7.6 |56.9 II— 7} 10.1 72| 69] 65 | II |20.6 |16.5 |18.1 |2.5 | 9.8 |10.2 |10.0 |1.6| 8.1 |55.2 D .\t1-11} 10.8 |59/54|55]| 4|16.4 {16.6 |16.5 |0.3| 9.7; 8.7} 8.4|1.3| 8.1 |51.0 E .|11-20| 14.0 |62!51]|59! 7 |22.1 |19.7 |21.4 |1.7 |13.3 | 9.6 |11.6 |2.0| 9.8 |54.2 I-16 8.9 | 63| 66] 63) 6 |23.6 |2I.0 |22.2 |1.4 |16.4| 9.8 |12.2 |4.2 |10.0 |54.9 F. .|12-18 9.8 |72|69]71| 2|24.5 |23.2 |23.4 |I.1 |16.6 |14.0 |14.5 |2.1 | 8.9 |61.9 G .|12-19 6.9 83|71|77| 6|25.0 |20.6 |22.8 |2.2 |15.4 |14.4 |14.9 |0.5 | 7-9 |65.4 H .| 1-15 7.5 |74171174] 3 |27.6 |26.6 |27.2 10.6 |22.0 |19.0 119.0 |3.2 | 8.2 |69.9 Although the actual amounts of the blood sugar, oxygen content and capacity may vary somewhat when studied at dif- ferent times in one individual, the average level assumed by each of these factors seems to be characteristic for the individual. See dogs A, C, and £, Table I. On the other hand, when the level of any one factor is compared in different individuals, wide varia- tions are frequently found. Compare the sugars of dogs A and E, or the oxygen contents and capacities of A and C. Again vari- ability of blood sugar seems to be characteristic of some indi- viduals while constancy characterizes it in others. Because of this occasional variability, conclusions in the present paper are drawn only from averages of several determinations. The difference between the oxygen capacity and the oxygen content of the blood remains singularly constant from individual to indi- vidual under the conditions of our experiments. See column XIV, Table I. CASTRATION OF HEN-FEATHERED CAMPINES. 69 Our experiments do not indicate that any immediate relation- ship exists between the sugar and either the oxygen capacity or oxygen content of the blood. There is a direct relationship, however, between the ratio of the oxygen content to the oxygen capacity, 7.e., the percentage saturation, and the sugar of the blood. From this it will be seen that the sugar varies in the opposite direction to the course which it takes in asphyxial hyper- glycemia, which is a well-established phenomenon. This lack of agreement is, to our minds, apparent rather than real since the hyperglycemia of asphyxia is probably due directly, at least in part, to the increased amounts of adrenalin discharged under these conditions and only indirectly to the low content of oxygen. See columns IV and XV, Table I. In a few experiments the relative volumes of the corpuscles and serum have been determined by a precision hematocrit. The content of oxygen per unit volume of corpuscles was then calculated and was found to bear a direct relationship to the blood sugar. The oxygen capacity per unit volume for the same experiments showed only slight variations. In these experiments the corpuscular volume might be taken as an index of the oxygen capacity of the blood. This confirms the observations noted above that there is a relationship between the blood sugar and the degree of saturation of the blood with oxygen. See Table II. TABLE II. O2 Capacity Oz Content ‘ IV Hes: Corp. Vol. Corp. Vol. Panes: Di. Ti * 7° FOR CR 53 38 73 19 bP ES oa ee 51 28 63 22 } Pee ee 57 36 41 20 Gaia tos 55 36 77 21 70 SCIENTIFIC PROCEEDINGS (104). 4I (1501) Castration of hen-feathered Campines. By THOMAS H. MORGAN. [From Columbia University, New York City.] A few races of domestic fowls, such as the Campines, have two kinds of males, one showing the ordinary cock-feathering, and the other the so called hen-feathering. Three young Campines of a hen-feathered strain were castrated, one before the adult feathers had appeared, one when they had begun to appear, and a third that was like the last but exceptional in certain respects. This bird will not be considered here. Both of the former birds developed cock-feathering after castration. It is evident that removal of the testes in this race produces the same effects as is produced in the hen feathered Sebrights, as reported three years ago before this Society. 42 (1502) The vermilion gene and gynandromorphism. By A. H. STURTEVANT. [From Columbia University and Carnegie Institution of Washington.] Morgan and Bridges (1919, Carnegie Inst. Wash. publ. 278) have recently described and discussed a large number of gynan- dromorphs of Drosophila melanogaster. They conclude that all female parts in gynandromorphs of this animal contain two X-chromosomes, and that all male parts contain only one X. The peculiarities of a given part are thus due to its own constitution, and are not dependent on the rest of the body for their differen- tiation. ‘The same principle was found to hold for the characters determined by the sex-linked genes, which are carried by the X-chromosomes. I have recently obtained evidence indicating that the sex- linked character vermilion forms an exception to this rule. A VERMILION GENE AND GYNANDROMORPHISM. 71 gynandromorph was obtained in which the male parts showed the sex-linked characters scute, echinus, cut, garnet, and forked. These male parts included the whole head, in which region effects of all five of these sex-linked genes could be identified with certainty. Only one of these genes was present in the mother of the gynandromorph, which was heterozygous for the sex-linked genes eosin, ruby, forked, ‘‘vermilion lethal,’ and perhaps for vermilion. All five of them were known to be present in the single X of the father of the gynandromorph, which must therefore have been the X present in the male parts of the gynandromorph itself. But the X of the father was known to carry also the gene for vermilion, and the eyes of the gynandromorph were not vermilion. The not-vermilion color was, then, apparently determined not by the genetic constitution of the eye-pigment itself, but by that of some other portion of the body. I have obtained two other gynandromorphs in which vermilion seems to have reacted in the same fashion; but these are not so certain as the case described. One of them can be accounted for by the rare type of ‘‘double-nucleus’’? gynandromorphism de- scribed by Morgan and Bridges, and in the other the eye-color, while not vermilion, was still not quite normal. Among the large number of gynandromorphs described by Morgan and Bridges there are only three in which the male parts were known to be genetically vermilion while the female parts were not genetically vermilion, and in which these male parts included eye tissue. In all of them, however, eosin eye color was also present with the vermilion. Whether the eosin is re- sponsible for the difference between these cases and the one re- ported here, or whether that difference is due to a difference in the genetic constitution of some other part of the body cannot be determined as yet. 72 SCIENTIFIC PROCEEDINGS (104). 43 (1503) Age of parents and vitality in Uroleptus mobilis. By Gary N. CALKINS. [From the Department of Zoélogy, Columbia University.] Previous studies on the vitality of Uroleptus mobilis have shown that all ex-conjugants start with an initial high vitality as measured by the division rate. From this initial high rate, vitality gradually decreases during the life cycle until the race dies a natural death. After the first fifty days of a cycle endogamous conjugation may occur at any time with a resulting renewal of vitality in the ex-conjugants. Sufficient data have now been obtained to indicate that there is a difference in vitality of offspring given off at different age periods of the parental series, and a summary of such data is shown in the accompanying table. Line I gives the total number of generations, or divisions, in in each series; line 2 the total number of days required for those divisions; line 3, compared with line 5, indicates the extent of the renewal of vitality resulting from conjugation; line 4 gives the average division rate for any ten days of the entire life cycle on the assumption that divisions are equally distributed; line 6 gives the age of parents in days, at the time of conjugation which re- sulted in the offspring series, and line 8 the age in generations; lines 7 and 9 give the number of days and the number of divisions subsequent to the time of conjugation which resulted in the off- spring series and represent the vitality remaining in the parental protoplasm. High vitality in a series is indicated (1) by a high division rate which represents high metabolic activity; (2) by the large number of generations, or divisions, which the series undergoes, indicating a combination of high metabolic activity and endurance or lasting capacity; and (3) by endurance or long length of life of a series in days which may be combined with, or independent of, (1) and (2). High vitality was shown in all three ways by series C, F, J, P, Ui, U2, and V. Low vitality was shown in all three ways by series Q, R, and a, while series W and H were low in regard to 73 AGE OF PARENTS AND VITALITY. — |9f |ze |g 9 z oF =| Loe | Sg! So- 1 LS | oS1/ Sos) er Phe) Cor) Of.) Seer a ee ee eee mee ea an -YO JO aUl]} J0zJe SUOISIAIP Jo JoquINN]| 6] 9 yp — | 16z|Szz|Sbz} orf | 11€ | LEz | ozr | goz | goz | 911 | gor | Sgr | 661 | EV1 |} 9g | LEI} gL |° °° Sutsdsyo jo out} 7e SUOISIAIP UI O3Y! 8 | WH ae ROS LE oor Be i h6 | L6t Os Ob Ord, | O61 WEL <1 00. |. O85. PPS tte Ta ee ae ee a -JJO JO UIT} J07Je OTT] Jo Skep jo roquinny| 4] § &. — |orz|o€1 | og1 | ooz | oS$z|oL1 | 06 | ovr |orrj|oL |o09 |ozzr}ozrjoor|oS |oor}o£ |°***** *suridsyo jo out} ye Shep ul o8y| 9 G & — lov | rs |rz [Ser | rr | gh | Sorl £°6 | €°6 | err} o'r1| O°6 | L°€r| r°r1} g’or| 1°z1| O11)" ** “surmdsyo jo oul} wiory 97eI UOISIAIG| S$ L:11| L:01| 0°6 | L°6 | 9°9 | g*zz| Gor} O'Z1| Q°I1| B*zI| HIT] O'ET| Q°ET| O'LI| HET) E71) VST] Q°IrT|‘asft] G1QUa 10} SACP OF Jod 9}e1 UOTSTAIG| V = | Pet GOL. errs’ Oo) PLT LOTS Er) BET, OCR) SS a sr O'Or) Sor) SOL) MEL CPT Ger) a i i ee ea omeatage a, Hn UOISTAIP 4S] 07 SAep-OF Jad a3ze1 uoTsIAIq] £| OF Loz | gbv1r | 6zz| Sgr |S€ | L61 | Svz | Soz | SSz | o€z | Lez | L61 | 6g1 | oz | obz | LSz | S1z | P6z |***** * “UOTSIATp ySe] 07 SAP Jo JOqUINN]| z Bo. o7ep ac. 0} zB & €1£ | 6S1| Loz |ogr | Ez | €Sz| goz| LE] rol | Sgz | Lzf | goz | LSz| Elz | eze| Li] 1Lz2 | OVE |***+* +++ * + “SUOTSTATp Jo Jaquinu jejOL} 1} ° CO: | CAUCHY POLI (CD CHIRP Cal) CT) | CZ TY A) C79 CP) 8 Ct CO CPE eee dae a “2 71 “W ‘O le oy og *A 3% ‘12 RE ‘O ‘N iy 7 “fT "TC 25) Peeeeneererreesareeenesnee srereseeee SuLIdsyO 74 SCIENTIFIC PROCEEDINGS (104). division rate and number of divisions, but high in endurance. The other series were intermediate as regards vitality some of them being high in (1) and intermediate in (2) and (3) as in series Di... O, anny. Without exception the series showing high vitality in all three ways came from young parents. Thus the C series started when the parent A series was 70 days old and in the 78th gener- ation; the F series started when the parent C series was 50 days old and in the 86th generation; the J series was started when the parent F series was 100 days old and in the 143d generation; the P series and both U series came from the L series which was 70 days old and in the 116th generation when the P series started, and 140 days, or twice as old, and in the 208th generation when U, and U2 were started; the V series, finally, was started from the P series when the latter was 90 days and 120 generations old. Without exception, also, the series showing low vitality in all three ways came from old parents. Thus the Q series was started when the parent I series was 200 days old and in its 316th gener- ation and should be contrasted with the LZ series which came from the same parent when younger; the R series was started when the parent J series was 160 days old and in the 245th generation, and should be contrasted with the WN series which came from the same parent when it was 120 days old and in the 188th generation; the a series was started when the parent P series was 210 days old and in the 29Ist generation and should be contrasted with the V series which came from the same parents when they were 90 ~ days old and in the 120th generation. The W series and the H series both of which were weak in respect to division rate and number of generations came from parents that were similarly old, the W series from N when in its 225th generation; the H series from A in its 237th generation. ‘The J series was exceptional, for, coming from very old parents (A series when 250 days old and in the 311th generation) it was weak only in endurance. *. =e " ee ea HEREDITY OF TWIN BIRTHS. 75 44 (1504) Heredity of twin births. By C. B. DAVENPORT. [From the Eugenics Record Office, Cold Spring Harbor, L. I.] About I per cent. of human births are twin births. However, there are certain families in which the proportion rises to 5, 10, or even 15 percent. There can be little doubt then that, as in sheep, so in man, there are strains having a special tendency toward the production of twins. It is commonly believed that this tendency toward the pro- duction of twins must be wholly a maternal quality, depending upon the inherited tendency to double ovulation. The study of the heredity of twins is accompanied by certain difficulties, such as the fact that the occurrence of twins is fre- quently isolated, apparently haphazard, occurring perhaps in only one case in a fairly large fraternity, in which other represen- tatives are single births. It will simplify the matter a little if we consider only those cases in which two or more sets of twins have arisen from a given mating. The study of twins is still further complicated by the fact that they are of two types, namely twins derived from a double ovula- tion and twins derived from a single ovulation, there being a sub- sequent fission or budding of the fertilized egg. Such single-egg twins are easily distinguished clinically by being both enveloped in the same chorion. They are also always of the same sex. The statement that the mother alone determines the tendency to twins is not, however, supported by the facts. Of the births giving rise to the fraternities of twin-repeating mothers, 4.5 per cent. are twin births. Of the births giving rise to fraternities of twin-repeating fathers, 4.2 per cent are twin births. These figures depend upon 355 and 289 labors respectively. The sisters of twin-producing fathers have twins in 8.2 per cent. of labors, while the sisters of twin-producing mothers have twins in 5.5 per cent. of labors. Among the children of brothers of twin-producing fathers, 6.5 per cent. are twins, among the 76 SCIENTIFIC PROCEEDINGS (104). brothers of twin-producing mothers 4.5 per cent of the children are twins. These figures indicate that the twin ratio is increased 4 to 7 times in twin-producing families and that the ratio of twin production is about as high on the father’s as on the mother’s side of a fraternity which contains 2 or more twins. This result dis- poses of the statement that fathers play no role in twin production, but at the same time raises the query—How can this be? It is relatively easy to understand how the sperm may in- fluence the early division of the fertilized egg so as to produce identical or 1-egg twins, and we find indeed that the rate of occur- rence of twin production is high, both on the paternal and on the maternal side of such fraternities containing identical twins. The rate in both cases is about 13 per cent. But if one takes only those fraternities producing 2 or more sets of twins of unlike sex, we still find an equality of influence on the paternal and on the maternal side. The difficulty in the way of understanding the equality of the part played by the father and the mother in the production of 2-egg twins arises from the assumption that such twins are due merely to double ovulation. Were they due merely to double ovulation, then there would be as many sets of twins born as there are double ovulations that occur at the time of conception. That this assumption is false is shown by several lines of evidence. 1. Not all of the eggs that are ovulated are fertilized. In the case of 4 sows’ uteri examined the number of embryos (well advanced in development) found was 22 and the number of recent corpora lutea 34. This indicates that fully one third of the eggs laid at these periods of conception were not fertilized. This number is, I think, rather too high for a general statement, probably not more than 10—25 per cent. of eggs ovulated at favor- able periods fail of fertilization. In humans also there is evidence based upon counts that have been made on the corpora lutea of non-pregnant and pregnant women that the rate of occurrence of 2.3 corpora lutea in the ovary is much greater than the occur- rence of twins. Thus, to cite only a single study, in 33 sets of ovaries with corpora lutea or follicles about to burst, 5 (or 15 per cent) showed that double ovulation was occurring. It seems, therefore, certain that many more cases of double ovulation occur than of twins. | | . { . HEREDITY OF TWIN BIRTHS. re Again, of eggs that are fertilized, a certain proportion are aborted at an early age. This fact is striking in mammals also, where occasionally one finds, as John Hammond, 1914, points out and as I can confirm by numerous examinations of the uteri of swine, one or more embryos degenerate at an early stage. Also the medical literature has abundant references to blighted twin fetuses, and the large number of miscarriages under 3 months is a familiar fact. It is probable, since genetic work has revealed a great number of lethal factors, that a large proportion of these atretic embryos is due to such lethal factors. The foregoing statements lead to the conclusion that our preconception that twinning is due merely to double ovulation needs revision. It is due to double ovulation combined with some other factors that induces a large proportion of such double ovulations to produce viable twins. The other factor is the paternal one. It depends upon the capacity of the male to fertilize all of the eggs ovulated with sperm which contain no lethal factors. Now, both the capacity for complete fertilization (high fecundity) and the absence of lethal factors in the male germ cells are hereditary factors. Attention may be called in passing to the fact that in human matings about 10 per cent. are sterile and, among these sterile matings, a certain proportion are physiologically such or at least no imperfections in the reproductive organs of either member of the pair can be detected. Summary.—The influence of the male in twin production is determined by the circumstance that twin production does not depend merely upon double ovulation but upon such a quality of the sperm as shall result in a high proportion of fertilization of eggs ovulated and a small proportion of fertilized eggs containing lethal factors. 78 SCIENTIFIC PROCEEDINGS (104). 45 (1505) The nutritive value of some nuts. By F. A. Cajori (by invitation). [From the Sheffield Laboratory of Physiological Chemistry in Yale University, New Haven, Conn.] In earlier experiments on the utilization of protein-rich nuts by man, it was found that the “coefficient of digestibility’? was practically equivalent to that of the protein coefficient of a mixed diet. The conclusion was drawn that such nuts, when used in the diet with due recognition of their concentrated make-up, are of a physiological value comparable with that of our more common foods. The investigation has been extended to a study of the nutritive value of specific constituents of nuts. Johns’ analysis of the nitrogenous components of the peanut and coconut suggest that they are sources of complete protein.?: *. This has been verified by the feeding experiments of Johns with the coconut‘ and Daniels with the peanut. Osborne and Mendel have maintained rats over long periods on excelsin, the principal protein of the Brazil nut, as the sole source of protein. Following the same technique, we have observed satisfactory growth on diets furnishing the almond, English walnut, filbert, and pine nut, respectively, as the essential source of protein in the ration, to the extent of 18 per cent. of the total diet. Experiments with the pecan nut have as yet been less successful. The presence of abundant quantities of water-soluble vitamine in the coconut and peanut has already been demonstrated,‘ 5. We have found that normal growth can be secured when rats are fed upon otherwise adequate diets containing the almond, English walnut, Brazil nut and chestnut as the sole source of water soluble vitamine and that animals which have declined on a diet devoid of 1 Cajori, F. A., 1918, J. of Home Econ., 10: 304. 2 Johns and Jones, 1917, J. of Bio. Chem., 30: 33. 3 Johns, Finks and Gersdorff, 1919, J. of Bio. Chem., 37: 149. 4 Johns, Finks and Paul, 1919, J. of Bio. Chem., 37: 497. 5’ Daniels and Loughlin, 1917, J. of Bio. Chem., 33: 295. * Osborne and Mendel, 1912, Zeit. f. Physiol. Chem., 80: 307. LocaL ANESTHETIC PROPERTIES OF BENZOYL CARBINOL. 79 water soluble vitamine promptly recover when either the almond, English walnut or filbert is introduced in the diet. Investigations in this field are being continued. 46 (1506) The local anesthetic properties of benzoyl carbinol. By AxEL M. HjortT and CHARLES E. KAUFMANN (by invitation). [From the Department of Pharmacology, Yale University School of Medicine.|' Since Macht? observed that benzyl alcohol possessed local anesthetic powers, several of its homologues have been studied for like properties in this laboratory. These consist of a? and 64 phenethylol and benzoyl carbinol. The structural relationship of these substances is: ao CH:20H on CHOHCH:; CH2CH:0H COCH:0H | ) | ) | | a \ NS Benzyl Alcohol @ Phenethylol 8 Phenethylol Benzoyl Carbinol Hirschfelder® recently reported his observations on a series of compounds closely related to the above. £8 phenethylol and phenyl glycol were amongst these, the latter being the reduction product of benzoyl carbinol, the subject of this paper. The relative efficiency of the substances investigated, with respect to their local anesthetic properties, is shown by the fol- lowing table: 1 This research has been supported by a grant from the Committee on Scientific Research of the American Medical Association. 2 Macht, D. I., Journ. Pharm. & Exp. Therap., 1918, xi, 263. 8’ Hjort, A. M., and Kaufmann, C. E., Ibid. In press. 4 Hjort, A. M., and Eagan, J. T., Ibid., 1919, xiv, 211. 5 Proc. Soc. Pharm. & Exp. Therap. (Dec., 1919), J. Pharm. & Exp. Thera?., 1920. 80 SCIENTIFIC PROCEEDINGS (104). Minimal Effec- | Minimal Effec- tive Anesthetic | tive Anesthetic Minimal Lethal | Quantity Non- Substance. Dose per 20-30 | lethal Intraven-| Concentration | Concentration G. White Mice | ously in Dogs, | 6p Rabbits’ in Homan Megms. Gms. per Kilo, Cornea, 4. Skin, %. Benzyl Alcohol.......... 50! 0.2! 1.253 1/30? a Phenethylol........... 20? 0.1? 0.752 1/402 8 Phenethylol........... 408 0.23 1.008 1/403,4 Benzoyl Carbinol........ 40? 0.2? 0.502 1/40? 1 Macht. 2 Hjort and Kaufmann. 3’ Hjort and Eagan. 4 (1/40) 13 wheals in 21. Sollmann‘ found that procaine in the strength of 1/32 per cent. was the minimal concentration which anesthetized human skin when injected intradermally and tested according to the wheal method of Hoffmann.’ From the above table, therefore, it is seen that benzoyl carbinol is at least as efficient on the human skin as any known anesthetic. Benzoyl carbinol is not as irritant on the tongue as the benzyl alcohol and its above mentioned homologues. In the skin, like- wise, it does not induce as extensive a local reaction of the tissues. It is soluble in water to the extent of about one half per cent. at 20° C., but when heated to 50° C. a one per cent. solution can be made which does not alter within two hours. Benzoyl carbinol is, therefore, soluble within practical limits. Summary.—1. Benzoyl carbinol possesses local anesthetic properties which in general are superior to those of its homologues herein discussed. 2. Benzoyl carbinol is less irritant to the body tissues than its congeners. 3. Its solubility in water is sufficient to make it a practicable local anesthetic. 4. It is the most stable of the series. 6 SolJmann, T., Ibid., 1918, xi, 69. 7 Hoffmann, u. Kochmann, Beit. Klin. Chir., 1914, xci, 489. wit Ss po, ACTION OF G-STROPHANTHIN AND DIGITALIS. 8I 47 (1507) A comparison of the action in patients of g-strophanthin and digitalis. By ALFRED E. CoHN and RoBERT L. LEvy. [From the Hospital of The Rockefeller Institute for Medical Research, New York.]| In the same patients we have compared the action of g- strophanthin given intravenously with the action of digitalis (digipuratum) given by mouth. Both drugs were standardized by the cat method. One cat unit equalled 0.1 mgm. g-strophan- thin; one cat unit was equal to 0.1 gm. of digipuratum. The patients selected for treatment were as far as possible sufferers from fibrillation of the auricles. Both drugs were usually given in divided doses, the duration of administration being in several instances comparable. The effect of the drugs on the speed of action, on the electro- cardiogram, and on the duration of the effect on the rate of the ventricles when the auricles fibrillate, we report in brief now. The speed of the action is often faster with strophanthin than with digitalis, though when strophanthin is given in divided doses it may require nearly two hours to obtain aneffect. In other instances an effect may be obtained, as is well known, in twenty minutes or less. An effect with digitalis has been observed in a little more than two hours. With digitalis the effect on the rate of the ventricles outlasts that of strophanthin. It is rare for strophanthin to keep the rate low for more than five days; it did so once for nine days. Digitalis effect endures usually beyond ten days and has lasted as long as twenty-three days. The effect on the T-wave of the electrocardiogram is absent or slight with strophanthin, and its duration when present tran- sient. The effect with digitalis endures in the manner now familiar. With doses equal, in cat units, to the strophanthin given, the effect on the T-wave is not maximal. Larger doses of strophanthin than I.I mgm. were not given, so that the dose of 82 SCIENTIFIC PROCEEDINGS (104). digitalis in this series was usually low (1.1 gm.). When on occasion we gave larger doses of digitalis, more striking effects on the T-wave were observed. 48 (1508) The isoagglutinins and isohemolysins of the rat. By G. L. ROHDENBURG (by invitation). [From Columbia University, George Crocker Special Research Fund, F. C. Wood, Director.] The possibility that some of the irregularities in immunity against transplanted tumors might be correlated with the isoag- glutinins or isohemolysins of the respective hosts prompted an investigation of these substances in the blood of rats. The animals tested were derived from three different sources showing rather marked external as well as biological differences. One group, pure white in color, was resistant to the growth of the Jensen rat sarcoma, showing a very high percentage of natural immunity; a second group, red and white in color, was equally resistant to the growth of the Flexner-Jobling rat carcinoma; a third group, hooded black and white, was equally susceptible to both tumor strains. Fifty animals were tested in the following manner for the presence of either isoagglutinins or isohemolysins. Nine drops of serum were mixed with one drop of a 5 per cent. suspension of washed red cells in a test tube. In the first series of ten animals the serum of each animal was tested against the cells of every other animal, and the cells of each animal were tested against the serum of every other. Five series were carried through in this fashion, each series consisting of three animals of two of the groups and four of a third group. Tests between animals of the same strain or between animals of different strains showed that neither agglutinins or hemolysins were demonstrable, this being contrary to the well-established phenomena in man, where four distinct groups have been found. LATENT INFECTION IN EXPERIMENTAL SPIROCHATOSIS. 83 49 (1509) Latent infection in experimental spirochaetosis. By Joun L. Topp. [From McGill Uniersity, Montreal.] The following observations were made while studying im- munity to Spirocheta recurrentis in white rats. Since it had been suggested that an immunity might be maintained by per- sistent latent infection, it was necessary to make careful examin- ations for spirochetes in all animals exhibiting resistance to in- fection. The work was done with strains of spirochetes which usually produced well-marked infection with one or morerelapses. During the observation of several hundred animals it was noticed that parasites occasionally reappeared in the peripheral circulation after unusually long absences. It often happened that spiro- chztes were found in blood films, on one or two days, after they had been absent for from ten to fifteen days; once spirochetes were seen after an absence of twenty-one days. Blood, taken from the tail, was examined for spirochetes in thick dehemoglobin- ized and stained films. Examinations, throughout the work, were made daily. It often happens that a normal infection follows when a rat is inoculated with blood from an animal, once infected, although repeated examinations have failed to find spirochetes in it. In many instances, infection resulted when rats were inoculated with blood taken from other rats which were killed from fifteen to twenty days, in one instance twenty-seven days, after spiro- chzetes were last found in them by microscopical examination. Although the inoculation of blood, and other material, obtained by killing an experimental animal, is sometimes a more efficient method of detecting spirochetal infection, it is not always so and, since it involves the destruction of the animal supplying the material inoculated, it is a method which cannot be repeated. Inoculation of material in which living spirochetes have been seen occasionally fails to infect one of two rats inoculated at the 84 SCIENTIFIC PROCEEDINGS (104). same time; sometimes, also, the infection produced remains unseen by microscopical examination and is only detected by killing the inoculated animal and by sub-inoculating another rat with blood from it. Inoculations are less likely to fail, or result in unobvious infection when considerable quantities of blood are inoculated. In order to obtain considerable quantities of material for inocu- lation from rats in which spirochetes could not be found by blood examinations, blood was aspirated from hearts and spleens were excised. Of these two methods the aspiration of heart’s blood was the more successful. Under chloroform, it is easy to aspirate from I to 2.5 ccm. of blood from the heart of an adult rat without injuring the animal. By so doing, spirochaetes were shown, in one instance, to be present in an apparently immune rat thirty-two days after the parasites were last seen by the daily examination of blood films. Previously, this rat had twice been infected by inoculation and had acquired an immunity shown by two unsuccessful inoculations. Two subsequent, also unsuccessful, inoculations showed that the rat was still immune to the strain employed in this experiment. It should be remarked that several similar experiments did not reveal latent infections in immune rats. 50 (1510) The influence of water-soluble vitamine on the nutrition of dogs. By W. G. Karr (by invitation) [From the Sheffield Laboratory of Physiological Chemistry, Yale University, New Haven, Ct.] Studies on a few species of animals have indicated that the water-soluble vitamine is of importance to nutrition during all stages of life and that prolonged absence of it from the diet will lead to a diseased condition. In the present investigation the role of water-soluble vitamines in the nutrition of the dog has been under consideration. The animals were fed a diet devoid of water-soluble vitamine and consisting of lard, sucrose, inorganic salts, and protein in the ae eee) ee ee ee ee a en eee VITAMINE ON THE NUTRITION OF Docs. 85 form of casein or wheat gluten. It furnished about 70-80 calories per kg. body weight. Such food mixtures were consumed readily by the dogs during a period from three to nine weeks; thereupon they began to refuse part or all of their food. Characteristic symptoms, similar to those described by Voegtlin and Lake! were developed by some of the animals. The ultimate failure to eat was always noted. Ingestion of as little as 1 gm. of brewer’s yeast, which had been previously dried, will cause a quick recovery of the desire to eat. 5 gm. of the dried yeast will bring about the disappearance of the polyneuritic symtoms in 8-12 hrs.and a quick recovery of the animal to its normal condition. The utilization of the protein nitrogen is not effected by the lack of water-soluble vitamine. Studies in metabolism on diets with and without vitamine are being conducted. The fat-soluble vitamine is apparently of less importance than the water-soluble in the nutrition of the adult dog. 1 Voegtlin and Lake, Am. J. Physiol., 1918, xlvii, 558. * * ae SCIENTIFIC PROCEEDINGS ABSTRACTS OF COMMUNICATIONS. One hundred fifth meeting. College of the City of New York, New York City, February 18, 1920, Vice-President Wallace 1n the chatr. 51 (1511) Pituitary feeding and egg production in the domestic fowl. By SUTHERLAND SIMPSON. fi [From the Physiological Laboratory, Medical College, Cornell University, Ithaca, N. Y.] Clark! found, in 1915, that the egg production of chickens (White Leghorns) was markedly increased by feeding dried pitui- tary (anterior lobe) in amounts representing 20 mg. of the fresh gland to each hen per day. The effect became evident four days after the feeding began and lasted for several days after the last dosage. The experiments were carried out in May, when the laying curve was on the decline, nevertheless the egg production reached 100 per cent. for the experimental pen (35 hens on two consecutive days laid 35 eggs) or double what it had been before the feeding was begun. He lays emphasis on the fact that the glandular material he used was taken from young, growing ani- mals—calves and lambs. I have repeated the experiments of Clark when the egg produc- tion was low and declining—in June and July, 1917, when low and increasing—December and January, 1917-18, when high (about the maximum)—April, 1918—and again in March and April, 1919, and have been unable to observe any increase from pituitary feeding. The adult gland (ox) was used and also that of the growing animal (calf), and the method of preparation adopted 1 Clark, L, N., Jour. Biol, Chem., 1915, 22, 485-491. 87 88 SCIENTIFIC PROCEEDINGS (105). by Clark was followed as closely as possible. To begin with, the amount given by him was administered, viz, the equivalent of about 20 mg. of fresh pituitary substance (anterior lobe), to each hen individually, in a gelatine capsule, and when no result was obtained the dose was doubled and later trebled. In no case was any distinct effect produced, the egg-laying curves running prac- tically parallel with those of the control pens. Single Comb White Leghorns were employed. 52 (1512) Some conditions affecting thyroid activity. By W. B. CANNON and P. E. SMITH. [From the Physiological Laboratory, Harvard Medical School.| 1. Gentle massage of the thyroid gland in the cat for two or three minutes will cause an increased rate of the denervated heart amounting in some instances to 33 per cent. over the basal rate. The development of the maximal increase of rate is usually slow, requiring from thirty to sixty minutes and passing off in a similarly slow manner. 2. Massage of another gland, e.g., the submaxillary, does not cause this effect. | 3. The augmentation of heart rate caused by thyroid massage occurs in the absence of the adrenal glands. 4. Stimulation of the cervical sympathetic trunk as it leaves the stellate ganglion induces a similar augmentation of the rate of the denervated heart: this does not occur if the thyroid gland has previously been removed. 5. If the cardiac fibers from the stellate ganglia are severed, as well as the vagus nerves, and an afferent nerve such as the sciatic or brachial is stimulated under a degree of anesthesia which will permit reflex retraction of the nictitating membrane and dilation of the pupil, there is a primary increase of rate due to adrenal secretion, followed by the slowly developing increase characteristic of the thyroid effect. 6. If the vagi and the cardiac fibers of the stellate are cut, a ee ee ee eee ee STUDIES IN THE ABSORPTION OF Fats. 89 and the animal is asphyxiated under conditions which permit the eye changes described above, there is a similar primary rise due to adrenal secretion, followed by the secondary thyroid effect. 7. If the thyroid glands have been previously removed, sensory stimulation and asphyxia induce only the increase of rate due to adrenal discharge. 53 (1513) Studies in the absorption of fats. By T. F. Zucker. (By invitation). [From the Depariment of Pathology, College of Physicians and Surgeons, Columbia University.] The general impression of workers in the field of fat metabolism, with regard to the path of absorption, seems to be that while most of the fat enters the circulation by way of the thoracic duct, a smaller portion can be absorbed directly into the blood stream. The most recent discussion of the subject is by Bloor.t The results of previous workers are, briefly, as follows: Walther? recovered in anesthetized animals only a small fraction of the absorbed fat from the lymph of the thoracic duct. Munk and Rosenstein? recovered from the discharged lymph 60 per cent. of the fat fed to a patient with lymph fistula. Frank‘ tied off the thoracic duct after feeding fat and still found the fat of the blood to increase during absorption. Hamburger® tied off three equal-sized loops of intestine and in the central loop ligated all lymph vessels. Then after injecting an oil and soap emulsion, he noted that this was absorbed from the central loop despite the tying off of the lymph vessels, although it was distinctly less in amount than in the control loops. Munk and Friedenthal,® in a preliminary communication, describe experiments similar to those of Frank, but use more precautions, such as tying off the 1 Bloor. Jour. Biol. Chem., 1916, XXIV, 457. 2 Walther, Archiv. f, Physiologie, 1890, p. 328. 3’ Munk and Rosenstein, Virchow’s Archiv, 1891, CX XIII, 230. 4Frank, Archiv. f. Physiologie, 1892, p. 497; 1894, Pp. 297. 5 Hamburger, Archiv. f. Physiologie, 1900, p. 554. 6 Munk and Friedenthal, Zeniralb. Physiologie, 1901, XV, 297. 90 SCIENTIFIC PROCEEDINGS (105). ductus lymphaticus communis dexter, besides the thoracic duct, to avoid possible anastomoses. In general, their findings are the same as those of Frank and they also conclude that the blood can absorb products of fat digestion directly from the intestine. d’Errico! approached the subject from a different angle. He determined the amount of ether extract in simultaneous samples of portal and jugular blood of an animal near the height of fat absorption after a meal. He finds more ether extract in the portal than the jugular blood and concludes that direct absorption by the blood takes place. In criticizing d’Errico’s work we must say that ether extract from blood and blood fat are not the same,” as d’Errico’s own re- sults show, since he finds even after feeding fat, not more than a maximum of 412 mg. of ether extract per 100 c.c. of blood, while the “total fat” of fasting blood is usually 600 mg. or more. d’Er- rico’s ether extract was probably principally the cholesterol, plus small quantities of other lipoids. The work of the other authors mentioned above all has this in common, that the conditions of the experiment are far removed from the normal, and in several cases the analytical technique leaves much to be desired. In connection with another research, our attention was called to the fact that this point had never been settled and we under- took a few experiments similar to those of d’Errico, but using the newer analytical procedures of Bloor! for the determination of cholesterol, fatty acids and phosphatides. In the phosphatide determination, Kober’s strychinine molybdate reagent was used. Hemoglobin was determined by the acid hematin method using a Dubosq or Kober colorimeter. In the earlier experiments, ether was used as anesthetic, but since ether inhibits the absorption of fats quite markedly, we dispensed with the anesthetic and in- stead stunned the dog by a quick blow on the occiput. This, when properly executed, is quite humane and very suitable for the requirements of our experiments. The following is the protocol of one of the experiments of which there were five in all. 1 d’Errico, Arch. fisiol., 1907, IV, 513. 2See Klein and Dinkin, Zeitschr. physiol. Chem., 1914 (XCII), 302; Kumagawa and Suto, Biochem. Zeitschr., 1908, VIII, 212; Hiirthle, Zischr. physiol. Chem., 1895. XXI, 331. STUDIES IN THE ABSORPTION OF Fats. QI A sample of jugular blood was taken from a dog of 14 kg. weight at 12:45, and then 50 c.c. of olive oil were given by stomach tube. Another jugular sample was taken at 3:45. At 4:45, the dog was stunned, the abdomen was opened and samples of portal and mesenteric blood taken. Then another sample of jugular was taken. The analyses of these samples are given in the table in gms. per 100 c.c. of blood. The cholesterol remains constant, The administration of 50 c.c. of olive oil raised the phosphatides 36 per cent. and the fatty acids 49 per cent. There is no dif- ference, within the limits of error of the method (5 per cent.), be- tween the jugular, mesenteric and portal. The absorption in this particular experiment was moderate. In another experiment, the fatty acids were increased go per cent. and still there was no difference between the portal and jugular. | Cholesterol. | Phosphatides.| Fatty Acids, | Hemoglobin. 12:45. Jugular before giving oil... .23 42 65 125% Sei —MONPEAES Oils oe tule weiias Lee « .22 52 93 123% 4:45. Animal stunned, portal..... | 57 .98 126% oT 2 SA Ae ee «22 .56 -97 122% Per Rete, Sia 2h) Se 2 23 58 1.02 125% To sum up the evidence then, we can safely say (1) that d’Erri- co’s findings cannot be accepted because of the methods employed, and that they are not corroborated by our own data, (2) that in the experiments in which tying-off of lymph vessels was done, absorption may have been due to the lymph stasis, and (3) that the data here presented preclude the assumption of any very marked participation of the blood vessels in the absorption of fat leaving open the question of absorption of small amounts beyond the detection of the methods used. 54 (1514) ~iaiuapy in the diastatic activity of the blood and blood sugar curves indicating a decreased carbohydrate tolerance in hyperthyroidism. | By Joun A. KILLIAN [From the Laboratory of Pathological Chemistry, New York Post Graduate Medical School and Hospital.] In studying the carbohydrate tolerance in hyperthyroidism and other conditions, three methods have been employed, the 92 SCIENTIFIC PROCEEDINGS (105). diastatic activity of the blood, the fluctuations in the level of the blood sugar, and the sugar excreted in the urine after the administration of a definite amount of glucose by mouth. The diastatic activity was determined by the technic of Myers and Killian,! the blood sugar was estimated by the Lewis-Benedict method as modified by Myers and Bailey;? the Benedict-Oster- berg? technic was utilized in the determination of the urinary sugar. It was early appreciated that fasting for twelve hours increased the tolerance for carbohydrate, so that glucose tolerance curves carried out on a fasting stomach did not truly represent the individual’s carbohydrate assimilation limit. In our series the patients received in the morning, a standard meal which con- sisted of one egg in any form, two slices of bread, a cup of coffee, without sugar or milk, or a glass of water. Two hours later, the patient voluntarily emptied the bladder, and then received about 100 c.c. of water to drink. One hour after this, a specimen of blood and a specimen of urine were obtained. These represent the control specimens. The glucose was then administered by mouth in a 50 per cent. solution, 1.75 gm. per kilogram of body weight. At hourly intervals the specimens of blood and urine were obtained, and in these and in the control the sugar was estimated. The diastatic activity was determined only on the control specimen of blood. The 23 cases presented are from a series of 275 patients ex- amined, representing normal and various pathological conditions. The control specimen of blood represents the level of blood sugar 3 hours after the standard meal. In normal cases it varied from 0.09 to 0.10 per cent.; in dyspituitarism of the Frélich type, in acromegaly and in Addison’s disease a hypoglycemia was noted. In hyperthyroidism, however, the blood sugar ranged from 0.11 to 0.13 per cent. In normal cases the urinary sugar excretion was found to be 20 to 30 mgs. for the control hour; the output in dyspituitarism, acromegaly and in Addison’s disease was dimin- ished; the hyperthyroids excreted from 24 to 95 mgs. Following the intake of glucose, the blood sugar in normal 1 Myers and Killian, Jour. Biol. Chem., 1917, X XIX, 179. 2 Myers and Bailey, Jour. Biol. Chem., 1916, XXIV, 147. ® Benedict and Osterberg, Jour. Biol. Chem., 1918, XXXIV, 1095. LocaL TIsSUE REACTIONS. 93 cases reached the maximum from 0.13 to 0.15 per cent. at the end of one hour and in two hours time returned to normal. In hypo- functions of the endocrine system, there was noted practically no increase in the blood sugar; on the contrary hyperfunction of the thyroid produced a pronounced hyperglycemia after the glucose ingestion which persisted for 4 to 5 hours. In the hourly specimens of urine from these cases there was an evident glyc- uresis, which for a period of 3 hours totaled 1.4 per cent. of the glucose given. These specimens of urine gave positive reactions for sugar with Benedict’s qualitative copper solution. The normal cases excreted during 3 hours from 0.1 to 0.2 per cent. of the glucose given and gave negative reactions with the copper reagent. The diastatic activity of the blood was found to be decreased in dyspituitarism, acromegaly and Addison’s disease, but in hyperthyroidism there was a distinct increase ranging from 20 to 34, except in 2 very early cases. 55 (1515) The influence of systemic changes on local tissue reactions. By JoHN AUER. [From the Laboratories of The Rockefeller Institute for Medical Research. In order to explain the occurrence of a massive, brawny edema at the site of operative wounds in sensitized, reinjected dogs, the following working hypothesis was formed: Sensitized animals which have circulating ineffective amounts of the antigen, may react locally with anaphylactic changes if through any mechanism (for example, by inflammation and edema) an effective dose of the antigen accumulates in those tissues. This conception was then tested experimentally in the rabbit. Rabbits were sensitized by four muscular and intraperitoneal injections of 4 c.c. horse serum at 4 to 5 day intervals. 15 to 21 days after the last injection, 10 c.c. of horse serum were given intraperitoneally. 30 to 45 minutes after the reinjection, none of the rabbits having shown any collapse, the hairy surface of the 94 SCIENTIFIC PROCEEDINGS (105). ear was moistened with 1 c.c. of commercial xylol in order to produce irritation and edema. Two groups of control experiments were carried out. In the first group, each normal rabbit received 10 c.c. of horse serum in- traperitoneally and after 30 to 45 minutes the ear was treated with the same kind and amount of xylol. In the second group of controls no horse serum was administered, merely one ear was treated with xylol. The results were strikingly different and support the working hypothesis. In a great majority of the 36 controls edema of a fair to good degree developed in six hours; it was generally less in 24 hours, and after 48 hours had largely disappeared, leaving a practically normal ear. No dermatitis with blisters and crusts was observed; nor were hemorrhages or gangrene seen except once among the controls. In this instance the loss of substance was not more than one half millimeter of the ear tip. In the sensitized series (17 rabbits) the edema of the ear de- veloped more slowly and less frequently than among the controls. The maximum was reached generally in 24 hours, and the sub- sidence was slow, lasting 5 to 9 days. Within 22 to 48 hours, numerous small hemorrhages, blisters and subsequent crusts appeared. In these rabbits (10 out of 17) the ear after a few days showed the picture of an exfoliative dermatitis. This dermatitis involved 1/3 to 1/2 of both surfaces of the ear, healed slowly as the deeper tissues were affected, and always caused dry gangrene of the ear tip. The loss from gangrene varied from I to 3 centi- meters. Healing was usually complete in three to four weeks. The ear stump was bald at first, but slowly became covered with a new growth of white hair. 56 (1516) The selective effect of the accelerator nerves on ventricular systole. By C. J. WIGGERS and L. N. Katz. [From the Department of Physiology, School of Medicine, Western Reserve University, Cleveland, Ohio.]} Object of Investigation Acceleration of the heart in man is chiefly due to a varying balance of control exerted through the ss VENTRICULAR SYSTOLE. 95 vagi and the accelerator nerves. The only hopeful method of determining which mechanism is at least predominantly concerned is suggested by the observations of Baxt, Pavlow, Frank and Reid Hunt and others! that the accelerator nerves exert a predominant effect on the length of systole, whereas the vagi nerves affect chiefly diastole. This, on superficial examination, of course ap- pears contrary to any mechanical conception of the cardiac regulation, such, for example, as the “law of Uniformity of Be- havior’? advanced by Henderson and his coworkers.’ It is quite obvious that, if, as appears from volume curves re- corded by many different investigators, the rate of ejection dimin- ishes late in systole, then, on the basis of the uniformity law enunci- ated by Henderson, the length of systole will be only slightly altered during the longer cycles but will be progressively more and more abbreviated in a mechanical way as the heart cycles become shorter and shorter. Inasmuch as vagus section and vagus stimu- lation ordinarily do not alter the heart rate beyond the range where slight variations might be expected, whereas accelerator stimulation quickens the beat so that pronounced shortening of systole might be anticipated, the mere demonstration that acce- lerator stimulation shortens the systole 1s proof neither of any specific influence of these nerves over ventricular contraction, nor does it prove that the heart deviates in its beat from a mechanical scheme. Only 4f it can be shown that the periods of systole during accelerator nerve stimulation vary materially from those which may be accounted for on the basis of volume curves, can any inference be drawn as to a selective action of the accelerator nerves on the ventricle. Such proof has, however, not been presented by previous investigators, hence a reconsideration of the subject seemed desirable. Methods.—As a criterion of the length of systole, we used the interval elapsing between the first and second heart sounds re- corded from animals by the direct method described by Wiggers and Dean. Sounds were recorded consecutively during the 1 For literature see Hofmann in Nagel’s ‘‘Handbuch der Physiologie des Men- schen,”’ 1905, I, 269. 2? Henderson, Amer. Jour. Physiol., 1909, XXIII, 351. 3 Wiggers and Dean, Amer. Jour. Med. Sciences, 1917, CLIII, 666; Amer. Jour, Physiol., 1917, XLII, 476 also Wiggers, Arch. Int. Med., 1918, XXII, 28. 96 SCIENTIFIC PROCEEDINGS (105). following experimental conditions, the general order being varied only to suit particular occasions: (1) during normal cardiac action under morphine and chlore- tone anesthesia; (2) during stimulation of stellate ganglion or accelerator nerves; (3) following section of both vagi; (4) during stimulation of stellate ganglion with vagi severed and (5) during combined stimulation of vagus and accelerator mechanisms. Using these results, we have found it possible, on the basis of the duration of systole and diastole of a long vagus beat to con- struct for each animal a theoretical volume curve and to calculate from this the theoretical relations of systole and diastole at all heart cycles in that animal provided the ‘‘law of uniformity”’ applies. With these ‘‘theoretical systoles’’ in different cycle ~ lengths we compared (best in the form of a plot) the actual systole lengths of different cycles during the above mentioned experi- mental activities. Results.—We find as follows: 1. The lengths of normal systoles agree very closely with those of theoretical systoles. 2. Vagus section, causing cardiac acceleration, decreases the length of systole slightly but in accordance with its theoretical duration. 3. Vagus stimulation, causing only a moderate slowing, in- creases the length of the actual systole, practically in accordance with the theoretical values at different rates. 4. Accelerator stimulation shortens the length of actual systole far more than that of the corresponding theoretical systole, in- dicating that im some way the accelerator nerves are capable of shortening systole more than can be accounted for by the mechanical operation of the law of ‘‘ Uniformity of Behavior.” We append a few data of a single experiment typical of many others to support these conclusions. Ee ee ee Ee Oe ee ee ae INFLUENCE OF ANXIETY ON GASTRIC DIGESTION. 97 Theoretical Actual Systole Cycle STM mas Determined. i! ee oe .23 -.24 BEE ar ay) “Senay controls. bg Pee .23 —.24 a.) Pear .135-.14 Accelerators stimulated. Vagus intact. ee 2 “ te -155-.16 PL ) ee .22 .215—.22 Vagi cut. > eee .225 .22 MES. ds he%s .235 .28 Vagus stimulation. Oy <2! .238 -275 PaO Ss. s 25 25 PPh -175 .125-.135 Accelerator stimulation, vagi cut. cc See .19 .14 pi eee .228 .20 Combined vagus and accelerator stimu- EC acs ate -235 .20 lation. SES acs -235 -205 6 Ss .238 .20 Eee sas 24 .20 57 (1517) The influence of anxiety on gastric digestion. By RAYMOND J. MILLER, OLAF BERGEIM, and PHILIP B. HawK. [From the Laboratory of Physiological Chemistry, Jefferson Medical College, Philadelphia, Penna.]| The study of the influence of emotional strain on digestion in man offers some difficulties due to the fact that the emotions cannot be readily controlled, nor are the subjects of extreme emotion readily amenable to experimentation. We were, however, able to obtain an interesting illustration of the profound effect of mental anxiety on gastric digestion in the case of one of our subjects. The man was a first-year medical student who had pre- viously served as a subject of gastric tests. He was given one hundred grams of fried chicken on the morning of an important examination in chemistry, and was asked to write out his answers during the course of the test. He was plainly worried over the outcome of the examination and of his year’s work. The resultant effect upon gastric digestion in prolonging evacuation for over two hours with high intra-gastric acidity is charted in the figure. The 98 SCIENTIFIC PROCEEDINGS (105). Case. . 66- Hie; Examination - 82- re, Normal. Total Aculcty, Exam. ree" * Nucmiek same chart gives the normal digestion curve for fried chicken on this subject as obtained a week later under the best mental conditions. 58 (1518) Is unpalatable food properly digested? By CLARENCE A. SMITH, RALPH C. HOLDER, and PHILIP B. HAWKE. [From the Laboratory of Physiological Chemistry, Jefferson Medical College, Philadelphia, Penna.] It is well known that different psychic stimuli promote or re- tard the secretion of digestive juices. The following experiment was conducted to determine whether the ultimate return to the body from unpalatable food was different from the return from the same food palatably served. The experimental procedure was simple. A 7-day period during which the subjects were on a uniform diet, served palatably and amid pleasant surroundings, was followed by a 2-day period AMINO-ACID SYNTHESIS. 99 during which the same diet was fed in an unpalatable condition and in dirty and unpleasant surroundings. The food was rendered unpalatable and unappetizing by the following treatment. All the food ordinarily used for each meal (meat, biscuits, jelly, corn- starch pudding, oleomargarine, etc.) was stirred together in a large, flat porcelain dish. The dish itself was smeared with animal charcoal, as was the beaker used as a drinking glass. The table was dirty and strewn with dirty dishes. A little indol was sprinkled about under the table. The subjects were kept in ignorance of the constituents of the unpalatable mixture. The food was so unpalatable that one subject vomited his first meal shortly after he had eaten it. | The following table shows the findings on the other subject. Nitrogen. i NHA NH are ‘ | As,0; OSO,H O— CO CHART I. “Apparently o-aminophenol is to a comparatively small extent converted into the isomeric, highly toxic para compound, while the remainder is conjugated either with H.SO, with the introduction of an acetyl group, or with urea with the subsequent elimination of 2 N Hs; radicals.”’ Sieburg also studied the behavior of para-arsenobenzoic acid and of 3-amino-arsenobenzoic acid in the body of the calf and concludes that in all cases a cleavage of the toxophoric— As = As — linking in ‘arseno’ compounds occurs in the living organism, whereby not only is the toxicity reduced, but transformation takes place into compounds which are more soluble in water and less easily soluble in other media. The characteristic effects are due to the small proportion of the substance which escapes oxida- tion to the arsinic acids, and to the subsequent liberation of the As,O3 and As.O;.”’ Sieburg’s discovery of conjugated phenol derivatives resulting from the metabolism of salvarsan as well as our independent investigations on the causes of the reactions following intravenous injections of arsphenamine and neo-arsphenamine! led us to direct our attention to the mechanism of detoxication which the body possesses,—the power of rendering innocuous various aromatic toxic substances by conjugation with sulphuric acid, d-glucuronic acid, urea, glycocoll, bile acids, etc. It seemed plausible to venture the hypothesis that in those patients in whom severe reactions are observed within a few hours or days after intravenous injection of arsphenamine or neo-arsphenamine, 1Schamberg, J. F., Kolmer, J. A., Raiziss, G. W., and Weiss, Charles, Arch. Dermat. & Syph., 1920, 1, 235-256. i i PHENOL ELIMINATION IN THE Doc. 105 detoxication is subnormal, owing to injury sustained during the course of the infection by the hepatic, renal and other cells con- cerned in this mechanism. Of course, normal variations in this process cannot be overlooked as a possible factor. We have, therefore, studied, in a preliminary way, the elimi- nation of free and conjugated urinary phenols after the intravenous injection of a single large tolerated dose (50 mg. per kilo) of neo-arsphenamine into the dog. Method.—A normal healthy female dog (16.82 kilo) was employed and was kept on a constant diet. The technique followed was that of Folin and Denis! with the modifications described by Dubin.? Results.—The results are summarized in Table 1 and indicate the following: TABLE I. PHENOL ELIMINATION AFTER A SINGLE INTRAVENOUS INJECTION OF NEO- ARSPHENAMINE (0.841 Gr.).! Control | rst Neo | 2d Neo | 3d Neo Period Preipas Period Beriod oaey Daily Elimi- = nation — — — — e — eo ‘ef vo os vo ve ta] o > © & é & 5 f 2 & 3 Periods of Exp E k 8 ° Ist day.....|106}156/117|158| 95/147] 90/127] Normal 528| 734'206| 72} 28 2 oe ee a I17|187| 96)125|/115/173/108/143] Ist neo 506] 673/167) 75| 25 5 RR Ade Oe aa 78|110| 82|109|105|140/119|173}] 2d neo 581) 875/294! 66! 34 3d neo SII} 728|217| 70} 30 4th “ .....{106)/149|106)143/152/238| 88/138] Total 3 neo periods I,598|2,276|678) 70| 30 5th “ .....)/121/132/105/138,114/177'106/147] 15 days , Bxtra”’ Total 5 days |528/734|506|673/581 ea awh 728] phenols 14} 74} 60] 19] 81 I5 days 1 All figures refer to miligrams unless otherwise indicated. - I. The elimination of ‘‘normal”’ free and conjugated phenols is quite uniform from day to day, if the animal is kept on a con- stant diet. The work of Folin and Denis is thus confirmed. 2. After a single intravenous injection of neo-arsphenamine, no appreciable variation in phenol elimination is observed during the first period of five days. During the next period there is an increase in both the free and conjugated phenols. During the 1¥Folin, O., and Denis, W., Jour. Biol. Chem., 1915, 22, 305-308. 2 Dubin, Harry, Jour. Biol. Chem., 1916, 26, 69-91. 106 SCIENTIFIC PROCEEDINGS (105). third period of five days there is a return to practically normal excretion. 3. The injection of a dose of 0.841 gr. of neo-arsphenamine results in the elimination of a total of only 74 mg. “‘extra’’ phenols during a period of 15 days; 81 per cent. of this is in the conjugated state. Of the ‘“normal’’ phenols only from 10 per cent. to 30 per cent. is conjugated. Since in the colorimeter 1 mg. of neo- arsphenamine reacts like 0.466 mg. phenol, we have the striking observation that only 9.2 per cent of the dose of the drug injected is eliminated in the urine in the form of a phenol or phenol deriva- tive. This suggests the following possibilities: (a) Some of the drug is eliminated as phenols in the feces; (6) a portion of it is burned; (c) a portion is retained or “‘fixed”’ by the various tissues, notably the liver, for a period much longer than 15 days; (d) most of the drug is oxidized to the stage of inorganic arsenic and elimi- nated as such; (e) a portion is eliminated as nitro-phenols which do not react with the Folin reagent. The latter suggestion seems the most improbable but not impossible. TABLE 2. PHENOL ELIMINATION AFTER INJECTION OF NEO-ARSPHENAMINE AND AMMONIUM PERSULPHATE.! Normal | Persul. | rst Period; 2d Period Control | Control | Neo. and | Neo and Summary. Period. | Period.| Persul. Persul. Daily | Elimination. i Bice a Pe be & ; e g |-> i es Oe gs hy | Pa g a || 9 fc & "3 ‘a o & cs & Periods hg c 3 ef = Ist day...| 99/154|137|188,136] 190) 235) 265} Normal period...... 523) 775|252,.68/32 : 5 days with 2d “ ...] O1/154)128|181)191} 272) 263) 301] persul. only .| 604) 834|/230/73|27 Ist period after neo 3d 06“ ...|106)152|106'135|137| 188) 320 365] and persul...| 885/1,163/278)|76/24 2d period after neo 4th “ .../113/153/124/173\212| 265) 275! 331] and persul.../1,426)1,630|204/88|12 Total ro days neo Sth “ .../114)162}109)157\209| 248) 333 368] and persul...|2,311/2,793|482 83/17 | “sire” Total | phenols 5 days ...|523/775 604/834 885 1,163]1.426 1,630] 10 days..... I,103|1,123 2198 2 1 All figures refer to miligrams. — w-—~ = PHENOL ELIMINATION OF THE Doc. 107 Since the administration of sulphates is known to counteract the effects of phenol poisoning, and particularly in view of the work of Bufalini,| we studied the phenol elimination in the same dog after a single intravenous dose of neo-arsphenamine during the time that she was receiving daily subcutaneous injec- tions of ammonium persulphate. Method. The animal was kept on the same constant diet. Phenol elimination was studied for five days (control period) ; ammonium persulphate was now given subcutaneously twice daily in 2.5 per cent. solution in physiological saline in doses of 10 c.c. and phenol excretion was measured during this period. A single intravenous dose of 0.841 g. of neo-arsphenamine was now given and persulphate injections and estimation of urinary phenols was continued for ten more days. Results are summarized in Table 2, and indicate the following: 1. The subcutaneous injection of ammonium persulphate which theoretically would furnish the organism with additional sulphuric acid for phenol conjugation, does not result in the elimi- nation of any “extra’”’ conjugated ‘‘normal’”’ phenols. During a period of five days, only a slight increase in the free ‘normal”’ phenols was observed. 2. After the intravenous injection of a single dose of 0.841 g. of neo-arsphenamine and the daily subcutaneous administration of the persulphate, an enormous increase in the output of “‘extra”’ free phenols was observed. Simultaneously, a marked diminution in the excretion of “‘extra’’ conjugated phenols was noted. The animal also showed definite symptoms of intoxication. We must, therefore, conclude that the ‘‘extra’’ free phenols were due entirely to tissue destruction resulting from the toxic effects of the persulphate. The writer is indebted to Dr. John A. Kolmer, Head of the Department of Pathology, for his encouragement and unfailing kindness throughout the course of this work. 1 Bufalini, G., Archives ital. de Biol., 1903, 40, 131-140. 108 SCIENTIFIC PROCEEDINGS (105). 63 (1523) The influence of hunger and temperature upon the utilization of food substances. By EDUARD UHLENHUTH. [From the Laboratories of the Rockefeller Institute for Medical Research.| In the larve of amphibians when the thyroid glands begin to excrete the thyroid hormone, metamorphosis of the larve into adult animals takes place. Since larve with: fully developed thyroid glands frequently do not metamorphose, and since in the thyroid gland of the normal larve, large quantities of ‘‘colloid”’ are present in the follicles for a considerable time before metamor- phosis, it is very probable that the thyroid gland cannot begin to excrete its hormone unless a second factor is present in the larve. It seems that this factor is elaborated during the process of growth and must be present in a definite quantity in order that the thyroid function may begin. This is shown in two tables in which for several series of the marbled and the tiger salamanders, the age at which metamorphosis took place and the rate of growth were recorded. ‘The greater the rate of growth the shorter the length of the larval period. As a consequence the product of the rate of growth into the duration of the larval period gives a fairly constant value K. The maximum deviations observed can be traced back to certain causes which will be discussed immediately. Hence it is evident that metamorphosis not only depends upon the thyroid hormone but also on a second substance, the quantity of which increases in the same ratio as growth. This second substance must be present in a certain quantity in order that metamorphosis can take place. The rate at which it is formed from the same kind of food and for the same rate of growth is distinctly influenced by two factors; by the quantity of food available to the larve, and by the temperature. A series of the marbled salamanders (A 1916) required 186 days to metamorphose, when they grew at a rate of 0.21; K was 39. Another series (C 1916), which was fed the same food but kept UTILIZATION OF FOOD SUBSTANCES. 109 at a temperature lower by 10 degrees, needed at the same rate of growth 243 days to metamorphose. K was therefore higher than for series A (51). The same was true for other series of marbled as well as tiger salamanders; in all of these series the length of the larval period was increased more than the rate of growth was decreased, and consequently the product K increased. In short, during the same amount of growth less of the substance was produced at low temperature than at high temperature, though the same kind of food was given to all the larve. If one diminishes the quantity of food instead of lowering the temperature, the rate of growth decreases as in the case of lowered temperature, and the length of the larval period increases, as is shown in the record of four series of marbled salamanders. All four series were kept at the same temperature and fed the same kind of food, but Series D received only one half, Series E only one quarter the amount of food which was given to Series C, and A received still less food. Consequently, as in lowered temperature, the rate of growth was decreased with the diminution of food, and the length of the larval period was increased. But while in lower temperature the length of the larval period was increased more than the rate of growth was decreased, hunger increased the length of the larval period less than it decreased the rate of growth, and consequently K decreased instead of increasing as in lowered temperature. When the same kind of food is available to the amphibian organism, a certain substance required for metamorphosis is elaborated, at the same rate of growth, the more easily the less food there is available, and the less readily the more the tempera- ture is lowered. awh on pn art, Melle bat ete) Jay ae ai SPN a ee ee ee ay oe PY ot Cy eg eek i, 5 ee —s —— we ‘ << 7”, * ' = SCIENTIFIC PROCEEDINGS. ABSTRACTS OF COMMUNICATIONS One hundred sixth meeting College of the City of New York, Presbyterian Hospital, N. Y. City, March 17, 1920 64 (1524) A case of lipuria associated with chronic nephritis. By L. BAUMAN and G. H. HANSMANN. [From the Depariment of Internal Medicine, State University of Towa.| Clinical, pathological and chemical data of a case of lipuria associated with chronic nephritis which terminated in uremia. The lipuria was influenced by the amount of fat in the diet. The absence of coagulated protein, the scarcity or absence of cells in the urine and the apparent absence of a fistulous communication between the urinary passages and the lymphatics at autopsy indicate that the lipuria was due to an altered permeability of the renal cells. The available evidence makes it probable that there are at least two types of lipuria, the one associated with a fistulous communication, the other entirely due to an abnormal condition of the kidney cells. 65 (1525) Calcium in the blood in diseases of the skin. By WILLIAM C. THRO and MARIE ESN. [From the Laboratory of Clinical Pathology, Cornell University Medical College, New York City.] Before proceeding with experiments on the efficacy of calcium in the treatment of furunculosis it was thought advisable to deter- II5 116 SCIENTIFIC PROCEEDINGS (106). mine the amount of calcium in the blood of patients with boils. We proceeded on the idea that the calcium might be decreased in such patients. As it happened, we first examined a number of patients with acne and we were much surprised to find the amount of calcium markedly increased in such. After testing about ten patients with acne we examined our first patient with boils and found the calcium was slightly below normal in amount. In one other patient with a large boil on the arm the amount of calcium was extremely low; that is, 5 mg. per 100 c.c. of plasma. Thus it is seen that in most of the patients with acne the amount of calcium in the blood was markedly increased while in some of the patients with furunculosis the amount of calcium was very low. So far as we have searched the literature we have not found any results reported on the amounts of calcium in the blood of patients with acne and furunculosis. The amounts of calcium found by us in the blood of patients with acne were so extremely large that we determined to subject the gravimetric method, which we used, to a careful investigation. We ran through six blanks by the ashing method, which we used, with-negative results. In one test we used 20 c.c. of our distilled water and found it free from calcium. In case No. 2, 22 c.c. of plasma were obtained at one time and divided into two portions of eleven c.c. each. In one portion the calcium was determined by Mr. Osterberg and in the other by Thro. The results disagreed by only 1.8 mg. per 100 c.c. of plasma. Incase No. 6, Mrs. Ehn divided the whole blood into two lots of 25 c.c. each and her determinations of calcium agreed exactly. Six times the titration method, as given by Halverson and Bergeim', was used as acontrol. While the results were not in strict agreement with those obtained by ashing, it was noted that if high results were obtained with one they were also obtained with the other. It is our opinion that the ashing method is more accurate with large amounts of plasma than with small amounts. It is also believed that it is better to use blood plasma or serum than whole blood, since results are obtained sooner and variations in corpuscular volume do not affect the results. In every one of our patients the volume of corpuscles was greater than the volume of plasma. 1 Halverson, J. O., and Bergeim, O., Jour. Biol. Chem., 1917, 32, 159-170. —_ CALCIUM IN THE BLOOD IN DISEASES OF SKIN. 117 As is well known, the giving of a diet low in carbohydrate!? tends to eliminate the calcium from the body. ‘This was tried twice on patients with acne, resulting in the lowering of the calcium in the blood. TABLE I. Mg.Cain1o0 C.c. Case ; : : hee | hh, a ee Ho. Name. Diagnosis. Quantity Used, C.c. Titra- Ashing.| ‘tion. 2 N. W. C Acne Plasma, Ti 19.9! a *. Jan.) 7,:1920.7 a II 18.1 5 | Miss C. a Whole blood, 25 24.5 nf ** v Jan. 19; 1920. i = ays, trae 13.6 Oo eG. CC. " oe ry 17.68 sé ae sé sé 25 17.6 9 eC. - oo tet, an 12.0 8 | Mr. A. i Plasma, 15 17.8 II Mary F. . rt 19 17.8 I2 Mr. R. a5 bk 12 10.7 ty | Mirs, V. iy x 19 25.4 18 | Miss M. i: ry 10 25.7 Pe oe Same ME vg 18 25.6 aq° i) Mr. L. od i 12.5 14.4 267 1 mir, S. a e 10 15.0 | 17.4 5. ers, SV Furunculosis Whole blood, 25 13.6 1?) De. B baa Plasma, 10 8.2 23 fi ee 2 14 5.0 6.0 22 | R ee i 18 11.8 | 14.4 27 | Mr. McA i is 20 II.5 Pe Be! a Folliculitis barbae cy 13.5 22.24 | 20.83 Ima j}|—-—_— Epilepsy Whole blood, 25 12.4 16 |—— i) rh 25 8.0 26). G, C. Diabetes Plasma, 12 20.4 | 17.6 4Mr. Osterberg ascertained the Ca in one lot of 11 c.c. and Thro in another portion of II c.c. 5 Between the two dates the carbohydrate intake was low. 6 so c.c. of blood divided into two portions of 25 c.c. each. H. J. Schwartz has found that in acne there is a carbohydrate fermentation of the feces when examined by the Schmidt method. He has also reported that in skin diseases there is an increase in the blood sugar* The blood of one diabetic was tested for the calcium content but the amount was not decreased. 1Thayer, W.S., and Hazen, H. H., Jour. Exper. Med., 1907, 9, I-17. 2 Mendel, L. B., and Benedict, S. R., Am. Jour. Physiol., 1909-10, 25, 25. 3 Schwartz, H. J., Heimann, W. J., and Mahnken, H. C., Jour. Cutan. Dis., 1917, March. 118 SCIENTIFIC PROCEEDINGS (106). The pathologic lesion of acne is in the sebaceous glands of the hair follicles, while in furunculosis the sweat glands become infected. It is of interest, then, to note that in one patient with folliculitis barbae, in which the hair follicle is involved, the amount of calcium in the blood was increased. GRAVIMETRIC METHOD USED. The blood was collected in a flask containing 90 mg. of sodium citrate. After centrifugalization the plasma was drawn off and evaporated in a platinum dish; ashed with addition of nitric acid. The ash was taken up with dilute hydrochloric acid. The solution was made alkaline with ammonia and then faintly acidulated with acetic acid. The iron was precipitated with 20 per cent. sodium acetate. The calcium was precipitated from the filtrate with 4 per cent. ammonium oxalate. The solution was allowed to stand over night. Gooch crucibles packed with asbestos were used. The calcium was weighed as calcium oxide. 66 (1526) The preparation of animal nucleic acid. By EMIL J. BAUMANN. [From the Chemical Laboratory of the Montefiore Home and Hospital, New York City.] The proposed method depends on the following properties of nucleic acid: (1) that it is separated from its protein combination by treatment with sodium hydroxide, (2) that it is soluble in dilute acetic acid and, (3) that it is precipitated by hydrochloric acid, or in the case of dilute solutions, by hydrochloric acid in the presence of magnesium sulphate. Fresh, trimmed glandular tissue, finely hashed, is mixed well with twice its weight of tap water and 100 c.c. of 50 per cent. sodium hydroxide is then added for each kilogram of tissue. The material is then heated until all except the connective tissue is dissolved (40° C.-70° C.). Neutralize hot and at once with strong acetic acid; the reaction should finally be distinctly acid to litmus. Bring to a boil and filter hot on large folder filters. When the filtrate has cooled an aliquot of 200 c.c. is taken and SoME HuMAN DIGESTION EXPERIMENTS. 119 concentrated hydrochloric acid diluted with an equal volume of water is added from a burette until precipitation is complete, carefully avoiding an excess. If the nucleic acid does not flock out after the solution has been quiescent for several minutes, the same process should be repeated upon another aliquot after first adding 5 per cent. of magnesium sulphate (MgSO.4.7H.O). Mag- nesium sulphate renders nucleic acid more insoluble. A pro- portionate quantity is then added to the bulk of the solution. The nucleic acid, which forms large flocks and slowly settles, is washed successively with 60, 80, and twice with 95 per cent. alcohol by decantation, filtered on hardened filter paper, washed again with 95 per cent. alcohol and finally with ether and then > rapidly dried at about 70° C. The yields from different glandular tissues vary from 0.8 per cent. to 1.5 per cent. 67 (1527) Some human digestion experiments on raw white of egg. By Mary SWARTZ ROSE and GRACE MACLEOD. [From the Department of Nutrition, Teachers College, Columbia University.| Experiments to determine the relative digestibility of raw and cooked white of egg in the human subject were carried out with four young women students in two periods of five days each. As they all had practically the same food requirements, they took the same diet, quantitatively and qualitatively, throughout the experiment, the only variation being the change from raw whites to cooked whites for half the time, and some differences in the ways in which the whites were prepared. The egg whites fur- nished 48 grams of protein per capita per day out of a total of 67. grams. Besides the eggs the diet consisted of rice, cream, saltines, butter, olive oil, fruit juice and a small amount of lettuce. The cooked eggs were never subjected to a temperature or method of cooking which would toughen them unduly; the raw whites were taken unbeaten by one person, all beaten light by another, and about half and half by the other subjects. The coefficients of digestibility for the two diets have been calculated for the total protein and for the egg white protein alone. 120 SCIENTIFIC PROCEEDINGS (106). COEFFICIENTS OF DIGESTIBILITY FOR RAW AND COOKED EGG WHITES. For Protein of Whole Ration. |For Protein of Whites of Eggs Only. Subject Eggs Eggs Diff. (in Eggs Eggs Diff. (in Remarks, Cooked, Raw, Favor of | Cooked, Raw Favor of %- %- Cooked). %. he Cooked. G. S. ’ 85.7 82.6 +3.1 85.4 81.0 + 4.4 4 beaten | Rs ee ¥% beaten M. s 83.2 83.9 —0.7 81.8 82.8 — 1.0 | All beaten M.F..| 83.3 75-7 +7.6 81.9 7i.2 +10.7 All unbeaten In these experiments the raw white was as well digested as the cooked if beaten light, and the difference between the two was not striking when taken half beaten and half unbeaten. The greatest difference was observed when the whites were swallowed with no subdivision whatever, and even then the dif- ference between the cooked and the raw was only II per cent. when as many as ten or twelve whites were taken per day. The effect of beating on the coefficient of digestibility is under further investigation. 68 (1528) A study of the sugar and oxygen relationships in the blood of dogs during exercise. By ERNEST L. ScoTT and A. BAIRD HASTINGS. [From the Department of Physiology, Columbia University, New York.| As a phase of our investigation of the chemical changes in the organism resulting from exercise, the following study of the sugar and oxygen relationships in the blood of dogs was under- taken. Samples of blood amounting to about 1 per cent. of the body weight were drawn from the external jugular vein. Determina- tions of the blood sugar by the MacLean method, of the oxygen by the Van Slyke technique, and of the volume of the corpuscles by a precision hematocrit were made every two hours during the course of six-hour working periods. During these periods the dogs ran on an electrically-driven, horizontal treadmill at the rate of about five miles per hour. For each such experiment, we made a corresponding series of control observations on the same SUGAR AND OXYGEN RELATIONSHIPS IN BLOoop. I2I dog similar in all respects save that the exercise factor was elimi- nated. The figures given in the table are the averages of the data obtained from five dogs. The fact that the averages of the initial samples are so nearly identical for each constituent, indicates that our series is sufficiently long to permit the attaching of significance to subsequent varia- tions. Since it is desirable to compare figures which were obtained under conditions identical except for the added element of exercise, we wish to point out that in studying the table the figures given for exercise should be compared with the figures for the corre- sponding periods of rest, rather than with the initial values for the exercise experiments. This is indeed necessary in the inter- pretation of the effect of work because variations of significant magnitude in the control series make comparisons between initial and successive periods unjustifiable for this purpose. Experi- | Number of Elapsed Time in Hours, mental |Experiments Conditions. | Performed. °. 2. ve 6. Oxygen content c.c. per 100 c.c. RMN Gre at Wot ota oA Wk. oie ioa Rest 12 16.5| 13.7| 14.5] 14.0 Work 13 16.1 eS ts TSO |<. 36.6 Difference due to work........ —0.4| +3.6] +2.5 | +2.6 Oxygen capacity c.c. per 100 c.c. 1 OSES AP ER le aoe Sere Rest 12 24.6 | 23.8.| 23.6]. 23.2 Work 13 24.8} 25.6] 26.4] 26.4 Difference due to work........ +o.2 | +1.8| +2.8|] +3.3 Volume of corpuscles expressed =f og ee ee ee Rest 8 49.9| 47.1| 46.9] 47.9 Work 10 49.1 | 50.6] 51.0] 51.5 Difference due to work........ —o0.8 | +3.5| +4.1 | +3.6 Sugar as glucose mg. per 100 c.c. ES BOS SP Ors es See eee Rest I2 69 68 70 66 Work 13 69 66 64 60 Difference due to work........ o —2 —6 —6 From the table it will be seen that the oxygen content of the blood rises during the first period of work as compared with the first period of rest and falls slightly thereafter. These results may be due among other things, to increased aeration in the lungs, increased oxygen capacity of the blood, and to an increased rate of blood flow through the tissues. The oxygen capacity of the blood rises progressively, but at a decreasing rate, throughout the period of work. This phenomenon 122 SCIENTIFIC PROCEEDINGS (106). is correlated with a parallel rise in corpuscular volume as indicated in the table. The concentration of sugar falls steadily throughout the work period. It should be noted, however, that the variations during the first period in work and those during the same period in rest are almost identical, but from this point on there is a divergence which becomes quite pronounced in the later stages of the expe- riment. It is perhaps worthy of notice that there is a rather distinct change in the magnitude of the effects of work after the first period. This we have interpreted as indicating that the day’s work falls into at least two phases; first, one in which certain augmenting effects of exercise apparently predominate, second, one in which fatigue phenomena are relatively more prominent. 69 (1529) Further studies in the measurement of vitamine content. By WALTER H. EDDY and HELEN C. STEVENSON. [From the Department of Physiology Chemistry, Teachers College, Columbia University and the Department of Pathology, New York Hospital.] During the past year two papers have appeared in the Journal of Biological Chemistry, one by Dr. Bachman! and the other by Dr. Williams’, which postulate the requirement of the water- soluble B vitamine in yeast growth. Both these papers suggest the use of yeast cells as a means for the measurement of vitamine content and present a technique that may be used to that end. In an earlier number of the PROCEEDINGS’ we reported ona study of the suitability of the Bachman test for vitamine measure- ment and some of the difficulties encountered. Since that time we have carried out similar studies of the Williams technique and as a result have devised a method which employs features of both 1 Bachman F., Vitamine requirements of certain yeasts. Journ. Biol. Chemistry IQIQ, xxxix, 235. 2 Williams, Roger J., The vitamine requirement for yeast. A simple biological test for vitamine. Journ. Biol. Chem., 1919, xxxviii, 465. 3 PROCEEDINGS Soc. ExpER. BIOL. AND MED., 1919, xvii, 52. MEASUREMENT OF VITAMINE CONTENT. 123 authors but seems to permit of better control and to be at the same time more easily handled than the Williams method. This method is presented herewith, not as a finished product for we are still experimenting with certain details of standardization, but with a view to stimulating criticism and suggestion. The Bachman test measures the vitamine activity in terms of gas generated; it is really a measure of enzyme activity. This fact makes it difficult to be sure that the stimulus is a growth stimulus or merely an enzyme control in any given test. The Williams test measures the effect of the vitamine in growth of yeast cells and this seems to us a more reliable indicator than the gas production. On the other hand the hanging drop method of Williams makes difficult the control of concentration during incubation and we found the preparation of drops containing single yeast cells far from easy to prepare. We believe that our method obviates both these difficulties while retaining the yeast cell count as an indicator. The Method.—The illustration shows the tools used, being essentially those of the opsonin technique. The first step is the preparation of two capillary pipettes by drawing out in the flame a 5 mm. glass tube as shown in A. This tube is marked at the center with a pen point (1) and with the aid of a drop of mercury accurately calibrated into units of equal volume on each side of the center point (2) and (3). The two halves are then separated at the point (1) into two pipettes. Each pipette is then heated at the large end and a constriction made to permit a flame seal later, the end plugged with cotton and the pipette sterilized. After sterilization and by fitting a rubber bulb to the end as in B the pipette is ready for use. After filling as described below the tip and constriction point are sealed in the flame and the tube is ready for incubation, C. The materials necessary for the test are the pipette described above, a dilute suspension of yeast cells in Nageli solution, and the vitamine extract to be tested. These are prepared as follows: A pure culture of Fleischman round yeast is maintained on an agar slant. Two days before the test a transfer is made to a fresh slant and in all our tests, cells of 48 hour growth are used. From such a slant the smallest amount of yeast that can be taken up on a 124 SCIENTIFIC PROCEEDINGS (106). needle point is transferred to 10 c.c. of Nageli solution in a sterile test tube and the tube shaken from 134 to 2 hours on a shaking machine. At the end of that time the uniformity of the emulsion is tested by removing ten units of the suspension with the pipette, blowing them out on a microscope slide, staining and counting. If the counts are uniform the suspension is used, if not it is shaken further. ‘This uniformity of the emulsion is extremely important in comparing results. Having secured a uniform emulsion the pipettes are prepared as follows: Two units of a sterile extract of vitamine are first drawn up into the pipette (the unit we have used is about 1/800 c.c.). We have prepared our extracts in various ways but in all cases they have been sterilized for two 30-minute intervals in the Arnold Sterilizer with a 24-hour inter- val. Next two units of yeast suspension are drawn up. By manipulating the bulb a bubble of air separates the yeast and vitamine in the tube and permits accurate measurement. When both materials are in the tube they are drawn up to the large part of the tube and mixed. The tube is then sealed as described above. The control tube contains the yeast cells in the Nageli solution (two units) but no vitamine. Our Nageli solution is the same as used by Dr. Bachman, being a sterilized solution of the following components: 100 c.c. distilled water; 10 gms. dextrose; I gm. ammonium nitrate 0.05 gms. calcium phosphate, 0.5 gms. potassium acid phosphate; 0.25 gms. magnesium sulfate. After the tubes are filled and sealed they are incubated at 35° Cent. for the time necessary, usually 18-24 hours. At the end of that time the ends are broken off, the rubber bulb adjusted and the contents blown out on a specially prepared counting slide, fixed and stained and can then be counted at leisure. For this purpose we prepare common microscope slides by etching 7 mm. squares on them, experience having shown that such a square will hold the contents of a tube. In this way it is possible to get a series of the contents of ten or twelve pipettes on one slide. Count- ing is done under the high power with the aid of a mechanical stage. Results with the Method.—The methodology reported above has been arrived at by experiment. The necessity for accurate calibration, the need for uniform suspensions of yeast, the size of TT , MEASUREMENT OF VITAMINE CONTENT. 125 the units etc. are the result of preliminary trials and tests that need not be reported here in detail. Certain questions however arise at once and some of our results to date are presented here as partial answers to these questions pending the accumulation of more complete data. The first question that arises is naturally as to the specificity of the test. Bachman and Williams have gone into this matter somewhat but we believe that our results with the above technique demonstrate additional evidence that this test is specific for the so-called antineuritic vitamine or water-soluble B. Dr. Funk kindly supplied us with a specimen of his purified antineuritic vitamine prepared after his well-known method in 1913. A water solution of this material sterilized in the Arnold sterilizer was tested with the results given below. We also made comparative studies of an extract of navy bean made after the McCollum method‘ before and after shaking with Lloyd’s reagent. This reagent is supposed to remove quantitatively the B and apparently with little effect upon the other constituents of the extract. The results follow: TABLE I. wads Na: Veust Calls a ae Cell Count of Incubated Tubes. Vitamine Preparation Tested. in Unit. Period. Vitamine Tube. | Control Tube. Water sol. Navy bean (a). 7-8 20 hours 11,237 | II3 Water sol. Navy bean (0). 2 20 672 15 Water sol. Navy bean (c) . 2 20 633 I5 Same after shaking with Lloyd reagent (a)...... 7-8 20 100 113 Same after shaking with Lloyd reagent (b)...... 2 20 2 I5 Same after shaking with Lloyd reagent (c)...... 2 20 fe) I5 Funk 1913 preparation... 7-8 20 hours 404 II 2 20 31 2 | 2 20 | 68 2 These results seem to demonstrate that what Funk calls the antineuritic vitamine as prepared by him and what we call water- soluble B as prepared by the McCollum method both respond to the test whether they are identical in character or not. The absence of salts and other substances present in the crude navy * McCollum and Simmonds, A study of the dietary essential, water-soluble B. Journ. Biol. Chem. 1918, xxxiii, 55. 126 SCIENTIFIC PROCEEDINGS (106). bean extract resulting from the Funk method seems to indicate that it is the vitamine and not the impurities that is responsible for the action. Many more experiments are of course necessary to settle this point fully but these preliminary ones justify faith that we are here dealing with a specific test for vitamine. We hope to confirm them in much greater detail later. DIAGRAM OF APPARATUS. {L) A (2) (3) SEN es Mee We ee ES Rubber Cotton Plug Counting Slide. Rubber Cotton C D Counting Slide Bulb Plug A (2) (1) (3) B The second question that arises is as to whether it is possible to compare solutions by this test and actually measure vitamine content quantitatively. Experiments in this direction have consumed most of our time to date and have led to the technique presented, since the earlier tests showed the necessity for rigorous control in each feature of the test. We have not yet attained satisfactory results in this respect but believe it is attainable by improvement in technique. In this connection it must be borne in mind that aside from uniform yeast suspensions, accurate cali- bration and other mechanical details the test is biological. Yeast cells like rats vary in metabolic activity, the vitamine necessary to stimulate growth does not vary directly with concentration but exhibits an optimum and minimum effect which must be MEASUREMENT OF VITAMINE CONTENT. 127 established in the comparative standard used. These factors complicate the problem. The following results show some of these difficulties and are given rather to suggest the problems than as conclusions. All these features are under study and we hope soon to present a report of progress toward their solution: TABLE II. FNS eS ae Count of Incubated Tubes. Vitamine Preparation Used. | Concentration. bation Weta sk TC tp ee J Vitamine Tube. | Control Tube. Water sol. vitamine from Navy bean, F24....... c/I 24 hours 5312 5 OB a ore c/2 24 3192 28 PO Pict te c/I 18 6161 14 c/2 18 472 58 c/8 18 59 20 LE eae a c/I 18 9724 123 c/2 18 362 : 60 c/8 18 85 9 Peto 55 2 c/I 18 6248 48 c/2 18 729 60 PO ans wo c/I 18 8531 07 eae c/I 6 24 2 c/2 6 147 6 c/4 6 119 3 The above were made with the first pipettes before accurate calibration or stan- dardized suspensions were made. The following were results with calibrated pipettes but the suspensions were not entirely uniform as shown by a count of 25 units of the suspension used.- These units were taken after an hour shaking and varied as follows: 18-3 7—12—1I-—3—2—2—0—I 5-3 5—46—78—5 7—6—2 7-3 I-33—-1 2-1 I-1 7-3 4-2 1-60-29 cells. Hae 6... c/I 6 27 15 c/2 6 9 one control c/4 6 14 for series c/I 6 20 6 c/2 6 16 one control c/4 6 60 for series c/8 6 12 The following were made with calibrated pipettes and a suspension for which eight counted units resulted as follows (2 hours shaking) : 4-2-3—1—3—5—2-3 cells. Le amie a c/I 22 hours 492 I c/2 22 660 one control c/4 22 102 for series c/8+ 22 II ~ 128 SCIENTIFIC PROCEEDINGS (106). It is unnecessary to suggest the various fields of application for the test in testing the effect of heat, alkali, quantity, etc. The above represents merely a preliminary communication and we hope soon to report such modifications as will meet the defects that have shown in the preliminary study outlined here. 70 (1530) A study of local anesthetics in respect to their antiseptic properties. By D. I. Macut and Y. SATANI. [From the Pharmacological Laboratory, Johns Hopkins University, and the James Buchanan Brady Urological Institute, Baltimore, Md.| An inquiry into the antiseptic properties of local anesthetics is interesting for two reasons: Firstly, in relation to the healing of post-operative wounds and secondly in connection with genito-uri- nary practice, where these drugs are introduced into the bladder cavity and may thus directly influence its bacterial flora. The following local anesthetics were studied by the authors: cocain, novocain, alpha-eucain, beta-eucain, stovain, holocain, alypin, apothesine and benzyl alcohol. Bacteriological tests were made in three ways. Firstly, the growth of bacteria in bouillon con- taining small amounts of the anesthetics was studied. Secondly, bacteria were suspended in weak solutions of the anesthetics for definite periods of time, then washed, and planted in various media in order to determine whether they will growjor not. Thirdly, bacteria were planted on agar impregnated with the various drugs and their growth in this medium was noted. It was found that some local anesthetics possess definite antiseptic properties, while others are entirely devoid of such. It was interesting to find that the chief local anesthetics in use, namely, cocain and novocain, possess no antiseptic properties. On the other hand, some of the other bodies studied and in par- ticular, benzyl alcohol, beta-eucain and holocain, showed distinct antiseptic properties. The fuller data will appear in due time in the Journal of Pharmacology and Experimental Therapeutics. CATALASE CONTENT OF TISSUES. 129 71 (1531) Comparison of the catalase content of the tissues of the mother and of the offspring. By W. E. BurGE (by invitation). [From the Physiological Laboratory of the University of Illinois.] Hasselbalch! found that oxidation or metabolism is very low in the infant during the first month of life, and Magnus-Levy and Falk? that it is high during childhood. Asa result of the work of these three observers and of Bailey and Murlin’, Murlin and Hoobler‘*, Howland *, Benedict and Talbot *, Benedict, Emmes, Roth and Smith’, Palmer, Means, and Gamble’, and others, it is now considered that oxidation or metabolism is low during the first month of life, high during childhood, and low after the onset of old age. Warburg® found that during the process of fertilization, oxidation was greatly increased in the sea-urchin egg. It is also known that oxidation is greatly increased in the greening of tubers and the germinating of grain. The present work is an attempt to find an explanation for the variation in the intensity of oxidation under the conditions named. Since we! had found that whatever increased oxidation in the body, the ingestion of food, for example, produded an increase in catalase, an enzyme possessing the property of liberating oxygen from hydrogen peroxide, by stimulating the alimentary glands, particularly the liver, to an increased output of this enzyme, and that whatever decreased oxidation, narcotics for example, dimin- ished catalase by decreasing its output from the liver and by direct destruction, we naturally turned to catalase for an expla- 1 Hasselbach, Bibliotek for laeger, Copenhagen, 1904, 8, 219. 2 Magnus-Levy and Falk, Arch. f. Anat. u. Physiol., 1899, Suppl. 315. 3 Bailey and Murlin, Am. Jour. Obst., 1915, Ixxi, 1. #Murlin and Hoobler, Am. Jour. Dis. Child., 1915, ix, 81. 5 Howland, Ztschr. f. physiol. Chem., 1911, Ixxiv, I. 6 Benedict and Talbot, Carnegie Institution of Washington, Pub. 201, 1914. Am. Jour. Dis. Child., 1914, viii, 1. 7 Benedict, Emmes, Roth and Smith, Jour. Biol. Chem., 1914, xviii, 139. 8 Palmer, Means and Gamble, Jour. Biol. Chem., 1914, xix, 239. 90. Warburg, Zeitschr. f. physiol. Chem., 1908. Ivii, 6, 10 Burge, Am. Jour. Physiol., 1918; xlv, 4, 1918; xlvii, 1, 1918. xlvii, 3, 130 SCIENTIFIC PROCEEDINGS (106). nation of the increase or decrease in oxidation under the conditions enumerated. On examination of the literature, it was found that Winternitz ! had already shown that the unfertilized hen’s egg showed no catalytic activity even after prolonged incubation, whereas the incubated fertilized egg rapidly acquired the power of decomposing hydrogen peroxide. We repeated and confirmed these observa- tions of Winternitz. Doubtless if Winternitz had determined the intensity of oxidation in the fertilized hen’s egg, he would have found that this increased parallel with the increase he observed in catalase, and if Warburg had determined the catalase content of the fertilized sea-urchin egg, he would have found this enzyme increased parallel with the increase he observed in oxidation. J. Loeb” attributes the development of the fertilized sea- urchin egg to the increase in oxidation, and the increase in oxida- tion to a change in the cortex of the egg which makes the entrance of oxygen, and hence oxidation possible, while R. Lillie holds that the cortical layer of the unfertilized egg prevents the diffusion of CO, from the egg and this CO, prevents oxidation, and hence development. A more plausible explanation for the increased oxidation or metabolism in the fertilized egg, and hence for the development of the egg, would seem to be that the spermatazoén furnishes a substance which stimulates the egg to an increased for- mation of catalase. Further evidence in support of this view is afforded by the fact that the very same chemicals (amines, alkalies, acetates, butyric acid, etc.) which Loeb found would bring about increased oxidation and artificial parthenogenetic. development of the egg, we found when introduced into the ali- mentary tract of animals, stimulated the alimentary glands, particularly the liver, to an increased output of catalase with resulting increase in oxidation. Battelli and Stern“ found that the catalase content of most of the tissues and particularly of the liver of newly born pigs is lower than the corresponding tissues of the mother, but that the Loeb, Artificial Parthenogenesis and Fertilization. University of Chicago Press, 1913. 8 Lillie, Am. Jour. Physiol., 1910, xxvii, 289. 14 Battelli and Stern, Arch. di Fisiol., 1905, ii, 471. CATALASE CONTENT OF TISSUES. 131 catalase activity rapidly increased until at the end of the seventh or eighth day it was as high as that of the adult. We repeated and confirmed these observations using the dog and newly-born puppies. We also determined the catalase content of the tissues of puppies that were about ten weeks old and found that the tissues generally were richer in catalase than those of the mother. The catalytic activity of the liver, for example, of the ten week old puppies was about thirty per cent. greater than that of the liver of the mother. The catalase was determined by adding one gram of the hashed tissue to hydrogen peroxide and the amount of oxygen liberated in ten minutes was taken as a measure of the amount of catalase. Appleman found that there was an increase in catalase parallel with the increase in oxidation in the greening of potato tubers, but that the oxidase activity was not increased and, in fact, was slightly decreased. He also found that the exposure of potatoes to ethyl bromide gas increased the catalase of the potato parallel with the increase in oxidation, while it had no effect on the oxidases. The low metabolism or oxidation in the newly-born is attri- buted to the low catalase content of the tissues, due undoubtedly to the small ouput of this enzyme from the liver, while the high metabolism in youth is attributed to the richness of the tissues in catalase brought about by a large output of this enzyme from the liver. Likewise, the increase in oxidation or metabolism in the sprouting of grain or of potatoes is attributed to an increase in catalase. The increase in oxidation or metabolism and hence the development of the fertilized egg is attributed to the increase in catalase brought about by the stimulation of the egg by the spermatazo6n to an acceleration in the formation of this enzyme. 1 Appleman, The Maryland Agricultural Experiment Station, 1915, Bull. ror. 132 SCIENTIFIC PROCEEDINGS (106). 72 (1532) The effect of compression on tissue enzymes. By BERT HOLMES HITE and WITHROW MORSE. [From the West Virginia Experiment Station Laboratories of Chem- istry and the School of Medicine, Department of Biochemistry, Morgantown.| The senior author (B.H.H.) has studied various enzymes when subjected to great compression in a special hydraulic press, capable of delivering pressure of over (10)> pounds per square inch. As a rule, these enzymes are inhibited or destroyed. At the same time, there is generally lee-way between the destruction of enzymes and the total destruction of bacteria included in the preparations. It is well known from the work of investigators, such as Wolbach, Sakai and Jackson,! that it is difficult if not impossible to obtain aseptic autolytic digests by employing the most rigid asepsis in operations of removing organs from mammals, for included bacteria are practically always present. The junior writer (W.M.) has abundantly verified this conclusion, although there exist reports of investigations, as those of Magnus-Levy,? where the work was controlled by aérobic and anaérobic cultures and aseptic organ suspensions seem to have been obtained. We have no suggestion to make regarding this discrepancy. In order to determine whether the lee-way mentioned above is of sufficient extent to warrant an attempt to study tissue enzyme action apart from bacteria, the following experiments were con- ducted, rabbits being used as subjects, the livers being excised, sieved, weighed to 20 per cent. digest, Io g. and 25 g. portions being transferred to special blocked tin tubes, resembling vaseline tubes, which were tightly stoppered by means of screw-caps, each tube then being introduced into the cylinder of the press surrounded by a jacket of water. ‘The following table gives the number of the sample, the pressure involved, the time of exposure to this pressure and the Kjeldahl data utilized to follow the rate of digestion, if any*; the term “‘initial’”’ refers to the non-protein 2 Magnus-Levy, Hofmeister's Beitrage, 1902, 2, 261. ’ For details see Bradley and Morse, Journ. Biol. Chem., 1915, 21, 209. FORMATION OF ANTIBODIES. 133 nitrogen analyzed immediately after removal of the tubes from the press and ‘48 hrs.,’’ that nitrogen found in aliquots after remaining in a thermostat at 37° for that period. Sample Number. Pressure. Time. Initial. 48 Hours. Control (séptic).......... pa. xx 0.50 c.c. 0.1 N. | 0.80 c.c. 0.1 N. By Ber as SS eAloteee fe 75(10)8 5 mins. .90 Wt BUCMEIE hee ets sais i 16(10)4 2 hrs. .50 BP. Meeribe. 22% woods Bos (10)5 | 16 hrs. 1.40 DN BERNE 2. ois a win vette (1ro)5 I min. -50 The explanation of number III being at variance with number IV, where both are at the same degree of compression, doubtless lies in the fact that during the longer period of exposure (16 hrs.), some enzyme action may proceed before inhibition sets in, espe- cially as the temperature rises somewhat‘ during this prolonged time of exposure. It is evident that the method can be used only with the greatest care to adjust the pressure so that complete inhibition does not occur, while at the same time, all bacteria® are killed, which is of doubtful practicability. At these pressures, egg albumen is coagulated (B.H.H.), but in the case of the tissue from the rabbit’s liver, there is little evidence of change in the colloidal dispersion, although the Ringer’s Solution used to make up the liver mass to 20 per cent. suspension separates in the tubes during compression from the semi-solid mass. 73 (1533) The relation between the disappearance of foreign proteins from the circulation and the formation of antibodies. By WARFIELD T. LONGCOPE and GEORGE M. MACKENZIE. [From the Presbyterian Hospital, New York City.] Some observations which Dr. Rackemann and one of us made a few years ago indicated that when serum disease followed the injection of horse serum in human beings anti-bodies, such as 4 The cylinder is cooled during the period of compression. 5 This probably lies near (10). 134 SCIENTIFIC PROCEEDINGS (106). precipitin, anaphylactic antibodies, and skin hypersensitiveness to horse serum appeared as a rule toward the termination of the serum sickness. It was considered that the excessive production of antibodies followed the cellular reaction which was made evident by the serum sickness. In two of the cases that were studied, serum disease did not follow the injection of horse serum and anti-bodies did not appear in the circulation. ‘The reason for this variation in susceptibility, a condition that has long been puzzling, has not been satisfactorily explained. The object of the present investigation was to determine, if possible, whether the presence of antigen, namely, horse serum, bore any relation to serum disease or to the production of anti- bodies. It is known that by the method of specific precipitation a reaction for horse serum may be obtained in the blood, both of animals and of human beings, for many days after the injection of horse serum. Since reactions for horse serum cannot be ob- tained in the urine, it is improbable that horse serum is secreted as such by the kidneys. The method employed has been to estimate by means of specific precipitin reactions the presence of horse serum at given intervals after its injection into human beings for therapeutic purposes in pneumonia and at the same time to follow the appearance and curves of precipitin for horse serum. Preliminary observations on rabbits showed that when 5 c.c. of horse serum per kilo body weight was injected intravenously, reactions for horse serum could be obtained in the blood of the rabbit over a period of from 7 days to three weeks. Precipitin for horse serum appeared in from 6 to 10 days and persisted for from 4 to 7 weeks. It is well known that animals differ widely in their ability to form antibodies, and observations on six rabbits showed considerable variation in the rate of precipitin formation, and in the duration of the re- action for horse serum in the circulation. By this same method we have studied 14 individuals who have received from 100 to 500 c.c. of antipneumococcus horse serum intravenously. An analysis of these cases shows that they fall into two groups: In one group of eight cases the curve of precipitin formation and the persistence of horse serum was much the same as that observed in rabbits. The persistence of FORMATION OF ANTIBODIES. 135 the reaction for horse serum was somewhat longer, 18 to 39 days, and the appearance of the precipitins slightly slower than in rab- bits, since they were not obtained in human beings until the 6th to the 15th day after injection. The injection of horse serum in all these cases was followed by severe serum disease, lasting from 11 to 28 days, and as a rule the precipitins appeared first or their concentration increased markedly towards the end of the serum disease. On the other hand, the reaction for horse serum diminished rapidly towards the termination of the serum disease and disappeared shortly thereafter. The intensity of the serum disease and the persistence of the reactions for horse serum in the circulation bore no direct relationship to the amount of serum injected. In the second group were five cases to whom 300 to 500 C.c. of horse serum were given. In this group the results differ entirely from those obtained in rabbits and in the patients of the first group. One characteristic of these cases is the persistence in the blood of the reaction for horse serum over a very long period of time which ranged from 49 to 67 days. Practically all of these patients were lost to observation before negative reactions for horse serum were obtained. Secondly, the precipitin formation was either of extremely short duration or entirely absent. In the third place, these patients either had very mild serum disease lasting from one to five days or had none at all. There was one other case receiving 630 c.c. of serum who showed the persistence of the reaction for horse serum in the circulation for over 75 days; he also showed precipitins and he had severe serum disease lasting 12 days. This is an exception to the cases in both groups. An analysis of these observations shows that the reaction for horse serum disappeared fairly promptly from the blood of indi- viduals who had severe serum disease and who formed precipitins towards horse serum, and that the disappearance of the antigens, that is the horse serum, came shortly after the subsidence of the serum disease, and when the precipitins were present in the largest concentrations. On the other hand, in the second group the reaction for horse serum persisted in the blood over long periods of time. In these patients the precipitin reaction was absent or slight and serum disease was absent or of very mild type. 136 SCIENTIFIC PROCEEDINGS (106). It is known that individuals vary within very wide limits in their reactions to injections of foreign proteins. These observa- tions suggest that those individuals who are relatively insuscep- tible have some protective mechanism either in the serum or in the cells of the body which prevents or delays the union of the antigen, in this case horse serum, with the cells of the body. ABSTRACTS OF THE COMMUNICATIONS, PaciFic CoAst BRANCH. Twenty-fourth meeting. San Francisco, California, February 11, 1920. 74 (1534) The determination of calcium in blood and plasma. By Guy W. CriarK (by invitation). [From the Department of Biochemistry and Pharmacology, University of California.] The method consists in a direct precipitation of the calcium without removal of the blood porteins. The only reagents necessary are ammonium chloride (to prevent the precipitation of magnesium), and ammonium oxalate, both adjusted approxi- mately to P,, 7.4. Blood.—Citrated blood (5 c.c. in a 50 c.c. centrifuge tube), is hemolyzed by the addition of four volumes (20 c.c.) of warm water and after standing 15 to 20 minutes is centrifuged to remove the stroma. An aliquot (20 c.c.) of the clear, red liquid is transferred to a 50 c.c. centrifuge tube. One c.c. of 5 per cent. ammonium chloride is added and, after mixing, 3 c.c. of 3 per cent. ammonium oxalate are added. The oxalate should be added slowly and the contents of the tube must be well mixed. After standing at least 16 hours (the much shorter time recently reported by de Waard! results in an incomplete precipitation of the calcium), the mixture is centrifuged and the supernatant liquid siphoned off. The tube is washed by the addition of 25-30 c.c. of cold water 1de Waard, D. J. Biochem. Z., 1919, 97-08, p. 186. DETERMINATION OF CALCIUM IN BLOOD AND PLASMA. 137 and immediately centrifuged. After removal of the wash water the precipitate is dissolved in 5 c.c. of approximately normal sulfuric acid and, after heating in a water bath to 75° C., the contents are titrated with 0.01 normal potassium permanganate. (After precipitation of the calcium the method follows, with minor modifications, that described by Halverson and Bergeim.’ Plasma.—Citrated plasma (5 c.c. in a 50 c.c. centrifuge tube) is diluted with an equal volume (5 c.c.) of one per cent. ammonium chloride and 10 c.c. of one per cent. ammonium oxalate are slowly added. After standing 16 hours the precipitate is centrifuged, washed and titrated as in the procedure for blood. The method has been checked by: 1. The determination of known amounts of calcium in solutions having approximately the same mixture of salts as is found in the blood. 2. The ashing of whole blood and plasma in platinum dishes and precipitation of the calcium by a modification of McCrudden’s? method. 3. The addition of known amounts of calcium to plasma and recovery by the above method. The advantages of this method are: 1. The small amount of sample necessary, 5 c.c. blood and 3 to 5 c.c. of plasma. 2. The few reagents necessary and the relative ease of preparing them free from calcium. 3. The precipitation of the calcium in the presence of the proteins. Methods” *now in common use require either ashing or removal of the proteins by precipitation, operations which greatly add to the chances for mechanical losses. The accuracy of this method lies between five and seven per cent. ? Halverson, J. O., and Bergheim, O. J. Biol. Chem., 1917, 32, p. 159. 3 McCrudden, F. H. J. Biol. Chem., 1909-10, 7, p. 83; J. Biol. Chem., 1911- 52, 10, px 187. 4Lamers, A.J. M. Z. Z. Gebiirts. u. Gynék., 1912, 71, p. 393- Lyman, H. J. Biol. Chem., 1917, 29, p. 169. Marriott, W. McK., and Howland, J. J. Biol. Chem., 1917, 32, p. 233- Jansen, W. H. Z. physiol. Chem., 1917-18, 101, p. 176. Heubner, W., and Rona, P. Biochem. Z., 1919, 93, p. 187. 138 SCIENTIFIC PROCEEDINGS (106). 75 (1535) Outline of a classification of the lipoids. By W. R. BLoor. [From Department of Biochemistry and Pharmacology, University of California.] At the meeting of the American Society of Biological Chemists in December, 1919, the matter of a classification of the fats and related substances was brought before the members for discussion. It was decided that the available knowledge on the subject was insufficient to justify a classification at that time. While ad- mitting the truth of the conclusion it has seemed to me since, that something might be gained by an attempt at classification,— in clarifying our ideas and in bringing the newer developments in the field into connection with the old—even though the scheme might later have to be radically changed or even abandoned. It is becoming more and more apparent, for example, that the fats and the substances ordinarily grouped under the name of lipoids are so intimately related both chemically and in meta- bolism—all being directly connected with the metabolism of the fatty acids—that they should be considered together, and when so considered they form a group which is believed to be as distinct and well defined as that of the carbohydrates and proteins. The following is an outline of proposed classification: THE LIPoIDs. The higher fatty acids, their naturally occurring compounds and certain substances found naturally in chemical association with them. The group is characterized in general by insolubility in water and solubility in ‘fat solvents’—ether, chloroform, benzol, etc. Simple Lipoids. Esters of the fatty acids with various alcohols. Fats.—Esters of the fatty acids with glycerol. (fats which are liquid at ordinary temperatures are called oils.) Waxes.—Esters of the fatty acids with alcohols other than glycerol. Beeswax, lanolin, cholesterol oleate. OUTLINE OF A CLASSIFICATION OF THE LIPOIDs. 139 Compound Lipoids. Compounds of the fatty acids with alcohols but containing other groups in addition to the alcohol. Phospholipoids.—Substituted Fats containing phosphoric acid and nitrogen—lecithin, cephalin, etc. Glycolipoids—Compounds of the Fatty acids with a carbo- hydrate and nitrogen but containing no phosphoric acid.—Cere- bron, etc. ) (Aminolipoids, Sulpholipoids etc.—Various groups which may be added as soon as they are sufficiently well characterized.) Derived Lipoids. Substances derived from the above groups by splitting, which have the general properties of the lipoids. Fatty Acids of various series. Sterols—Alcohols, mostly large molecular solids, found naturally in combination with the fatty acids and which are soluble in ‘‘fat solvents’-—Cetyl Alcohol (CigsH330H), Myricyl Alcohol (C3)9Hs10H), Cholesterol (C27;H4;0H), etc. NOTES ON THE CLASSIFICATION. The group is specifically limited and defined in two ways; 1. Only substances are included which are chemically and metabolically related to the fatty acids. 2. Only naturally occurring substances are included. The definitive chemical entity of the group is the fatty acid and it is intended to include only those substances which are closely concerned with the metabolism of the fatty acids. The second limitation—“naturally occurring”’ is intended to exclude organic compounds which have no relation to the metabolism of the fatty acids but which would otherwise be included owing to composition or physical properties. The name ‘‘Lipoid”’ has been chosen for the group because 1. By derivation it is suitable. | 2. When limited as above it is sufficiently definite for the purpose and yet general enough to include all necessary sub- stances. 140 SCIENTIFIC PROCEEDINGS (106). 3. It is already in considerable use on the continent in approxi- mately this (Bang, Czapek and most French investigators in the field). The term ‘‘Lipin,”’ introduced by Gies, although by derivation equally suitable, is believed to be less desirable because of the present practice among biochemists of making little or no distinc- tion between—in and—ine (signifying the presence of nitrogen). For example MacLean uses the term lipin to mean lipoid sub- stances containing nitrogen. Very little originality can be claimed for the above classification and the writer freely acknowledges the help obtained from a study of earlier classifications, notably those of Bang, Leathes, Gies and MacLean. At the same time it is believed to be an improvement on preceding classifications in that it provides a definite chemical and metabolic basis which has hitherto been lacking. 76 (1536) Sodium citrate and scurvy. By HAROLD K. FABER. [From the Stanford Medical School, San Francisco, California.] An Italian child ten months old was brought to the Children’s Clinic, Stanford Medical School, in October, 1919, suffering from severe scurvy. The child had been fed from birth on raw certified milk to which had been added sodium citrate in the proportion of one grain to each ounce of modified milk. A series of seventeen guinea pigs was fed on oats and milk. In the case of nine of these guinea pigs sodium citrate was added to the milk in the following proportions: 2 animals, 0.25 per cent.; 2 animals, 0.50 per cent.; 3 animals, I per cent.; I animal I.3 per cent.; I animal 2.0 per cent. The period of feeding in this series was ten to forty-six days. Of the animals to which sodium citrate was given all except one, to which 0.50 per cent. was given for forty-six days, developed scurvy. Of the eight animals to which milk and oats only were given two developed scurvy. These control animals were observed for periods of forty to forty-six days. The average milk intake was 39.1 c.c. of milk per day for the contro] animals and 41.6 c.c. for the sodium citrate animals. SODIUM CITRATE AND SCURVY. 141 The incidence of scurvy in the latter series was 88.9 per cent. and in the control series, 25 per cent. The results of the control series are not in accordance with those of Chick, Hume and Skelton who state that guinea pigs receiving less than 50 c.c. milk per day invariably develop scurvy but indicate that with Inspected Milk used in this laboratory the lower limit is about 32 c.c. The average intake in the two series reported was approximately the same. A series of electrometric determinations performed through the eourtesy of Dr. E. C. Dickson showed the following changes in fresh Inspected Milk when sodium citrate in varying amounts was added: IA arr ae aS ba pclae ae ea Uw eres we ots pH 6.45 Opty tier Coma, STN CATALS PEGE 2 oes ss cb be eae oc 6.50 0.50 per cent. + “i Sabena 2 eee sed SE a cae eins acids 6.95 I.0 per cent. s ~ eS Se SEER OCS Ce a ee ee 7.05 1.25 per cent. “ - ye SAW A SR te eee edn dy Fs 2.0 per cent. ss = Maa eo Are ee Sas Ge BEN Oo w le Se 7.35 The results of these experiments indicate that sodium citrate even in concentrations of 0.25 per cent diminishes or destroys the anti-scorbutic substance normally present in small amounts in raw cow’s milk. ot ie ae ioe~ hes . gee ate 7 SCIENTIFIC PROCEEDINGS. One hundred seventh meeting. ABSTRACTS OF COMMUNICATIONS. University and Bellevue Hospital Medical College, April 21, 1920. Vice-President Wallace in the chair. 77 (1537) Effect of underfeeding on ovulation and the cestrous rhythm in guinea-pigs. By GEORGE N. PAPANICOLAOU and CHARLES R. STOCKARD. [From Cornell University Medical College, New York City.] Under well-regulated food conditions the cestrous cycle in the guinea-pig is almost uniformly 16 to 17 days in duration. Underfeeding with a diet of 20 grams of carrots per day pro- duces a prolongation of the dicestrum and, at the same time, a congestion in the ovary and uterus and a degeneration of develop- ing graafian follicles. The extent of prolongation of the dicestrum depends upon the stage at which an animal is underfed. Underfeeding during the first 5 to 7 days of the dicestrum has only a slight effect, postponing the next cestrus for one or two days, while underfeeding during the later part of the dicestrum gives much more marked results. When an animal is underfed for 5 days, from the 12th to the 17th day after an ovulation and cestrus, the next ovulation and cestrus is delayed for about 7 days, being expressed at the 23d to 25th day instead of at the 17th. Should an animal be underfed for 7 days, from the 1oth day to the 17th day after cestrus, the next ovulation and cestrus is post- poned for 10 to 11 days, arriving at the 27th to 28th day, instead of the 17th day. 143 144 SCIENTIFIC PROCEEDINGS (107). This variation in the effect of the underfeeding when applied at different periods of the dicestrum is associated with the fact that the conditions of the ovary differ at the different times. Shortly after an ovulation the ovary contains almost entirely small primary follicles. These follicles are not so unfavorably affected by food conditions as are the large graafian follicles, which begin their growth and development during later stages of the dicestrum. A large follicle at the height of its development seems to require much better nutrition than a small primary follicle, and the lack of proper food arrests its progress very readily. Thus a late underfeeding has a more injurious effect than an early one, and the postponement of the next cestrus is correlated with a post- ponement of the development of new ripe follicles in the ovary. The entire cestrus activity depends chiefly upon the conditions prevailing in the ovary. The fact that following a late and long underfeeding the next ovulation is delayed about 11 days after the underfeeding has been stopped is in accord with the results of operation experi- ments which Papanicolaou has performed on the corpora lutea in guinea-pigs. These experiments show that after removal of all young corpora lutea following an ovulation, the next ovulation arrives in about II days instead of 16 to 17 days as would be expected. This acceleration of 5 to 6 days is due to the absence of the corpora lutea, which if present evidently inhibit the maturation, or prolong the time necessary for the development, of ripe follicles in the Ovary. These experiments all demonstrate the sensitiveness of the follicles within the ovary to environmental conditions and when considered in more detail than is here possible, they throw light on many peculiar reproductive phenomena observed in nature. The extreme variations in the oestrous cycles recently recorded for the rat by Long and Evans (Proc. Am. Ass’n of Anatomists, Anatomical Record, April 1920) may be in part, at least, due to the variations in the diet taken by the individuals. When rats are fed a mixed diet no doubt certain individuals receive a ration quite different from that eaten by certain other members of the olony. ABDOMINAL MUSCULATURE IN CAT. 145 78 (1538) The effect of varying pressures upon the abdominal musculature in the cat. By HELEN C. COOMBs. [From the Department of Physiology, Columbia University.] Sherrington! has summarized rather completely our know- ledge of tonus for smooth and striated muscle. He believes that muscle fiber is not to be considered as an elastic string, for it has the property of exhibiting different lengths with one and the same degree of tension. This doctrine is of special interest in the case of the abdominal musculature, which is of necessity subject to many changes in pressure due to the many variations which occur in the abdominal contents. We should therefore expect to find the muscle fibers of the abdomen showing different lengths with the same degree of tension, or, to put it conversely, to exhibit a fairly constant pressure with varying increments of volume. The object of these experiments has been to determine whether this regulation of inter-abdominal pressure is essentially a function of the nervous mechanism of the abdominal walls or of the intrinsic musculature itself. Cats were used in these experiments under the several condi- tions of light and deep anesthesia and decerebration. A cannula introduced into the abdominal cavity was connected by a 3-way stopcock with a manometer and a burette filled with 0.9 per cent. sodium chloride solution kept at a temperature of 38 degrees centigrade. Costal respiration was recorded throughout the experiment. The warmed saline was admitted to the abdominal cavity at the rate of 10 c.c. a minute. With each increment of fluid the pressure was read from the manometer and plotted against the volume. A curve was thus obtained for the entire experiment, which, in about fifty cases was found to be typical. There was a slow increase of pressure in proportion to volume until a certain point was reached, from which pressure rose much more rapidly. At this point also, costal respiration increased very greatly in depth to effect a compensation for the lack of adequate abdominal 1 Sherrington, Brain, 1915, xxxviii, 191. 146 SCIENTIFIC PROCEEDINGS (107). respiration. In many cases the experiments were continued until there was a failure of respiration which was likely to occur when the pressure had risen to from 250 to 300 millimeters of saline. In other cases they were intermitted when respiration was observed to be labored. The curve always had much the same form indicating a slow rise in pressure in proportion to the volume up to a critical point, after which the pressure increased much more rapidly, as though such a point indicated the end of the ability of the musculature to lengthen with a minimum increase in pressure and thereafter exhibited only elasticity. The anesthesia, when neither very light nor very deep, had no effect upon the pressure, volume remaining constant. Very light anesthesia sent the pressure up, and very deep anesthesia sent it down a little. Decerebration had no effect upon it. The next step was to determine whether ablation of the motor nerves of the abdominal musculature would have any effect upon the pressure curve. After the control curve had been taken, therefore, the fluid was removed from the abdominal cavity and no further procedures were undertaken for an hour. The three branches of each phrenic nerve were then removed and the filling of the abdominal cavity was repeated. In a similar manner, after obtaining the normal curve in other cats, the ventral roots of the spinal nerves from the mid-thoracic through the lumbar region were destroyed (laminectomy having previously been done) and another curve obtained. Removal of the phrenics with paralysis of the diaphragm was in all cases found to cause a slight increase in pressure proportionate to volume, and the same was the case with removal of the spinal nerves. The curves, however, were so closely parallel to the normal curve as to be hardly significant. In order to obtain the effect of isolation of the muscle from all motor impulses, curare was injected into the femoral vein of a number of cats and artificial respiration was maintained. There was little variation from the control curve. After the death of the animal, curves were taken of the pressure at intervals of one and three or two and four hours in order to determine the réle played by the intrinsic elasticity of the mus- culature in the maintenance of this curve. With each succeeding ABDOMINAL MUSCULATURE IN CAT. 147 hour after death the pressure curve more closely approximated the straight line exhibited by any elastic body. The condition of pregnancy is of interest in connection with these experiments. Many cats in varying stages of pregnancy were examined. They all, in proportion to their weight, exhibited a greater degree of extension of the abdominal musculature, with a lesser amount of pressure in proportion to volume, than the non-pregnant cats. Even when the abdominal musculature was greatly distended, these cats appeared capable of the accommoda- tion of a relatively large volume of fluid. wwohuays UoHesucdsas diaypr suid 182440 & Grey? has pointed out that time is an essential factor in the expression of this lengthening of the muscle fiber with no greater degree of tension. It is probable that the conditions of preg- nancy, with the lengthy time factor involved, are far more ideal for demonstrating this particular form of muscular activity than a laboratory experiment can ever be; it is significant that the abla- 2 Grey, Ernest G., Amer. Jour. Physiol., 45, 272. 148 SCIENTIFIC PROCEEDINGS (107). tion of the motor nerves of the abdominal musculature in preg- nancy is not more effective in causing alterations of the pressure curve than in the non-pregnant cat. The function appears to be part and parcel of the musculature itself rather than of the extrinsic nerves, although some slight nervous regulation and codrdination does undoubtedly exist. 79 (1539) Coli fever and blood volume in dogs. By H. G. BARBouR and A. J. HOWARD. [From the Pharmacological Laboratory of the Yale University School of Medicine.] For the continuation of work on the mechanism of fever reduc- tion by drugs we have been seeking a satisfactory method of producing fever in dogs. In these animals a predictable curve of neurogenic fever is very difficult if not impossible to obtain. A few injections of peptone have given us a maximum rise of less than 1° C. with a rapid return to normal within two or three hours (maximum dose employed: 7 c.c. per kilo of 67 per cent. ‘‘bactopeptone.’’) Turning to injections of killed cultures of colon bacilli we made nineteen experiments with subcutaneous injections of a vaccine containing 325,000 million bacilli per c.c. and in fourteen of these obtained a temperature the following morning (that is, after 15 hours), varying from 0.4° to 1.7° C. above normal. In the other five, no elevation of temperature was seen. The next procedure was to inject in the morning, following the curve throughout the day. For this purpose a more concen- trated vaccine was selected, containing 1,625,000 million bacilli per c.c. In five uncomplicated experiments in which this vaccine was used, maximum temperature increases of 2.4° to 1.5° C. (with hourly readings) were obtained with doses of I c.c. per kilo. With % c.c. per kilo the maximum increase was 2.4°. A smaller dose (0.2 c.c. per kilo) gave, however, an increase of only Cott FEVER AND BLOOD VOLUME IN DoGs. 149 0.6° C. A third strength of vaccine used contained one million bacilli per c.c.; doses of 1% and I c.c. per kilo, gave maximum increases of 1.0° and 1.2° C. respectively. In all of the experi- ments with the last two vaccines the maximum temperature was attained by either the second or the third hour after the sub- cutaneous injection. At the end of the day (6-8 hours after in- jection) the temperature elevation was usually reduced by about one half. The last two strengths of vaccine mentioned were therefore considered suitable for further studies. On the second day the temperature was usually found somewhat elevated, but thereafter not at all. The dogs could not as a rule be used for more than one injection each as repetition of the same or larger doses after four days gave a less pronounced effect, indicating that some degree of immunity had been produced. In all experiments the injections gave sterile abscesses de- veloping to a considerable size and breaking down several days after the injection. About half of our dogs were fasted for two days before being injected, the others being kept on a constant adequate diet of meat, lard, and bread; water ad libitum was always allowed. No essential differences were noted between fed and fasted animals. og Bl. Solids e € 40 220 ¢— -—e~ my PER, 4 ig a en ee... A T cons (#5) 1 cc. per kilo 58 180, j i i 1 j Hows 10 12 2 4 Fic. A. Dog 47. Jan. 3. (Fasted 2 days.) eS blood solids. ———————— rect. temp. 150 SCIENTIFIC PROCEEDINGS (107). Changes in the blood volume of coli fever dogs were followed by means of hourly determinations of hemoglobin and blood solids. In uncomplicated experiments the blood solids showed constantly an increase running parallel with the increase of body temperature. This increase varied at the maximum point from 3.3 to 7.6 per cent. in five different animals. It is illustrated in the accom- panying figure (A) as well as in Figures 2 and 3 of the following paper on dextrose plethora. Similar hemoglobin changes were quite definite in some cases. The increase in the percentage of the blood solids is interpreted as a diminution in the volume owing to loss of water. We are not yet prepared to discuss the fate of this water, but have noted no significant increases in the amount of urine. The loss of water from the circulation is probably the chief factor in the decreased heat dissipation which accompanies the initial rise of temperature in infectious fevers. 80 (1540) Dextrose plethora and its antipyretic effect in coli fever.! By H. G. BARBOUR and A. J. HOWARD. [From the Pharmacological Laboratory of the Yale University School of Medicine.| The antipyretic action of dextrose was pointed out by one of us in the PROCEEDINGS of this Society about one year ago in connection with observations upon rabbits with peptone fever, and febrile human individuals to whom the sugar had been admin- istered by mouth.2 In view of the experiments reported some 1 The assistance of an appropriation from the Bache Fund of the National Academy of Sciences in support of the researches reported in this and in the preceding paper is gratefully acknowledged. The authors desire to thank Dr. George H. Smith, of the department of bacteriol- ogy, for valuable assistance in the preparation of the greater part of the coli vaccines used in this work. 2“*The Antipyretic Action of Dextrose,’’ Proc. Soc. Exp. Biot. AND MED., 1919, XVI, 136. In connection with the demonstration that antipyretic drugs increase the blood sugar this has been interpreted as a potent factor in their action. See Barbour and Herrmann, ‘‘On the Mechanism of Fever Reduction by Drugs,"’ Proc. Nat'l. Acad. of Sci., 1920, VI, 136. DEXTROSE PLETHORA. I5I years ago from Dr. Lusk’s laboratory by Fisher and Wishart! it was believed that our observations could be correlated with the dextrose plethora described by the last mentioned investigators. To this end dextrose was first given to dogs by mouth in doses varying from 1 to 10 gms. per kilo with quantities of water, usually sufficient to make about a 30 per cent. solution. Observations were made upon dogs which had just been fasted for two days subsequent to receiving an adequate daily ration of meat, bread, and lard. Two dogs showed a maximum in- crease of 15-20 per cent. in the hemoglobin and 6.6-8.6 per cent. in the total blood solids. In no case was there any indication that the blood volume was increased. The maximum rises of temperature in these two experiments were 0.6° and 0.2° C. respectively, the minimum temperatures noted being 0.2° respectively above and below normal, (hourly observations after injection of dextrose). In two similar experi- ments with normal dogs, no variation greater than 0.2° C. was observed in the course of the day. In two dogs which had been given a coli injection (325,000 million bacilli per c.c.) on the previous day, reductions in tem- perature of —o0.2° and —o.5° C. respectively were noted (at the end of three hours). From the above it was concluded that it is difficult to give dextrose by mouth in such a dose as constantly to affect the temperature either of normal or of fever dogs. The changes in the blood above described must be attributed to a tendency of the sugar to enter the circulation slowly or else to leave it rapidly, in either case abstracting water. ‘To confirm the observations on this blood concentration may be cited three experiments in which dextrose was given per os simultaneously with a coli injection. In one of these was noted an exaggeration of the usual (3-7 per cent.) increase in blood solids produced by such coli injections. In the hemoglobin content, 23, 29 and 15 per cent. increases were noted. Knowing that a plethora could be induced by dextrose if introduced into the circulation in the proper amounts and at the proper rate, intravenous injections? were next instituted. Two 1 Fisher, G. and Wishart, J. Biol. Chem., 1912, xiii, 49. 2 Starling, E. H., Jour. Physiol., 1899, 24, 317. 152 SCIENTIFIC PROCEEDINGS (107). normal dogs and two dogs with coli injections were carefully studied, being kept in a comfortable recumbent position through- out the day without anesthesia. The following table illustrates the results: TABLE I. SINGLE INTRAVENOUS INJECTIONS OF DEXTROSE IN QUIET UNANESTHETIZED DoGs. Dex- Normal, With Fever, With With Secon- : trose, | Water, Plethora, dary Rise, 3 Dog.| Condition Dose | C.c. & * | Gms. | per Bl, | Bl. Bl. Bl. per ilo, Temp, Solids, Temp.| Solids, Temp. Solids, Temp. Solids, Kilo, ee sos We ee ed ah || ee SOC See AP) ee 2 31| 49 ormal...| 2.2 ; — 1/24 | 47 | Coli fever .| 3.6 | 7.2 | 39.0 | 18.1 | 40.6 | 19.3 | 40.2 | 16.7 | 41.1 | 20.7 1/31) 41 | Coli fever . | 4.3 : 21. From the table it is obvious that dextrose given intravenously in fever dogs produces a plethora. The volume change is approxi- mately two to three times as extensive as that produced by corre- sponding injections in normal dogs. The normal dogs (Fig. 1) showed no diminution in temperature, but after about half an hour the curve began to rise, maximum increases of 0.4° and 0.3° respectively, having been noted. ‘The return to normal occurs after about three hours. In the fever dogs (Fig. 2), on the other hand, the temperature was depressed within fifteen minutes in the one case falling 0.4° and in the other case 0.6°. This was followed by a prompt return to and above the former level. In °c Bl. “db. Solids 39 200 100 t Dextrose (40 c.c.of 50%) 58 180 50 ] tt j , ! ! Hours 12 2 4 Fic. 1. Dog 41. January 24, 5.9 kilos. — — — — blood solids. ----- hemoglobin. rect. temp. DEXTROSE PLETHORA. 153 both of these experiments the curves for hemoglobin and total solids ran qualitatively parallel to the temperature curve, both in changing from normal to fever, from fever to antipyretic effect, and from antipyretic effect back to the new high level. 6'*/ Bis Hb. e Solids . 1 40 220 150 7 39 200 100 whe ee —a« 2 ' / e- N i e. . Coli (L), 5.8 cece t Dextrose (50 c.c. of 50%) | 1 | [ 1 | ! 1 Hours 9 11° ~ = Fic. 2. Dog 42. January 31, 5.8 kilos. — — — — blood solids. ------ hemoglobin. Feet, Lemp. °. ‘“ f AN og: PRR Solids 40 220 150 ee eens Pratt aad BS = se Ou —— @ e -_——_ 39 200 100 Retake eh eek soe ndiaw een cae ees = ‘ ee er ie ogee ‘er’ T coli (L) 4.5 cec. 7 0.9% Nach, 72 c.c. 38 160 50) " t | H | | Hours 45 12 2 4 Fic. 3. Dog 53. April 2, 8.9 kilos. — — — — blood solids. ------ hemoglobin. rect. temp. 154 SCIENTIFIC PROCEEDINGS (107). As a control to the above experiments, intravenous injections were made of 0.9 per cent. sodium chloride, of which was given 8 c.c. per kilo to each of two dogs exhibiting a typical coli fever. in the first fifteen minutes there was observed no change whatever in the temperature which subsequently ascended to points 0.7° C. and o.4° C. respectively higher than the former febrile level. The blood solids on the other hand showed a slight diminution after half an hour, and the hemoglobin fell within the first quarter of an hour. (See fig. 3). The control experiments therefore showed that isotonic NaCI in amounts comparable to the dex- trose injections is not able to reduce the temperature of coli fever dogs, nor is the increase in blood volume as marked as when the dextrose was given either to the normal or the fever dogs. It is therefore concluded that the antipyretic effect (not noted in health) of intravenous dextrose injections is due to osmotic action by which in fever dogs an unusually profound increase in the fluids of the blood results for a short time. This increase in fluid is of value to the animal in promoting heat elimi- nation. The tissues in coli fever appear to contain a higher percentage of “available water”’ than is normally present. Sen- sitivity to antipyretic drugs can thus be accounted for. 81 (1541) Preparation and refining of diphtheria toxin-antitoxin. By EDWIN J. BANZHAF. [From the Bureau of Laboratories, Department of Health.] The mixture has been readjusted so that after standing eight months and longer, late paralysis will occur when the mixture is injected into guinea pigs. This is done by adding to each L+ dose one unit of a properly aged antitoxin. When five mils of this mixture is injected into guinea pigs acute death occurs within four or five days. The mixture is then stored in a refriger- ator for a month to six weeks for stabilizing. On reinjecting five mils, after storage, the guinea pigs die of late paralysis after twenty to twenty-five days. It is then properly REFLEX GASTRIC HYPERMOTILITY. 155 balanced and safe for distribution. Prepared as stated it retains its balance eight months and possibly much longer and its practi- cally full immunizing value at least a year. What slight deterior- ation occurs takes place equally in the toxin and antitoxin and therefore there is no danger of the mixture becoming toxic. Constitutional disturbances are frequent. They are usually observed in adults, especially in those who give a pseudo Schick reaction. These disturbances are mostly due to the bacillary substances which are present in the diphtheria toxin. Methods for refining the mixture for the complete removal of the bacillary substances have not been found. Considerable of these reacting substances can be eliminated through the use of ammonium sulphate, sodium chloride and alcohol. 82 (1542) Further observations upon reflex gastric hypermotility. By W. HowarpD BARBER and GEORGE D. STEWART [From the Department of Surgery, New York University and Bellevue Hospital Medical College.| Increase in the force or rate or change in the direction of gastric contractions have followed irritation of the gallbladder, duodenum, or appendix, experimentally, and these motor changes have been associated with pathological gallbladders, duodenums, and appendices, clinically.1_ It may be assumed, subject to further experimental proof, that these organs constitute three of the possible foci of reflex gastric stimulation. Were the nerve paths known along which these impulses travel, it might be possible to explain these motor responses and group other possible causes of gastric motor unrest. Other observations of abnormal reflex gastric activity in which the pyloric and fundic parts functionate separately are the fol- lowing: 1. Prostalsis of the pars pylorica, alone, occurring in the course of irritation of the above organs and after thoracic vagus section. 2. Anastalsis of the pars pylorica, alone, associated with traumatization of the gallbladder. 1 PROCEEDINGS Soc. Exp. BIOL. AND MED., Vol. XVI., No. 7. 156 SCIENTIFIC PROCEEDINGS (107). 3. Pro- and anastalsis of the pars pylorica, alone, after extra- gastric traumata before the stomach appears to settle down to definite rhythmical contractions and is produced mechanically by dividing or blocking the stomach at the junction of the pyloric part and the fundus. It also follows thoracic division of the vagi. 4. Pylorospasm, diffuse, with fundic relaxation resembling a pylorofundic intussusception (see diagram). This fourth type has been observed repeatedly under experi- mental traumatization of the gallbladder, duodenum, or appendix and once in the human with evidence of appendical and gall- bladder disease. It can be produced by direct stimulation of the lesser curvature at the junction of the descending and horizontal arms. The subjective evidence, associated with this motor state, is anorexia, vomiting, and epigastric pain. The objective signs are mass and tenderness over the stomach, present at times and absent at other times. This form of motility, as appears to be the rule with the reflex types, disappears with parietal peritoneal irritation. Experimental data, to date, indicate the total hypermotility to be of probable vagus and the pyloric hyperactivity, alone, to be of probable vago-sympathetic origin. SCHEMATIC REPRESENTATION OF DIFFUSE PyLoric SPASM AND FUNDIC RELAXA- TION RESEMBLING A PYLORO-FUNDIC INTUSSUSCEPTION. BEHAVIOR OF CROWN GALL ON RUBBER TREE. 157 83 (1543) The behavior of crown gall on the rubber tree (Ficus elastica). By MICHAEL LEVINE (by invitation). [From the Montefiore Hospital, Cancer Research Laboratory, New York City.] Smith (1911-12) in his extensive studies on crown gall and its resemblance to animal cancer shows that the physiological effects of these tumors vary from species to species and also within the species and are generally less pronounced and speedy than one might expect. He holds that it is difficult to show conclusively that the substances produced in the tumor by the parasite are absorbed and act as slow poisons. ‘This is expecially difficult in view of the fact that the galls are often soaked by rains and become infected with other parasitic and saprophytic organisms. Levin and Levine (1918) in a preliminary report on the malig- nancy of the crown gall and its analogy to human cancer pointed out that a number of the phenomena in both diseases are analogous. They contend that the neoplasms in plants produced by Bacterium tumefaciens are sometimes benign, though some are true malignant growths. ‘The latter generally dwarf the plant so affected and cause the necrosis of the tissue above and below the gall. These studies and those of other workers were carried out on annuals, biennials or deciduous trees in which the period of growth of the host as well as the crown gall is normally interrupted. The difficulty in determining the effects of crown gall, is made greater by the intervention of natural death, caused by changes in temperature and its concommitant factors, and second, by the occurrence of infections caused by fungi and even insect grubs, the eggs of which are deposited in the soft tissue of the young crown gall. The purpose of this report is to bring forward further evidence on the malignancy of the crown gall experimentally induced on mature evergreen perennials such as the common rubber tree, Ficus elastica. In such plants where the growth is rather active all the year round, when kept under uniform, green house 158 SCIENTIFIC PROCEEDINGS (107). conditions the effect of the crown gall organism and the neoplastic growth on the host can be kept under observation for an extended period. Drenching rains and destructive insects are avoided and very often other parasiticand saprophyticfungi. In this way and in such plants as Ficus elastica it is possible to show definitely whether and in what degree the crown gall has an effect upon the adjacent normal tissue of the host. It was found that Bacterium tumefaciens inoculated into the apical internode of the branches, into the leaves or main stem of the rubber tree, Ficus elastica, stimulates the development of a neoplasm in the region of inoculation of a benign or malignant nature. The crown galls so formed, in this plant, are of two kinds, one in which growth is uniform and appears to be a swelling, the other is the characteristic convoluted type indicating a peri- pheral growth of isolated nodules. The early stages in the develop- ment of the crown gall in Ficus elastica does not interfere with the nutrition of the plant as a whole nor does it interfere with the growth of the inoculated branches. The crown gall in Ficus elastica after a number of months of active growth becomes hard and dry and finally dies. This is associated with the differentia- tion of the tissue which converts the gall into a mass of paren- chymatons cells and nodules of woody fibers. The central portion of the crown gall which generally lies near the wood cylinder disintegrates. The invasion of the stem by the new growth does not destroy the entire conducting system of the stem, yet that portion of the stem above the gall dies as well as considerable portion of the stem below. Cultures made from pieces of the crown gall and stem above and below the gall yield only a shizomycete which in appear- ance is not unlike Bacterium tumefaciens and which when inocu- lated into the stems of young geraniums and rubber plants produce crown galls in the region of inoculation. It is altogether possible that substances of the disintegrating crown gall or products of the crown gall forming organism are carried into the circulation of the stem and are responsible for the progress of the death of the stem from the gall upward and downward. The death of the plant due to crown gall is at least suggestive of the death caused by the invading and disintegrating malignant growths in animal cancer. BILIARY STASIS. 159 84 (1544) A. Vicious activity of the gall bladder during biliary stasis. B. The determining factor in the causation of white stasis bile. By PEYTON Rows and PHILIP D. McMASTER. [From the Rockefeller Institute for Medical Research.| Ligation experiments in dogs, cats, and monkeys show that under stasis conditions the gall-bladder and bile-ducts act very differently. The gall-bladder, continuing to exercise functions that are normal to it, effects a great concentration of the stasis bile and adds mucus thereto in quantity. As result, when an obstruction is produced below the entrance of the cystic duct all of the extralobular biliary channels come at length to be filled with a thick, greenish-black fluid. The ducts, on the other hand, have no concentrating faculty, and their lining secretes but little mucus. In an obstructed duct system blocked off from the gall bladder, or connecting with one so changed as to be incapable of functioning, there regularly accumulates a limpid, watery fluid devoid of pigment and bile salts even when the animal is heavily jaundiced. This is the ‘‘white bile’’ of the surgeons. The passages soon become so distended with it that true bile ceases to enter them. The facts as given relate to uninfected and uninflamed bile passages. So far as they go they point to cholecystectomy as a wise measure in gall stone cases, and notably when the gall- bladder is but little damaged. For an abnormal concentration and thickening of the bile such as the gall bladder can effect must act both to promote the formation of stones and to render ob- struction by them more complete. The frequent rapid increase in size of stones partially obstructing the common duct (Naunyn) is attributable to the concentration of stasis bile by the gall bladder. 160 SCIENTIFIC PROCEEDINGS (107). 85 (1545) The effect of therapeutic doses of digitalis on the contraction of heart muscle. By ALFRED E. COHN and ROBERT L. LEvy. [From the Hospital of the Rockefeller Institute for Medical Research, New York, N. Y.] That digitalis causes alterations in the heart resulting in changes in the form of the electrocardiogram is now well known. Robinson and Wilson have estimated in experiments on cats that the quantity of digitalis which can induce a change is about 30 per cent. of the calculated lethal dose. So far, however, no evi- dence has been presented to show that this amount of digitalis is beneficial—except in cases of fibrillation of the auricles in which block of auricular impulses, mainly through stimulation of the vagus nerves, takes place. A beneficial action must be based on the ability of the drug to increase the volume output of the heart, and it must be able to do this in therapeutic doses, that is to say, in doses which influence the T wave of the electrocardiogram or reduce the rate in auricular fibrillation. We have accordingly injected this amount into the veins of dogs, 11 of which received the tincture of digitalis and 19 of which received g-strophanthin; and into cats, 5 of which received g-strophanthin, and 9, the tincture of digitalis. Alterations in volume output were studied in curves obtained by the use of the Roy and Adami myocardiograph. ‘The curves represent longitudinal linear alterations in the form of ventricles and may, under the conditions of cardiac contraction, represent changes in volume of the cavities and consequently of volume output. The results are reported as changes in the degree of contraction. The animals were anesthetized with ether only. The chest was opened in the median line. Other details of the technical procedure, which are important, will be given in the full report of these experiments. The significant results concern the effect of these two drugs on the T wave and on the degree of contraction (Table I). In EFFECT OF DIGITALIS ON CONTRACTION OF HEART. I6I TABLE I. No. Drug. Rate. | bao a iat thay B-P, Result. Dogs, 19. .|_ g-Strophanthin...| 9 oO ) I I Decrease. 8 4 II I4 4 Increase or change. 2 15 8 4 fe) No change. Thal; ot Woigitalis. 653. 4 2 (s) e) fe) Decrease. 5 I 6 10 2 Increase or change. 2 8 5 I fe) No change. Cats, 5..| g-Strophanthin...| 3 I fe) I I Decrease. 2 8) 4 2 3 Increase or change. fe) 4 I 2 fe) No change. Sick Aes EMR EANG fins os 7 s) fe) 5 3 Decrease. (e) 2 7 2 6 Increase or change. 7 2 2 oO No change. 30 dogs, the T wave changed 17 times, and remained uninfluenced 13 times; the degree of contraction increased 24 times; decreased in I; and remained unchanged 5 times. The degree of contraction changed, then, in the greater number of animals; the T wave, in more than half. In 14 cats, the T wave changed I1 times and remained uninfluenced in 3; the degree of contraction increased 4 times, decreased 6 times, and remained unaltered 4 times. That is to say, the T wave usually changed; the degree of contrac- tion decreased in more than half. The effect on contraction differed, therefore, in cats and dogs. In 7 dogs and 13 cats the record of the blood pressure in the femoral artery was added to the other records. Except in one dog and 4 cats, the blood pressure usually rose. With the tincture of digitalis a significant fall of pressure often preceded a rise. Anesthesia and operative procedures, it was thought, might disturb the electrocardiogram. Experiments were therefore done on dogs in which electrocardiograms and blood pressure records were taken without anaesthetic and without operation. The electrocardiograms were taken in the usual way. The blood pressure curves were obtained in dogs previously prepared by a method described by van Leersum. By this method a long stretch of one carotid artery was enclosed in a stretch of skin included between two parallel incisions. The tube containing the artery lay free of the neck, and surrounded by a small rubber cuff. Water transmission to a mercury manometer permitted the taking of records. Minimum and maximum oscillations after the manner of Erlanger indicated systolic and diastolic pressures. It has 16335 SCIENTIFIC PROCEEDINGS (107). been found in the few experiments which have so far been done that T wave changes occurred uniformly and that the blood pres- sure usually rose, the increase varying from 20 to 66 mm. Hg. Data on the effect of the drugs on rate and on conduction are reserved for later publication and likewise detailed descriptions of differences between the two drugs. CONCLUSIONS. With doses of therapeutic range equal to 30 per cent. of the calculated lethal dose, digitalis and strophanthin (1) increased the contractile power of the cardiac muscle, and by so doing in- creased the volume output. This effect supplies a firm basis for the statement that these drugs may exercise a beneficial action. (2) At the same time, the T wave is usually altered, and (3) there is a transient elevation of blood pressure. 86 (1546) A method for the estimation of lactic acid in blood. By GEORGE A. HARROP, JR. (by invitation). [From the Chemical Division of the Medical Department of Johns Hopkins Hospital, Baltimore.| The procedure is based upon the observation of Denigés! that lactic acid, in the presence of concentrated sulphuric acid, is converted into acetaldehyde, and can then be detected by certain reagents, particularly phenols and morphine alkaloids. 5 c.c. of untreated whole blood or serum is delivered directly into 15 c.c. of acidified copper sulphate solution, the flask being in the meanwhile gently shaken. It is heated 4-5 minutes on the water bath, cooled, and an excess of powdered calcium hydrate is added. It is then allowed to stand for 30 minutes and filtered. A water-clear solution is obtained which is free from sugar and other aldehyde forming substances, and which does not char appreciably during the subsequent treatment with sulphuric acid. One part of filtrate is added cautiously to 4 parts of pure concen- trated sulphuric acid, the mixture being meantime shaken and 1G. Denigés, Ann. de Chem. et de Phys. (8), 18, 149. EFFECT OF OPIATES ON MEMORY AND BEHAVIOR. 163 cooled in a dish of ice water. It is then placed in the boiling water bath for 2 minutes and immediately cooled in ice water, after which 3 drops of 5 per cent. solution of guiacol are added. With pure lactic acid solutions a rose color is developed which remains stable and clear for some time. The maximum color is developed in the blood filtrates in about 20 minutes and it must then be read against standards prepared with known amounts of lactic acid (conveniently prepared from zinc or lithium lactate). An appre- ciable. color is produced by 0.01-0.02 mg. of lactic acid. The colors may be compared in small flat-bottomed Nessler tubes, or the concentrated acid solutions may be compared in the Duboscq colorimeter without injury to the cups. On prolonged standing a turbidity develops in the blood filtrates which renders it impossible to read them accurately. 87 (1547) ‘Effect of opiates on memory and behavior of albino rats. By D. I. Macat and C. F. Mora. [From the Pharmacological and Psychological Laboratories of the Johns Hopkins University.| Studies were made by the authors on the behavior of white rats in Watson’s circular maze. A total number of eighteen rats was used. The animals were trained to find their way through the intricate labyrinth of the maze in the shortest period of time without making any error. They were then injected with the various drugs studied, and their behavior, both immediately after injection and for some time afterwards, was observed. ‘The experiments on the rats in the maze gave data concerning the memory habit of the animals, their activity, and the codrdination of their movements after the administration of the narcotics. A large number of experiments was performed on different rats, and the effects of the following opiates were studied: Morphin, codein, thebain, narcotin, narcein and papaverin. In addition to the individual alkaloids, the following combinations were also administered: pantopon (total opium alkaloids) and narcophin (morphin plus narcotin). 164 SCIENTIFIC PROCEEDINGS (107). It was found that morphin, in both large and small doses, impaired the memory and the behavior of the rats. In the case of only one of the animals was there an excitation or stimulation noted after small doses of the drug. After large doses of morphin, the impairment was long-lasting, continuing for several days. In most cases, however, the animals eventually completely recov- ered from the effects of the narcotic. In regard to the comparative effects of morphin alone and morphin given in combination in the form of pantopon or narco- phin, the following interesting observations were made: Out of 26 experiments with pantopon and morphin, in 22 the effects of morphin were found to be more depressant than those of pantopon. Out of 9 experiments with morphin and narcophin on the same animals, 7 experiments showed that morphin was more depressant than narcophin. In all the experiments with the opium alkaloids, the effects were noted on both the time in which the rats went through the maze and the total distance traversed or the number of errors made. Codein, in 9 experiments out of II, produced impairment, as indicated by both the time and the distance. Narcotin, in 5 experiments out of 7, produced also a depression. Narcein, out of a total number of 10 experiments, produced a slight depres- sion in 4, and no effect in 6. Thebain indicated a slight retarda- tion in 10 experiments out of 11. In the case of papaverin, it was found that very little effect was produced by small doses of the drug, but that after large doses (10 or 15 mgs.), a depression was noted. The complete data of this research will appear in due time in the Journal of Pharmacology and Experimental Therapeutics. 88 (1548) On the generalization of treponema pallidum in the rabbit fol- lowing local inoculation. By LovUISE PEARCE and WADE H. Brown. [From the Laboratories of the Rockefeller Institute for Medical Research, New York.]} That a widespread dissemination of Treponema pallidum may be produced in the rabbit by local inoculation has been shown TREPONEMA PALLIDUM IN THE RABBIT. 165 by the recovery of the organisms in isolated instances from the blood, lymphnodes or other organs as well as by the occasional occurrence of generalized lesions in infected animals. However, there is no evidence to show either the time or frequency with which this dissemination occurs or whether the organisms thus distributed over the body are capable of sustaining the infection in these animals. With these questions in mind, a series of experiments was undertaken, the object of which was to determine the frequency of invasion of the regional lymphatics and the general circulation following inoculation in the scrotum or testicles and how soon a self-sustaining generalized infection might be established. Time and Frequency of Invasion of Regional Lymphatics.— An examination was made of the inguinal lymphnodes in a series of 29 rabbits which had been inoculated by the introduction of a bit of infected tissue beneath the skin of the scrotum. ‘The nodes were excised under ether anesthesia at intervals of from 61 days down to 48 hours after inoculation and the presence or absence of Treponema pallidum determined by dark field examination or by animal inoculation. The first group of nodes studied included those showing well marked enlargement and induration and these gave positive results in all cases. Nodes were then taken 5 days after inocula- tion and after the lapse of only 48 hours. Positive results were again obtained in all cases. Invasion of the Blood Stream.—A similar series of experiments was carried out to determine the time and frequency of blood stream invasion and something of the character of the blood stream infection with relation to processes of reaction in the pri- mary lesions. With a few exceptions, the animals used for these experiments were inoculated in the testicles. The mode of deter- mining the presence of Treponema pallidum in the blood of infected animals was by bleeding from the heart, defibrinating and injecting 0.5 c.c. of blood into each testicle of 2 normal rabbits. A total of 81 bleedings was made on a series of 37 rabbits at intervals of from 7 to 99 days after inoculation. The earlier bleedings were all spaced with reference to some phase of the testicular infection and from these it was found that organisms 166 SCIENTIFIC PROCEEDINGS (107). could be recovered from the circulating blood from the time an infection could first be detected clinically (12 to 14 days) until regression of the primary lesions took place. The number or the virulence of the organisms as indicated by the incubation period and the constancy of infection in subinoculated animals varied, however, according to the stage of development and the state of the reaction in the primary lesions. A small series of animals was then bled arbitrarily one week after inoculation and it was found that even as early as this, the number of organisms in the circulating blood was sufficiently great for each 0.5 c.c. of blood to constitute an infecting dose. In view of these facts, there seemed to be no immediate object in further reducing the time limits. The Establishment of a (True) Generalized Infection.—When it had been shown that Treponema pallidum appeared to be widely distributed through the body within a very short time after inoculation, it was considered necessary to determine whether a true generalized infection had been established in these animals or whether the organisms proved viable only because they were transferred to such a favorable medium as the testicles of normal rabbits. For this purpose, Io rabbits were inoculated in the right scrotum only (using implants), and 48 hours later, the entire right scrotum and testicle were amputated under ether anesthesia. In spite of the complete removal of a wide zone surrounding the area of inoculation, 9 of the 10 rabbits showed well-marked infections by the end of the seventh week and the tenth developed lesions at the end of 2% months after inoculation. 7 Conclusions.—These experiments show that following a local inoculation of well adapted strains of Treponema pallidum, there is an immediate invasion of the tissues of the animal and that within a very short time, organisms may be recovered from both the regional lymphatics and the circulating blood. They also show that the blood stream infection tends to pursue a course parallel with that of the primary lesions. Finally it was shown that within 48 hours or less, a true generalized infection had been established which was capable of maintaining the infection in the animal independent of that at the site of inoculation. GENERALIZED SYPHILIS IN RABBIT. 167 89 (1549) On the production of generalized syphilis in the rabbit by local inoculation. By WabDE H. BRowN and LOUISE PEARCE. [From the Laboratories of the Rockefeller Institute for Medical Research, New York.| Two of the most striking features of the infection usually produced in rabbits by testicular or scrotal inoculations of well- adapted strains of Treponema pallidum are the marked reaction at the site of inoculation and the total absence of generalized lesions. In fact, these features of the reaction to infection are so conspicuous as to suggest a casual connection between the two, especially when it has been shown that the failure to produce generalized lesions can in no wise be attributed to the absence of a generalized infection or to an insusceptibility on the part of the animal’s tissues to react to such organisms. Specifically, it appeared to us that in all probability, the failure to produce generalized lesions was due in a large measure to an inhibitory influence arising from the reaction at the primary focus of infection and that the reduction or suppression of this reaction might be sufficient in itself to permit the development of generalized lesions. In order to test this hypothesis, three types of experiments were carried out which were intended to compare the effects produced by unilateral and bilateral inoculations, the effects of castration and the effect of suppression of the primary lesions by the use of therapeutic agents. The castrations were done under ether anesthesia. Effects of Unilateral and Bilateral Inoculation and of Castration. —In the first series of experiments, there were 27 rabbits inocu- lated in one testicle and 20 inoculated in both testicles, giving a total of 47 rabbits. These were divided into two groups, one of which was castrated soon after the appearance of the primary lesion and the other held as controls. Both groups were kept under observation for a period of 4 months after inoculation. Of the 27 rabbits inoculated in one testicle, 14 were castrated 168 SCIENTIFIC PROCEEDINGS (107). and 13 were held as controls. Generalized lesions developed in 8 of the 13 controls and 13 of the 14 castrated animals. Of 20 rabbits inoculated in both testicles, 6 were held as controls and 14 were castrated. Generalized lesions occurred in I of the 6 controls and in 13 of the 14 castrated animals. Several other experiments of a similar character gave essen- tially the same results. In one of these, 46 rabbits were given a heavy inoculation with a testicular emulsion—half of them uni- laterally and the other half bilaterally. With these animals, the influence of castration at different periods of the infection was studied and the effects of suppression of the local infection by the use of a therapeutic agent. Only the results of the therapeutic experiments can be given here. Effects of Suppression of Primary Lesions by Therapeutic Agents.—In carrying out these experiments, a drug was chosen from among those studied by us in collaboration with Dr. W. A. Jacobs and Dr. Michael Heidelberger whose effect in inducing resolution of lesions was much greater than its spirocheticidal action. This substance was arsenophenylglycyl dichloro-m- aminophenol. Twelve rabbits, 6 of them inoculated unilaterally and 6 bilater- ally, were given a single intravenous injection of this drug 14 days after inoculation and the results were controlled by 6 untreated rabbits from each of the respective groups. In the unilateral series, the lesions present were almost com- pletely resolved and the local reaction suppressed for between 2 and 3 weeks. At the end of 3 months, all of these animals had developed generalized lesions as contrasted with 3 of the 6 con- trols. The effect of the drug upon the animals inoculated in both testicles was less marked and lasted for only 7 to Io days. © At the end of 3 months, generalized lesions had developed in 4 of the 6 treated animals and in I of the 5 surviving controls. Effects of Complete Prevention of a Primary Reaction and Early Removal of the Medium of Inoculation.—An experiment originally carried out for the purpose of determining the time at which a true generalized infection became established in the rabbit proved to be a remarkable demonstration of the effects which might be obtained from complete prevention of the develop- BLoop CHANGES IN ETHER ANESTHESIA. 169 ment of a primary lesion plus an effect which appeared to be attributable to the early removal of even the small bit of syphilitic tissue used in the process of inoculation by the implantation of a small piece of infected testicle beneath the skin of the right scrotum. 48 hours later, the entire scrotum and testicle were amputated under ether anesthesia. By the end of the 7th week, 8 of the Io rabbits had developed marked generalized syphilis while the other 2 showed a definite lymphadenitis. One of these developed slight generalized lesions at the end of 2 months and the other 214 months after inoculation. As a whole, however, the generalized infection was the most pronounced which we have seen in any single group of animals. Conclusions.—The conclusions to be drawn from these experi- ments are: That the marked character of the reaction which takes place in the rabbit following local inoculation of old strains of Treponema pallidum is in a large measure responsible for the absence of generalized lesions; that an inhibitory influence is exerted upon the development of other lesions which is proportion- ate to the reaction taking place at the site of inoculation and that the reduction, suppression or prevention of this reaction will remove this influence to a sufficient extent to permit the develop- ment of a generalized disease analogous to that which occurs in man. 90 (1550) Blood changes in ether anesthesia. By DONALD D. VAN SLYKE, J. HAROLD AUSTIN and GLENN E. CULLEN. 3 [From the Hospital of the Rockefeller Institute for Medical Research, New York.]| | During light ether anesthesia the bicarbonate content of the arterial blood falls, the carbon dioxide tension (determined directly by the tonometric method on the blood) rises, as does the hydrogen ion concentration. These phenomena indicate a state of uncompensated acidosis. The oxygen saturation increases, indicating that ventilation is accelerated in response to the stimulus 170 SCIENTIFIC PROCEEDINGS (107). of increased carbon dioxide tension. ‘The acceleration does not, however, as under normal conditions, reach the height necessary to keep CQO, tension and hydrogen ion concentration down to normal limits. It therefore appears that even in light etherization the respiratory center is markedly deadened. In deep etherization the carbon dioxide tension rises still higher (over 80 mm. has been observed) and the Py may fall to below 7.2. Respiration not only fails to be accelerated in response to the increased CO, tension but may even be so retarded that the oxygen saturation of the arterial blood falls below that normally found in venous. The blood tends to become concentrated. Conductivity and chloride determinations on the serum indicate only minute changes. The only striking electrolyte changes appear to be the increase in hydrogen ions and the replacement of part of the bicarbonate HCO; anions by the anions of acids as yet unidentified. SCIENTIFIC PROCEEDINGS. ABSTRACTS OF COMMUNICATIONS. One hundred eighth meeting. Osborn Memorial Laboratory of Zoélogy, Yale University, New Haven, Conn. May 22, 1920. Vice-President Wallace in the chair. QI (1551) Additional experiments showing the production of fat from protein. By GRAHAM LUSK. [From the Phystological Laboratory, Cornell University Medical College, New York City.] A dog, which had been fed for two days with 1,000 gm. of meat daily at 8 a.m. and which had had at 5 p.m. on the same two preceding days the usual standard diet containing about 70 gm. of carbohydrate, was given 1,000 gm. of meat at 8 a.m. The respiratory quotients for the fifth, sixth and seventh hours after giving this quantity of meat were 0.842, 0.845 and 0.845. Com- puted on the basis of the metabolism of protein which is the equivalent of 1.44 gm. of urinary nitrogen per hour, there ap- peared to be a retention of material derived from protein which, if it had been burned, would have shown respiratory quotients in the successive hours of 0.708, 0.688 and 0.685. This indicates that the carbon-containing material derived from protein which was retained in the body had a respiratory quotient approximately the same as that of fat. | For the three hours by direct calorimetry 85.32 calories were found. By indirect calorimetry, if the retained carbon be cal- culated as having been deposited in the form of fat, 83.58 calories may be calculated as the heat which should have been expected to 17I 172 SCIENTIFIC PROCEEDINGS (108). arise under those circumstances. Had the carbon been retained as glycogen the calculated heat production would have been greater (about 4 calories per hour). This work confirms in three successive hourly periods the work previously published by Atkinson and Lusk! concerning the for- mation of fat from protein under experimental conditions similar to the ones here described. The experimental data follow: Doc XVIII. Experiment 68—Basal metabolism—A pril 14, 1920. Indirect = 14.25 calories Direct = 14.3% 5°“ R:Q. = 0.82 Experiment 70—After 1,000 grams meat. Sth hour: N—COz........ 13.46 N—Onrc........ 12.097 > N—enls. .¢ va Ae 38.17 Resp.—CO2z..... 10.10 Resp.—Or2z..... 8.72 Deposit cals...... II.32 3.36 3.45 Indirect cals...... 26.85 R. Q. of deposit = 0.708 Direct. cals... ... «<2 9.52 6th hour: N—COns........ 13.46: N—Os lois ss 02.07 |) N—colei 2. eee 38.17 Resp.—CO2..... 10.45 Resp.—Os2z..... 8.99 Deposit cals...... 10.43 3.01 3.48 Indisect'eais....25 o's 27.74 R. Q. of deposit = 0.688 Direct cals. 343. 20-07 “th hour: N—CO:........ 13.46 N—Or........ w2.27 ..di—cales. 3h cee 38.17 Resp.—COz2z..... 10.56 Resp.—O2..... 9.09 Deposit cals...... 10.10 2.90 3.08 Indirect cals...... 28.07 R. Q. of deposit = 0.685 Direct cals...... ..28:73 Total indirect = 82.66 calories ** direct = 85.32 “ ** indirect as calculated: on C retained as fat = 83.58 ; on C retained as glycogen = 96.8 > Urine nitrogen = 1.44 gm. per hour 1 Atkinson, H. V., and Lusk, Graham, ‘‘On the Problem of the Production of Fat from Protein in the Dog,”’ Proc. National Academy of Sciences, 1919, V, 246. PROTEIN MEAL AT END oF 8-Day Fast. Li fe 92 (1552) An observation of the effect of a protein meal given to a man at the end of an 8-day fast. By WILLIAM S. McCann (by invitation). [From the Russell Sage Institute of Pathology and the Second Medical Division (Cornell) of Bellevue Hospital.] In an observation of the effect of a protein meal on a man at the end of an eight day fast a rather unexpected result was obtained which throws light on the fate of the carbohydrate portion of the protein cleavage products. The subject was a normal man, 23 years old. Height 180 cm., weight 70.4 kgm. At the time of observation he had a marked odor of acetone on his breath. Respiration was normal. The CO, combining power of the blood plasma was 50 volumes per cent. He was placed in the respiration calorimeter for a basal obser- vation of 2 hours. He was then given a meal which consisted of 350 gm. of lean beef and Io gm. of butter. One hour after taking the meal a second observation was started and continued for 3 hours. During the first two hours of this latter observation respir- atory quotients were obtained of .687 and .681. During the third hour the quotient was .740. The first two quotients are similar to those found in severe diabetes, or in phlorhizin glycos- uria. During the basal periods the quotients were normal for the fasting state, .733 and. 723 respectively. Following the meal there was very little increase in heat production. The nitrogen excretion was not increased. As evidence that the subject was not diabetic his tolerance for 100 gm. of glucose was normal. The fasting level for blood sugar was .086 per cent. After glucose it rose to .137 per cent. There was no glycosuria. One week after the first observation the experiment was repeated. During the interval the subject took a normal diet. The basal heat production was lower, as was the basal nitrogen excretion. After the meal a marked rise in heat production oc- 174 SCIENTIFIC PROCEEDINGS (108). curred (17-28 per cent.), but the hourly heat production is practi- cally the same after the meal as it was in the first observation. The nitrogen excretion was much increased after the meat. The quotients are normal throughout and higher than after fasting. The explanation of the diabetic respiratory quotient has been made by Lusk! who estimates the amount of oxygen and carbon dioxide exchanged in respiration for each gram of urinary nitrogen in severe diabetes (D : N = 3.65). In the case of a dog with phlorhizin glycosuria, and the cases of two diabetic men, diabetic quotients were obtained, but non-protein quotients were found to closely approximate that for fat. In the present case the non- protein quotients are below that for fat, due to a failure of all of the nitrogen metabolized to appear in the urine. It seems probable that the diabetic quotients in this case were obtained by the same mechanism as in diabetes, except that the glucose instead of being excreted in the urine, was stored as glycogen. TABLE I. EFFECT OF 350 GM. MEAT ON A FASTING MAN, AGE 23, WT. 70.4 KG., HT. 180 Cm. Before—Basal. Eating. After. Hours if Dae a at zr. 2. Re 4. Bs 6. COW Gall 3 acien we ee 22.92 22.46 23.02 23:93 24.34 ON BW is 08 eae bse 22.73 22.60 PROTEIN 24.38 25.36 23.91 [A SE ET ae ak Oe OP 733 723 70 gm. .687 .681 -740 Ni ELT iie ea oe -639 -639 FAT .637 .637 .637 Cals. per hor... . 1 73.85 73.30 28 gm. 78.77 81.98 77.89 Observation 391 made at the end of an 8 day fast. TABLE II. BEHAVIOR OF THE SAME INDIVIDUAL ONE WEEK LATER, HAVING BEEN ON A NORMAL DIET IN THE INTERVAL. Basal. Eating. After. Hours. ae oy Ae Pe TEE Ne I 2 3 4 | 5 | 6 th a ee | 20.67 29.88 23.89 26.60 25.97 Ein so nue ¥0 19.09 19.53 22.82 24.96 24.91 (1 De are Se -788 -778 -762 -775 -758 Pe Fak boise Sia 1 .599 -599 1.020 1.020 1.020 Cals. per hour...... 62.81 64.11 74.08 81.38 80.98 The same meal was given after a basal observation. 1Lusk, Graham, ‘“‘On the Diabetic Respiratory Quotient,’’ Arch. Int. Med., 1915, 15, part II, p. 939. TEST FOR ANTI-BERIBERI VITAMINE. 175 TABLE III. | COs gm, .Oo gm, Calories. In diabetes I gm. urinary N = 4.00 4.56 13.04 In periods 4-5, 0.637 gm. N = 2.55 2.90 8.31 Period 4. Ohtle Sots eee in alee tes 23.02 24.38 m. Demuce for Weis cies dw Aide ca oe 2.55 2.90 -687 INGE PEOUREIG! bo, soe Wi cat el ae oe 20.47 21.48 .693 Period 5. EE Pern he eeu te bhoed Ue te 23.93 25.36 681 DIGaBeE Tam IY poi so ver tee tt cs PA 2.90 Pia OC RNIN sorts as cec ore kee e ar oes 21.18 22.46 .686 Compare with Peay R.Q. Non-protein R.Q. Etienne COR. hs < ong Wd ee Oe ved ewrelas 3.54 .687 -704 TUR PaLAE TRAE rc wi srofe ais, Ohta a kee ker ake a 25 .697 -700 ACRE SNRIR AS eet rd a8 G'S id bate > Sow WSs ares ahs 2.07 .687 .699 93 (1553). Experiments on a quantitative and qualitative test for anti- beriberi vitamine. By CASIMIR FUNK and Harry E. DUBIN. [From the Research Laboratory of H. A. Metz, New York City.| In 1912 one of us (C.F.) experimented with the anti-beriberi vitamine to see if it could not act as coferment in alcoholic fer- mentation. The experiments failed at that time, but now the subject through the work of Williams, Bachmann and Eddy has attained a new interest. Their results seem to prove that vitamine activity can be measured by growth of yeast cells and would permit of dispensing with animal experimentation in the initial stages of vitamine fractionation. Because of the uncertainty of the Bachmann fermentation test, and the complicated procedure of Eddy’s test, it was thought desirable to develop a simple macro method. Our procedure is as follows: A yeast suspension is prepared according to the method of Eddy by shaking a loopful of 48 hours yeast culture in a definite amount of Naegeli’s solution for 4 hours on a shaking machine. Duplicate tubes are then prepared containing: (1) yeast suspen- 176 SCIENTIFIC PROCEEDINGS (108). sion + Naegeli, (2) vitamine sol. + Naegeli, and (3) vitamine sol. + yeast suspension + Naegeli. The tubes are incubated for 20 hours at 30° and the fermentation then interrupted by heating to 70° for a few minutes. The contents of the test tubes are then transferred to special centrifuge tubes, the bottom part of which ends in a capillary 2% cm. in length and divided in mm. The tubes are centrifuged at 2600 r.p.m. for 15 min. and the reading made without much delay as there is a tendency for the yeast cells to swell up slightly after a time. Sterility during the entire process, excepting centrifugation, is of paramount importance. The results obtained with this method were as follows: The pro- cedure was so well standardized that the controls with yeast suspension alone were practically constant. Duplicate experi- ments checked very well so that the method was applicable for a quantitative determination. In developing the test, autolyzed yeast was used as a source of vitamine. It was noticed, however; that the amount of cells obtained, as shown by the reading on the centrifuge tube, does not increase double if the vitamine amount is doubled. By trying varying amounts of vitamine a curve was established, using autolyzed yeast as standard, which permits of comparison between an unknown vitamine content and our stan- dard. The sensitivity of the method is .ooo1 c.c. of autolyzed yeast, and the amount to be tested is best made so that it corre- sponds to our curve between 0.01 and 0.1 c.c. of autolyzed yeast. In this interval the curve climbs up abruptly and small differences can be read with facility. It was found also that contrary to the method of Eddy, where the number of cells incubated varies the controls to an undesirable extent, this is not the case in our method. The variation of cells in our yeast suspension has very little effect on the final result. First of all, we were interested to know in how specific this test is for B-vitamine. Among the substances found negative in their action were allantoin, hydantoin, nicotinic acid, several purine and pyrimidine bases and several aminoacids. Pilocarpine found by Dutcher to be curative in pigeon beriberi was found inactive, while thyroid gland was active, as it contained the celullar con- stituents. An extract of pituitary gland was also found active for the same reason. In a second series the substances isolated INTRAVENTRICULAR HEART BLOCK. 177 from the vitamine fraction from yeast in 1912 and 1913 were tested. Those of 1913 were analyzed at that time and their formulas established. These substances showed an attenuated but definite activity. Polished rice was entirely negative as opposed to the statement of H. H. Green. Saliva was found inactive and urine as already reported by Muckenfuss was active. The latter test may prove of clinical value later on, but possibly blood could be used to greater advantage for diagnostic purposes. A few discrepancies were found however; for instance, yeast treated with Lloyd’s reagent still retained a large portion of its activity and also it was found that corn and wheat, separated from the germ by the method of Voegtlin and Myers, still exhibited a large activity, although several times less than the germ con- taining portion. Further experimentation is under way to clear up these matters. By comparing the method of extraction of Osborne and Wake- man from yeast with an extract prepared from autolyzed yeast and then heated over 50° to coagulate proteins (we found some heat coagulable protein in autolyzed yeast) we found that Osborne’s preparation contains only one-fifth of the nitrogenous substances but exhibits only one-fifth of the activity of the autolyzed yeast extract. It remains to be seen therefore whether it is advisable to use very purified extracts for vitamine fractionations, since it appears that the concentration of vitamines in such crude extracts as above varies with the amount of impurities present. 94 (1554) The site of the cardiac lesion in two instances of intraventricular heart block. By B. S. OPPENHEIMER and H. E. B. PARDEE. [From the Physiological and the Pathological Laboratories, Columbia University.| The hearts of two cases were examined in order to determine the site of the lesion associated with electrocardiograms suggesting partial block of either the right or the left branch of the auriculo- 178 SCIENTIFIC PROCEEDINGS (108). ventricular system. As the main deflections in the two sets of electrocardiograms were in opposite directions, it was to be ex- pected that the lesions, if any, in the two instances, would be found on opposite sides of the heart. There has been consider- able theoretical discussion as to which type of electrocardiogram is associated with right-sided and which with left-sided block. The first case showed electrocardiographically a main deflection inverted in lead I, upright in leads II and III, a marked widening of the foot-points of the Q.R.S. complex, and only moderate voltage; in addition there was auricular fibrillation. Microscopic examination of serial sections of the A.-V. system, showed that the right bundle branch became attenuated almost immediately after its origin from the main stem, and was surrounded by con- nective tissue. This diminution became more pronounced until at a distance of 7.5 mm. from the bifurcation, scarcely one or two doubtful muscle fibers could be seen. Below this the right branch increased in size again until at 4 cm. below the bifurcation it was of normal dimensions. There was marked fibrous myo- carditis of the septum, involving chiefly the left side, especially the sub-endocardial region. The left branch presented no lesion. The second case showed, on two examinations at an interval of six weeks, electrocardiograms in which the main deflection was upright in lead I, inverted in leads II and III, was notched, and its foot-points abnormally separated. Wave P was present throughout. Serial sections showed the A.-V. node, stem and right branch intact. The left bundle branch was imbedded in dense fibrous tissue throughout its course, and at a distance of 3.5 cm. below its origin, its posterior (dorsal) half was replaced by con- nective tissue continuous with an adherent, organized mural thrombus. There was also a thrombus within the apex of the right ventricle, partially adherent to its right lateral wall, but notinvolving theseptum. In addition, there was a general fibrous myocarditis which predominated in the left side of the septum, and an intense thickening of the endocardium on the left side only. The direct application of the published electrocardiograms associated with experimental bundle branch lesion in dogs to the interpretation of bundle branch block in man is rendered some- what doubtful by certain anatomical pecularities of the dog’s STUDIES IN PYRIMIDINE METABOLISM. 179 heart and its relation to the thorax. If the results in these two human cases are corroborated repeatedly by similar findings in other instances, it is possible that the usually accepted electro- cardiographic interpretation of right and left bundle branch block may have to be revised. 95 (1555) Studies in pyrimidine metabolism. By D. WRIGHT WILSON (by invitation). [From the Laboratory of Physiological Chemistry, Johns Hopkins University, Baltimore.]| By partial hydrolysis of yeast nucleic acid, preparations con- taining pyrimidines as the only nitrogenous constituents were prepared and administered to rabbits. Uracil nucleoside when administered per os, subcutaneously or intraperitoneally caused an increased excretion of urea often much more than enough to account for the nitrogen administered. The undetermined nitrogen (the difference between the total nitrogen and the urea nitrogen) was always increased. A part of the increase in the undetermined nitrogen was due to the excretion of free uracil which was isolated in pure crystalline form. As much as 20 per cent. of the uracil fed as the nucleoside was recovered free in the urine. When a mixture of cytosine and uracil nucleosides was admin- istered to rabbits, there was an increased excretion of urea and usually no increase in the undetermined nitrogen. Uracil was isolated from the urine of one animal and was barely detected by a color reaction in another. No increase of creatine, creatinine, or purines was detected after feeding either preparation. Not even a color reaction for pyrimidines was obtained by using the same procedures on the urine obtained after feeding yeast nucleic acid. Mendel and Myers were unable to find a trace of pyrimidine in the urine after feeding yeast nucleic acid but found that uracil, when fed, was excreted unchanged. Taken together, the data show that increasing quantities of uracil appear in the urine as simpler complexes containing the uracil group are fed. The con- 180 SCIENTIFIC PROCEEDINGS (108). clusion may therefore be drawn (at least in respect to uracil), that, in the metabolism of yeast nucleic acid before the pyrimidine is liberated and even before the nucleoside is formed, the pyrimidine is altered in such a way that it may be further broken down and its nitrogen converted into urea. 96 (1556) The variable acidity of hemoglobin and the distribution of chlorides in the blood. By FRANKLIN C. McLean, H. A. Murray, Jr. and L. J. HENDERSON. | [From Boylston Chemical Laboratory, Harvard University.| We have undertaken an investigation of the shift of chlorides between the serum and corpuscles of the blood described by Koeppe and by Hamburger, and have studied this phenomenon particularly in its relation to the heterogeneous acid-base equili- brium between hemoglobin, oxygen, carbon dioxide, bicarbonate, and the concentration of hydrogen ions. Such a shift in chlorides may be easily produced, in vitro, by disturbing, in any way, the acid-base equilibrium, and is observed, under physiological conditions, between arterial and venous blood. For the purposes of the investigation we have used fresh defibrinated ox blood, expelling the oxygen from combination with hemoglobin by first passing through carbon dioxide at 38° and then by boiling in vacuo at the same temperature. This can be accomplished with only very slight hemolysis. The blood has then been brought into equilibrium, at constant temperature and at atmospheric pressure, with various tensions of carbon dioxide, first in an atmosphere free from oxygen and then in an atmosphere with oxygen present at the tension at which it is present in atmospheric air. The whole blood has then been analyzed for oxygen and carbon dioxide, free and combined, and the serum, obtained by immediate centrifugalization under oil, analyzed for carbon dioxide and chlorides. The atmosphere | : VARIABLE ACIDITY OF HEMOGLOBIN. 181 with which the blood has been brought into equilibrium has in each instance been analyzed for carbon dioxide and oxygen after equilibrium has been reached. From the data obtained the hydrogen ion concentration has been calculated from the ratio of free dissolved carbonic acid to bicarbonate. When the concentration of bicarbonate has been plotted against the hydrogen ion concentrations, curves similar to those given by L. J. Henderson in a recent paper! have been ob- tained, showing an isohydric shift of base between hemoglobin and the other constituents of the blood, according to whether the hemoglobin was oxygenated or reduced. When the bicarbonate was plotted against the logarithm of the figure obtained for the hydrogen ion concentration a linear relationship was found. On plotting the serum chlorides against the hydrogen ion concentration an isohydric shift of chlorides has been noted in every experiment—+.e., at the same hydrogen ion concentration the concentation of chlorides in the serum is higher in the case of oxygenated blood than in the case of reduced blood, which is the reverse of the condition in the case of bicarbonate. The total isohydric shift of chloride amounts to about two thirds that of base under the same conditions. On plotting the serum bicarbonate against the serum chlorides it was found that they have a linear relationship, in the case of both oxygenated and of reduced blood, and that an increase in the concentration of bicarbonate in the serum is accompanied by a decrease of chlorides, corresponding to somewhat more than one half of the increase of bicarbonate. These curves appear to be straight lines, and are apparently parallel, indicating a constant difference between oxygenated and reduced blood at all hydrogen ion concentrations. This constant difference, accompanying the change in the state of hemoglobin, and not dependent on the change in concentration of bicarbonate, we are inclined to attribute to the change known to occur in the relative volumes of the red cells and serum. Of the total shift of chlorides at an isohydric point, this constant difference makes up only a small part. Since the shift of chlorides, from or to the cells, accounts for 1L, J. Henderson, ‘‘The Equilibrium between Oxygen and Carbonic Acid in Blood,’”’ J. Biol. Chem., 1920, XLI, 4or. 182 SCIENTIFIC PROCEEDINGS (108). two thirds of the shift of base, and since other anions, chiefly HCO,’ but also SO,’, shift in the same direction as Cl’, it seems improbable that there is any considerable migration of kations in and out of the cells, with the exception of hydrogen ions. This is in accord with the older ideas on this subject. We are at present investigating also the extent of migration of HCO,’ ions. As a general conclusion it may be stated that when the hetero- geneous acid-base equilibrium is disturbed, from any cause, the new equilibrium is established by the migration of acids in and out of the cells, and that about two thirds of this acid is hydro- chloric acid. Further confirmation of the mechanism of the chloride shift is obtained from the fact that the shift is produced by varying the tension of oxygen as well as by varying the tension of carbon dioxide. The fact that the chloride shift occurs isohydrically provides additional evidence as to the change in the acidity of hemoglobin at varying tensions of oxygen. In all of our experiments we have obtained data pointing to an extremely constant relationship between the various factors studied—7.e., hemoglobin, oxygen, carbon dioxide, bicarbonate, chlorides, and hydrogen ion concentration—for different samples of oxblood. Weare proceeding with the investigation in the hope of reducing these relationships to precise mathematical form. We have obtained evidence to the effect that the buffer action of the serum under physiological conditions as compared with isolated serum is increased at least ten times by the change in acidity of hemoglobin. It should be noted that we have studied the hydrogen ion concentration on the alkaline side of the isoelectric points of hemo- globin and serum proteins. At these hydrogen concentrations, as has been shown by Loeb,! the amphoteric colloids dissociate entirely, or almost entirely, as acids, so that we have disregarded, for the present, the possibility of combination of chloride and other anions with the colloid substances present. 1 Jacques Loeb, ‘‘Emphoteric colloids. I. Chemical influence of the hydrogen ion concentration,’’ J. General Physiol., 1918, I, 39. MAMMALIAN EMBRYOS. 183 97 (1557) The determination of small quantities of sugar in urine, including observations on the polysaccharide content of human urine. By STANLEY R. BENEDICT. [From the Department of Chemistry, Cornell University Medical College, New York City.| Conditions have been found for carrying out the reaction be- tween sugar and picric acid which render it possible to determine sugar in the presence of three or four times its weight of creatin or creatinine without affecting the results. When this reaction is applied directly to urine, figures for sugar are obtained which are only a few hundredths of a per cent. higher than those obtained after precipitation with mercuric nitrate as described by Benedict and Osterberg. After treatment of the urine with a suitable purified bone-black the figures obtained duplicate very closely those found with the mercuric nitrate method. The procedure recommended will be described in detail in the near future. Using the new technique, observations have been made on the increase in reducing substance of normal urine which results from mild hydrolysis with hydrochloric acid. The increase in reducing substance thus obtained amounts to about 0.5 gm. per day calculated as glucose, for the 24 hour elimination of a normal human adult. Very much higher figures (several grams per day) have been observed in cases of diabetes mellitus which were under treatment, and where the glucose eliminated amounted to from 3 to 20 grams per day. 98 (1558) Disturbances in the development of mammalian embryos caused by radium emanation. By J. F. GUDERNATSCH and H. J. BaGe (by invitation). [From the Department of Anatomy and the Memorial Hospital, Cor- nell University Medical College, New York City.] As has been shown by various observers, the exposure of living tissues to the influence of radium rays leads to a severe injury and 184 SCIENTIFIC PROCEEDINGS (108). ultimate destruction of these tissues. In our work an attempt was made to study this destructive influence on mammalian embryos in utero, in the hope that a partial or complete destruc- tion of one or more tissues might lead to definite abnormalities or malformations in these fetuses. Bagg had lately used a method of applying radium, which was described in the Journal of Cancer Research, Vol. V, 1920. Radium emanation, carried in a very small amount of saline solu- tion, was injected in measured quantities into adult rats, either subcutaneously or intravenously. This solution contained all the properties of the radium metal itself, and, no doubt, the re- sulting physiological changes were due mainly to the activity of a-rays. Such an injection produced peculiar destructive changes in the inner organs of the animals. The same method was used in our experiments. After long experimentation we found that a dose of 5 mc. (= milli-curies, a standard unit in radium experimentation) was about the optimal dose. Such an amount was injected into female rats, pregnant and non-pregnant, with the purpose of either injuring the ovarian or uterine tissues, or, in case of pregnancy, the embryonic tissues. While the results were not those which we expected, viz., the production of various types of monstrosities, yet a definite in- fluence of radium on the fetal and placental tissues was noticeable. Radium-treated rats were killed at different periods of pregnancy, so as to procure a series of fetuses of various ages. The most destructive results of radium emanation, injected subcutaneously, were seen in a number of pregnant females, in which the embryos were killed in the uterus and, instead of being aborted, remained attached to the uterine wall and were gradually absorbed (group I). Whether the embryos were killed primarily, or their death was due to the destructive influence of the radium on the maternal, placental tissues, cannot, of course, be deter- mined. Probably the first assumption is correct, since other findings (group II) showed, that the toxic agent does pass the pla- centa and affects the embryos directly. A number of such partially absorbed embryos were found, the age of which, naturally, could not be determined. Judging from the sizes of their respective placente, however, development must —————— ee le MAMMALIAN EMBRYOS. 185 have proceeded to some extent before the radium was applied. The remnants of the embryos were small, nodular bodies attached to the placente (figures were shown) and had lost all resemblance to properly developed fetuses. In one case a small, ovoid shaped sac was found, attached by a thin stalk to the uterine wall (figure shown). ‘This apparently represented the remnants of a former embryo and placenta, although neither one could be recognized any longer. In the sac extravasated blood and cell detritus were found. A great many large cells of an epithelioid nature probably belonged to the former embryonic syncytium. The wall of this cyst was formed by fibrous connective tissue. In a number of other cases (group II), the fetuses were not killed by the radium emanation, but peculiar macroscopic lesions appeared in their skin vessels. When the fetuses were removed from the uter!, peculiar hemorr- hagic areas were noticeable, in some cases just along the dorsal midline, in other cases, spreading over the entire body with the exception of the ventral surface. These extravasations took place in the vessels of the subcutaneous connective tissue and along the meningeal sinuses. In all cases, one or more hemorrhages appeared in the midline, mainly in the head and thoracic region. It seems that the vessels in this dorsal median zone are especially liable toinjury. In one instance, there was a large area of hemor- rhage extending over the thoracic and lumbar region. Its outline Was just symmetrical to the dorsal midline (figure shown). In other cases, a great number of such hemorrhagic areas, some extremely small, were found over the lateral aspects of the head and body. Probably these affected fetuses would have died, if left longer in the uterus, and would have undergone absorption. In many animals which we killed in the early parts of the experi- ments we failed to find any fetuses, although we definitely believed that these animals had been pregnant before. We probably waited too long after treatment, so that the embryos were completely absorbed, when the animals were opened. Not all of the fetuses of one litter are affected in the same degree. In one case, for instance, we found among 7 fetuses 3 showing hemorrhagic lesions, 2 beginning to macerate and 2 in the process 186 SCIENTIFIC PROCEEDINGS (108). of absorption. This difference in resistence may be due either to the higher or lower vitality of the embryos themselves or to the amount of radium which passes the placenta. In another case the fetuses, although injured, were carried to full term and among 6 young of one litter we found two normal and four showing hemorrhagic spots on head, face and along the dorsal midline. In one very remarkable instance the female had been treated 22 days previous to conception and yet the fetuses, approximately 16 days old, showed areas of extravasation (one of considerable size shown in figure). These lesions were much more widely distributed than in previous cases, extending over both lateral and dorsal surfaces (figure shown). These results cannot be ex- plained at present. It would seem as if the treatment of the mother previous to conception had lessened the faculty of the later embryos to form proper endothelial walls. The wide distri- bution of the lesions would seem to substantiate such a view. This is in accordance with findings in adult animals treated with radium in which the extravasations in the organs are due not only to increased blood pressure, as would seem at first, but to the actual breaking down of the endothelial tubes. In other words, the effect of radium on endothelium might be selective. When the radium was injected intravenously (group III) instead of subcutaneously, the same lesions resulted along the vascular channels. Females of about 19 days pregnancy were injected intravenously and the young, born dead 24 hours later, showed the hemorrhagic lesions along the dorsal midline (figures shown). In one case we found a striking difference in the size of the placente of different fetuses. One fetus, for instance, had a markedly enlarged placenta completely filled with blood, so that it had the appearance of a large hemorrhagic sac. This fetus did not show any hemorrhagic lesions, while their pla- cente were of normal size and moderately filled with blood. It would seem as if in the first case the placenta functioned as an effective ‘‘shock-absorber,’’ while in the other cases the radium emanation passed through the placentz to the fetuses. Lately Bagg exposed pregnant females, near full term, directly to the action of y-rays (group IV). This radiation of the fetuses in utero, through the abdominal walls produced hemorrhagic PHENYLCINCHONINIC ACID. 187 lesions of the same nature as described above. However, the lesions did not appear until about 10 days after exposure. The young were born 2 days after treatment and appeared normal. After about a week they began to fail considerably, hemorrhagic areas appeared along the mid-dorsal line, especially in the head region and death followed. The hemorrhages in these animals were mainly along the meningeal sinuses (figures shown), in some cases frontal and occipital hemorrhages were just beginning, in others they extended considerably over the cerebral hemispheres. Additional lesions on the dorsal side of the thorax were found. The interval of 10 days after treatment strictly corresponds to the time at which a primary skin erythema develops in radium treated patients. Again it seems as if the endothelial walls had been injured at the time of exposure and gradually gave way to the blood pressure. In the course of the experiments, we also found numerous hemorrhagic areas in the uteri and especially in the ovaries (figures shown). Congestion of the uterine vessels always was pronounced. While in experiments on adult animals reported by Bagg before, the injection of radium emanation led to considerable injuries in the internal organs, in our experiments the weaker doses did not produce any macroscopically visible effects on the maternal tissues. However, the embryonic differentiating tissues were easily affected. This fact might be of some biological signi- ficance, when one remembers that radium rays have a decided effect on fast growing tumor and cancer tissues. 99 (1559) The influence of phenylcinchoninic acid and its methyl derivative on the uric acid and urea content of the blood. By V. C. Myers, J. A. KILLIAN and G. E. SIMPSON. [From the Laboratory of Pathological Chemistry, New York Post- Graduate Medical School and Hospital.] It has been recognized now for some time that the adminis- tration of salicylates and phenylcinchoninic acid (cinchophen) 188 SCIENTIFIC PROCEEDINGS (108). resulted in an increased excretion of uric acid in the urine. Quite recently it has been shown by Folin and Lyman,! and Fine and Chace,’ in particular, that this action was accompanied by a marked drop in the uric acid content of the blood, and later the same was shown to be true of salicylates by Fine and Chace,? and Denis.4 It has been assumed that these drugs induce an increased output of uric acid by endowing the renal cells with an increased power for eliminating uricacid. Fineand Chace® have pointed out that, in the last stages of interstitial nephritis cinchophen has little influence on the excretion of the uric acid, indicating that the renal cells can no longer be stimulated to increased activity. INFLUENCE OF PHENYLCINCHONINIC ACID AND ITS METHYL DERIVATIVE ON THE UrRIC AND UREA CONTENT OF THE BLOOD. Uric Acid Urea N Creatinine Case. Date 1920. Mg. to roo C.c. Mg. to roo Cyc. Mg. to roo Cyc. Fy cesses Le 3/26 6.0 29 I.5 3/28 5.2 29 2.4 3/31 trace 16 2.0 4/5 3.0 24 2.4 4/12 3.5 18 3-0 4/13 3-9 15 2.4 ay |b a eet 3/16 2.8 26 2.6 3/18 2.8 22 2.6 3/22 trace 13 a:7 3/25 2.6 20 2.4 3/29 2.6 19 a9 4/9 2.8 16 2.4 4/12 0.7 13 2.6 4/15 2.0 17 2.4 Case 1, male, aged 58; clinical diagnosis, chronic interstitial nephritis, arterios- clerosis; 50 grains tolysin daily 3/28 to 4/3. Case 2, female, aged 53; clinical diagnosis, neurasthenia, visceroptosis; vegetable diet; 50 grains tolysin daily 3/19—-22; 50 grains cinchophen daily 4/9—-12. We have been endeavoring to obtain further light on the mechanism of the action of these drugs by experiments upon both man and animals. Setting aside cases of advanced interstitial nephritis where the action of cinchophen is comparatively slight, we have been struck by the fact that this drug and its methyl 1 Folin and Lyman, Jour. Pharmacol. and Exper. Therap., 1912-13, IV, 539. 2 Fine and Chace, Jour. Pharmacol. and Exper. Therap., 1914, VI, 219. * Fine and Chace, Jour. Biol. Chem., 1915, X XI, 371. 4 Denis, Jour. Pharmacol. and Exper. Therap., 1915, VII, 255, 601. 5 Fine and Chace, Arch. Int. Med., 1915, XVI, 401. PURIFICATION AND CONCENTRATION OF ANTIGENS. 189 derivative show quite different ability to lower the blood uric acid in different individuals, the action being very pronounced in some cases and comparatively slight in others. In their experiments Folin and Lyman noted the interesting fact (in two cases) that cinchophen not only brings about a dimunition of the uric acid in the blood but also seems to lead to a dimunition of the nonprotein nitrogen and urea whenever these are present in the blood in unusual amounts. Unfortunately these two experiments were not followed by a control after period. Data are given in the table above on two cases in which we believe there is unmistakable evidence of a drop in the blood urea as well as in the uric acid. It will be noted that the action on the uric acid was very pronounced in both cases, the first case having a high and the second a normal initial uric acid. 100 (1560) The purification and concentration of antigens by new methods of adsorption. By AuGustus B. WADSWORTH and FRANK MALTANER. [From the Division of Laboratories and Research, New York State Department of Health, Albany.] The results of recent studies of complement fixation in tuber- culosis suggest that the test might be of considerable diagnostic value if satisfactory antigens could be prepared. Three strains of tubercle bacilli were selected: one a virulent, another a non-virulent, human strain and a third bovine strain. Immune serum sufficient for titration with all the antigens was obtained from inoculated horses. The antigens were prepared from cultures by various methods of fractioning and extraction similar to those used by other ob- servers. The culture filtrates were so anticomplementary that they could rarely be used. The glycerine and distilled water extracts gave the most active antigens. These active antigens and also the culture filtrates for comparison were selected for further study by methods of adsorption. 190 SCIENTIFIC PROCEEDINGS (108). Animal charcoal and globulin (horse serum) were used to adsorb the substances possessing antigenic action from the filtrates. When animal charcoal was added, the antigenic properties were removed, and could not be recovered by extraction of the charcoal. The adsorption with globulin was a more complicated procedure. The technique was briefly as follows: The original antigen was dialyzed and horse serum, one part to twenty of antigen, was added and allowed to stand in the incubator half an hour. The globulin was then precipitated by passing purified CO, gas free from HCl through the mixture for one half an hour at 37° C. The globulin precipitate was collected and by shaking it with alcohol the antigenic substances were extracted. The alcoholic extract was concentrated in a vacuum. The preliminary dialysis of the culture filtrate eliminated practically all of its anticomplementary action. The adsorption with globulin removed the antigenic substances from the culture filtrates and the aqueous extractions so that they were easily obtained in greatly purified and concentrated form. In preliminary studies the antigens which are used in the diagnosis of syphilis by complement-fixation were also purified and concentrated by similar methods. This method thus allows more precise study of many phases of infection and immunity than has hitherto been possible. IOI (1561) Observations on the immunization of rabbits with single strain and combined multiple strain vaccines. By W. C. NoBLE, JR. and RuTtTH A. THOMAS. [From the Department of Bacteriology, New York University (Uni- versity and Bellevue Hospital Medical College).] Previous to 1916, the Army and the National Guard were immunized with typhoid vaccine. During the late summer and early autumn of that year, numerous cases of paratyphoid fever developed among the troops along the Mexican border, and the Army medical authorities therefore felt it desirable to substitute a triple vaccine, of typhoid bacilli combined with the paraty- IMMUNIZATION OF RABBITS. I9I phoids A and B, for the single strain typhoid vaccine then being issued. Similar experience with paratyphoid in the British Army in Flanders in the same year had also resulted there in the adoption of a combined vaccine. This immediately brought up the question as to the basis for the use of combined vaccines for prophylactic inoculation and led to the initiation, in the winter of 1917, of the experiments here presented. Other workers in this field had preceded us. Castel- lani! in 1903, showed that on injecting an animal with two different organisms at the same time, agglutinins were produced for both, and that the amount of agglutinin for each of the two organisms was about the same as in those animals inoculated with but one type of organism only. Subsequently he stated that as many as six different types of bacteria might be combined in a single vaccine with this same result, but that if more than six types were combined, a diminished amount of agglutinin for each type resulted. The problem has been attacked by other investigators, who have followed the methods of Castellani, by comparing the agglutinin formation when single and combined vaccines were employed. As we have long known that agglutinin formation is not a real index of the degree of immunity, it has seemed desirable that fur- ther study of the problem should be made, in observing the relative amounts of bacteriolysins produced. Four series of from four to five rabbits each, were immunized; one series was immunized with combined triple vaccine of typhoid and paratyphoid bacilli A and B; three other series with single strain vaccines,—one series each with typhoid, paratyphoid A, and paratyphoid B. At the beginning and end of the experimental period, the blood serum of each rabbit was tested for specific and group bacteriolysins and also for agglutinins. In the series immunized with the single strain typhoid vaccine, all the animals ‘show a sharp rise in immunity, but an interesting paradox is to be observed in the excessive production of the (group) para A lysin beyond the specific typhoid lysin (the highest lytic dilution for para A being 1-700,000, for typhoid 1-75,000; and for para B 1-400.) In the series immunized with para A vaccine, the highest lytic dilution for the homologous strain is I-20,000; for typhoid it 1 Castellani, A., Zeitschrift f. Hyg., 1902, 40, p. I. 192 SCIENTIFIC PROCEEDINGS (108). is I-3,000; and for para B 1-300. The third series immunized with para B shows its highest lytic dilution with a heterologous strain, typhoid (1-950); while for para A and para B the corre- sponding dilutions are 1-380 and I-—350 respectively. The fourth series of rabbits was immunized with combined triple vaccine, and the degree of immunity reached for each organ- ism (as measured by the highest lytic dilution) would appear to be lower than the degree of immunity obtained for the same or- ganism by inoculation with the single strain vaccine. This result was not wholly anticipated, from the previous work of Castellani! and also of Davison.? The latter in a series of very completely worked out experiments with agglutinins, reports that the combined vaccine when injected, gives for each organism as good and usually a greater immunity response than if it had been injected alone. Davison, and likewise Bull’ have also observed some tendency in respect to the heterologous strains to lead to an added formation of the specific agglutinin, a phenomenon some- what comparable to that of ‘‘summation”’ in muscle contraction. Our results with bacteriolysins, in a small series of animals, if confirmed, would appear to differ from Davison’s observations with agglutinins. Further experimentation along these lines is desirable, and it is our hope to amplify our own work with improved methods. 102 (1562) Studies on intestinal implantation of Bacillus acidophilus. By Harry A. CHEPLIN and LEO F. RETTGER. — [From Yale Uniersity, New Haven, Conn.]| B. acidophilus (Moro) is a common inhabitant of the intestinal tract of the albino rat and of man. Ordinarily it is present in very small numbers, however, and often may escape detection. Diet exerts a profound influence on the character of the intestinal flora. Lactose and dextrin, when fed in sufficient amounts, bring about a marked transformation in bacterial types. 2 Davison, W. C., Arch. of Inter. Medicine, 1918, XXI, 437. 3 Bull, C. G., Jour. Exp. Med., 1916, XXIII, 410. INTESTINAL IMPLANTATION OF BACILLUS ACIDOPHILUS. 193 The present investigation has shown, also, that the ingestion of pure cultures or suspensions of B. acidophilus results in a similar change. } The administration of 2 grams of either lactose or dextrin to white rats, in connection with a basal diet of bread and meat, caused within three to six days a complete transformation of the fecal flora from the ordinary mixed type to one strongly domi- nated by B. acidophilus, while the same amounts of maltose, sucrose and glucose failed to exert any transforming influence. The ingestion of 1 gram of lactose or dextrin brought about a partial change, whereas the addition of I c.c. (nephelometer 5) of living cultures of B. acidophilus to the I gram of carbohydrate effected a complete simplification of the intestinal flora. Iden- tical results were obtained by the administration of 2 c.c. of the B. acidophilus culture or suspension alone. Post-mortem exam- inations of the different sections of the alimentary canal of rats harboring a simplified flora of the aciduric type revealed a general distribution of B. acidophilus throughout the length of the in- testine. The implantation of B. bulgaricus was not effected by the ingestion of even 5 c.c. of B. bulgaricus suspension of the same concentration as those of B. acidophilus, either with or without accompanying lactose. However, when B. bulgaricus and lactose were given simultaneously a transformation of the intestinal flora took place corresponding to that obtained with the feeding of 2 grams of lactose alone. In feeding experiments with human subjects it has likewise been shown that either lactose or dextrin, when added in sufficient amounts to the ordinary diet, causes a pronounced transformation of the intestinal flora within four to six days, with a marked predominance of B. acidophilus. With a single exception thus far, the ingestion of 300 grams of either of these carbohydrates led to such a proliferation of B. acidophilus that it entirely domi- nated the flora and effected an almost complete suppression of all other viable bacterial types. The simple character of the flora tended to remain permanent so long as the ingestion of these carbohydrates was continued. The administration of 400 grams Was necessary in the one exception noted. 194 SCIENTIFIC PROCEEDINGS (108). No radical suppression of the commonly prevailing types was observed when but 150 grams of lactose or dextrin were ingested, barring two notable exceptions. The addition of 150 c.c. of living cultures of B. acidophilus to the 150 grams of lactose or dextrin caused a very pronounced transformation of the fecal flora from the usual mixed type to one strongly dominated by B. acidophilus. The daily ingestion of 300 c.c. of B. acidophilus suspension alone effected simplification of the flora, reducing it to the aciduric type. The striking results obtained from the use of milk soured with B. acidophilus bring to view a new avenue of approach to the field of B. acidophilus implantation within the alimentary canal. The administration per os of one liter of B. acidophilus milk, in conjunction with the ordinary daily diet, exercised in every instance a telling effect upon the complex flora, which was clearly manifested within one to six (usually less than three) days in the establishment of a non-gas-producing intestinal flora dominated by B. acidophilus. On the other hand, the ingestion of asimilar amount of B. bulgaricus milk offered no encouragement whatever to the aciduric type of organisms. In a few instances the consumption of only 500 c.c. of B. acidophilus milk was sufficient to establish a B. acidophilus flora. The addition of 100 grams of lactose or dextrin to 500 c.c. of B. acidophilus milk resulted in a rapid simplification of types in those subjects who did not respond readily to the ingestion of the 500 c.c. of milk culture alone. There appears to be a definite correlation between the rate of absorption in the alimentary canal of a utilizable carbohydrate and its tendency to effect a transformation of the intestinal flora. This relation was indicated in the observation that the feces of lactose- and dextrin-fed rats contained reducing substances at the times when B. acidophilus was present in preponderating numbers, while the feces of the animals receiving maltose, sucrose or glucose gave negative results with Benedict’s solution and presented no change in the types of bacteria. With the human subjects results were obtained after lactose ingestion similar to those furnished by the albino rats. No definite relation could be established, on the other hand, BACTERIAL INVASION OF RESPIRATORY TRACT. 195 between the hydrogen ion concentration of the feces and the bacterial flora. That is to say, the hydrogen ion concentration limits remained essentially the same during the preliminary and the transforming periods of the different experiments. 103 (1563) An unrecognized pathway for bacterial invasion of the respiratory tract. By M. C. WINTERNITZ, G. H. SMITH and E. S. ROBINSON (by invitation). [From the Brady Laboratory of Pathology and Bacteriology, Yale University School of Medicine, New Haven, Conn.| Normally, the ciliated, mucus-secreting epithelium is a mech- anism competent to protect the lungs against infection by way of the upper respiratory tract. When this epithelium is injured by toxic gas or when the mechanism is otherwise incapacitated, as for example, in aspiration pneumonia, the lumen of the trachea undoubtedly is the pathway traveled by the agent responsible for the pulmonary inflammation. Pneumonia may occur, however, and this is especially true of the lobar type, without demonstrable gross lesion of the upper respiratory tract, and in these circum- stances some route other than the above must have been provided. Experimentally, the introduction of pneumococci by intra- tracheal instillation or by needle puncture of the tracheal wall through the neck, may result in pneumonia. With either of these methods of inoculation, local damage to the mucosa of the trachea occurs. When the needle method is employed, an opportunity is at hand, not only for infection of the submucosa, but of the ‘peritracheal tissue as well. When the organisms are introduced by insufflation into the rabbit, damage to the mucosa of the larynx or upper trachea can hardly be avoided. In either case, an atrium of invasion for the submucosa of the trachea is provided, and histologically infection of the submucosa is evident at the point of inoculation. From here it may be traced throughout the submucosa of the trachea and larger bronchi to the hilum of the 196 SCIENTIFIC PROCEEDINGS (108). lung by way of the peribronchial and periarteriolar structures into the pulmonary tissue. Infection of the lung occurs under these conditions even though cultures from the lower trachea are sterile. An abundant lymphatic system can be demonstrated by the injection of India ink into the submucosa of the trachea. This plexus extends from the epiglottis downward as far as the bifur- cation of the trachea and connects directly with similar plexi in the submucosa of each bronchus. With further subdivisions of the bronchi, the condition noted above is duplicated. At the points of bifurcation throughout the cartilage-bearing bronchi, anastomotic branches connect the plexi with the periarteriolar lymphatics, and other branches pass directly to the regional lymph glands. Thus a short circuit around the valves of the deeper pulmonary lymphatics is provided. The distribution and extension of these tracheal lymphatics can be demonstrated equally well by Gram-stained sections pre- pared from an animal inoculated through the submucosa of the trachea by injection or insufflation of virulent pneumococci. The presence of the organisms shows the distribution of the infec- tion through the lymphatics of the submucosa of the trachea, past the hilum of the lung, into the pulmonary parenchyma. Thus a direct pathway of infection is provided. On the other hand, the manner in which the lymphatic plexi are sharply demar- cated at the bifurcations of the trachea and bronchi suggests that this lymphatic system may also serve as a protective mech- anism, since, undoubtedly, many of the invading bacteria are, at these points, diverted to the protective regional lymph glands. 104 (1564) The life of the white mouse. By W. B. KirKHAM (by invitation). [From the Osborn Zoélogical Laboratory of Yale Universily, New Haven, Conn.|} White mice may give birth to young every month in the year, but most of the litters come during the warmer months. Males THE LIFE OF THE WHITE MOUSE. 197 and females are both sexually mature when about six weeks old, and in females of that age, ovulation occurs regardless of whether or not there has been a previous pairing. The gestation period of non-suckling mice is from 18 to 21 days, so that the first litter may be born when the parents are about two months old, and in all cases of normal, paired animals a litter has appeared before they were three months of age. This first litter may be large (12) or small (2). If the first litter includes more than five young (the average size of litters among white mice) the succeeding litters from that female are usually above the average number of young, but the reverse of this proposition issometimes true and at other times not. Sixteen litters seems to be the limit for one female to bear, and some stop before reaching that number. The total number of young produced may reach 80, and appears to bear no relation to the size or the total number of litters. Females which are allowed to suckle their young, cease to bear at 18 to 22 months of age, after producing 12 to 16 litters, while those females whose young are removed as soon as found usually die before they cease to bear, but in the rare instances where such females survive, their litters come nearer together, but reproduction stops at an earlier age, so that the total number of litters produced is within the limits stated above. Females appear to be somewhat shorter lived than males, but animals of both sexes if healthy at birth, given reasonably good care, and protected from contagious diseases, have an expectation of life of about two years. Females suckling young frequently fail to at once again become pregnant, due to some influence from the mammary glands which inhibits ovulation, and in all cases where more than two young are being suckled and the female becomes pregnant, the implantation of the embryos is retarded for about nine days, until the young cease to suckle, and a corresponding prolongation of the gestation period occurs, as compared with that of non-suckling females. Both of these phenomena appear to be protective for the parent organism, since females whose young are removed at birth usually bear litters in such rapid succession that they die of exhaustion before the termination of the reproductive period. Males and females which were never allowed to pair have lived the same length of time as other animals, and have shown no peculiarities of behavior or appearance. 198 SCIENTIFIC PROCEEDINGS (108). Healthy animals, for experimental work, are only to be secured by individual selection, as not all members of a litter are equally healthy, but too rapid breeding of healthy animals is sure to pro- duce weak offspring. Extreme heat will kill mice quicker than cold. Pulmonary diseases must be guarded against by rapid removal of infected animals. Sarcomas have also caused the death of a number of animals in the course of this investigation, usually not appearing in animals less than a year old. Tape-worm cysts have been found in the livers of some mice, but seemed to have been without effect on the general health and reproductive activity of their hosts. A few female white mice have shown a peculiarity common in yellow mice, sterility accompanied by extensive laying down of fat, after having four to six litters. The cause of this behavior is at present unknown. 105 (1565) Reaction of cells to the galvanic current in tissue cultures. By Sven INGVAR (by invitation). [From the Osborn Zoélogical Laboratory, Yale University, New Haven, Conn.| By applying a weak constant galvanic current (strength 2-4 billionths of an ampere, density approximately 1/1000—1/2000 6, nonpolarizable electrodes) to tissue cultures made according to Harrison’s method, the following observations were made: The galvanic current has a directing influence upon the cell and fiber outgrowth in the cultures so that this occurs almost entirely along the lines of force in the galvanic field. Whereas in the control preparations the outgrowth occurs in all directions, cell movements under the influence of a galvanic current take place toward the anode and the cathode. The cell processes growing toward the anode show morphological differences from those growing toward thecathode. A new biological cell character may in this way be revealed. If a weak electric current by means of a single conductor is drawn through the culture, the outgrowth of the fibers and cells EXPERIMENTS ON LENS IN AMBLYSTOMA. 199 always takes place perpendicular to the conductor. Bok has called attention to the fact that in the living organism the nerve fibers grow out from the spinal cord perpendicular to the long fiber paths growing down from the brain stem. Kappers has tried to explain this as a galvanotropic phenomenon. To this observation, an interesting analogy is thus found in tissue cultures. The hypothesis of Kappers, as the main result of this author’s work on ‘‘neurobiotaxis,”’ that electrical forces are determining factors in the outgrowth and distribution of the different constitu- ents of the nervous system, has been proved to be a fact in pieces of the central nervous system of the chick cultured in vitro. As several authors (Hyde, Mathews, Pfeffer) have pointed out, electrical currents flow in developing organisms. The currents successfully employed in our experiments correspond in range in electromotive force with those found in various embryos. From this it may be concluded that electrical forces play a réle in the formative processes in morphogenesis. 106 (1566) Experiments on the lens in amblystoma. By Ross G. HARRISON. [From the Osborn Zoélogical Laboratory, Yale University, New Haven, Conn.| The embryo of Amblystoma punctatum has been reported as one of those in which the ectoderm normally giving rise to the lens is dependent upon the continued influence of the optic vesicle to effect its differentiation! It was surprising, therefore, to find that in certain experiments, directed toward the study of the gills, lenses developed from the proper ectoderm when transplanted to regions far from the eye. There are obviously two ways of testing the independence of lens differentiation: one is to take away the eye rudiment as has been done in previous experiments (Spemann, Lewis, Le Cron); the other is to transplant the lens-forming ectoderm to another 1 Wilbur L. Le Cron, ‘‘Experiments on the Origin and Differentiation of the Lens in Amblystoma,”’ Am. Journ. Anat., 1906-7, VI. 200 SCIENTIFIC PROCEEDINGS (108). region of theembryo. The present experiments upon Amblystoma show that the results may be different in the two cases. Excision of the eye rudiment in the medullary plate stage is followed by suppression of the lens. Likewise, if the optic vesicle is removed immediately or shortly after closure of the medullary folds, the lens fails to develop, as shown by Le Cron. If, however, this same lens ectoderm is transplanted to other regions of the head, a well differentiated lens will develop, pro- vided the ectoderm is taken from the eye region after closure of the medullary folds. Contact between optic vesicle and ectoderm has at this time been established, though the two are not adherent and may be readily separated without cells from one layer sticking to the other. If the lens ectoderm is taken in earlier stages, small and not fully differentiated lenses are sometimes but not always formed. Barring one or two questionable cases, there is no evidence that, in Amblystoma, ectoderm from other parts of the head or from the trunk can give rise to a normal lens. When such ecto- derm is transplanted to the eye region, even before closure of the medullary folds, abnormalities in the optic cup, due to irregular infolding, frequently arise and no lens develops. When a circular piece of ectoderm, having the diameter of the optic vesicle, is removed from the eye region, the surrounding ectoderm, which pushes in and covers the wound, usually gives rise to a lens, as Spemann found to be the case in 7viton. When larger pieces of ectoderm are removed, the lens usually does not develop. These experiments show that Amblystoma must be added to those forms in which the lens ectoderm is capable of self-differen- tiation. Why this power is manifested only when it is removed from its normal position and not when it is left in place after removal of the optic vesicle is problematical. The difference in behavior can scarcely be referred to differences in the degree of injury to the cells, but it is apparent that at times secondary cir- cumstances of some unknown character may dominate more fun- damental ones and thus lead to mistaken conclusions. ee a Code) peat ee PERMEABILITY OF THE CAPILLARY WALLS. 201 107 (1567) The effect of solutions of certain salts and colloids on the permea- bility of the capillary walls. By ARTHUR H. SMITH and LAFAYETTE B. MENDEL. [From the Sheffield Laboratory of Physiological Chemistry, Yale University, New Haven, Conn.| In order to study the permeability of the capillary walls for various substances, the material was injected, in isotonic solution, intravenously into rabbits, the diffusion out of the circulation being estimated by the relative concentration of the blood as indicated by the hemoglobin. The rate of injection was such that a volume of fluid equal to the estimated blood volume was intro- duced in two minutes. When the chloride, bromide, sulfocyanate, nitrate or acetate of sodium was used, the blood volume returned to normal in thirty minutes. When the tartrate, citrate or sulfate was used the blood volume remained somewhat higher than normal through- the experiment which usually lasted more than two hours. The injection of M/15 hydrochloric acid, calcium chloride or colloidal silver (Solargentum, Squibb) produced no delay in the removal of fluid from the circulation. Acacia, however, markedly sustained the increased blood volume at a point one third above the normal for more than two hours. Under the conditions of these experiments, there was no evi- dence of edema or of increased water content of the muscles even after repeated injections. The excess of fluid was eliminated through the gastro-intestinal tract, the kidneys and to a slight extent, into the serous cavities. 202 SCIENTIFIC PROCEEDINGS (108). 108 (1568) Observations on the physiology of the otic labyrinth. The influence of prolonged rotation on the duration of post- rotatory nystagmus. By ALEXANDER L. PRINCE. [From the Physiological Laboratory, Yale School of Medicine, New Haven, Conn.| Rotation of the head in space is accompanied by ocular move- ments of a definite type. These ocular movements (nystagmus) present two phases: a slow deviation of the eyes opposite to the direction of rotation, followed by a quick return to the initial position. On cessation of rotation similar movements are observed. In the post-rotatory nystagmus, however, the slow deviation phase is in the direction of the preceding rotation. The deviation phase as shown by Wilson and Pike! is dependent on labyrinthine stimulation whereas the quick return is due to a proprioceptor reflex arising from the stretched antagonist ocular muscle during the deviation phase. These ocular movements can be especially well studied in the post-rotatory period. The duration of the post rotatory nystagmus is dependent on the intensity of the labyrinthine stimulus and therefore on the speed of rotation and up to a certain point on the number of revolutions at any given speed. Using a standard of ten turns in twenty seconds, the duration of post rotatory nystagmus in man has been utilized as an index of labyrinthine efficiency. This test has been applied extensively in the examination of candidates for the U. S. Aviation Service. The post rotatory nystagmus under the standard rotation test has an average duration of 23 seconds. ‘This figure is based on a large number of determinations made by investigators in the oto- logical department of the Air Service.2, On the other hand, accord- ing to members of the psychological department of this Service? 1 Wilson and Pike, Philosophical Transactions of the Royal Society, London, 1912, Series B, CCIII, 127. 2 Manual of the Medical Research Laboratory, War Department, Air Service, Division of Military Aeronautics, Washington, D. C, PHYSIOLOGY OF THE OTIC LABYRINTH. 203 the nystagmus time decreases progressively with practice. They find that in four persons subjected to frequent periods of rotation over an extended period (20 days) the average post-rotatory nystagmus time fell from 24.9 to 6.3 seconds. The question then arises as to whether the nystagmus time can be taken as an absolute index of labyrinthine efficiency. In experiments on animals it has been shown that the nystagmus following unilateral extirpation of the otic labyrinth disappears a few days after operation and that the ocular symptoms (deviation) recur after ablation of the cerebrum. This seems to indicate that the disappearance of nystagmus under these circumstances is dependent on the activity of a compensatory mechanism localized in the cerebrum.! In order to decide whether such compensation occurs after repeated rotation I have tested the matter as follows: Cats were subjected to interrupted horizontal rotation over a prolonged period and the nystagmus time observed. The rotation tests (20 turns in 20 seconds) were applied 50 to 60 times daily at intervals of two minutes and were repeated for twenty consecutive days. The duration of post-rotatory nystagmus under these condi- tions was found to vary slightly in different animals. With 20 revolutions in 20 seconds the maximal variations in the post-rota- tory nystagmus time observed in six animals were from I2 to 19 seconds. The ocular responses to rotation for each animal remained fixed within narrow limits and were unaffected by prolonged rotation. It may be concluded therefore that rotation tests give an adequate index of labyrinthine activity. 1 Prince, A. L., PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE, 1917, XIV, 133. 204 SCIENTIFIC PROCEEDINGS (108). 109 (1569) Variations in the affinity of hemoglobin for carbon monoxide in health and disease. By ALEXANDER L. PRINCE. [From the Physiological Laboratory, Yale School of Medicine, New Haven, Conn.| It is known that the affinity of hemoglobin for carbon monoxide varies slightly in different individuals and even more in the blood of various species. But the possibility of variations in human blood under conditions of health and disease has not received special consideration. Bloods from normal individuals and selected hospital cases in whom the percentage of hemoglobin varied from 38 to 110 per cent. were examined. In each case the blood was equilibrated in vitro at 20° C. with known concentrations of carbon monoxide in atmospheric air and the carbon monoxide-hemoglobin dissociation curve of the blood determined. From the curves the oxygen-carbon monoxide affinity ratio was computed by the following formula: TO. X CO per cent. X affinity Oz (100—CO per cent.) X TCO when TO, = the tension of O: (2,100 when expressed in parts per 10,000 of air). | TCO = the tension of CO (expressed in parts of carbon monoxide in 10,000 of air) at any given point in the carbon monoxide dissociation curve of the blood. CO per cent. = percentage saturation of the blood at a given tension of CO. Affinity O, and affinity CO = the relative affinity of O2 and CO for the blood when affinity for O2 = I Affinity CO = The results obtained are shown in the following table. It will be noted that (1) the carbon monoxide variations in the oxygen- ELIMINATION OF CARBON MONOXIDE. 205 carbon monoxide affinity ratio of all bloods examined fall between I : 254 and 1 : 378, and (2) these variations bear no relation to the hemoglobin percentage of the blood nor to the age and condition of the subject. Subject, Age. Sex Condition, a ye Per cent. MEE LS 2 esc ie os 59 | Male Pernicious anemia 38 I: 310 _ Cee 27 | Female | Pernicious anemia, recent 60 E3323 transfusion ) ee 38 | Male Diabetes mellitus 60 I : 310 » aS. en 28 ms Pulmonary tuberculosis 80 I : 286 Mw, Ei. 3 | Female | Broncho pneumonia 80 I : 298 ON ee 31 | Male Chronic endocarditis 85 Bs 323 SRP ie eae I2 Lobar pneumonia 85 E2254 A” A re ae 14 | Female | Diphtheria (convalescent) 85 E2275 ae : S-Eeey eeetie 48 rs Tumor of spinal cord 85 I : 364 Se Se pee ee 53 f Mucous colitis 85 E2323 Oe a ee 8 = Tuberculosis of hip joint 90 ti 275 Sa. Ges oc otres é 30 | Male Diabetes mellitus 100 E °323 Bo Was ee oe ke 20 = Normal 100 I : 286 EE Oe | eae 30 * os 100 & 2378 Bee Be WES Sores 23 | Female i 100 I: 378 7G SS OP ea 36 | Male zs 105 I 3: 364 Rk oe b aileke 46 ™ a IIo 2295 Pewee it. se 28 is a IIo t 2-323 IIO (1570) The elimination of carbon monoxide and a method of acceleration. By Howarp W. HAGGARD (by invitation). [From the Physiological Laboratory, Yale University Medical School.| Carbon monoxide combines with hemoglobin with an affinity about 300 times as great as that of oxygen for hemoglobin. Blood is deprived of its oxygen-carrying power by combining with CO and the organism suffers from a corresponding degree of anoxemia. The severity of the damage done to the victim is dependent upon the degree of anoxemia and especially upon the duration. Evi- dently the rate of elimination is extremely important. A study has been made of the normal rate of elimination in dogs gassed to 60 to 80 per cent. saturation of the hemoglobin with 206 SCIENTIFIC PROCEEDINGS (108). CO. If the animals survived, the blood was practically free of CO in from two to three hours. The curve so obtained was relatively flat for the first half of the period of elimination. A dog with 80 per cent. of the hemo- globin combined with CO, at the end of an hour still showed 60 to 70 per cent. During the remainder of the elimination period the drop in the curve was rapid. It is evident that considerable damage may be wrought even after the inhalation of CO has stopped. The elimination of CO was studied in animals inhaling oxygen, carbon dioxide, and oxygen-carbon dioxide mixtures following gassing. 3 With the inhalation of oxygen the rapidity of elimination was increased to approximately double. The curve of elimination of CO from the blood still maintained its normal shape. Deaths from respiratory failure still occurred. Inhalations of 6 per cent. carbon dioxide in air, increased the pulmonary ventilation, and thus accelerated greatly the period of elimination to one half or less of the normal. The curve of CO-hemoglobin in the blood tended to approach more nearly a straight line. Inhalations of oxygen containing 6 per cent. carbon dioxide resulted in complete elimination of the CO from the blood in from 15 to 20 minutes. ‘The curve obtained was a straight line. With the inhalations of carbon dioxide death from respiratory failure was prevented. III (1571) The influence of oxygen in expelling CO. from the blood. By YANDELL HENDERSON and H. W. HAGGARD (by invitation). [From the Physiological Laboratory, Yale University Medical School.| Considerable theoretical significance attaches to the interaction of oxygen and CQO, in blood. That variations of CO, tension influence the capacity for oxygen is generally accepted. There has not been, however, universal agreement among observers EXPELLING CO: FROM THE BLOOD. 207 as to the influence of variations in oxygen tension upon CO, capacity. Our experience has led us to believe that the quality of exhi- biting this reaction depends upon the previous treatment of the blood. Thus in our hands freshly drawn dog’s blood shows it when defibrinated, but not when oxalated. The following is typical of our results. Equilibrating Gas Mixture Defibrinated Blood Oxalated Blood Air plus 5.6% COz...53.5 volumes per cent. COz...55.0 volumes per cent. CO2 Nitrogen plus 5.6% ‘6 58.5 sé sé sé 55-0 ‘6 “ec sé Experiments are under way to determine whether the influence of CO, upon oxygen capacity is likewise lacking in oxalated normal blood. Variations within the body in respect to the capacity of the blood for interaction of the two gases would afford a possible ground for reconciling some otherwise difficult discrepancies. II2 (1572) Observations on the connective tissue ground substance in living amphibian embryos. By GEORGE A. BAITSELL. [From the Osborn Zodlogical Laboratory of Yale University.| Results obtained from a study of the development of con- nective tissue in amphibian embryos, as presented in a previous communication,! show that the process is essentially in agreement with the intercellular theory of connective tissue formation. The previous observations were made on preserved amphibian material. It has been found possible this spring to demonstrate the presence of a primary ground substance in various stages of living amphibian embryos. Living embryos, ranging from a late gastrula stage up to the free-swimming embryo, have been dis- sected under the binocular microscope and it has been possible in all stages to show, as was previously demonstrated in the prepared 1A report of this work was presented at a meeting of the National Academy of Sciences held at New Haven, November, 1919, and an abstract appears in the Proceedings of that Society, 1920, VI, 77. 208 SCIENTIFIC PROCEEDINGS (108). material, that an homogeneous, intercellular, gelatinous material, permeates various regions of the embryo. This substance as seen in a living embryo is perfectly transparent and is, therefore, difficult to detect under the binocular or under a compound microscope equipped for ordinary illumination. It can, however, be readily studied with a microscope equipped for dark field illumination and numerous observations on this material have been made in this manner. The present observations on the ground substance in living amphibian embryos, therefore, confirm those previously obtained with prepared material and show conclusively that the primary substance in connective tissue formation is a secreted, inter- cellular material. 113 (1573) Effects of pilocarpine upon salivary fistula dogs before and after coli injection. By H. G. BARBOUR and B. P. FREEDMAN. [From the Department of Pharmacology, Yale University School of Medicine.| In each of two healthy dogs was made a fistula of Wharton’s duct. After several weeks a standard dose of 0.5 mgm. per kilo pilocarpine hydrochloride was injected at intervals of two or three days. After each dose the curve of secretion was followed for two hours by weighing the saliva on cotton pledgets by five minute periods. Contrary to expectation, a normal curve was not readily established. The secretion increased with each experiment until both the total for two hours and the figure for the maximum indi- vidual five-muinte period were approximately doubled in one case and trebled in the other. After six to eight injections the results became more constant. This gradually increasing sensi- tivity to pilocarpine has not yet been explained. After a fairly constant degree of sensitivity to the drug had been established each of the above animals received an injection of % c.c. per kilo of a coli vaccine (one million million killed coli TEMPERATURE CHANGES INDUCED By GUM ACACIA. 209 per c.c.). At the height of the ensuing fever (7.e., after three or four hours) was injected the standard dose of pilocarpine. In both cases the secretion curves exhibited an unusually slow onset and a much diminished maximum as well as total secretion. The saliva was of a much thicker consistency than normal. These experiments were made at a stage of fever in which Barbour and Howard! have demonstrated a thickening of the blood. It is suggested that the latter is the chief causative factor in numerous cases of diminished secretion which have been reported in fevers. 114 (1574) Temperature changes induced by gum acacia injections in normal and fevered animals. By H. G. BARBOUR and L. H. BARETZ. [From the Department of Pharmacology, Yale University School of Medicine.| The effects of gum acacia upon the body temperature have been studied in both normal and fevered rabbits and dogs. Solu- tions were made in water redistilled from glass and given intra- venously. In the following experiments upon rabbits 20 c.c. of fluid per kilo were injected unless otherwise stated. Intravenous injections of control fluids (Locke’s solution or physiological saline) gave an increase in temperature of 1° C. or more, subsequent to a brief depression of 0.2° C. This temperature increase could be superimposed upon the rise induced by bacto- peptone injections. Similar amounts of 7 per cent. acacia (also Io c.c. of 20 per cent.) gave a slight depression in normal rabbits by a few tenths of a degree centigrade, never an increase. In five bactopeptone rabbits in which the temperature had reached a level of about 1° C. or more above normal within 4 or 5 hours, an injection of acacia (7-10 per cent. or 10 c.c. per kilo of 20 per cent.) brought the temperature back to approximately 1 Barbour, H. G. and Howard, A. J., Proceedings of the Society for Experi- mental Biology and Medicine 1920 XVII. 210 SCIENTIFIC PROCEEDINGS (108). the normal level in 20-40 minutes. A rabbit in which fever was induced by puncture of the corpus striatum gave a very rapid fall (average 1.1° C. in 20 minutes) of temperature on three dif- ferent days as a result of 7 per cent. acacia injections. In rabbits, therefore, acacia injections induce a mild tempera- ture depression in health but a marked antipyretic effect in fever. Four dogs responded to 4 c.c. per kilo of 20 per cent. acacia injected intravenously by increases in body temperature varying from 0.9 to 1.8 °C. In one of these, however, a preliminary depression of one half degree was observed. The normal tem- perature was regained within from 3 to 8 hours after injection. Two dogs were given coli fever (method of Barbour and Howard) and the usual increase in blood solids was noted. Fol- lowing an intravenous injection of 4 c.c. per kilo of 20 per cent. acacia in each dog reductions of 0.4 and 0.7° C. respectively were noted within 20 minutes, with a corresponding diminution in the total solids of the blood. This was followed however by a renewed temperature rise in both cases. Intravenous acacia injections therefore raise the temperature of normal dogs but in fever dogs exert a brief though marked antipyretic action, accompanied by increased blood volume. The latter phenomenon is obviously parallel to the antipyretic effect of intravenous injections of 50 per cent. dextrose. 115 (1575) Studies on salt action. III: The effect of hydrogen ion concen- tration upon salt action. By I. S. FAK (by invitation). [From the Department of Public Health, Yale School of Medicine.] Our recent studies of the effect of electrolytes upon the via- bility of bacteria have indicated the important influence of even slight variations in hydrogen ion concentration upon such phe- nomena. Working with Bact. coli we find that hydrogen ion concentrations above pH = 6.0 or below pH = 7.0 give a much more rapid death rate than occurs when the pH is maintained within these limits. Our experiments suggest that a very careful BLoop OxyYGEN ANALYSES. 211 control of hydrogen ion concentration is absolutely essential before valid conclusions can be drawn as to the influence of electro- lytes, alone or in combination. Furthermore, it is essential to follow with care the changes which go on in a suspension of living and dead cells as well as to determine the initial conditions which are provided. We find that a bacterial suspension in 5 isotonic NaCl solution quickly reverts to a pH of about 7.2 whether its initial hydrogen ion concentration be above or below this value. Asimilar change takes place in a balanced solution of 5 isotonic NaCl + isotonic CaCl, but at a much slower rate as indicated by the table below. HYDROGEN ION CONCENTRATION OF SUSPENSIONS OF Bact. coli IN THE PRESENCE OF ELECTROLYTES. 5 Isotonic NaCl, 5 lsotonic NaCl+ Isotonic CaCle. Initial. Initial 44 Hrs 9 Hrs 30 Hrs 43 Hrs. 9 Hrs, 24 Hrs. 4.0 7.0 7.0 2 4.0 4.8 4.8 5.8 5.0 7.3 7.2 Be 5.0 6.5 6.5 7.2 6.0 7.5 a2 7.2 6.0 7.2 7.2 i 7.0 Yee 7.0 7.4 7.0 7.2 7.2 7.5 8.0 f Pe ve 752 8.0 8.0 8.0 8.0 116 (1576) Discrepancies in blood oxygen analyses by the methods of Van Slyke and Henderson-Smith.! By Arthur H. Smith, J. A. Dawson and Barnett Cohen. [From the Sheffield Laboratory of Physiological Chemistry, Osborn Zoélogical Laboratory and the Laboratory of Public Health, Yale University, New Haven.| Loosely bound oxygen is liberated from the hemoglobin in blood by the addition of potassium ferricyanide. In the Van Slyke method,’ all the gases are exhausted by means of a Toricellian vacuum from a laked blood-ferricyanide mixture and measured directly. In the Henderson-Smith method,’ the oxygen is evolved 1 This work was initiated in the Laboratory of Intermediary Metabolism, Chemi- cal Warfare Service, Yale Station, under Lt.-Col. F. P. Underhill. 2 Van Slyke, D. D., Jour. Biol. Chem., 1918, X XXIII, 127. 3 Henderson, Y., and Smith, A. H., Jour. Biol. Chem., 1918, XXXIII, 39. 212 SCIENTIFIC PROCEEDINGS (108). into a fixed volume of air, a portion of which is analyzed directly for oxygen by absorption with alkaline pyrogallate. After applica- tion of all the corrections suggested by the authors of these methods, the results are not identical,—analyses by the Van Slyke method yielding 4 to 10 volumes per cent. more oxygen than those by the Henderson-Smith method. The divergence between the results from each method may represent a variation of 17 to 64 per cent. __., —_ nis na Cent. Oz. Per Cent. On. Amount, | Per Cent. 18a. Venous content......... 12.9 10.4 2.5 | 19.4 18b. Venous capacity......... 18.5 12.8 5-7 30.8 18c. Arterial content......... 20.1 * aA 5.5 27.4 18d. Arterial capacity........ 20.0 15-5 4.5 22.6 It is clear that there is a constant factor or factors at play, inherent in the methods of analysis employed, which ought to account for this discrepancy. The gas evacuated by the Van Slyke procedure is not all oxygen but probably contains in addition to nitrogen, minute amounts of carbon monoxide, hydrogen, methane and the rare atmospheric gases. Bohr! states that blood contains 1.23 volumes per cent. of nitrogen (incorrectly quoted by Van Slyke as 0.9 vol. per cent.) and 0.22 volumes per cent. of the other gases—a total of about 1.45 volumes per cent. of gas not oxygen. We have absorbed with alkaline pyrogallate, the oxygen from the gas extracted in the Van Slyke procedure and have found in all cases a residue of 0.055 to 0.082 c.c. from 2 c.c. of blood—an average of about 3.3 volumes per cent. This residue does not contain COx, and we have reason to believe that it is practically all nitrogen. It occurs to the same extent in aerated blood as in venous. Since this residue is almost constant within narrow limits, its value may be subtracted from the total Van Slyke gas volume as an average correction. We have applied this correction in Table II, and have thereby reduced the level of the Van Slyke figures by about two volumes per cent. A further refinement would be to deduct 1 Bohr, C., in Nagel, W., ‘‘Handb. Physiol. d. Mensch.,’’ 1909, I, 117. BLoopD OXYGEN ANALYSES. 213 the amount of oxygen physically held in the blood if the analysis is intended to show the amount of oxygen loosely held in the oxyhemoglobin. In the Henderson-Smith method a small correction should be applied for water vapor pressure. This is however of theoretical interest for it is only 0.1 to 0.3 volumes per cent. in amount. Much more important is the fact that the oxygen content of the air in the diffusion tube has been considered without regard to the amount physically held in the 3 c.c. of blood-ferricyanide mixture. The volume of air in the diffusion tube is constant at about 10 c.c. When 0.5 c.c. potassium ferricyanide is injected through the rubber stopper, this volume is reduced, and the pres- sure within the tube correspondingly increased by about 5 per cent. As a result of the chemical reaction oxygen is liberated, and the pressure further increased by one percent. Consequently, the 3 c.c. of fluid within the tube will absorb oxygen in proportion to its increased partial pressure. This physically held oxygen should be added to that determined analytically to give the total contained in the sample of blood. We have calculated the amount of oxygen that the blood- ferricyanide mixture would hold on the basis of the gas laws (assuming that the mixture has 90 per cent. the absorption capacity of water), and it will be seen from Table II that the level of the Henderson-Smith figures has been raised about 1.5 volumes per cent. TABLE II. Van Slyke Method. Henderson-Smith Method. Difference Be- Sample aT Mees eis eet sys. I kat) oe | tween VS and Volumes Per Cent. O2 Volumes Per Cent. O2 | H-S Corrected Old Value. {Corrected Value.| Old Value, {Corrected Value. Values. 18a 12.9 10.7 10.4 II.7 —I.0 18d 18.5 16.7 12.8 I4.1 2.6 18¢ 20.1 18.0 14.6 16.0 2.0 18d 20.0 18.4 15-5 16.9 Es From Table II it is evident that our corrections have brought the Van Slyke and Henderson-Smith values for blood oxygen much closer together than they were originally. The figures are however not identical, and we are now engaged in testing out other factors that may be involved. 214 SCIENTIFIC PROCEEDINGS (108). 117 (1577) On the comparative toxicity of some alcohols with especial refer- ence to isomers. By Davip I. MAcurt. [From the Pharmacological Laboratory of the Johns Hopkins Uni- versity, Baltimore.| While the comparative toxicology of various alcohols belonging to the aliphatic series has been studied by various authors, very little is known concerning the comparative pharmacological properties of some of their isomers. The present author, during the past winter, had occasion to determine the relative toxicity of methyl, ethyl, propyl, butyl and amy! alcohols, and, in that con- nection made a comparative study of isopropyl, isobutyl, and isoamyl alcohols, together with the normal propyl, butyl and amyl alcohols. The lethal dosage of the various alcohols was first determined by the cat method, that is, by injections intra- venously of 5 or I per cent. solutions at regular intervals, and determining the amount of drug per kilo required to kill the animal. It was found, in these acute experiments, that the lethal dose of the normal alcohols followed the well-known Richardson’s law, that is, decreased with the increase in molecular weight of the alcohol, as has been shown already by many observers. On comparing the toxicity of propyl, butyl and amyl] alcohols, how- ever, with their isomers, the secondary alcohols, it was found that in every case the secondary alcohols were less poisonous than the primary ones. Following the determination of the lethal dosage, experiments were made on isolate frogs’ hearts, and here again the same relationship was noted. A third series of observa- tions on the effects of the different alcohols on surviving plain muscle (ureter), also showed that the secondary alcohols were less toxic than the primary ones. CONCENTRATING ACTIVITY OF GALL BLADDER. 215 118 (1578) The concentrating activity of the gall bladder. By PEYTON Rous and PHILIP D. MCMASTER. [From the Rockefeller Institute for Medical Research.| In a previous paper we have noted the fact that the fluid which collects in bile ducts experimentally obstructed is an in- spissated, tarry bile when the ducts communicate with the gall- bladder, whereas in ducts unconnected with this viscus the fluid is thin and soon becomes free from pigment and cholates. It has long been recognized that the gall-bladder must have a concentrat- ing function, since bladder bile is more concentrated than duct bile from the same animal; and continued functioning during stasis will explain the tarry bile then found. The inspissation occurs so rapidly as to raise the question whether concentration of the bile in periods of intermittent or partial stasis may not be an important favoring element in the formation of gall-stones. To determine the rate of concentration advantage has been taken of the arrangement of the hepatic ducts in the dog. There are three of these, which unite to form a common duct, with the cystic duct emptying high up into the central one. Through an opening near the lower end of this last a catheter was pushed into the neck of the gall-bladder, which was emptied and washed with salt solution; and the duct was ligated after the catheter had been withdrawn. The bile from the middle lobes of the liver had now no way of escape save into the gall-bladder. That from the lobes to either side still reached the common duct, but from this it was collected into a rubber balloon placed in the peritoneal cavity. The laparatomy incision was completely closed. The dogs tolerated the operation well. Control experiments in which a second balloon was substituted for the gall-bladder showed that the separated portions of bile differed little in their pigment content, which was taken as the index to concentration. On examination after twenty-four hours the gall-bladder, still undistended, was regularly found to contain only one sixth to one tenth as much fluid as should on calculation have reached 216 SCIENTIFIC PROCEEDINGS (108). it, but this, thick and dark, was six to ten times as concentrated in pigment as the control specimen in the rubber balloon. The results were the same when, without other variation in the exper- iment, the gall-bladder was filled to the normal distension with sterile bile of known character prior to withdrawal of the catheter. The contents of the branches of the hepatic duct connecting with the gall-bladder were always examined at autopsy. Here a thin bile, like that in the balloon, was obtained, a direct proof that the thick contents of the gall-bladder had not come as such from the liver. It is evident that the normal gall-bladder can concentrate bile with very great rapidity. 119 (1579) Osmosis as a factor in the local accumulation of leucocytes in the animal body. By FRANK MALTANER and E. N. Hoppe (by invitation). [From the Division of Laboratories and Research, New York State Department of Health, Albany.] Chemical forces have generally been held responsible for the chemotaxis of leucocytes. Some of the early classical experiments which led to this belief were repeated. The results showed that the work had been misinterpreted, and also indicated that the forces which were active in producing the phenomenon were physical in character. | These physical forces are the forces responsible for osmosis and diffusion. Inasolution not at concentration equilibrium they will act in directions counter to each other. In the aqueous solutions examined, leucocytes are shown to move in the direction of the osmotic force and opposite to the direction of the diffusing substances in solution. This motion is explained as being due to the greater perme- ability of leucocytes for water and the fact that their total mass is negligible as compared to their content of water. ALTERATIONS OF INTRACRANIAL TENSION. 217 120 (1580) Alterations of intracranial tension by salt solutions in the alimen- tary canal. By HARVEY CUSHING and FREDERICK E. B. FOLEY. [From the Laboratory of Surgical Research, Harvard Medical School, and the Surgical Clinic, Peter Bent Brigham Hospital]. In two interesting and suggestive papers! Weed and McKibben demonstrated the significant physiological fact that it is possible to reduce the cerebrospinal fluid pressure and diminish the bulk of the brain by injecting a hypertonic solution into the blood stream. Conversely they found that hypotonic solutions had the opposite effect: a rise of cerebrospinal fluid pressure and an increase of brain bulk. In the course of our studies their work has been repeated and their general conclusions confirmed. The clinical bearing of these facts is obvious. Particularly they concern the states commonly referred to as “ pressure symp- toms.’’ By similar methods it has been found possible to secure these same results in patients with increased degrees of intracranial tension. It was felt that the undesirable effects on pulse, respir- ation and blood pressure of such intravenous injections might contraindicate their use. For this reason the effects of gastroin- testinal doses of hypertonic solutions were studied. In a large series of animal experiments it has been found that practically the same effects may be obtained by the gastrointest'nal route of administration. By this method the intracranial changes are not attended by disturbances of pulse, respiration or blood pressure, also the possible alterations of the cellular elements of the blood are avoided. Twenty to thirty cubic centimeters of a saturated sodium chloride solution introduced into the duodenum or rectum of an average-sized cat produced a maximal fall of cerebrospinal fluid pressure precisely comparable to that which occurred when the solution was given intravenously. Following such doses the 1 “Experimental Alterations of Brain Bulk,” Am. Journ. Physiol., 1919, XLVIII, Pp. 531; “Pressure Changes in the Cerebrospinal Fluid following Intravenous Injection of Solutions of Various Concentrations,”’ Ibid., p. 512. 218 SCIENTIFIC PROCEEDINGS (108). average fall of pressure in a large series of experiments was 258 mm, normal saline. The changes were roughly proportionate to the concentration of the salt and the size of the dose. Two per cent. sodium chloride solution in large doses or a saturated solution in doses as small as 5 c.c. gave corresponding though less marked effects. Non-absorbable salts gave similar reductions though slower in occurrence and less in extent. Dextrose solution caused still less striking results though identical in nature. The converse results seen after the ingestion of hypotonic solutions (water) were not great in extent nor well sustained. A previous dose of a hypertonic solution rendered more extensive the rise of pressure in these cases. The changes observed were independent of blood pressure and were not attended by significant alterations of pulse or respiration. These effects on cerebrospinal fluid pressure and brain volume were investigated in patients. An excellent opportunity for this was afforded by patients with brain tumor and cerebral herniz subsequent to decompression operations. In these patients there was a lowering of tension when hypertonic salines were given by mouth. Occasionally very striking results could be obtained in which case the tense convex protrusion became a soft concave area over the decompression site. I2I (1581) \ Some results with a new technique in vitamine measurement. By WALTER H. Eppy and HELEN C. STEVENSON. [From the Department of Physiological Chemistry, Teachers College, Columbia University and the Department of Pathology, New York Hospital, N. Y. City.] By means of a technique described in detail in the March, 1920 number of the PROCEEDINGS certain results have been at- tained that seem to indicate the specifity of the test for the “B”’ vitamine. These results are briefly as follows: (a) Application of the test to three specimens of Funk’s antineuritic vitamine prepared in 1912 and '13 showed that the VITAMINE MEASUREMENT. 219 two 1913 preparations were active, though one was markedly more so than the other, and that the 1912 preparation was in- active. These specimens were supplied by Dr. Funk and represent the products of greatest purity as obtained by his fractional precipitation method. The only known impurity present was nicotinic acid and pure synthetic nicotinic acid failed to respond to the test. (b) Lloyd’s reagent was shown by Seidell and Williams to remove the B vitamine and by Harden and Silva to have little if any action upon the C vitamine. This point was tested with the new method and orange juice obtained by sterile puncture was shown to be deprived of its powers of responding to the test by shaking with the Lloyd reagent. This showed that the reagent removes the cause of the test reaction. Through the kindness of Mr. La Mer working in Professor Sherman’s laboratory orange juice shaken with the Lloyd reagent and then filtered, was used in the treatment of a guinea pig suffering from scurvy. The filtrate was curative in fifteen days. It was also used as a pro- tective agent in the diet of a pig started on a scurvy-producing diet. The symptoms had not appeared in twenty days. From these two experiments we can conclude that the test is not affected by the C vitamine and that the cause of the response is removed by Lloyd’s reagent. The power of the Lloyd reagent to remove the cause of the test was confirmed by experiments with other vitamine extracts. Such experiments are not conclusive evidence but since the effects are so striking and the Lloyd has been demonstrated to remove the B type they seem to justify the belief that the caus- ative agent in the test is the anti-neuritic or water voluble B vitamine. (c) Results were also presented showing that the test is appli- cable to blood. Specimens of blood plasma furnished by Dr. N. R. Blatherwick representing bloods from the jugular and the mammary veins of a cow were poured into alcohol, the alcohol filtered off, evaporated to dryness and the residue taken up with enough water to restore it to the original plasma volume (10 c.c.). Repeated tests showed the mammary vein plasma residue to be markedly richer in the B vitamine than that from the jugular veins, indicating drainage from a mobilizing (?) point of the vitamine. 220 SCIENTIFIC PROCEEDINGS (108). Aside from the particular interest in the bloods examined, the sensitiveness of the test to amounts present in blood suggests the use of the test in localizing the distribution of the substance in the blood streams and possibly in pellagra investigations etc. (d) The value of the test in comparisons of vitamine concentra- tions was demonstrated and the point made that there is an opti- mum amount which must first be established by diluting theextracts studied. Below this optimum amount the test varies approximately with the concentration. Amounts above the optimum cannot be detected except by this method of applying the test. By such a method it was shown that autoclaving for three hours at 15 lbs. and approximately 120° temperature produces some destruction of the vitamine. Alkali was shown to be definitely destructive in concentrations of N/20 to N/40 NaOH. Concentration lower than N/40 seemed to have little effect. (e) The results of applying the test to water extracts of some ten sources of the B vitamine confirmed the feeding experiments with these substances. The detailed counts upon which these conclusions are based were presented but need not be recorded here. 122 (1582) Dietaries of infants in relation to the development of rickets By ALFRED F. HEss and LESTER J. UNGER. [From the Home for Hebrew Infants, New York City.] For the past two years we have been observing the effect of various diets on the development of rickets in infants. The babies received orange juice to exclude the possibility of latent scurvy. All have been on the diet for at least six months, and were followed clinically as we as controlled by means of the X-ray. It was found that many diets supposed to be eminently conducive to rickets resulted in normal nutrition. Condensed milk, for example, only exceptionally induced rickets. The one food which almost regularly led to marked rickets was a “protein milk”’ prepared by precipitating buttermilk with heat (not with rennin). ae ee - a ee ae a DIETARIES OF INFANTS IN RELATION TO RICKETS. 221 This preparation contained about 3.3 per cent. protein; 2.5 per cent. fat; and 6.6 per cent. carbohydrate. Its ash was about 0.44 per cent., of which the calcium and phosphorus stood about mid- way between that of human milk and cow’s milk; its sodium content was even higher than that of the latter. Its fat-soluble vitamine content was high, its water-soluble vitamine low. Judged from the clinical standpoint, this must be regarded as a diet markedly productive of rickets. RECAPITULATION OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE COMMUNICATIONS. VOLUME XVII. Allen, B. 1464. See Sherman, H. C. Altenburg, E. 1465. See Muller, H. J. Atkinson, H. V. 1466. See Lusk, Graham. Auer, John. 1515. The influence of systemic changes on local tissue reactions. Austin, J. H. 1494. [with Edgar Stillman and D. D. Van Slyke.] The excretion of urea. 1550. See Van Slyke, Donald D. Bagg, H. J. 1558. See Gudernatsch, J. F. Baitsell, George A. 1572. Observations on the connective tissue ground substance in living amphibian embryos. Baldwin, Mabel E. 1462. See Thomas, Arthur W. Banzhaf, Edwin J. 1541. Preparation and refining of diphtheria toxin- antitoxin. Barber, W. Howard. 1542. [with George David Stewart.] Further observa- tions upon reflex gastric hypermotility. Barbour, H. G. 1539. [with A. J. Howard.] Coli fever and blood volume in dogs. 1540. [with A. J. Howard.] Dextrose plethora and its antipyretic effect in coli fever. 222 i a ee sw ee | . on ei, NAMES OF AUTHORS. 223 1573. [with B. P. Freedman.] Effects of pilocarpine upon salivary fistula dogs before and after coli injection. 1574. [with L. H. Baretz.] Temperature changes in- duced by gum acacia injections in normal and fevered animals. Baretz, L. H. 1574. See Barbour, H. G. Bauman, L. 1524. [with G. H. Hansmann.] A case of lipuria asso- ciated with chronic nephritis. Bauman, Emil. J 1526. The preparation of animal nucleic acid. Benedict, Stanley R. 1557- The determination of small quantities of sugar in urine, including observations on the polysaccharide content of human urine. Bergeim, Olaf. 1517. See Miller, Raymond J. Bloor, W. R. 1535. Outline of a classification of lipoids. Bowen, Robert H. 1493. New methods for the analysis of cytoplasmic struc- tures. With demonstrations. Bridges, Calvin B. 1461. The developmental stages at which mutations occur in the germ tract. Brown, Wade H. 1548. See Pearce, Louise. 1549. [with Louise Pearce.] On the production of generalized syphilis in the rabbit by local inoculation. Bronfenbrenner, J. 1472. [with M. J. Schlesinger.] Serologic method for - detecting infection in foods. 1473. [with D. Soletsky and M. J. Schlesinger.] On methods of isolation and identification of the members of the colon-typhoid group of bacteria. Further studies on C R indicator. Brownell, Edith D. 1499. See Rose, Mary Swartz. 224 SCIENTIFIC PROCEEDINGS (108). Burge, W. E. 1531. Comparison of the catalase content of the tissues of the mother and of the offspring. Cajori, F. A. 1505. The nutritive value of some nuts. Calkins, Gary N. 1467. Rejuvenescence without encystment and without nuclear fusion in Uroleptus? 1503. Age of parents and vitality in Uroleptus mobilis. Cannon, W. B. 1512. [with P. E. Smith.] Some conditions affecting thyroid activity. Chambers, Robert. 1483. Some studies on the surface layer in the living egg cell. Cheplin, Harry A. 1562. [with Leo F. Rettger.] Studies on intestinal im- plantation of Bacillus acidophilus. Clark, Guy W. 1534. The determination of calcium in blood and plasma. Cohen, Barnett. 1576. See Smith, Arthur H. Cohn, Alfred E. 1507. [with Robert L. Levy.] A comparison of the action in patients of g-strophanthin and digitalis. 1545. [with Robert L. Levy.] The effect of therapeutic doses of digitalis on the contraction of heart muscle. | Coombs, Helen C. 1538. The effect of varying pressures upon the abdominal musculature in the cat. Cullen, Glenn E. 1497. See Levy, Robert L. 1550. See Van Slyke, Donald D. Cushing, Harvey. 1580. [with Frederick E. B. Foley.] Alterations of in- tracranial tension by salt solutions in the alimentary canal. Davenport, C. B. 1504. Heredity in twin births. NAMES OF AUTHORS. 225 Dawson, J. A. 1576. See Smith, Arthur H. Dubin, Harry E. 1553. See Funk, Casimir. Eckman, Rena S. 1499. See Rose, Mary Swartz. Eddy, Walter H. 1491. [with Helen C. Stevenson.] The suitability of the ‘‘Bachman Test’’ for water-soluble B. 1529. [with Helen C. Stevenson.] Further studies in the measurement of vitamine content. 1581. [with Helen C. Stevenson.] Some results with a new technique in vitamine measurement. Ehn, Marie. 1525. See Thro, William C. Faber, Harold K. 1536. Sodium citrate and scurvy. Falk, I. S. 1575. Studies on salt action. III. The effect of hydrogen ion concentration upon salt action. Field, Cyrus W. 1477. Blood sugar curves with glucose, lactose, maltose, mannite, and cane sugar. Foley, Frederick E. B. 1580. See Cushing, Harvey. Freedman, B. P. 1573. See Barbour, H. G. Funk, Casimir. 1553. [with Harry E. Dubin.] Experiments on a quanti- tative and qualitative test for anti-beriberi vitamine. Gates, Frederick L. ; 1482. A method of standardizing bacterial suspensions. Greenberg, J. P. 1471. See Macht, David I. Greenwald, Isidor. 1489. [with Joseph Gross.] A method for the deter- mination of calcium, magnesium, potassium, sodium, chlorides and ‘‘acid-soluble’”’ sulfur and phosphorus in one sample (25 cc. ) of blood. 226 SCIENTIFIC PROCEEDINGS (108). Gross, Joseph. 1489. See Greenwald, Isidor. Gudernatsch, J. F. 1558. [with H. J. Bagg.] Disturbances in the develop- ment of mammalian embryos caused by radium emanation. Haggard, Howard W. 1570. The elimination of carbon monoxide and a method of acceleration. 1571. See Henderson, Yandell. Halsey, Robert H. 1480. Profound effects of digitalis on the vagus producing severe detrimental subjective symptoms, as shown by simul- taneous electro-cardiograms and pneumograms. Hansmann, G. H. 1524. See Bauman, L. Harrison, Ross G. 1566. Experiments on the lens in amblystoma. Harrop, George A. 1546. A method for the estimation of lactic acid in blood. Hastings, A. Baird. 1500. See Scott, Ernest L. 1528. See Scott, Ernest L. Hawk, Philip B. 1517. See Miller, Raymond J. 1518. See Smith, Clarence A. Hawley, Edith. 1499. See Rose, Mary Swartz. Henderson, L. J. 1556. See McLean, Franklin C. Henderson, Yandell. 1571. [with H. W. Haggard.] The influence of oxygen in expelling CO, from the blood. Hess, Alfred F. 1488. [with Lester J. Unger.] The rdle of fat-soluble vitamine in the dietary of infants. 1582. [with Lester J. Unger.] Dietaries of infants in relation to the development of rickets. a” a aa ee —- =~ F a ee NAMES OF AUTHORS. 227 Hite, Bert Holmes. 1532. [with Withrow Morse.] The effect of compression on tissue enzymes. Hjort, Axel M. 1506. [with Charles E. Kaufmann.] The local anesthetic properties of benzoyl carbinol. Holder, Ralph C. 1518. See Smith, Clarence A. Hoppe, E. N. 1579. See Maltaner, Frank. Howard, A. J. 1539. See Barbour, H. G. 1540. See Barbour, H. G. Ingvar, Sven. 1565. Reaction of cells to the galvanic current in tissue cultures. Isaacs, S. | 1471. See Macht, David I. Jencks, Zalia. 1485. A note on the carbohydrates of the root of the cat-tail (Typha latifolia). Kahn, Max. 1474. The protein and lipin content of blood serum of nephritic patients. Karr, W. G. 1510. The influence of water-soluble vitamine on the nutrition of dogs. Katz, L. N. 1516. See Wiggers, C. J. Kaufman, Charles E. 1506. See Hjort, Axel M. Killian, John A. 1514. Studies in the diastatic activity of the blood and blood sugar curves indicating a decreased carbohydrate tolerance in hyperthyroidism. 1559. See Myers, V. C. Kirkham, W. B. 1564. The life of the white mouse. 228 SCIENTIFIC PROCEEDINGS (108). Lancefield, D. E. 1492. Two sex-linked lethals of simultaneous appearance in Drosophila obscura. Larimore, Louise D. 1498. See Rous, Peyton. Levine, Michael. 1543. The behavior of crown gall on the rubber tree ( Ficus elastica). Levy, Robert L. 1497. [with Glenn E. Cullen.] On the deterioration of crystalline strophanthin in aqueous solution. 1507. See Cohn, Alfred E. 1545. See Cohn, Alfred E. Lewis, Howard B. | 1519. [with Lucie E. Root.] Amino-acid synthesis in the organism of the white rat. Longcope, Warfield T. 1533. [with George M. Mackenzie.] The relation be- tween the disappearance of foreign proteins from the cir- culation and the formation of antibodies. Lusk, Graham. 1466. [with H. V. Atkinson.] The influence of lactic acid upon the metabolism of the dog. 1551. Additional experiments showing the production of fat from protein. McCann, William S. 1552. An observation of the effect of a protein meal given to a man at the end of an 8-day fast. McLean, Franklin C. 1556. [with H. A. Murray, Jr. and L. J. Henderson.] The variable acidity of hemoglobin and the distribution of chlorides in the blood. McMaster, Philip D. 1544. See Rous, Peyton. 1578. See Rous, Peyton. MacDougal, D. T. 1479. [with H. A. Spoehr.] Hydration effects of amino- compounds. NAMES OF AUTHORS. 229 Macht, David I. 1471. [with J. P. Greenberg and S. Isaacs.] Concerning the influence of antipyretics on the acuity of hearing. 1484. Concerning the toxicity of acetanilid and bicarbonate combinations for muscle-nerve preparations. 1496. [with C. F. Mora.] Effect of the anesthetization on the subsequent behavior and intelligence of albino rats. 1520. A pharmacodynamic analysis of Straub’s morphine reaction. 1521. [with S. Matsumoto.] The action of prostatic extracts on isolated genito urinary organs. 1530. [with Y. Satani.] A study of local anesthetics in respect to their antiseptic properties. 1547. [with C. F. Mora.] Effect of opiates on memory and behavior of albino rats. 1577. On the comparative toxicity of some alcohols with especial reference to isomers. Mackenzie, George M. 1533. See Longcope, Warfield T. MacLeod, Grace. 1527. See Rose, Mary Swartz. Maltaner, Frank. 1560. See Wadsworth, Augustus B. 1579. [with E. N. Hoppe.] Osmosis as a factor in the local accumulation of leucocytes in the animal body. Matsumoto, S. 1521. See Macht, David I. Mendel, Lafayette B. 1486. See Osborne, Thomas B. 1567. See Smith, Arthur H. Miller, Raymond J. 1517. [with Olaf Bergeim and Philip B. Hawk.] The influence of anxiety on gastric digestion. Moore, A. R. 1481. The action of camphor on the central nervous system of the squid. Mora, C. F. 1496. See Macht, David I. 1547. See Macht, David I. 230 SCIENTIFIC PROCEEDINGS (108). Morgan, Thomas H. 1501. Castration of hen-feathered Campines. Morse, Withrow. 1532. See Hite, Bert Holmes. Muller, H. J. 1465. [with E. Altenburg.] The rate of change of heredi- tary factors in Drosophila. Murray, H. A., Jr. 1556. See McLean, Franklin C. Myers, V. C. 1559. [with J. A. Killian and G. E. Simpson.] The influence of phenylcinchoninic acid and its methyl derivative on the uric acid and urea content of the blood. Noble, Willis C., Jr. 1561. [with Ruth A. Thomas.] Observations on the immunization of rabbits with single strain and combined multiple strain vaccines. Oppenheimer, B. S. 1554. [with H. E. B. Pardee.] The site of the cardiac lesion in two instances of intraventricular heart block. Orr, Paul F. 1487. Some observations on the biological characteristics of Bacillus botulinus. Osborne, Thomas B. 1486. [with Lafayette B. Mendel.] Do fruits contain water-soluble vitamine? Papanicolaou, George N. 1537. [with Charles R. Stockard.] Effect of underfeeding on ovulation and the oestrous rhythm in guinea pigs. Pardee, H. E. B. 1554. See Oppenheimer, B. S. Paton, Julia B. 1495. Enzymes of pollen. Pearce, Louise. 1548. [with Wade H. Brown.] On the generalization of Treponema pallidum in the rabbit following local inoculation. 1549. See Brown, Wade H. Pease, Marshall C. 1478. The bacteriology of infectious gaseous gangrene. NAMES OF AUTHORS. 231 Prince, Alexander L. 1568. Observations on the physiology of the otic laby- rinth. The influence of prolonged rotation on the duration of postrotatory nystagmus. 1569. Variations in the affinity of hemoglobin for carbon monoxide in health and disease. Rettger, Leo F. 1562. See Cheplin, Harry A. Robinson, E. S. 1563. See Winternitz, M. C. Rohdenburg, G. L. 1508. The isoagglutinins and isohemolysins of the rat. Root, Lucie E. I519. See Lewis, Howard B. Rose, Mary Swartz, 1463. On the utilization of the salep mannan. 1499. [with Rena S. Eckman, Edith D. Brownell, Edith Hawley and Ella Woods.| The utilization of the calcium of carrots by man. 1527. [with Grace MacLeod.]| Some human digestion experiments on raw white of egg. Rosenbloom, Jacob. 1476. On the certain dietary factors to be considered in the treatment of cases of hyperthyroidism. Rous, Peyton. 1498. [with Louise D. Larimore.] The relation of the portal blood to liver maintenance. 1544. [with Philip D. McMaster.] cabccds ei teeks sium deen eReRee 25 East 62nd St., N. Y. City. WOAMAEY; TARGUS... Na: aiid on ELE ire bee Cornell University Medical College. PASTORS, i Ss bs ine ee aS he eRe PRL See a Cae Yale University. BALZO, Pei Pea te i watinks & bao 24a ca eee on Fleischmann Co., Peekskill, N. Y. BANTA, A. M....... Carnegie Institution, Station for Experimental Evolution, Cold Spring Harbor, Long Island, N. Y. Bastar, BOWIN Jos sch vin te adecees ceed Department of Health, New York City. Banwen, We Tass iit akc Ass Sack pia dae ire one ke Re eee New York University. Bannon; FeOe Gh ovida du obs « civtdoweninc heb babu aa kee Yale University. Bani; COAGLBOR A. os sca seceded ses chscaneneteeas University of Wisconsin. Banrveers, GRINS Doc vee vck Manwatwedsemewnt ean Leland Stanford Jr. University. BAUMARK, S40ER, 4 owiienaet chine dada tetas PAE SEE Presbyterian Hospital. BREDICT, STAT WR, igs vie eisnie gas hb) Sain Cornell University Medical College. BERG, WUATAM Noise sccnacauss Bureau of Animal Industry, Washington, D. C. BORG, Oi soci iat wadav wn catears Jefferson Medical College, Philadelphia, Pa. PE, AVES vies cc da keer boas Reamer ee reeks University of Pennsylvana. RATIONED MEMMARD oop i pc ccie@eeieads ctbewsenbivl thaunh heap eee Germany. on FO ee Sere Dominion Laboratory, Winnipeg, Man., Canada. BLAKESLEE, ALBERT F.....Carnegie Institution, Station for Experimental Evolu- tion, Cold Spring Harbor, Long, Island N. Y. BLO0R, VE Riss sio'c'c con uae ho pee Mek ded et at caine University of California. BORER: | FE EAAN Sei wiceidcoed babotioueades Hygienic Lab., Washington, D. C. BRONPENBREMMER, JOCODscccciccesvavevantidcuccéeds Harvard Medical School. BROOKS, TIARLOW .. ose nccttal dee eecvrdsatend laws ss ....-New York University. 238 ROLL OF MEMBERSHIP. 239 BROW WAI Blind ddd 6 bts isne'cs Rockefeller Institute for Medical Research. PrEEs A EAD Wei eS tok bea ewe pws ewiee's gales College of City of New York. I a le Le od case inivincare pe bieiaeeecelele Johns Hopkins University. NNN AR OR re Dyed la 2 a 5 ao. my, , 4, 6) 6) 0s hs, 808 ehegarele Kua. a, University of Wisconsin. DeRRMORIER MEL OC SC Aki Gs J liawaee es faba t ba eae eyes ees University of California. PEROT NEL Le eas Lae Oak bale ce nees Washington University Medical School. ee eee Peres ER STEELE b's 8 A WIE hsb ype meas b oe Bale wiwo ow al ate Columbia University. Ue NEON 0 AUG EN ra sau! oe inate el ae weet te watereete oles os 135 E. 34th Street, N. Y. City. ee eben tks Niet haw Oss od bic Bis aie ieee aes Ae bmn de WiciahD ace Columbia University. aes Di AME Pig eB SOUSA alae ea ca eie mah ye oa Harvard University. RE eR eo ce ee ne Dts Oe Uy keane ad aay University of Chicago. DA ALBIS oho ae Sa Gale pid Picola Rockefeller Institute for Medical Research. SCO 00 ey | © (a ae ae er oe 13 Spadina Road, Toronto, Can. SiN MI catia Ae toed irre i BAS ae dad bw ne cee 4 eyes Bellevue Hospital, N. Y. City. On ky oh | ee ee New York Post-Graduate Medical School. SEA sy TR OISISE wk oss i5-p ob pin iene wn Cornell University Medical College. pera oo) Dane OSB sie Caine» AER ie dean a ae Rape St PaO a Bes West Virginia University. RR BE ie cd a kh nce coches Gare Qaeda ely uk ames es Yale University. A ge cee on ie lure ek Oa Uk wis eiaatale @ es gh btn ely Yale University. Grant, P. Bice aes TNE ATER ES SENG ONLY ERR Reo aredE RS University of Wisconsin. 2 Oe Soe, ee ee FR eiad Soh Fete Eli Lilly and Co., Indianapolis, Ind. eee MN eh heres ea aye Cornell University Medical College. COOEN, TeORMEET (02 si as ker ileccreenet an Hygienic Laboratory, Washington, D. C. SBE UMAR ETNN Soe bc ss o'er sie'sle ga 8 a New York Post-Graduate Medical School. oy OF BG a Rockefeller Institute for Medical Research. ee eRe HOEe Poa Liat wu twiar cls oA Sak pe lags WOR University of Wisconsin. eee EONS Solve cee ec eke gee swale Rockefeller Institute for Medical Research. CINE TON ho eS ili we te Ohad hae Aa he beleewme chebe New York University. A ELAINE Pee ges Ge Se AS a bbb barb ere ears University of Buffalo. pee Se IN Nag eG Lire sh ehh ens hh beble eS b ad bao Princeton University. Ces Boers cos Oe ek SKS ee Washington University Medical School, St. Louis. Mee Ee Ce eye thine ete ceca heed tateaede wales Columbia University. ee SCG NE SAS La Set aia o eiateaetpiela elves lela es wes Johns Hopkins University. a TAQ pa a Aaa REDE Bp eS RRS SE ALE Se Se oe A a Johns Hopkins University. Ie WV A et Ath ah te taladseatalsteacdds Harvard University. Ceaseron, ‘CC. WARDS. i.e es es Department of Education, New York City. OR eT Coin ceinie sooo o's kOe Caletnlend ows Leland Stanford University. RRs PAIRS WV wea chi ea ae oc tea pie ales Western Reserve University, Cleveland. SUSHING, HIABVEY. 54555055 see vee Peter Bent Brigham Hospital, Boston, Mass. Semele os 28 2m so Ne bie Nose 4 oad Oeee Scarborough-on-Hudson, N. Y. Be UT AGT Foie Ban 5 2k oo! ale letclelclotetnie's dale be late a wtavaternce’s es Johns Hopkins University. DAVENPORT, CHARLES B.. .Carnegie Institution, Station for Experimental Evolution, Cold Spring Harbor, Long Island, N. Y. ND SRN eS Clee the oss A Wie tah Weta ete oP thn e's Stanford University Medical School. IGRI ARR ee OL a isn C8 eee wR a bea Seren Johns Hopkins University. DIGMATDSONG Ed, Eis eke eh PES: Wistar Institute of Anatomy, Philadelphia. PORAPERS NGRORGERS fh 8 soa Vise i elotatel Presbyterian Hospital, Columbia University. PIRAPEA STL aot e eee eS ENS eee ae Seales 23 East 38th St., N. ¥,-City: 240 SCIENTIFIC PROCEEDINGS (108). Dewmmiace, Bi eos. cee JUSS eet Albany Medical College, Albany, N. Y. Dope nt 038 PETE eae veronica Cornell University Medical College. DpGGAR, (B. Met. SSP i cb cies Missouri Botanical Garden, St. Louis, Mo. Dusan, Enwarp E..: ooo. cle sc csicnn eS aeenseee 35 East 68th St., N. Y. City. Dervan, Cranes W..). Joc deoe oAddaree Tulane University, New Orleans, La. Keer, Wattee Siokk so Mo ese casas k aes eee eee Columbia University. EDMUNDS, Co Wo iid be © © Sides University of Pittsburgh. C8 EOC RSE Ae RE pach ae Een ek ay eee ane Cr University of California. ere. SelM OCS OG. alae Vices eb ea wah rs Few ene ee er Columbia University. Cc Maree rt 1, ocliS. o6 ona aye vs diode oaeuuas 1805 N. Carolina St., Baltimore, Md. 2 eh Se a Rockefeller Institute, Princeton, N. J. pe EM a Se a ee ee Johns Hopkins University. cen teaitass “ANE SS ns be See, PS a Se sc bkeld Ss bie bea Johns Hopkins University. Pe Re Re ee PAS eras cron WRN aint aS ab wierateie d University of Michigan. BRS Ege, SSRs SP Pcie ere ys oe ee a Harvard University. Cs RBA aa St a ae ee ae University of Toronto. (ge OP EL: Oe ae ee oe a eee ae University of California. NIIP tad Sapa cre Se A are University of Cincinnati. ener SIME MINS Me eS ass acc eit ad vleieaie due es oe New York University. eM WE URE AS aoc so 5s wisleseis wae s vc Rockefeller Institute for Medical Research. IRIS AR EM ne Ratt 2 fave ules e< wet d'ccia alee as Memorial Hospital, N. Y. City. SADT, INELSON W... «5. «onic c.o ca wee ues Cottage Hospital, Santa Barbara, Calif. MEMES SOU vans a San os ab ewe th Ga aw 8 wate’ 0 Wo Johns Hopkins University. Ge EMME ANS toss Ooo alin Dk eo oE Nido w wcich PCOS be eh ees Columbia University. OMEN SS erate Sok be uaa 6 ae Rockefeller Institute, Princeton, N. J. CR PE SON OaE 2) ee a a aE ee eee .,.-Johns Hopkins University. Ce BS ee a University of Virginia. POSER S SMT Pe che oui TSE Seog os St. Louis University Medical School. EME BRA es oer n okies abd CRT Ca ietea ee Beth Israel Hospital, N. Y. City. EC asirare, Ge ees a a eae a bs Michigan Department of Health, Lansing, Mich. 242 SCIENTIFIC PROCEEDINGS (108). TAME, Bas Red oc SOAR ME UD USC SR ESLANSRABERSESS Lakeside Hospital, Cleveland. RAGE) RAMS OO Pee UT eR Ss CE New York Post-Graduate Medical School. Weenie, Va Ea ee or bk ir eee Stanford University, California. Bean, JOR AL 25 oo ro ese aoe New York Post-Graduate Medical School. Rees, Ate AL oot ack woenne seen St. Louis University Medical Schaol. Biustenem BART Fo. os. ove ch eee State Hygienic Laboratory, Albany, N. Y. Beem. 1. Solder akc se esp wes Le eea eee Flower Hospital, N. Y. City. Reeeee, B. 3.. 505 £25 uvsemoerseae Rockefeller Institute for Medical Research. Bee, as Sass os Ge oe ee cee ee Montefiore Home and Hospital, N. Y. City. BETS, OGRE. 6 ac iwc} hc higne hee < oes t ee University of Pittsburgh. ERWUEI, BUSBUR. . Nios bs ox pigeaeteet Albany Medical College, Albany, N. Y. Rimes Oo Fh: ks cob bins ad aee eee SQUIBB, E. R. AND Sons, New Brunswick, N. J. BAC EE Pla. oe hag side eek ae ees Trudeau Sanatorium, Saranac Lake, N. Y. moworm, (Canta Ay occa np ckinald we cathe we eee University of California. BOMB. Pi aks ass g ba R ET ETc MOU eae University of Pennsylvania. Umea DD, (SC. Sok ve Ge nda = bee tae eee eee University of Pennsylvania. LAMAR RICHARD Wie sibs oo e iwc se ceases 8 eee Eee ae ae ees University of Virginia. Tapas) Both a5 ish ss eee Peet BR Adare Sei beens Yale University. LAUGHLIN, H. H.. Eugenics Record Office, Cold Spring Harbor, Long Island, N. Y, LROURENG, TIRE OT Ob 6o eo venir nb be eee: abe bene Yale University. Peers 15s: Tai 0 a nk hone deka bidanoewere Sheffield University, England. Rie! SIR Bo in nk Sa Ab od ee ene Rew eeeb eee Columbia University. Sees Pe is eta bh ke be banee Rockefeller Institute for Medical Research. Pes: SOAS Dee Pie Wan cae eRe erm enteetateeeee Columbia University. GN, as. Sa CREE. od ak nbeac daeheaxh been cums banaeees University of Illinois. we A Ba a sn Ab Hae eae an es variews Phipps Institute, Philadelphia. Rage, CGS ao 5 ets ad en ee Bae cee oaks Ghana bhakees Columbia University. Live, Vee Bik 5b i Sendo d 26 a ne vai dle st mines University of Chicago. Lwieee, TALE Gre Fo Apes soe wredsn sen Clark University, Worcester, Mass, LittLe, C. C.....Carnegie Institution, Station for Experimental Evolution, Cold Spring Harbor, Long Island, N. Y. Lt SAAIORD, shir 5 oo ke whaithode Ken wee Rockefeller Institute for Medical Research. Lec LS Sab hobannwGsuiny bbe ene Washington University, St. Louis. LOR VIIIAIT, PARTE Gen cd 5 0 baat noe iaa en eh ices University of Wisconsin. - LOMBLRD, WARRIIN Pik iat tis beinteedd Chak seh va twee area University of Michigan. LONGCOES, Vis Riwacwtadddsane ewan Presbyterian Hospital, Columbia University. LAK, Wis a TAbSs van pies pie 6404 kao oidaut treads keke University of California. LOCERARIT,, AWG Ties an wos gends pe csivemdein cde ee eee University of Chicago. LAINDOGUAN, CSMIGTIAN oc dc ttkbeuhe eerie University of Copenhagen, Denmark. CRE COROTEASE : yo & nin giv praia ddd ean Cornell University Medical College. NG RR on a's ate! oii’ 6 9'o 0.0 b's bin eek 4 ORE 5 Raab ee Roosevelt Hospital, N. Y. City. Ry Behd snc acdnee pata babredeisak (sake Meeker University of Minnesota. DRRCAEE SING, hn Whi $ 2h bbc Gactiasetiectaneases aan McGill University, Montreal. ME ADCAUIO ERs. le tins ves rwrideeeburonsawetiieen Johns Hopkins University. MACTIOUGAE SEP. has vob's vic det ie pe hnnendes Desert Laboratory, Tucson, Arizona. MacDowELL, E. CARLETON... Carnegie Institution, Station for Experimental Evo- lution, Cold Spring Harbor, Long Island, N. Y. EACLE DO, FF o Miss bu bah pw whine has betes abalone datduaes ee University of Toronto, ee a - = - ee . | ROLL OF MEMBFRSHIP. 243 pe a se New York Post-Graduate Medical School. ERO WFP as Se nese v indus dadedoucevevenes University of North Carolina. BC IT Fas ee wba sdb c ba owls wees Robert Brigham Hospital, Boston, Mass. MeRomnths PRANMIAN Cai ek ee peek eben Peking Union Medical College. TS Be) Ste ee ee eee aes eae St. Francis Hospital, Pittsburgh. Re SEMEN Cis acl ic sag toile le bo oie loleini’s Mine Wh Johns Hopkins University. Se NEN SEW Se Soy ia a vs 0 08 oo hw eed ae bee New York University. UNNI A ERIN Se Wig 12 gods we how 4 te te he Wi hn era ne We See leh ee New York Univers!ty. RrerwaARnwia, W.. Tesi tes iiideasddsaacess Leland Stanford University, California. TEARING; DAVID? oi cide 04 pe daeeee Montefiore Home and Hospital, N. Y. City. Pes oe Sede sk ea eK RR ROL A SSR bitetivs babys University of California. MOAVGR,; ALFRED Geiss eee dea wkts Carnegie Institution, Washington, D. C. Tees LOWARD Bi, sn nd 8 1445 Rhode Island Ave., Washington, D. C. coy ER OS BS ee eee eee Rockefeller Institute for Medical Research. De oe oa what crn toh do oo ae al seh a ial bea ate! Dw Bhahs Yale University. re ese CS Sc eda kip dab lds cok ade hee Columbia University. DS APR Tk LAS ae Ge eee Cee me uwae hos Johns Hopkins University. Berens ae eee ed Se So is ie hd ASD oka ale cele ds Johns Hopkins University. Pare, GUSTAVE Mw... 5 eta eel Rockefeller Institute for Medical Research. Ee em ener Pek ok a tL fch wos bees bbls University. of California. Bre Ba Ps ds Ba = be x eas Bureau of Animal Industry, Washington, D. C. ES SR ee ee eee ee v3s2 oe, Rutgers College, New Brunswick, N. J. I 2 NES Rs sia we bcorcatee c¥odn bs & oo ewaiclaw aes & Columbia University. Ne eM POM 1 os civ baaiviardia 4 ana de ok alee University of West Virginia. Peers, Fireman, Ou. . 2.5 bcs ccu ccs = bee cen mabe 49 East 53rd St., N. Y. City. PR AD EMIASE Peo oe GL et in cine do oe bso bo wees University of Texas. IMI DUET OTS S02 507d 5 2. uiia tala mia dase e aca bs alee ie Sw ae University of Rochester. Merete, CAMBS Fos og oc bee we Rockefeller Institute for Medical Research. PRES BERL GMs SEEN ie ce Sb cs va eee ete eae cee ws 262 South 21st St., Phil., Pa. MUMMERS Roc. Se Uk AAs wb ees wo eee New York Post-Graduate Medical School. ey Wi aR TOR 5) crete shu twrotaralalalgtetcie’q'ersto's Cornell University Medical College. Pe SPT e re oe We ee eo AE aloe lnlevete tw ints une ate lero New York University. WO, ERIEVOL dan te AA Rockefeller Institute for Medical Research. pO Co gS ea ae cee oc Medical Director, New York City. NWorrneer,s JOHW His ooo 2 5 oo ca d. Rockefeller Institute for Medical Research. coats Sr renee ts es 0 a eb lek edu b asp eed eicee University of Michigan. ae PEE SP ees bee ceictn bk be bis htiahiase nwa’ McGill University, Montreal. SISRT, PRIGR Bo osc boven eens od Rockefeller Institute for Medical Research. Pee CIRM eh dee dae woods ee enecdaued be Stanford University Medical School. Renae ies VU RELAIS SE Shoes oo aa ak oh alpen tehalclepabai Gare @erere Leland Stanford University. RpnNnG EUNUGMINIE ls fa 2. Snide w crane beer ae hee ee ees Washington University, St. Louis. Ser eeO OME ES Sees ee aides ee ees ha bie vhecuddageeemare Columbia University. OSBORNE, THOMAS B............... Connecticut Agricultural Experiment Station, New Haven, Conn. COST ee a, Wakes eae Aa toa 08 ee te Os bddkesb nvewe Harvard University. OETENDERG, Boe Jo 2k 16 si eit ke eke eased Mount Sinai Hospital, N. Y. City. PRE MGR) TOMIPERIE ANS fo aoc Sale ods bata Sik eS eee oe hea oo Johns Hopkins Hospital. 244 SCIENTIFIC PROCEEDINGS (108). Parmginuiume: ALwin Mil ooo os pals sea ca ee aaa Columbia University. Pads. iol bnew leh cos ap mee seeee eee eee Johns Hopkins University. PAwmE; Wiiiahue Fic 5355 Jase et btestadtee Department of Health, New York City. PAsReR SsB0RG8 H..). sisted ook, CASA a Ree ee Harvard University. PEABODY, FRANCIS Wii. i cide in aed Peter Bent Brigham Hospital, Boston, Mass. PrAmGH,) LOUISE iid 0a 5 Fs a ba Rea Rockefeller Institute for Medical Research. Pees, | FLA VON iS i gs oa. eee ae ee Johns Hopkins University. PRM SM ARSHALE Goi ses 545 oor ee wee N. Y. Post-Graduate Medical School. PEMBERTON, RALPHA Ss Gsis ccs ch Saweelebs ate ieee 318 South 21st St., Phil., Pa. Prereasen WW. Mesccicd ak ets packets 31 East Elm Street, Chicago, IIl. Perrss, ©. F. PRRRs ce ees been eee University of Pennsylvania. PEARR, (Eiosiicndiale ok eels hehes COREE tee a eee eee Harvard University. PYRISRER( J: ; hd Baca Sak aes Mean oe abe 1421 Edmondson Ave., Baltimore, Md. Price, (Fis Fag 6 63: é tdomcdl tad. ee iodeatny Ses ee eee eee Columbia University. ETIEE EIARIE oS sn on poe PORES eee ee Dae ee Mt. Sinai Hospital, N. Y. City. Pores: Wairtdat. Tie a a basen a thee Harvard University. Pratr,: Jusmer bls. eects ee Rte Scene Satire eats eetatsate toaee Harvard University. PRInes AD Lsis ahi when as co Fob meee 4 Wilcox St., Wethersfield, Conn. FRAIZISS, TaRORG Wa, os bic dinates sie ek Research Laboratory, Phil., Pa. RAVENEL, Magers Po ons ccs elias ake toe eres University of Missouri. ICH ERT | TOWARD Ticino 005 6 as ek ini ae Se he eee University of Pennsylvania. BMITGER, LEO Bos cis ens odes ck een en te he hee eee Yale University. RICHARDS, ALBRED: TN. .:ci0u chs ss ous Mike eee ea eee University of Pennsylvania. BACHMARDS, FIURREGT D6 oo oyu ofan oa a ae Lea Cee ote Columbia University. +s Be a Ree Station for Experimental Evolution, Cold Spring Harbor, N. Y. MISES Ss Mane 4 ns 4c wo eRe Re iin a oe a 141 W. 78th Street, N. Y. City. ROBERTSON, T. BRAILSFORD...........- University of Adelaide, South Australia. TIN, es RMI sin os gis: d Wnt yee ote 211 W. Madison St., Baltimore, Md. ROSE, ANTON Teo. '3s ido ase eine hide wee ae New York Post-Graduate Medical School. ROSE, BEARS Tec! Ax vidin cies thi wes ak ped Cohn nie wee SRR eiaitcaee Columbia University. ROSHMAU; BERTON Bits biieiad Sows pew rnee eres Wa ee eee Harvard University. ROSENBLOOM, JACOB Si. ii ct tawsees Western Pennsylvania Hospital, Pittsburgh, Pa. Rore, Gropiene Be. ios ow oes'bs ds sll nee Hygienic Laboratory, Washington, D. C. ROTHSCEIED, Oi Aoi Eh ds ee bh cen bee ae tan Mt. Sinai Hospital, N. Y. City. ROUSE, PRIOR ook ode 2 kj ea bt obtlaeka Rockefeller Institute for Medical Research. RUM AG la ithk esas 4 acatedan Ga Scovill Manufacturing Co., Waterbury, Conn. SALAM, FV ALTAM. V9. oi tate 5 5s eee weeee aan 133 West 122nd St., N. Y. City. Semi ons, OSC6 is Ws ids ihe an Rated dekh tone A eae 39 East 61st St. N. Y. City. CMEISOR AE 5 I ns v3 ha vials BAKED ee a eee University of Minnesota. MNS CARE, Bos lad ein Fi eis ad ea ke teens University of California. Coo Re ge - RO ee GE re ee eR eee George Washington University. Peres, DRIES io bee cscceedhes Kastanienbaum, near Luzern, Switzerland. ete Te Bas 652k os baw iietton dc ocak Raa lc itis eee eee Columbia University. Ei Rhe Sais ino nM ce bcd hod bbieteaenea koe eee College of the City of New York. MOVER: Mi diel adecaapevae Western Reserve Medical College, Cleveland, Ohio. ees Bie Di bia sil baitive okie ea ORNS a 0nd 8 Ce eee New York University. STARVE, PMI Bins occ initia i cake aes Washington University, St. Louis BRAEILE, A Olina snkene tse a Baylor University Medical School, Dallas, Texas, ROLL OF MEMBERSHIP. 245 EN Boa. viehaiwsnieieleis miele vivicstinine sae avienssaas Columbia University. EE ie pio wd nah wake one oe he Pisjessieiee Fordham University. SHIVE, J. W...New Jersey Agricultural Experiment Station; New Brunswick, N. J. 0 OS Oe ne aoe Office of the Surgeon General, Washington, D. C. TE TI Se oak. or hccksasus ono mnie Ok ila Qua deans University of Maryland. PE CRITIC REAND) 0. cies nec cea wens oases Cornell University, Ithaca, N. Y. on PS GB SS, | te tp al tre geen ene tee Columbia University. EN UI MAIO aS Narn cs as 8, gi Os iar winced bo 2 Sy o ines Bhat mcedepegnr ee ol ay, ale w Yale University. MEN, bE TINMREES, Beck Weert etre ed cbinite Wit WES Fad caeea.e University of California. SMe PPORGBRES Ss o teed fe doves: Rockefeller Institute, Princeton, N. J. SORE Teme reese se Bue 3% Western Reserve University, Cleveland. SPastit, REYNOLD ALBBMCHE josie ccna sc cacnscees Johns Hopkins University. oS ES 2 er ee ae ee, Oe SE ee Desert Laboratory, Tuscon, Arizona. Pe EOE ee ods eke awe a caaac anes Flower Hospital, N. Y. City. Serpette) Bo is eis ead cece s Western Reserve University, Cleveland. Pe ENE Gore ee A et eg Bo Sr eS a wap Harvard University. SIrecmanm, CHABERS Beoyiii enc k oo ees ns eee Cornell University Medical College. Sroemey, Lv eAw se kc. oss University of Southern California, Los Angeles. DEORE; ; PHOMASIAG GC. 20 se. Gk 1800 Virginia Ave. N.W., Washington, D. C. Renee ees POS aos) hae ad neh re eae awe e ehh Harvard University. PERO, CHUEIAON 0 55 oils cin ce cae ai weies 104 S. Michigan Ave., Chicago, Illinois. NNER MER Clie 5% ba wes atone he eo age kobe ees a> Columbia University. SINE Stal ce as aa bin oie Pale kk be < Stanford University, California. EL EDR EM Ss eae bs vob nce ae ees paw he aw ee A _...University of Pennsylvania. MRS Se IRN Sk ah ssw iudein aber ahaitecore-o we WA arr de ne emake « Columbia University. REE UMURTOE AS Oona bu wt kaa Co ete ee Nin eB we ee hE eee ees New York University. NN ON EE ce ok wk eM ae Chew ccen ee beuns Johns Hopkins University. pee NS 2 0 OURS eae otter eae ka a University of Cincinnati. eM PURER TEC) oho aid! cia ave Seas hicks oe Ew Ae ape eK University of Pennsylvania. Rie Ee or a oe ahah Sais cee een N. Y. Post Graduate Medical School. IEAN PRMD er a a's ok Wood hee tee wes wee eae N es Columbia University. eee ee MOAB ect aw ees cas kena bee cee ee Peking Union Medical College. EERE aa oe oa awit es Coa hee ss Vanderbilt University, Nashville, Tenn. RUN PMT SINOE WN corn ane cae a cate ate ewes Ma bw as woe eu pews Columbia University. SR Ren CAS Rahs kiak Cm ck mae ioa ee wie Cornell. University Medical College. RE MEI Caso Soe crn aS Se kee oP ene McGill University, Montreal. SRE a ig PERE lah prcla oh twinln ites oaks Sinle Oot. we Cornell University Medical College. Pn eee in ep Sy he pio Rie inva Resale alyie a RAN Ge 8 WR Harvard University. UBLENBUTH, EDUARD. 6c... ccs cece ote Rockefeller Institute for Medical Research. MES MEP toot ow 2s va aaa 6 op OR RS eee ek Cake te Yale University. Va atwen, DONALD D:.. 5.2568. 25s: Rockefeller Institute for Medical Research. WADSWORTH, AUGUSTUS B............ State Department of Health, Albany, N. Y. WaxsMan, S. A...New Jersry State Agricultural Experiment Station, New Brunse wick, N. J. WeeeRAC a, Serre Ee ah oS eo ack aa Ske Se ae Soe ae es New York University. Re Pe aes ae oa ee ka sce woe ie Seahawk ee Nee University of California. SEPT Gs PUR NYNMEERS ne teresa oa a dik OMe ad a Gee Wiebe sos University of Michigan. 246 SCIENTIFIC PROCEEDINGS (108). NVARSIES ELL cc hk ekki cece aire an RAVER ERE EERE University of Toronto. Wramarane i ce chy eae et ba eos te ene University of California. Wy SABBCOMABE ES. 54 idl os shea ee Ok Ca Be University of Pennsylvania. Were rt, WV ELEITAM EL os so aw Oe BORER See Johns Hopkins University. WEEEER, WILLIAM Bo! once ee ask Gee CES ON EAE eR RE University of Illinois. WELLER, ‘CARL ‘VERNON. ics ovicte tne oP pet ala eee University of Michigan. WH ESE, Boe: Be hal Leite ek lekebase wht ehaketa ates 30 Charles River Road, Cambridge, Mass. Warrrcs, 4s: ES os eee ek swe eae heen e Ce paiement University of California. WHITE, BENJAMIN............: Antitoxin and Vaccine Laboratory, Boston, Mass. Werk; 0: Be esse ee Tee Brooklyn Boganic Garden, Brooklyn, N. Y. WiceErs Cos ies. oe eae ae ee Western Reserve University, Cleveland, Ohio. WVaL Liaw, Aaa WW, 5S ih enc Agamne Department of Health, New York City. Wirrass,; Be Bo Se) 2 Soa ee oh aa ene ae a Columbia University. WZIAMS, LLERBERT URIs 68s Ske he bin ioe ee University of Buffalo. WILSON, ESDMOND BB. 2 iS Oates Bae ee 2 os Ole in ote oan delaleha ear = Columbia University. Wascow, Co Ase. cere users CR eee eee heehee eats eee Yale University. WeLnacs,’ 6.) BURT. ENE BASES ie cb ons OS AB RE Harvard University. Wor, Charis Goold §. 5500 Ses ee a ie oe Se eee Cambridge, England. WOLLSTEIN, MARTHA: if. 62.6. 0 eS, Rockefeller Institute for Medical Research. Woon; PRANCIBIGE.. dashed pare hls Ube aaaieenaees Columbia University. Woonrnvusrs; Loranpe 1,058) 6 6 si te as dona chaos tacaaeemeee Yale University. WATS, INAGHUUE 6 he e's Paz 6 eee ad kOe et cee ee ae eee University of Japan. Wemers, ROBERT Mio. oie id oleae es ORR 1864 Park Road, Washington, D. C. Ce (=o ae, i eee ee eee eee er Department of Health, New York City. RIS RAIA ein 5m 6 fa'p'a nlalatn tata etn al la’ al ale os tee ets Ol ache Columbia University. Total number of members at the close of the academic year, 1919-20: 338. — —_— President Vice-President. .... Librarian Treasurer Secretary President Vice-President..... Treasurer Secretary President Mt B66 Ue id Se elce ce Vice-President. .... Sec’y-Treas........ Additional members of Council!...... 1 The Past Presidents are also members. 46a ee, a eee a Oe dah e Ss Seem e St 6 oe ee (6 oe fe teri, ee e } OFFICERS. 1903-1920. I1903-"04 1904-05 1905-'06 Meltzer Jackson Gies Auer Meltzer Ewing Lusk Calkins Gies IQIO—’10 Morgan Gies Lusk Opie IQI6—17 J. Loeb Gies Jackson Auer Dubois Wilson Dunham Lusk Calkins Gies IQII—I2 Morgan Levene Lusk Wallace 1917-18 Gies Auer Jackson Dubois Wallace 247 1906-07 Flexner Dunham Calkins Gies IQI2—13 Ewing Levene Norris Wallace 1918-19 Gies Auer Jackson Wallace Sherman 1907-08 Flexner Morgan Calkins Gies I9I3—14 Ewing Field Norris Jackson I9IQ—’20 Calkins Wallace Jackson Sherman Jobling 1908-09 Lee Morgan Lusk Gies IQI4—I5 Lusk Gies Murlin Jackson 1920-21 Calkins Wallace Jackson Jobling Hess CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, Thomas A. Storey. Columbia University.—C. H. Bailey, A. K. Balls, Russell Burton-Opitz, Gary N. Calkins, George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. T. Longcope, Charles W. Metz, Thomas H. Morgan, Herman J. Muller, B. S. Oppen- heimer, Alwin M. Pappenheimer, 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. Cornell University Medical College—Harold Bailey, 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. Torrey. Fordham University School of Medicine.-—A. O. Shaklee, Carl P. Sherwin. Hospitals, Beth Israel—Max Kahn. Flower.—I. S. Kleiner. Memoria. Hospital—H. H. Janeway. Montefiore Home.—George Fahr, B. S. Kline. Mil Sinai.—George Baehr, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Rothschild. New York.—Nellis B. Foster. Presbyterian.—Louis Baumann. Rooseveli.—K. G. Falk, I. Greenwald, W. G. Lyle. St. Lukes.—L. W. Famulener. New York City Depariments. Education—C. Ward Crampton. Health.— James P. Atkinson, Edwin J. Banzhaf, R. L. Cecil, Alfred F. Hess, William H. Park, Anna W. Williams, A. Zingher. Medical Director —Charles Norris. New York Post-Graduate Medical School——Arthur F. Chace, Martin Cohen, Ludwig Kast, John A. Killian, W. J. MacNeal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. New York University —W. H. Berber, Harlow Brooks, Warren Coleman, A. O. Gettler, Holmes C. Jackson, Arthur R. Mandel, John A. Mandel, W. C. Noble, H. D. Senior, Douglas Symmers, George B. Wallace. Rockefeller Institute for Medical Research.—Harold L. Amoss, John Auer, J. H. Austin, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, Simon Flexner, F. L. Gates, Walter A. Jacobs, I. J. Kligler, P. A. Levene, Jacques Loeb, S. J. Meltzer, Arthur L. Meyer, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. Brooklyn Botanic Garden.—C. Stuart Gager, O. F. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 819 Madison Avenue, N. Y. City.—H. D. Dakin. 248 CLASSIFIED LIST OF MEMBERS 249 I17E. 38th St., N. Y. City.—J. W. Draper. 126 E. 64th St.—Cyrus W. Field. 341 W. 45th St., N. Y. City.—Casimir Funk. 141 W. 78th St., N. Y. City.—A. I. Ringer. 133 W. 122nd St., N. Y. City.—William Salant. 35 E. 68th St., N. Y. City.—E. K. Dunham. 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, Albert F. Blakeslee, Charles B. Davenport, J. Arthur Harris, H. H. Laughlin, C. C. Little E. Carleton MacDowell, O. Riddle. (Desert Laboratory, Tucson, Ariz.).—D. T. MacDougal. (Washington, D. C.).— Alfred G. Mayer. State Boards of Health. New York (Albany).—Mary B. Kirkbride, A. B. Wads- worth. Hospitals. _General (Cincinnati, Ohio).—Martin H. Fischer. Cottage (Santa Barbara, Calif.).—Nelson W. Janney. Lakeside (Cleveland, Ohio).—H. T. Karsner. Massachusetts General (Boston).—David L. Edsall. Mercy (Pittsfield, Mass.).— Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francts W. Peabody. Robert Brigham (Boston).—F.M.McCrudden. Royal Victoria (Montreal). —Horst Oertel. St. Francis (Pittsburgh).—J.W. McMeans. Western Pennsylvania (Pittsburgh).—Maurice H. Givens, Jacob Rosenbloom. Institutes. Antitoxin and Vaccine Laboratory (Boston).—Benjamin White. Gratwick Laboratory (Buffalo).—H. R. Gaylord. Juvenile Psychopathic (Chicago).— Herman M. Adler. Phipps (Philadelphia).—Paul A. Lewis. Research Council (Ottawa, Can.)—A. B. Macallum. Rockefeller (Princeton).—Paul E. Howe, F. S. Jones, Theobald Smith, Carl Ten Broeck. Trudeau Sanatorium (Saranac Lake, N. Y.).—R. A. Kocher. Wistar (Philadelphia).—H. H. Donalvson, Shinkishi Hatai. U.S. Departments. Agriculture (Washington, D. C.).—CarlL. Alsberg. Bureau of Animal Indusiry.—William N. Berg, J. R. Mohler. Health (Lansing, Mich.).— R.L. Kahn. Hygienic Laboratory (Washington, D. C.).—George B. Roth. Surgeon General’s Office (Washington, D. C.).—J. F. Siler. Universities. Adelaide (South Australia).—T. Brailsford Robertson. Amherst.— Otto Glaser. Buffalo.—Katharine R. Collins, Herbert U. Williams. California. Walter C. Alvarez, W. R. Bloor, T. C. Burnett, H. M. Evans, F. P. Gay, S. J. Holmes, Charles W. Hooper, Samuel H. Hurwitz, Charles A. Kofoid, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker, C. K. Watanabe, G. H. Whipple. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati—D. E. Jackson, Shiro Tashiro. Clark.—Ralph S. Lillie. Copenhagen.—Christen Lundsgaard. Cornell.——Sutherland Simpson. Georgia.— Richard V. Lamar. George Washington.—W. H. Schultz. Harvard.—Jacob Bron- fenbrenner, Walter B. Cannon, W. T. Councilman, 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. Japan.—Noahidé Yatsu. Jefferson.— O. Bergeim, P. B. Hawk. Johns Hopkins.—John J. Abel, C. G. Bull, George W. 250 SCIENTIFIC PROCEEDINGS (108). Corner, Walter E. Dandy, A. R. Dochez, W. W. Ford, W. S. Halsted, William H. Howell, John Howland, H. S. Jennings, Walter Jones, W. G. MacCallum, David I. Macht, Adolph Meyer, Walter, W. Palmer E. A. Park, Raymond Pearl, Reynold Albrecht Spaeth, William H. Welch. Kansas.—Bennet M. Allen. Leland Stanford. —Thoma Addis, George D. Barnett, A. C. Crawford, E. C. Dickson, Harold K. Faber, A. W. Hewlett, W. L. Holman, V. L. Kellogg, W. H. Manwaring, W. Ophiils, R. E. Swain. Liverpool——J. G. Adami. Maryland.—Charles E. Simon. McGill (Montreal).—John L. Todd. Michigan.—C. W. Edmunds, G. Carl Huber, Ralph A. Kinsella, Warren P. Lombard, Frederick G. Novy, Aldred S. Warthin, Carl Vernon Weller. Minnesota.—R. A. Gortner, A. D. Hirschfelder, E. P. Lyon, F. W. Schlutz. Missouri.—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Carolina.— W. deB. MacNider. Northwestern.—Solomon Strouse. Oregon.—C. F. Hodge. Peking Union Medical.—FranklinC. McLean. Pennsylvania.—Alexander C. Abbott D. H. Bergey, J. A. Kolmer, E. B. Krumbhaar, O. H. Perry Pepper, Edward T. Reichert, Alfred N. Richards, J. Edwin Sweet, A. E. Taylor, Charles Weiss. Philip- pine.—R. B. Gibson. Pittsburgh.—C. C. Guthrie, Davenport Hooker, Oskar Klotz. Princeton.—Edwin G. Conklin, E. Newton Harvey. Rochestey.—John R. Murlin. Rutgers ——John F. Anderson, A. R. Moore. Sheffieldi—J. B. Leathes. Southern California (Los Angeles).—Lyman B. Stookey. St. Louis—DonR. Joseph. Toronto —J. G. Fitzgerald, A. Hunter, J. J. R. Macleod, H. Wasteneys. Tulane.—Charles W. Duval, E. M. Ewing. 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 Erlan- ger, Leo Loeb, Eugene L. Opie, G. C. Robinson, Philip A. Shaffer. Wisconsin.— Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenbery, Paul J. Hanzlik, David Marine, Torald Sollmann, G. N. Srtewat, C. J. Wiggers, West Virginia.—F. E. Chidester, Withrow Morse. Yale.—C. V. Bailey, George A. Baitsell, Henry Gray Barbour, R. H. Chittenden, J. W. Churchman, Barnett Cohen, Rhoda Erdmann, Ross G. Harrison, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, A. L. Prince, Leo, F. Rettger, Arthur H. Smith, Frank P. Underhill. C.-E. A. Winslow, Lorande Loss Woodruff. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer, 1805 N. Carolina St.— R. G. Hoskins. Cambridge, England.—C. G. L. Wolf. Cambridge, Mass., 30 Charles River Road.—C. J. West. Chicago, Illinois, 31 East Elm St.,—W. F. Petersen. Indianapolis, Ind.—Eli Lilly and Co.—G. H. A. Clowes. Kastanienbaum, Switzerland.—Fritz Schwyzer. Newark, N. J., Calco Chemical Co.—M. S. Fine. New Brunswick, N. J.—E. R. Squibb and Sons, P. A. Kober. Philadelphia, Pa., H. K. Mulford Co., 1524 Chestnut St.—T. S. Githens. Philadelphia, Pa., 318 South 21st St.—Ralph Pemberton. Toronto, Canada.—13 Spadina Road.—A. H. Caulfield. Tuckahoe, N. Y.—Isaac F. Harris. Washington, D. C., 1445 Rhode Island Ave.-—Edward B. Meigs, 1864 Park Road, —Robert M. Yerkes. Winnipeg, Canada, Dominion Grain Laboratory.—F. J. Birchard. Germany.—Reinhard Beutner. EE ———— 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.) Abdominal musculature, effect of varying pressures upon, 1538. Absorption, of fats, 1513. Accelerator nerves, effect of, on ventri- cular systole, 1516. Acetanilid, toxicity of, for muscle nerve preparations, 1484. Alcohols, toxicity of, 1577. Amblystoma, lens in, 1566. Amino-acid synthesis, in white rat, 1519. Amino-compounds, hydration effects of, 1479. Anesthesia, ether, blood changes in, 1550. Anestheitc properties of benzoyl carbinol, 1506. Anesthetics, local, antiseptic properties Oi, £532. Anesthetization, effect of, on albino rats, 1496. ' Antibodies, formation of, after disap- pearance of proteins from circulation, 1530. Antigens, purification and concentration of, 1560. Antipyretic effect of dextrose plethora in coli fever, 1540. Antipyretics, influence of, on hearing, 1471. Antiseptic properties of local anesthetics, 1533- Anxiety, influence of, on gastric digestion, I517. Bachman test, in vitamine, 1491. Bacillus acidophilus, intestinal implan- tation of, 1562. Bacillus botulinus, biological characteris- tics of, 1487. Bacterial invasion of respiratory tract, 1563. Bacterial suspensions, of, 1482. Benzoyl carbinol, anesthetic properties of, 1506. Bicarbonate, toxicity of, nerve preparations, 1484. standardization for muscle Biliary stasis, activity of gall bladder during, 1544; white, causation of, 1544. Blood, calcium in, 1524; 1528; changes in ether anesthesia, 1550; discrepancies in, 1576; estimation of lactic acid in, 1546. Calcium, in blood, 1528; in plasma, 1524; of carrots, utilization of, 1499. Camphor, action of, on central nervous system, 1481. Campines, castration of, 1501. Cane sugar, blood sugar curves with, 1477- Carbohydrates of the cat tail, 1485. Carbon dioxide, influence of oxygen in expelling, from blood, 1571. Carbon monoxide, affinity of hemoglobin for, 1569; elimination of, 1570. Carbonate content, of normal blood plas- ma, 1468; of plasma in chronic heart disease, 1469; of plasma in chronic pulmonary emphysema, 1470. Cardiac lesion, site of, in two instances of intraventricular heart block, 1554. Carrots, utilization of calcium of, 1499. Castration, of campines, 1501. Catalase, in tissues of mother and off- spring, 1534. Cat tail, carbohydrates of, 1485. Cells, reaction of, in galvanic current, 1565. Chlorides, effect of, on hydrogen ion concentration, 1462; in blood, effect of acidity of hemoglobin and the distri- bution of, 1556. Coli fever, effect of dextrose plethora in, 1540; in dogs, 1539. Coli injection, effect of pilocarpine in, 1573- Colloids, effect of solutions of, on the permeability of the capillary walls, 1567. Colon-typhoid group, method for identi- fication of, 1473. Connective tissue, in amphibian embryos, E572: 251 252 Crown gall, behavior of, on the rubber | tree, 1543. Cytoplasmic structures, methods for analysis of, 1493. Dextrose plethora, in coli fever, 1540. Diastatic activity, of blood in hyperthy- roidism, I514. Digitalis, action of, in patients, 1507; effect of, on vagus, 1480; on the con- traction of the heart muscle, 1545. Diphtheria toxin-antitoxin, refining of, 1541. Drosophila, appearance of, in, 1492; hereditary factors in, 1465; hereditary tumor in, 1490. Egg, white, human digestion experiments with, 1530. Egg cell, surface layer in, 1483. Egg production, effect of, on domestic fowl, I5II. Embryos, amphibian, connective tissue ground substance in, 1572; mammalian, effect of radium emanation on the development of, 1558. Emphysema, carbonate content of plas- ma in, 1470. Enzymes, compression on tissue, 1535; of pollen, 1495. Exercise, sugar and oxygen in blood during, 1531. Fat, production of, from protein, 1551. Fats, absorption of, 1513. Food, influence of hunger and tempera- ture upon utilization of, 1523; un- palatable, digestion of, 1518. Fruits, water soluble vitamine in, 1486. Gall bladder, concentrating activity of, 1578. Gangrene, infectious gaseous, ology of, 1478. Gastric digestion, influence of anxiety on, I517. Gastric hypermotility, reflex, 1542. Gene, vermilion, 1502. Glucose, blood sugar curves with, 1477. Growth, upon simplified food supply, 1464. Gum acacia, temperature changes fol- lowing injection of, 1574. Gynandromorphism, 1502. Hearing, influence of antipyretics on, 1471. Heart block, intraventricular, site of the cardiac lesion in two instances of, 1554. Heart disease, carbonate content of plasma in, 1469. Heart muscle, effect of digitalis on the contraction of, 1545. bacteri- SCIENTIFIC PROCEEDINGS (108). Hemoglobin, acidity of, affecting chlor- ides in blood, 1556; affinity of, for carbon monoxide, 1569. Hereditary factors in Drosophila, 1465. Hunger, influence of, upon food utiliza- tion, 1523. Hydrogen ion concentration, effect of chlorides and sulphates on, 1462; effect of, upon salt action, 1575. Hyperthyroidism, diastatic activity of blood in, 1514; dietary factors in, 1476. Immunization, with vaccines, 1561. Infants, dietaries of, in relation to the development of rickets, 1582; fat soluble vitamine in diets of, 1488. Infection, in foods, serologic method for detecting, 1472. Inorganic salines, method for determining part. Intracranial tension, alterations of, by salt solution, 1580. Iodine, distribution of, in thyroid, 1475. Isoagglutinins, of the rat, 1508. Isohemolysins, of the rat, 1508. Isomers, comparative toxicity of some alcohols with reference to, 1577. Labyrinth, otic, physiology of, 1568. Lactic acid, estimation of, in blood, 1546, influence of, upon metabolism, 1466. Lactose, blood sugar curves with, 1477. Lens, in amblystoma, 1566. Lethals, appearance in Drosophila, 1492. Leucocytes, osmosis as a factor in local accumulation of, 1579. Lipin, in blood serum of nephritis, 1474. Lipoids, classification of, 1525. Lipuria, in chronic nephritis, 1527. Liver maintenance, relation of portal blood to, 1498. Maltose, blood sugar curves with, 1477. Mannite, blood sugar curves with, 1477. Metabolism, pyrimidine, studies in, 1555. Morphine reaction, analysis of, 1520. Mother, catalase content of, 1534. Mouse, life of, 1564. Neoarsphenamine, phenol elimination of, after injection, 1522. Nephritis, blood serum in, 1474; chronic lipuria in, 1527. Nucleic acid, preparation of, from animal tissues, 1520. Nuts, nutritive value of, 1505. Nystagmus, post-rotatory, influence of prolonged rotation on duration 1568. Oestrous rhythm, effect of underfeeding on, 1537- INDEX. 253 Offspring, catalase content of, 1534. Opiates, effect of, on memory and be- havior of rats, 1547. Osmosis, effect of, in local accumulation of leucocytes, 1579. Ovulation, effect of underfeeding on, 1537. Oxygen, analysis of, in blood, 1576; in blood, during exercise, 1531; influence of, in expelling carbon dioxide from the blood, 1571. Parents, age of, in Uroleptus, 1503. Permeability, of capillaries, effect of colloids on, 1567; effect of salts on, 1567. Phenol elimination, amine, 1522. Phenylcinchoninic acid, effect of, on uric acid and urea in blood, 15509. Pilocarpine, effects of, after coli injection, 1573- Pituitary feeding, effect of, on domestic fowl, I5II. Plasma, calcium in, content of, 1468. Pollen, enzymes of, 1495. Polysaccharide, in human urine, 1557. Prostatic extracts action of, on genito urinary organs, I52I. Protein, production of fat from, I551. Protein feeding, effect of, a tend of fast, 1552. Protein meal, effect of, at end of 8-day fast, 1552. Proteins, foreign, disappearance of, and antibody formation, 1536. Pyrimidine metabolism, 1555. after neoarsphen- 1524; carbonate Rabbits, observations on the immuniza- tion of, with single strain vaccine, 1561. Rabbits, observations on the immuniza- tion of, with combined multiple strain vaccine, 1561. Radium emanation, effect of, upon de- velopment of mammalian embryos, 1558. Rat, amino-acid synthesis in, 1519; isoagglutinins and isohemolysins of, 1508. Rats, albino effect of anesthetization of, 1496; effects of opiates on memory and behavior of, 1547. Rejuvenescence, in Uroleptus, 1467. Reproduction, upon simplified food sup- ply, 1464. Respiratory, tract bacterial invasion of, 1563. Rickets, dietaries of infants in relation to the development of, 1582. Rubber tree, behavior of crown gall on, 1543: Salep mannan, utilization of, 1463. Salt solution, alteration of intracranial tension by, 1580. Salts, effect of, on permeability of capil- laries, 1567. Scurvy, sodium citrate in, 1526. Sodium citrate, in scurvy, 1526. Spirochetosis, infection of experimental, 1509. Straub‘s morphine reaction, analysis of, 1520. G-Strophanthin, action of, in patients, 1507. Sugar, determination of small quantities in urine, 1557; in blood, during exer- cise, I53I. Sulphates, effect of, on hydrogen ion concentration, 1462. Surface layer in living egg cell, 1483. Syphilis, generalized by local inoculation, 1548. Systole, ventricular, effect of accelerator nerves on, I516. Temperature, influence of, utilization, 1523. Thyroid, iodine of, 1475. Thyroid activities, conditions affecting, TST2. Tissue reaction, influence of systemic changes on, I5I5. Toxicity, comparative, of some alcohols with reference to isomers, 1577. Treponema pallidum, generalization of, after local inoculation, 1548. Tumor, benign, in Drosophila, 1490. Twins, heredity of, 1504. upon food Urea, effect of phenylcinchoninic acid on, 1559; excretion of, 1494. Uric acid, in blood, effect of phenyl- cinchoninic acid on, 1559. Urine, polysaccharide content of, 1557. Uroleptus, age of parents and vitality in, 1503. Vaccines, single and multiple strain, immunization by, 1561. Vagus, effects of digitalis on, 1480. Vitality, in Uroleptus, 1503. Vitamine, anti-beriberi test for, 1553; fat soluble in diet of infants, 1488; influence of water soluble, on nutrition, 1510; measurement of, 1532; results in measurement of, with a new tech- nique, 1581; water soluble B, Bach- man test for, 1491. White mouse, life of, 1564. a ee - -, A Cae = arextn Bake whores Sn et haa ot eS eo = « 9 | | = == - ners ee | | oe cael af i ¥, ti ay) ie ° J 7 5 if anh « U a i rh fa ul ye + ' Ay A a PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED FIRST MEETING CORNELL UN IVERSITY MEDICAL COLLEGE NEW YORK CITY OCTOBER 15, 1919 AND TWENTY-THIRD MEETING PACIFIC COAST BRANCH SAN FRANCISCO, CALIFORNIA OCTOBER 8, 1919 VoLUME XVII No. 1 NEW YORK 1919 CONTENTS. CALVIN B. BRIDGES (by invitation): The developmental stages at which mutations occur in the germ tract. 1 (1461). ARTHUR W. THOMAS AND MABEL E. BALDWIN: Contrasting effects of chlorides and sulphates on the hydrogen ion concentration of acid solutions. 2 (1462). Mary SwArtz Rose: On the utilization of salep mannan. 3 (1463). H. C. SHERMAN, M. E. Rouses, B. ALLEN AND E. Woops: Growth and reproduction upon simplified food supply. 4 (1464). H. J. MULLER AND E. ALTENBURG: The rate of change of hereditary factors in Drosophila. 5 (1465). ; GRAHAM LUSK AND H. V. ATKINSON: The influence of lactic acid upon the meta- bolism of the dog. 6 (1466). Gary N. CALKINS: Rejuvenescence without encystment and without nuclear fusion in Uroleptus? 7 (1467). R. W. Scott (by invitation): The total carbonate content of the arterial and venous plasma in normal individuals. 8 (1468). R. W. Scott (by invitation): The total carbonate content of the arterial and venous plasma in patients with chronic heart disease. 9 (1469). R. W. Scott (by invitation): The total carbonate content of the arterial and venous plasma in patients with chronic pulmonary emphysema. 10 (1470). D. I.°*Macut, J. P. GREENBERG AND S. Isaacs: Concerning the influence of anti- pyretics on the acuity of hearing. 11 (1471). J. BRONFENBRENNER AND M. J. SCHLESINGER: Serologic method for detecting in- fection in foods. 12 (1472). J. BRONFENBRENNER, D. SOLETSKY AND M. J. SCHLESINGER: Further studies on C Rindicator. 13 (1473). Max KAuwn: The protein and lipin content of blood serum of nephritic patients. 14 (1474). ARTHUR L. TATUM (by invitation): Method and results of a study of the distribu- tion of iodine between cells and colloid of thyroid glands. 15 (1475). JACOB ROSENBLOOM: On the certain dietary factors to be considered in the treat- ment of cases of hyperthyroidism. 16 (1476). 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 four dollars each, of Vol. IV to current volume is two dollars each, postage prepaid. The price of copies of the proceedings of any meeting is thirty cents each, postage prepaid. Subscriptions are payable in advance. PRESIDENT—Gary N. Calkins, Columbia University. VICE-PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—Henry C. Sherman, Columbia University, and J. W. Jobling, Columbia University, and ex-Presidents. MANAGING Epitor—The Secretary-Treasurer, 338 East 26th St., New York City. —oe —_— eS ee PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED SECOND MEETING NEW YORK POST-GRADUATE MEDICAL SCHOOL NEW YORK CITY NOVEMBER 109, I919 VoLUME XVII No. 2 NEW YORK 1919 CONTENTS. Cyrus W. FIELD: Blood sugar curves with glucose, lactose, maltose, mannite and cane sugar. 17 (1477). MARSHALL C. PEASE: The bacteriology of infectious gaseous gangrene. 18 (1478). D. T. MacDouGAL AND H. A. SPOEHR: Hydration effects of amino-compounds. 19 (1479). RoBERT H. HAtseEy (by invitation): Profound effects of digitalis on the vagus pro- ducing severe detrimental subjective symptoms, as shown by simultaneous electrocardiograms and pneumograms. 20 (1480). A. R. Moore: The action of camphor on the central nervous system of the squid. 21 (1481). FREDERICK L. GATES: A method of standardizing bacterial suspensions. 22 (1482). ROBERT CHAMBERS: Some studies on the surface layer in the living egg cell. 23 (1483). Davin I. Macut: Concerning the toxicity of acetanilid and bicarbonate combinations for muscle-nerve preparations. 24 (1484). ZALIA JENCKS (by invitation): A note on the carbohydrates of the root of the cat-tail (typha latifolia). 25 (1485). TuHomAS B. OSBORNE AND LAFAYETTE B. MENDEL: Do fruits contain water-soluble vitamine? 26 (1486). PAuL F. Orr (by invitation): Some observations on the biological characteristics of bacillus botulinus. 27 (1487). 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 four dollars each, of Vol. IV to current volume is twodollars each, postage prepaid. The price of copies of the proceedings of any meeting is thirty cents each, postage prepaid. Subscriptions are payable in advance. PRESIDENT—Gary N. Calkins, Columbia University. ViICE-PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—Henry C. Sherman, Columbia University, and J. W. Jobling, Columbia University, and ex-Presidents. MANAGING EpitoR—The Secretary-Treasurer, 338 East 26th St., New York City. = ee PROCEEDINGS SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE ONE HUNDRED THIRD MEETING ROCKEFELLER INSTITUTE FOR MEDICAL RESEARCH NEW YORK CITY DECEMBER 17, I919 VoLUME XVII No. 3 NEW YORK 1919 CONTENTS. ALFRED F. HESS AND LESTER J. UNGER: The role of fat-soluble vitamine in the dietary of infants. 28 (1488). IsIDOR GREENWALD: A method for the determination of calcium, magnesium, po- tassium, sodium, chlorides and “‘acid-soluble’’ sulfur and phosphorus in one sample (25cc.) of blood. 29 (1489). Mary B. Stark (by invitation): A benign tumor hereditary in Drosophila. 30 (1490). WALTER H. Eppy AND HELEN C. STEVENSON: The suitability of the ““Bachman Test”’ for water-soluble B in vitamine. 31 (1491). D. E. LANCEFIELD (by invitation) : Two sex-linked lethals of simultaneous appear- ance in Drosophila obscura. 32 (1492). RoBeErT H. BoweEN (by invitation): New methods for the analysis of cystoplasmic structures. With demonstrations. 33 (1493). DoNALD D. VANSLYKE, J. H. AUSTIN AND E. STILLMAN: The excretion of urea. 34 (1494). Jutra B. Paton (by invitation): Enzymes of pollen. 35 (1495). Davip I. MacuT AND C. F. Mora: Effect of the anesthetization on the subsequent behavior and intelligence of albino rats. 36 (1496). RoBERT L. LEVY AND GLENN E. CULLEN: On the deterioration of crystalline stro- phanthin in aqueous solution. 37 (1497), 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 four dollars each, of Vol. IV to current volume is two dollars each, postage prepaid. The price of copies of the proceedings of any meeting is thirty cents each, postage prepaid. Subscriptions are payable in advance. PRESIDENT—Gary N. Calkins, Columbia University. VicE-PRESIDENT—George B. Wallace, University and Bellevue Hospital Medical College. SECRETARY-TREASURER—Holmes C. Jackson, University and Bellevue Hospital Medical College. Additional members of the Council—Henry C. Sherman, Columbia University, and J. W. Jobling, Columbia University, and ex-Presidents. MANAGING Epitog—The Secretary-Treasurer, 338 East 26th St., New York City CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, Thomas A. Storey. Columbia University.—C. H. Bailey, A. K. Balls, Russell Burton-Opitz, Gary N. Calkins, R. L. Cecil, George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. T. Longcope, Thomas H. Morgan, Herman J. Muller, B. S. Oppenheimer, Alwin M. Pappenheimer, 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. Cornell University Medical College-—Harold Bailey, Stanley R. Benedict, Robert Chambers, E. F. DuBois, D. J. Ed- wards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, Robert A. Hatcher, Graham Lusk, John R. Murlin, Walter L. Niles, Charles R. Stockard, W. C. Thro, John C. Torrey. Fordham University School of Medicine —A. O. Shaklee, Carl P. Sherwin. Hospitals, Beth Israel—Max Kahn. Memorial Hospital—H. H. Janeway. Montefiore Home.—George Fahr, B.S. Kline. Mt. Sinai.—George Baehr, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Roths- child. New York.—A. F. Coca. Presbyterian.—Louis Baumann, Ralph A. Kinsella, Walter W. Palmer. Roosevelt.— K. G. Falk, I. Greenwald, W. L. Lyle. St. Lukes—L. W. Famulener. New York City Departments. Education.—C. Ward Crampton. Health—James P. Atkinson, Edwin J. Banzhaf, C. B. Fitzpatrick, Alfred F. Hess, William H. Park, Anna W. Williams, A. Zingher. Medical Director.—Charles Norris. New York Post-Graduate Medical School.—Arthur F. Chace, Martin Cohen, Ludwig Kast, John A. Killian, W. J. MacNeal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. = New York University —W. H. Barber, Harlow Brooks, Warren Coleman, J. W. Draper, Edward K. Dunham, A. O. Gettler, Holmes C. Jackson, R. L. Kahn, Arthur R. Mandel, John A. Mandel, W. C. Noble, H. D. Senior, Douglas Symmers, George B. Wallace. Rockefeller Institute for Medical Research—Harold L. Amoss, John Auer, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, F. L. Gates, Walter A. Jacobs, I. S. Kleiner, I. J. Kligler, P. A. Levene, Jacques Loeb, S. J. Meltzer, Arthur L. Meyer, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 819 Madison Avenue, N. Y. City —H. D. Dakin. 126 E. 64th St.—Cyrus W. Field. 142 W. 49th St., N. Y. City—Casimir Funk. 141 W. 78th St., N. Y. City.—A. I. Ringer. 302 Convent Ave., N. Y. City—William Salant. 50 Vanderbilt Ave., N. Y. City—Benjamin White. 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, 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. New York (Albany).—Mary B. Kirkbride, P. A. Kober, A. B. Wadsworth. Hospitals. General (Cincinnati, Ohio).—Martin H. Fischer. Cottage (Sania Barbara, Calif.)—Nelson W. Janney, Lakeside (Cleveland, Ohio)—H. T. Karsner. Massachusetts General (Boston).—David L. Edsall. Mercy (Pittsfield, Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Presbyterian (Phila.).—Ralph Pemberton. Robert Brigham (Boston).—F. M.McCrudden. Royal Victoria (Monireal).—Horst Oer- tel. St. Francis (Pitisburgh)—J. W. McMeans. Western Pennsylvania (Pittsburgh)—Maurice H. Givens, Jacob . Rosenbloom. = Institutes. Gratwick Laboratory (Buffalo).—G. H. A. Clowes, H. R. Gaylord. Juvenile Psychopathic (Chicago).— Herman M. Adler. Phipps (Philadelphia)—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F. S. Jones, Theobald Smith, Carl Ten Broeck. Wéistar (Philadelphia) —H. H. Donaldson, Shinkishi Hatai. U. S. Departments. Agriculture (Washington, D. C.)—Carl L. Alsberg. Bureau of Animal Industry.— William N. Berg, J. R. Mohler. Hygienic Laboratory (Washington, D. C.).—George B. Roth. Medical Corps (Fort Sam Houston, Texas).—J.F. Siler. Ohio River Investigation (Ohio).—W. H. Frost. Universities. Adelaide (South Australia).—T. Braiisford Robertson. Amsherst.—Otto Glaser. Buffalo—Katharine R. Collins, Herbert U. Williams. California.—Walter C. Alvarez, W. R. Bloor, T. C. Burnett, H. M. Evans, F. P. Gay, S. J. Holmes, Charles W. Hooper, Samuel H. Hurwitz, R. A. Kocher, Charles A. Kofoid, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker, C. K. Watanabe, G. H. Whipple. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Clark.—Ralph S. Lillie. Copenhagen.— Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.—Richard V.Lamar. George Washington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B.Cannon, W. T.Councilman, Otto Folin, Worth Hale, Reid Hunt, W. J. V.Oster- hout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, E. E. Southard, Percy G. Stiles, Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach. Iilinois.—H. B. Lewis, William H. Welker. Japan.—Noahidé Yatsu. Jefferson. —O. Bergeim, P.B. Hawk. Johns Hopkins.—John J. Abel, C. G. Bull, George W. Corner, Walter E. Dandy, W. W. Ford, W. S. Halsted, William H. Howell, John Howland, H. S. Jennings, Walter Jones, W. G. MacCallum, David I. Macht, Adolph Meyer, H. O. Mosenthal, E. A. Park, Raymond Pearl, Reynold Albrecht Spaeth, William H. Welch. Kansas. —Bennet M. Allen. Leland Stanford—Thoma Addis, George D. Barnett, A. C. Crawford, E. C. Dickson, Harold K. Faber, 1. W. Hewlett, V. L. Kellogg, W. H. Manwaring, W. Ophiils, R. E. Swain. Maryland.—Charles E. Simon. McGill (Montreal).—J. George Adami, John L. Todd. Michigan.—C. W. Edmunds, Nellis B. Foster, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, Aldred S. Warthin, Carl Vernon Weller. Minnesota.—R. A. Gortner, A. D. Hirschfelder, E. P. Lyon, F. W. Schlutz. Missouri.—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Caro- lina.—W. deB. MacNider. Northwestern.—R. G. Hoskins, Solomon Strouse. Oregon.—C. F. Hodge. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, D. H. Bergey, J. A. Kolmer, E. B. Krumbhaar, Richard M. Pearce, O. H. Perry Pepper, Edward T. Reichert, Alfred N. Richards, J. Edwin Sweet, A. E. Taylor, Charles Weiss. Philippine-——R. B. Gibson. Pittsburgh.—C. C. Guthrie, W. L. Holman, E. R. Hoskins, Oskar Klotz. Princeton.—Edwin G. Conklin, E. Newton Harvey. Rutgers.—John F. Anderson, F. E. Chidester, A. R. Moore. Sheffield—J.B.Leathes. Southern California (Los Angeles).—Lyman B. Stookey. St. Louis.—Don R. Joseph. Toronto.—A. H. Caulfeild, J. G. Fitzgerald, A. Hunter, A. B. Macallum, J. J. R. Macleod, H. Wasteneys. Tulane.— Charles W. Duval, E. M. Ewing. 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, G. C. Robinson, Philip A. Shaffer. Wisconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Western Reserve (Cleveland).—George W. Crile, A. B. Ejisenbrey, Paul J. Hanzlik, David Marine, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.— Withrow Morse. Yale.—George A. Baitsell, Henry Gray Barbour, W. H. Chittenden, J. W. Churchman, Barnett Cohen, Rhoda Erdmann, Ross G. Harrison, Davenport Hooker, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, A. L. Prince, Leo F. Rettger, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer. Cambridge, England.—C. G. L. Wolf. Chicago, Illinois, 31 East Elm St.—W. F. Petersen. Kastanienbaum, Switzerland.—Fritz Schwyzer. Newark, N. J., Calco Chemical Co.—M. S. Fine. Philadelphia, Pa., H. K. Mulford Co., 1524 Chestnut St.—T. S. Githens. Tuckahoe, N. Y.—Isaac F. Harris. Washington, D. C., 3315 Wisconsin Ave.—C. J. West, 1445 Rhode Island Ave.—Edward B. Meigs, 1864 Park Road.— Robert M. Yerkes. Winnipeg, Canada, Dominion Grain Laboratory.—F. J. Birchard. Germany.—Reinhard Beutner. London, England.—Cameron V. Bailey. Members present at the one hundred third meeting: Auer, Austin, Burton-Opitz, Calkins, DuBois, Eddy, Field, Funk, Gies, Githens, Greenwald, Hess, Jackson, Kahn, R. L., Meltzer, Mosenthal, Muller, Myers, Riddie, Rose, A. R., Salant, Sherwin, Thro, Torrey, Uhlenhuth, VanSlyke. Members elected at the one hundred third meeting: B. M. Duggar, H. A. Spoehr, S. C. Brooks. Dates of the next two meetings: January 21, 1920—February 18, 1920. CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, Thomas A. Storey. Columbia University.—C. H. Bailey, A. K. Balls, Russell Burton-Opitz, Gary N. Calkins, R. L. Cecil, George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, Isaac Levin, Cc. C. Lieb, W. T. Longcope, Thomas H. Morgan, Herman J. Muller, B. S. Oppenheimer, Alwin M. Pappenheimer, 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. Cornell University Medical College.—Harold Bailey, Stanley R. Benedict, Robert Chambers, E. F. DuBois, D. J. Ed- wards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, Robert A. Hatcher, Graham Lusk, John R. Murlin, Walter L. Niles, Charles R. Stockard, W. C. Thro, John C. Torrey. Fordham University School of Medicine.—A. O. Shaklee, Carl P. Sherwin. ‘Hospitals, Beth Israel—Max Kahn. Memorial Hospital—H. H. Janeway. Montefiore Home.—George Fahr, B.S. Kline. Mt. Sinat.—George Baehr, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Roths- child. New York.—A. F. Coca. Presbyterian.—Louis Baumann, Ralph A. Kinsella, Walter W. Palmer. Roosevelt.— K. G. Falk, I. Greenwald, W. L. Lyle. St. Lukes.—L. W. Famulener. . New York City Departments. Education.—C. Ward Crampton. Health—James P. Atkinson, Edwin J. Banzhaf, C. B. Fitzpatrick, Alfred F. Hess, William H. Park, Anna W. Williams, A. Zingher. Medical Director.—Charles Norris. New York Post-Graduate Medical School.—Arthur F. Chace, Martin Cohen, Ludwig Kast, John A. Killian, W. J. MacNeal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. New York University —W. H. Barber, Harlow Brooks, Warren Coleman, J. W. Draper, Edward K. Dunham, A. O. Gettler, Holmes C. Jackson, R. L. Kahn, Arthur R. Mandel, John A. Mandel, W. C. Noble, H. D. Senior, Douglas Symmers, George B. Wallace. Rockefeller Institute for Medical Research—Harold L. Amoss, John Auer, Wade H. Brown, Alexis Carrel, A. E. Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, F. L. Gates, Walter A. Jacobs, I. S. Kleiner, I. J. Kligler, P. A. Levene, Jacques Loeb, S. J. Meltzer, Arthur L. Meyer, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E. 34th St., N. Y. City.—E. E. Butterfield. 819 Madison Avenue, N. Y. City.—H. D. Dakin. 126 E. 64th St.—Cyrus W. Field. 142 W. 49th St., N. Y. City.—Casimir Funk. 141 W. 78th St.,.N. Y. City —A. I. Ringer. 302 Convent Ave., N. Y. City.—William Salant. 50 Vanderbilt Ave., N. Y. City—Benjamin White. 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, 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 Healih. New York (Albany).—Mary B. Kirkbride, P. A. Kober, A. B. Wadsworth. Hospitals. General (Cincinnati, Ohio)—Martin H. Fischer. Cottage (Sania Barbara, Calif.).—Nelson W. Janney, Lakeside (Cleveland, Ohio).—H. T. Karsner. Massachusetis General (Boston).—David L. Edsall. Mercy (Pittsfield, Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Presbyterian (Phila.).—Ralph Pemberton. Robert Brigham (Boston).—F. M. McCrudden. Royal Victoria (Montreal).—Horst Oer- tel. St. Francis (Pittsburgh)—J. W. McMeans. Western Pennsylvania (Pittsburgh).—Maurice H. Givens, Jacob Rosenbloom. Institutes. Gratwick Laboratory (Buffalo).—G. H. A. Clowes, H. R. Gaylord. Juvenile Psychopathic (Chicago).— Herman M. Adler. Phipps (Philadelphia)—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F. S. Jones, Theobald Smith, Carl Ten Broeck. Wistar (Philadelphia) —H. H. Donaldson, Shinkishi Hatai. U. S. Departments. Agriculture (Washington, D. C.).—Carl L. Alsberg. Bureau of Animal Industry.—William N. Berg, J. R. Mohler. Hygienic Laboratory (Washington, D. C.).—George B. Roth. Medical Corps (Fort Sam Houston, Texas).—J.F. Siler. Ohio River Investigation (Ohio).—W. H. Frost. Universities. Adelaide (South Australia).—T. Braiisford Robertson. Amherst.—Otto Glaser. Buffalo—Katharine R. Collins, Herbert U. Williams. California—Walter C. Alvarez, W. R. Bloor, T. C. Burnett, H. M. Evans, F. P. Gay, S. J. Holmes, Charles W. Hooper, Samuel H. Hurwitz, R. A. Kocher, Charles A. Kofoid, W. P. Lucas, S. S. Maxwell, K. F. tm Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker, C. K. Watanabe, G. H. Whipple. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Clark.—Ralph S. Lillie. Copenhagen.— Christen Lundsgaard. Cornell.—Sutherland Simpson. Georgia.—Richard V.Lamar. George Washington.—W. H. Schultz. Harvard.—Jacob Bronfenbrenner, Walter B.Cannon, W. T.Councilman, Otto Folin, Worth Hale, Reid Hunt, W. J. V.Oster- hout, G. H. Parker, F. Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, E. E. Southard, Percy G. Stiles, Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach. JIllinois.—H. B. Lewis, William H. Welker. Japan.—Noahidé Yatsu. Jefferson. —O. Bergeim, P.B. Hawk. Johns Hopkins.—John J. Abel, C. G. Bull, George W. Corner, Walter E. Dandy, W. W. Ford, W. S. Halsted, William H. Howell, John Howland, H.S. Jennings, Walter Jones, W. G. MacCallum, David I. Macht, Adolph Meyer, H. O. Mosenthal, E. A. Park, Raymond Pearl, Reynold Albrecht Spaeth, William H. Welch. Kansas. —Bennet M. Allen. Leland Stanford—Thoma Addis, George D. Barnett, A. C. Crawford, E. C. Dickson, Harold K. Faber, A. W. Hewlett, V. L. Kellogg, W. H. Manwaring, W. Ophiils, R. E. Swain. Maryland.—Charles E. Simon. McGill (Montreal).—J. George Adami, John L. Todd. Michigan.—C. W. Edmunds, Nellis B. Foster, G. Carl Huber, Warren P. Lombard, Frederick G. Novy, Aldred S. Warthin, Carl Vernon Weller. Minnesota.—R. A. Gortner, A. D. Hirschfelder, E. P. Lyon, F. W. Schlutz. Missouri—Mazyck P. Ravenel. Nebraska.—A. E. Guenther. North Caro- lina.—W. deB. MacNider. Northwestern.—R. G. Hoskins, Solomon Strouse. Oregon.—C. F. Hodge. Peking Union Medical.—Franklin C. McLean. Pennsylvania.—Alexander C. Abbott, J. H. Austin, D. H. Bergey, J. A. Kolmer, E. B. Krumbhaar, Richard M. Pearce, O. H. Perry Pepper, Edward T. Reichert, Alfred N. Richards, J. Edwin Sweet, A. E. Taylor, Charles Weiss. Philippine —R.B. Gibson. Pittsburgh—C. C. Guthrie, W. L. Holman, E. R. Hoskins, Oskar Klotz. Princeton.—Edwin G. Conklin, E. Newton Harvey. Rutgers.—John F. Anderson, F. E. Chidester, A. R. Moore. Sheffield.—J.B.Leathes. Southern California (Los Angeles).—Lyman B. Stookey. St. Louis.—Don R. Joseph. Toronto.—A. H. Caulfeild, J. G. Fitzgerald, A. Hunter, A. B. Macallum, J. J. R. Macleod, H. Wasteneys. Tulane.— Charles W. Duval, E. M. Ewing. 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, G. C. Robinson, Philip A. Shaffer. Wasconsin.—Charles R. Bardeen, C. H. Bunting, J. L. Cole, P. F. Clark, J. A. E. Eyster, Arthur S. Loevenhart. Western Reserve (Cleveland).—George W. Crile, A. B. Eisenbrey, Paul J. Hanzlik, David Marine, Torald Sollmann, G. N. Stewart, C. J. Wiggers. West Virginia.— Withrow Morse. Yale.—George A. Baitsell, Henry Gray Barbour, W. H. Chittenden, J. W. Churchman, Barnett Cohen, Rhoda Erdmann, Ross G. Harrison, Davenport Hooker, R. A. Lambert, Henry Laurens, Lafayette B. Mendel, A. L. Prince, Leo F. Rettger, Arthur H. Smith, Frank P. Underhill, C.-E. A. Winslow, Lorande Loss Woodruff. Baltimore, Md., 1421 Edmondson Ave.—J. A. F. Pfeiffer. Cambridge, England.—C. G. L. Wolf. Chicago, Illinois, 31 East Elm St.—W. F. Petersen. Kastanienbaum, Switzerland.—Fritz Schwyzer. Newark, N. J., Calco Chemical Co.—M. S. Fine. Philadelphia, Pa., H. K. Mulford Co., 1524 Chestnut St.—T. S. Githens. Tuckahoe, N. Y.—Isaac F. Harris. Washington, D. C., 3315 Wisconsin Ave.—C. J. West, 1445 Rhode Island Ave.—Edward B. Meigs, 1864 Park Road.— Robert M. Yerkes. Winnipeg, Canada, Dominion Grain Laboratory.—F. J. Birchard. Germany.—Reinhard Beutner. London, England.—Cameron V. Bailey. Members present at the one hundred second meeting: Auer, Calkins, Field, Gates, Gies, Githens, Greenwald, Harris Isaac, Jackso™, H. C., Jobling, Killian, Kligler, MacDougal, MacNeal, Moore, Morgan, Myers, Olitsky, Rose, A. R., Scott, Senior, Wallace, Wilson. Members elected at the one hundred second meeting: Robert A. Gesell, Jean Redman Oliver, Oscar M. Schloss, R. W. Scott. Dates of the next two meetings: December 17, 1919—January 21, 1920. CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE. Resident (Greater New York). College of the City of New York.—W. W. Browne, A. J. Goldfarb, G. G. Scott, Thomas A. Storey. Columbia University.—C. H. Bailey, A. K. Balls, Russell Burton-Opitz, Gary N. Calkins, R. L. Cecil, George Draper, Walter H. Eddy, Andrew Eggstein, William J. Gies, J. Gardner Hopkins, J. W. Jobling, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. T. Longcope, Thomas H. Morgan, Herman J. Muller, B. S. Oppenheimer, Alwin M. Pappenheimer, 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. Cornell University. Medical College—Harold Bailey, Stanley R. Benedict, Robert Chambers, E. F. DuBois, D. J. Ed- wards, C. Eggleston, William J. Elser, James Ewing, J. A. Hartwell, Robert A. Hatcher, Graham Lusk, John R. Murlin, Walter L. Niles, Charles R. Stockard, W. C. Thro, John C. Torrey. Fordham University School of Medicine—A. O. Shaklee, Carl P. Sherwin. Hospitals, Beth Israel—Max Kahn. Memorial Hospital—H. H. Janeway. Montefiore Home.—George Fahr, B.S. Kline. Mt. Sinai.—George Baehr, Charles A. Elsberg, Albert A. Epstein, R. Ottenberg, Harry Plotz, M. A. Roths- child. New York.—A. F. Coca. Presbyterian.—Louis Baumann, Ralph A. Kinsella, Walter W. Palmer. Roosevelt.— K. G. Falk, I. Greenwald, W. L. Lyle. St. Lukes.—L. W. Famulener. New York City Departments. Education.—C. Ward Crampton. MHealthJames P. Atkinson, Edwin J. Banzhaf, C. B. Fitzpatrick, Alfred F. Hess, William H. Park, Anna W. Williams, A. Zingher. Medical Director.—Charles Norris. New York Post-Graduate Medical School.—Arthur F. Chace, Martin Cohen, Ludwig Kast, John A. Killian, W. J. MacNeal, V. C. Myers, Marshall C. Pease, Anton R. Rose, R. M. Taylor. New York University —W. H. Barber, Harlow Brooks, Warren Coleman, J. W. Draper, Edward K. Dunham, A. O. Gettler, Holmes C. Jackson, R. L. Kahn, Arthur R. Mandel, John A. Mandel, W. C. Noble, H. D. Senior, Douglas Symmers, George B. Wallace. Rockefeller Institute for Medical Research. Siapeid L. Amoss, John Auer, Wade H. Brown, Alexis Carrel, A. E, Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, F. L. Gates, Walter A. Jacobs, I. S. Kleiner, I. J. Kligler, P. A. Levene, Jacques Loeb, S. J. Meltzer, Arthur L. Meyer, Gustave M. Meyer, James B. Murphy, Hideyo Noguchi, J. H. Northrop, Peter K. Olitsky, Louise Pearce, Peyton Rous, Eduard Uhlenhuth, Donald D. Van Slyke, Martha Wollstein. Brooklyn Botanic Garden.—C. Stuart Gager, O. E. White. 135 E..34th St., N. Y. City.—E. E. Butterfield. 819 Madison Avenue, N. Y. City—H. D. Dakin. 126 E. 64th St.—Cyrus W. Field. 142 W. 49th St., N. Y. City.—Casimir Funk. 141 W. 78th St., N. Y. City.—A. I. Ringer. 302 Convent Ave., N. Y. City.—William Salant. 50 Vanderbilt Ave., N. Y. City.—Benjamin White. 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, 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. New York (Albany).—Mary B. Kirkbride, P. A. Kober, A. B. Wadsworth. Hospitals. General (Cincinnati, Ohio).—Martin H. Fischer. Cottage (Santa Barbara, Calif.)—Nelson W. Janney, Lakeside (Cleveland, Ohio)—H. T. Karsner. Massachusetts General (Boston).—David L. Edsall. Mercy (Pittsfield, Mass.).—Thomas Flournoy. Peter Bent Brigham (Boston).—Harvey Cushing, Francis W. Peabody. Presbyterian (Phila.).—Ralph Pemberton. Robert Brigham (Boston).—F. M.McCrudden. Royal Victoria (Montreal).—Horst Oer- tel. St. Francis (Pittsburgh).—J. W. McMeans. Western Pennsylvania (Pittsburgh) Maurice H. Givens, Jacob Rosenbloom. Institutes. Gratwick Laboratory (Buffalo).—G. H. A. Clowes, H. R. Gaylord. Juvenile Psychopathic (Chicago).— Herman M. Adler. Phipps (Philadelphia)—Paul A. Lewis. Rockefeller (Princeton).—Paul E. Howe, F. S. Jones, Theobald Smith, Carl Ten Broeck. Wistar (Philadelphia) —H. H. Donaldson, Shinkishi Hatai. U. S. Depariments. Agriculture (Washington, D. C.).—Carl L. Alsberg. Bureau of Animal Industry.—William N. Berg, J. R. Mohler. Hygienic Laboratory (Washington, D. C.).—George B. Roth. Medical Corps (Fort Sam Houston, Texas).—J.F. Siler. Ohio River Investigation (Ohio).—W. H. Frost. Universities. Adelaide (South Australia).—T. Braiisford Robertson. Amherst.—Otto Glaser. Buffalo.—Katharine R. Collins, Herbert U. Williams. California.—Walter C. Alvarez, W. R. Bloor, T. C. Burnett, H. M. Evans, F. P. Gay, S. J. Holmes, Charles W. Hooper, Samuel H. Hurwitz, R. A. Kocher, Charles A. Kofoid, W. P. Lucas, S. S. Maxwell, K. F. Meyer, Carl L. A. Schmidt, Philip E. Smith, E. L. Walker, C. K. Watanabe, G. H. Whipple. Chicago.—A. J. Carlson, Frank R. Lillie, Arno B. Luckhardt. Cincinnati.—D. E. Jackson, Shiro Tashiro. Clark. Sd ee oe Ne de AE 2 rig tr te. ~ eee he): iin