Experimental Biology and Medicine

1. Tissue extracts from mice of the high-leukemia strain Ak injected into newborn mice greatly increased the leukemia incidence in Rf, DBA 1 and C₃Heb mice. and greatly hastened the development of leukemia in AkR mice. 2. Mortality without gross anatomic manifestation of leukemia was also enhanced in extract-injected mice of low-leukemia strains. 3. Although all leukemias in Ak mice were lymphoid with thymic involvement, many leukemias in the low-leukemia strains were of non-thymic types. 4. Transplantation assays of leukemias induced in the low-leukemia strains disclosed a heterogeneity in types, some being related immunogenetically with both host and donor strain. 5. Neither salivary gland nor other tumors were induced by the leukemia agent studied. 6. The available information suggests consideration of a process analogous with DNA-induced transformation (transduction) of genetic properties of microorganisms rather than to a natural type of viral infection.

Hypothyroid, hyperthyroid and euthyroid rats were subjected to acute starvation to extremis. Their oxygen consumption was measured daily. All groups showed an approximately equivalent early depression of oxygen consumption which is independent of the absolute level of the initial oxygen consumption.

Feeding weanling male rats a diet free of essential fatty acid (EFA), containing 1% cholesterol, 2% sulfasuxidine, and 1% hydrogenated coconut oil for 3.5 months induced an EFA deficiency accompanied by severe testicular degeneration. The degeneration was partially corrected by supplementation with linoleate or linseed oil for 3 weeks. However, sperm were not found in the epididymis in any case. Trans isomers of linoleic acid were unable to induce recovery of the testis. The trans isomers inhibited growth, worsened the skin condition, and were found to be deposited in the animal fat.

The islets of Langerhans of scorbutic guinea pigs and those injected with acetoacetate were hypertrophied. Injection of acetoacetate into scorbutic animals caused further degranulation and degenerative changes. Acetoacetate brought about hypertrophy of renal cortex and scurvy caused atrophy of glomeruli. Combined treatment aggravated these degenerative changes. Both scurvy and acetoacetate caused central degeneration of liver and this is enhanced by the combined treatment. In agreement with histologic changes, biochemical findings indicate the toxicity of acetoacetate. Acetoacetate may be a prime factor in clinical development of diabetes.

A method has been devised for preparation of highly purified pancreatic callicrein. The fractionation procedure involves three ethanolic fractionations at pH 5.5, 7.5. and 6.5 followed by one ammonium sulfate precipitation. Solutions of hypotensive enzyme at this level of purity proved highly unstable and further purification was abandoned. Preliminary purification of urinary callicrein revealed that the method could be adapted to this source.

Inclusion of N, N'-diphenyl-p-phenylenediamine in a vit. E-low diet fed to female rats through growth and reproduction has resulted in varying degrees of reproductive failure. Toxicity was encountered during growth at a level of 0.5% but not at 0.1% or below. Rats receiving 0.1% dppd exhibited uterine and vaginal hemorrhages at parturition accompanied by anorexia, anemia and emaciation. Concentrations of 0.025%, 0.01% and 0.005% resulted in a high incidence of stillbirths. No adverse effects were encountered from feeding 0.0006% dppd in the diet. The data show that dppd elicits symptoms of toxicity in the rat at concentrations below those used in commercial mixed feeds.

1. Quinidine sulfate at a concentration of 1 × 10^-3 M strongly inhibits the uptake of glucose and fructose by intact rat diaphragm under anaerobic conditions. 2. Quinidine is without effect on endogenous anaerobic lactic acid production by rat diaphragm. 3. Glucose at a concentration of 1 mg/ml or 4 mg/ml markedly stimulates anaerobic glycolysis while fructose at a concentration of 5 mg/ml produces only a slight stimulation. 4. Stimulation of glycolysis by glucose or fructose is abolished by 1 × 10^-3 M quinidine and partially inhibited by 5 × 10^-4 M quinidine. 5. It is suggested that quinidine inhibits glucose or fructose uptake prior to the step at which these sugars and glycogen share a common metabolic pathway.

Omega-methylpantethine was observed markedly to inhibit sulfanilamide acetylation at concentrations which were ineffective against citrate formation in pigeon liver homogenates. With aliquots of the same homogenate. bis- beta-pantoylaminoethyl) disulfide was observed to inhibit citrate formation at concentrations which were ineffective against sulfanilamide acetylation. In either case, higher concentrations of these compounds were effective in inhibiting both systems.

In analysed systems representing artificially reconstituted plasmas of known composition with respect to prothrombin. (pro)convertin, and AcG. One-stage clotting-times with Ca-thromboplastin, essentially in the manner of the “prothrombin time” (Quick) test, show: (1) prothrombin concentration is a major variable. (2) (pro) convertin concentration is a minor variable, which, except at very low(10) levels, affects the clotting-times significantly only at very low prothrombin levels. (3) High (pro)convertin levels, short of an unexplained inhibitory phenomenon (minor), do not compensate significantly for low levels of prothrombin or of AcG. (4) AcG levels are a major variable. (5) Excessively high (200-300% of'normal') AcG levels can compensate for moderately low prothrombin and proconvertin, causing the one-stage test to lose significance as a measure of these other two factors. (6) It seems valid to apply this last result to explain the normal “prothrombin time” (and lack of bleeding) in normal healthy newborn infants(1).

A study of the aortas of 44 female rabbits in 2 age groups (3 mos. and 46 mos.) corresponding approximately to human childhood and middleage, indicates elevation of the water, sodium, calcium, non-lipid phosphorus, and possibly of the cholesterol content of the vessel wall in the older group. The potassium content remains relatively constant. The generally lower levels of total lipid and of cholesterol in the rabbit aorta compared with that of man probably account at least in part for the freedom of the former from spontaneous atherosclerosis.

Twenty-three strains of fibroblasts derived from normal rabbit and chick embryo tissue were serially propagated on glass in a variety of media. All except 3 strains failed to proliferate for more than 8 to 12 subcultures. The 3 successful strains were derived from adult testis, embryonic skin-muscle and adult muscle respectively and continue to proliferate at a constant rate after 38 to 61 subcultures. Vaccinia virus multiplies in all 3 strains of fibroblasts resulting in the complete destruction of cells.

1. Addition of 0.5% sodium sulfate to a diet low in sulfur stimulated growth of the young chicken. 2. When Na₂S³⁵O₄ was fed for a period of 1 week, approximately 23% of the S³⁵ ingested was incorporated into the tissues. Most of the S³⁵ was recovered as sulfate-S³⁵. However, about 19% was present as taurine-S³⁵, 6.6% as unknown organic compounds and barely detectable amounts as sulfur amino acids. Two weeks after Na₂S³⁵O₄ feeding a larger proportion of S³⁵ was present in the carcasses as taurine and unidentified organic compounds and 1.5% of the S³⁵ in the chickens was accounted for as cystine and methionine. It was concluded that there is a physiological requirement for sulfate per se and that dietary sulfate promotes growth primarily by providing sulfur for this purpose. 3. Taurine was found to be the major sulfur constituent of the water-soluble portion of feathers.

The combination of 0.1% yeast extract and 0.1% proteose peptone #3 in Hanks'balanced salt solution plus 20% horse serum stimulated the rate of growth and produced a higher total count of L cells at the end of 5 days than did the standard reference medium (60% medium 199, 20% whole egg ultrafiltrate, and 20% horse serum). Lowering the serum content in the new medium to 10% still permitted better growth than the standard reference medium. The yeast extract-proteose peptone solution could be sterilized by autoclaving without affecting its growth-stimulating properties.

1. A rotating disc type reservoir-oxygenator is described. 2. Its effectiveness in maintaining circulation and oxygenation is discussed. 3. It has proven satisfactory in both its laboratory and clinical application.

The concentrations of carbutamide in the liver and plasma of rats receiving a wide range of doses of the drug have been determined. When carbutamide was added to a rat liver microsome preparation in these concentrations no inhibition of glucose-6-phosphatase activity was observed.

The benzyl analog of bufotenin, BAB. or 1-benzyl-2,5-dimethylbufotenin (1-benzyl-2-methyl-5-methoxy-N, N-dimethyltryptamine hydrochloride) has been found to be a highly active antagonist of serotonin. Not only was it effective on isolated tissues, but. unlike many others, was highly active as an antagonist to the pressor effects of serotonin in dogs, even when given by the oral route. Some differences in action from the corresponding primary amine (benzyl analog of serotonin) were found.

The normal serum levels of glycoproteins in 19 New Zealand white rabbits are reported. It was shown that there was no significant difference between the sexes and essentially little effect on total serum glycoprotein concentration when the rabbits were bled weekly in the quantity reported over an 11 week period.

1. Newly hatched larvae of Caenorhabditis briggsae can grow to adulthood and F₁ larvae can be produced on a completely defined synthetic medium (GM-8) consisting of 18 L-amino acids, D-glucose, 4 ribonucleotides plus thymine, choline, i-inositol, ascorbic acid, 17 “trace” vitamins and growth factors, and a mixture of salts. 2. Media (GM-9 and −11) like GM-8 except for certain omissions (and minor additions to restore acid-base balance and salt levels) also support maturation and reproduction. 3. In the case of GM-11, omitted substances include the 8 “non-essential” amino acids. 4. The presence of a trace amount of the standard liver medium used for the indefinite axenic cultivation of C. briggsae is markedly stimulatory to the synthetic media, and some F₁ larvae mature and produce F₂ larvae when liver medium is present at only 0.032% of its standard strength. 5. For sustained growth, however, C. briggsae requires the presence of much higher levels of liver medium—between 10 and 50% of the standard level used for stock cultures. 6. Evidence is here presented that the liver medium contains 2 requirements, or sets of requirements, not present in the synthetic media, at least in adequate amounts or proportions.

1) The fate and distribution of Fe⁵⁹ tagged ferrocyanide were evaluated in humans and dogs. 2) The tracer could not be detected in red blood cells, gastric juice, saliva or feces. 3) The apparent volume of distribution of the tagged ferrocyanide was frequently smaller than the assumed extracellular space, and in 3 instances was less than the simultaneously measured inulin space. 4) In vitro binding to plasma albumin was demonstrated by electrophoretic and protein precipitation technics. 5) Because of its binding to plasma protein and rapid excretion by the kidneys, radioactive ferrocyanide is considered unsuitable as a measure of extracellular fluid.

Chorioallantoic membrane tissue cultures (CamTC) in balanced salt solution (BSS) with pyruvate as the sole carbon source supported synthesis of influenza virus (strain WS) but not its neurotropic variant (strain NWS). Various other substrates were studied alone and together with pyruvate for their effect on synthesis of strain NWS in chorioallantoic membrane. Cells maintained in BSS containing glucose, xylose, or glycerol as the sole carbon source supported viral synthesis, while cells maintained in ribose, dihydroxyacetone, glycerophosphate, adenosine-5 phosphate, adenosine diphosphate, adenosine triphosphate, or diphosphopyridine nucleotide supported viral synthesis only in combination with pyruvate. It is suggested that synthesis of strain NWS in CamTC requires nutritional factors which are not supplied by cells maintained in a medium with pyruvate as the sole carbon source.

An attempt has been made to determine the cause(s) for variation in weight of suckling rats. It was found that the 14th day weight of rats did depend upon the number of “milking” glands of the mother rats. The average 14th day weight of young born in the first litter was lowest when compared with the weights at corresponding periods of those delivered in the subsequent litters. The average number of “functional” mammary glands of mothers nursing their first born was also the lowest as compared with those of mothers rearing their second, third and fourth litters.

Day-old chicks raised on a purified diet containing approximately 0.046% fat grew normally for 6-8 weeks, at which time weight gains began to decrease slightly. No marked symptoms of deficiency developed, but depigmentation of feathers and scaliness of skin were observed. Fat-deficient male chicks had smaller testes and some females had immature oviducts, when compared with normal chicks. Water consumption by the deficient birds did not increase markedly until the 11th week on the fat-free diet.

The after-discharge of the hippocampus is formed by a number of components which can combine to form several fairly discrete well organized patterns. These patterns seem to be related more to the state of the organism at the moment the after-discharge is produced than to the precise anatomic location of the electrodes or to the parameters of stimulation. Although the activity emanating from symmetrical electrode placements may show differences in pattern, there are systematic relationships between the right and the left which would indicate some bilaterally expressed control on the after-discharge. Extremely high voltage electrical after-discharge may be present in the hippocampal formation with only minimal disturbance in the dorsolateral cortex, at least by transverse recording, and without the precipitation of a major convulsion.

1) The results of 2 studies with chicks and one study using Broad Breasted Bronze turkey poults have indicated that addition of molybdenum to purified basal diets resulted in a growth advantage of 13.5-19.1% over the unsupplemented group. 2) The feeding of a reconstituted ash mixture which simulated the ash of distillers dried solubles resulted in significant increases in growth rate of both chicks and poults. 3) From the data presented it appears that under conditions of these studies a nutritional requirement for molybdenum by chick and poult may be demonstrated.

The feeding of large doses of nicarbazin to calves produced moderate nephrotoxicity associated with precipitation of crystalline deposits in the renal tubules. These crystals were identified as 4, 4′-bisacetamidocarbanilide. Additional evidence is presented that, after absorption, nicarbazin splits into its components, dinitrocarbanilide and hydroxydimethylpyrimidine, which are metabolized separately. The dinitrocarbanilide is first reduced to soluble diaminocarbanilide and then acetylated to insoluble 4,4′-bisacetamidocarbanilide. The renal tissue reaction and the metabolic degradation of 4,4′-dinitrocarbanilide resembles that known for the sulfonamides.

Fifteen rats with severed spinal cords, kept alive on a respirator, were given massive doses of epinephrine, that on controls produced consistently an intense acute pulmonary edema. None of these rats developed the usual acute edema, and in 10 cases the lungs had normal weight. This result is interpreted as a proof of the central nervous mediation of the acute pulmonary edema produced by administration of epinephrine.

Delayed type hypersensitivity to cat scratch disease can be passively transferred by blood leukocytes, but not by plasma from hypersensitive donors convalescent from the disease.

1. The influence of azetazoleamide upon fasting blood sugar and the glucose tolerance curve was studied in a group of diabetic and normal subjects. Azetazoleamide was given either as a single dose or for a period of 3 weeks. 2. No significant alteration of the fasting blood sugar or the glucose tolerance curve was noted following the administration of azetazoleamide.

1. In studying the mechanism of the “capsular reaction” one of the first questions to be answered is whether or not the capsule swells when antibody is added. Data presented here indicate that it does not, using C. neoformans as a test organism. Exposure of washed, encapsulated C. neoformans to antibody results in a prominent clarification of capsular outline, but no increase in capsular size as determined by packed cell volume after centrifugation and by direct microscopic measurement. When cells with large capsules are employed, there is an apparent decrease in capsular size during the reaction with antibody. 2. If capsulated cells are not washed free of soluble capsular polysaccharide present in the cultural supernate, the soluble antigen coprecipitates when antibody is added. This may lead to the erroneous conclusion that there is an increase in capsular volume.

Both yellow (A^y) and black (C57BL) mice produce grey hairs when weaned to a diet known to cause biotin deficiency. Histologically, the greyed hair follicles of biotin deficient mice bear a striking resemblance to the hair follicles of albino mice. The depigmented melanocytes of biotin deficient mice are similar in morphology to the follicular clear cells of the albino. Unlike clear cells, however, depigmented melanocytes react positively to tests for the presence of dopa oxidase.

A simple and rapid procedure is presented for the in vivo measurement of total body water in sheep using I¹³¹ 4-iodo-antipyrine as the test substance. Body water values for 62 young and mature sheep averaged 61.3 ± 6.5 and 55.3 ± 6.3, respectively, by the antipyrine method, and 62.3 ± 6.8 and 56.2 ± 6.6%, respectively by the labeled antipyrine procedure. Thyroid uptake of I¹³¹ from labeled test substance amounted to 3 to 5% and urinary excretion was less than 20% of administered dose during the critical 5-hour test period. Protein binding was evidenced, indicating that in sheep both test substances were reversibly bound to plasma protein. Consideration is given to the use of these body water volume values in establishing a predictable relationship to body constituents and to provide additional criteria for estimating animal response to experimental treatments.

The antibacterial thymus peptide has been characterized and strong confirmatory evidence has been presented to support the view that it has been derived from a histone present in thymus tissue.

Uterine response to estradiol in immature female mice is inhibited significantly by concurrent injection of estriol. Extent of inhibition is proportional to amount of estriol added. Impeding interaction occurs along the ascending segment of the estradiol dose-response curve and with higher doses as well. In no case does a combination of estrogens have a positive additive action. These conclusions are based on both wet and dry uterine weight observations. Day by day analysis shows that uterine weight gain of each 24 hour period during 3 day estradiol treatment is inhibited when estriol is given concurrently. These studies negate the possibilities that estriol-estradiol interaction is a masked-summation effect in which time of maximum effect has shifted, or that an additive effect occurs in which two estrogens combined exceed maximum response dose. Rather, it appears that estriol competes actively at a physiologic site essential in development of usual estradiol stimulation of the uterus.

1. Exposure of adult mice infected with the Conn. 5 strain of Coxsackie virus to a cold environment resulted in an infection with a persisting viremia, high levels of virus in the liver, pathologic lesions in other organs as well as in the pancreas, and a uniformly fatal outcome. 2. A lethal infection was prevented by neutralization of the virus or passive immunization of adult mice with specific antiserum. 3. The LD₅₀ for adult mice at 4°C was approximately ten times that for suckling mice at 25°C. 4. It was demonstrated that to initiate a fatal infection exposure to cold had to be continuous and begun within a few days after virus inoculation.

Butyrate, isobutyrate, propionate, caproate, and acetoacetate failed to reverse the block in acetate-1-C¹⁴ oxidation found in liver slices from rats fasted at 0-2°C for 24 hours. It appears that the ability of glucose and pyruvate to increase the acetate oxidation activity of these liver slices is related to a specific step in carbohydrate metabolism rather than solely to the fact that carbohydrates are excellent sources of energy.

Twenty-four dogs were subjected to an ambient temperature of sufficient magnitude to elevate rectal temperatures to 42.5†C for one hour. These animals were studied with regard to plasma 17-hydroxycorticosteroid concentration and plasma and erythrocyte electrolyte levels. Significant changes were observed in all measured variables with the exception of plasma Na concentration, which was not altered from the control value. The physiological implications of these data are discussed.

Increased activity of the enzymes involved in ammonia metabolism in the kidneys of guinea pigs was produced by prolonged treatment with acid or alkali. When the results were expressed on a per cell basis, Glutaminase I was increased in acidosis while in alkalosis Glutaminases I and II and the Glutamine synthesizing enzyme were increased. These increases in activity are suggestive of enzyme induction though all the criteria for this process have not yet been fulfilled.

(1) Lipide analyses of cellular fractions in liver necrosis produced by the administration of bromobenzene was studied in the rat. (2) A statistically significant decrease occurred in the total lipide, lecithin and cephalin phosphorus in the homogenate fraction of the necrotic livers as compared to the controls. However, the total cholesterol content was significantly increased. (3) In the nuclei of the necrotic livers a decrease in the concentration of the lecithin phosphorus and an increase of total cholesterol were noted.

The Thiobarbituric Acid Test for oxidative products of essential fatty acids (HUFA), primarily linolenic, present in tissues in vivo and after aerobic oxidation in vitro, with and without ascorbic acid, was applied to brain, liver and testes tissues of hamsters of various ages. Minute measurable quantities of oxidative products (probably peroxides) are formed in vivo. The quantity of HUFA present in brain tissue increased gradually with post partum development of the brain, with constant levels apparent after 10-16 days of age. The quantity of HUFA present in testes tissue correlates well with development and function of that organ with constant levels established, following a peak during age of sexual maturity. There is a significant rise in total HUFA content of liver tissue, associated with emergency of birth. Lack of catalytic activity of ascorbic acid upon in vitro oxidative reaction involved in embryonic tissue has been demonstrated.

1. Twenty-five compounds representing structural derivatives of benzoic acid were studied with regard to their effects on circulating 17-hydroxycorticosteroid (17-OHCS) concentrations in guinea pigs. 2. Benzoic acid had no effect on these concentrations. 3. Among the monohydroxybenzoic acids, the o- and m-substituted derivatives produced elevated plasma 17-OHCS concentrations. The latter compound exerted its effect earlier following administration than did salicylate. p-Hydroxybenzoate had no significant effect. 4. Among the mono-ether derivatives of benzoic acid only the o-substituted compounds produced significant elevations of plasma 17-OHCS concentrations. Significant elevations were produced by 3, 4, 5-trimethoxybenzoic acid. 5. Both o- and paminobenzoic acid produced steroid elevations of borderline significance. 6. The addition to the salicylate structure of a second hydroxyl group in the 3 or 6 position or of an ether group in the 3 position appeared to augment the effect on 17-OHCS concentrations. 7. Aspirin exerted the greatest effect on plasma 17-OHCS concentrations of any compound tested. 8. The salicylate metabolite, salicyluric acid, had no effect. 9. A comparison of the influence of these compounds on 17-OHCS concentrations with antirheumatic and anti-inflammatory properties and with effects on adrenal ascorbic acid concentrations, as reported by others, was attempted.

Adsorption losses of albumin-I¹³¹ to volumetric glassware may be sufficient to cause significant underestimates of plasma volume. Some factors affecting these losses are examined and methods of minimizing their effects are described.

Boiled filtrates of acidified plasma obtained from 7 patients with Cooley's anemia and one of 2 patients with sickle cell anemia stimulated erythropoiesis in intact rats. A similarly prepared extract of urine from one of the Cooley's subjects was also active. Plasma filtrates from normal subjects and one patient with chronic hypoplastic anemia were inactive. The implications of these findings are discussed.

1. Both cortisol and corticosterone were effective in protecting rats against endotoxin when the steroid hormones were given intracardially at the time of administration of endotoxin. 2. Cortisol was more protective than corticosterone when both were given subcutaneously 24 hours before endotoxin; as the interval between the time of injection of steroid and the time of challenge with endotoxin was decreased, corticosterone was increasingly protective. 3. The rates of disappearance of corticosterone and cortisol from the blood of rats after intracardiac administration of steroid were similar. 4. The evidence indicates that corticosterone is more rapidly mobilized from local tissue sites of deposition than is cortisol and that the difference in the activity of these 2 substances after subcutaneous administration is due in large part to this difference. By inference, the many published differences in the biologic activities of cortisol and corticosterone may be subject to revision when control of this variable has been achieved. 5. Corticotropin and lipoadrenal extract were protective against endotoxin but desoxycorticosterone was not.

1. In the normal thymus the relatively few mesenchymal reticular cells give a positive reaction for acid phosphatase in acetone fixed sections. There are fewer reactive cells in the cortex than in the medulla. 2. As early as 2 hours after irradiation the mesenchymal reticular cells are mobilized to become active macrophages which stain deeply in the acid phosphatase technic. These become more numerous until at 24 and 36 hours after irradiation the involuted cortex is loaded with macrophages containing ingested lymphocytes or full of vacuoles. At these hours the browned cytoplasm of the epithelial cells is seen as the stroma of the depleted cortex. At 3. 5 and 7 days there is a gradual diminution in number and size of reactive cells. At 9 days the appearance of the thymus is essentially normal.

Ten patients with hypercalcemia and hypercalcuria and 1 patient with normocalcemia and hypercalcuria were treated with 6-diazo-5-oxo-L-norleucine alone or in combination with 6-mercaptopurine. Full correction of hypercalcemia with treatment occurred in 8 of 13 instances in these patients. Correction of hypercalcuria was observed in 9 of 14 instances. Various possible mechanisms underlying these observed effects are considered.

The local or initial response of crustacean single motor axons has been recorded at the stimulating cathode. The deflections caused by a short duration shock of positive and negative polarity are asymmetrical. Normally the deflection caused by the negative pulse is considerably larger. In the absence of sodium, with either dextrose or Choline Chloride substituted, the local potential disappears. The deflections caused by positive and negative pulses become symmetrical. The effect is instantaneous and reversible. It is concluded that the sodium ion is essential for the production of the local or initial response.

The pH of tissue culture medium was altered purposefully to measure quantitatively the effects upon virus yield and host cell metabolism. By use of strain HeLa cells and poliovirus Type 1 (strain Mahoney) for growth in media of pH from 6.3 to 8.0 it was established that virus yield was affected by the pH of the culture medium. Decreased virus yield at decreased or increased pH levels was related to a decrease in metabolic rate of host cell cultures.

A method has been developed for chronic catheterization of one renal artery, with collection of urine from each kidney separately. The presence of the catheter has not altered renal function over a period of months. Preliminary studies with infusion of hypertonic saline into the renal artery of the dog in congestive heart failure indicate a marked increase in tubular reabsorption of sodium.

It was found that adenosine at 150 mg kg of ration failed to stimulate chick growth when supplementing a 30% protein, soy protein type purified diet. Supplementation of such rations with 2% fish meal ash gave only small and variable responses, while supplementation with 10% fish meal uniformly gave marked growth response.

The activity of the thyroid gland appears to be directly related to the incidence of dental caries in the rat, for thyroxine injected significantly decreases the caries rate and radiothyroidectomy significantly increases it. The thyroid hormone appears to be primarily involved since the increased caries rate resulting from radiothyroidectomy is directly counteracted by the administration of thyroxine. Testosterone similarly appears to play a role in this mechanism, since the dental caries rate is also significantly reduced when it is injected into radiothyroidectomized animals even in the absence of thyroxine.

Three salts of penicillin G and one of penicillin V were found to be essentially equal when administered orally in sparing the rat's dietary requirement for thiamine, as determined by rates of growth and thiamine levels in the liver. Growth was not stimulated to a statistically significant degree upon administering these antibiotics intraperitoneally every other day, although thiamine levels in the livers were elevated by such treatment. The effects of penicillin on thiamine levels in ileal and cecal contents were inconclusive.

Blood pepsinogen levels were measured in cats, dogs, rats, and man following acute and chronic administration of adrenocorticotrophic hormone and/or cortical steroids. Despite demonstration of adrenal cortical stimulation, pepsinogen activity did not increase. The data indicate that blood pepsinogen activity does not constitute an index of adrenal cortical activation.

A chromatographic system of high resolving power was devised for the purification of growth hormone and was adapted for use in the separation of other peptides.

1) Female albino mice of two different breeds were treated intraperitoneally with a mixture of bovine serum and pertussis vaccine. In confirmation of findings of other investigators, it was found that a large percentage of the animals so sensitized died within 30-60 minutes following the intravenous injection of very small amounts of serum 2-4 weeks after sensitization. The sensitizing effect appeared specific as no deaths occurred when either rabbit serum or egg albumin was used for the challenge injection. 2) Mice pretreated by intraperitoneal injection of bovine serum and pertussis vaccine, or of pertussis vaccine alone, were infected intravenously several weeks later with staphylococci resuspended in 1/2000 bovine serum. When they were sacrificed 24 hours after infection, it was found that many more staphylococcal colonies could be recovered from the kidneys, liver and lungs of the animals that had been sensitized to bovine serum, than from the organs of animals of the control group.

1. Intravenous administration of 0.1 γg Piromen/kg body weight to young, normal male and female subjects causes a significant rise in body temperature and plasma hydrocortisone levels. 2. Increased concentration of plasma free 17-OHCS following Piromen administration has been shown to be the result of increased secretion of hydrocortisone. 3. The rise in plasma hydrocortisone distinctly precedes the elevation of body temperature above 37°C. Fever is, therefore, not the cause of the elevated plasma hydrocortisone levels. 4. The increases in plasma hydrocortisone levels, eosinopenia, temperature, and pulse rate caused by Piromen administration are significantly lessened by the concomitant administration of aspirin. 5. Administration of reserpine in a dose of 4 mg/24-hr period caused no hypothermia in normal human subjects. 6. Pyrogenic, eosinopenic, and adrenocortical stimulatory effects of Piromen are not altered by the prior and concomitant administration of reserpine.

1. In man, the intravenous injection of synthetic lysine-vasopressin causes a rise in plasma hydrocortisone concentration comparable to that following the intravenous administration of Pitressin. The effect of the latter is therefore not due to impurities. 2. The pressor effect of synthetic vasopressin is of the same order as that noted following the injection of Pitressin.

1) Parathyroid extract increased the urinary excretion of calcium and strontium as well as increased deposition of these minerals in the kidney tissue. 2) Cortisone prevents accumulation of strontium and calcium in kidney tissue but does not influence urinary excretion or diuresis.

1. Increased amounts of nuclear DNA were found in basal cells during periods of augmented growth in the estrous cycle. 2. These changes in nuclear DNA parallel the mitotic activity of the basal cells. 3. Exfoliated basal cells containing intermediate and tetraploid amounts of DNA were found only during early proestrus. 4. The paucity of a tetraploid nuclear population during estrogenic stimulation may indicate rapid cellular division once DNA synthesis has occurred.

In Zonotrichia leucophrys gambelii there is a striking deposition of fat in subcutaneous and visceral depots following prenuptial molt and preceding beginning of vernal migration. During the last 2 weeks of April, in natural populations, mean lipid indices increase from about 5% to more than 20%. A similar premigratory fat deposition occurs in captive birds under natural conditions of photoperiod and temperature. Similarly in these captive birds this fat deposition is followed by development of migratory behavior (Zugunruhe). Measurements of metabolizable energy intake in these birds show that fat deposition is accompanied by a marked increase in energy intake to levels of 30 to 50% above the preceding minima. This temporary hyperphagia represents an altered physiologic state rather than simply the effect of more daylight for feeding since, during hyperphagia, the amount of daylight increases only 9%. The temporary hyperphagia is the principal factor responsible for positive energy balance required for premigratory fat deposition. The role of daily photoperiod in development of temporary hyperphagia was demonstrated experimentally by subjecting birds in mid-winter to 20 hours of light per day. These developed a hyperphagia and an accompanying fat deposition very similar to those observed in spring under natural conditions.

In the epithelial metaplasia of the rat uterus that occurred following vit. A deficiency, glycogen was observed in the stratified epithelium. An earlier report showed that the normal uterine epithelium of the rat uterus contains no glycogen.

1) Two patients with choriocarcinoma and one with chorioadenoma destruens with proven progressive metastases were treated with methotrexate, a folic acid antagonist, according to intensive regimen of repeated courses of dosages producing substantial but reversible toxicity. Initially high urinary gonadotropin present in all 3 cases approached normal levels following this regimen and such low levels were sustained following cessation of treatment. 2) Unequivocal clinical improvement and radiological evidence of regression of pulmonary metastases was observed in all 3 cases. Nevertheless, extreme variations in pattern of clinical behavior of this type of tumor necessitate more observations before any definitive therapeutic implications may be drawn from these studies.

The anti-inflammatory activity of egg yolk has been studied in weanling guinea pigs. 1. The active material resided in the alcohol soluble fraction and was absent from the protein and acetone soluble fractions. 2. The anti-inflammatory effect did not appear to be mediated by the adrenals. 3. Feeding egg yolk and yolk alcohol solubles resulted in an elevation of serum total lipids with no increase in cholesterol to phospholipid ratio. Inactive yolk fractions caused elevations of serum total lipids, phospholipids, cholesterol, and cholesterol to phospholipid ratio. 4. Serum levels of total N and non-protein N were unaffected by yolk alcohol solubles.

The effects of orally and intravenously administered Δ¹-hydrocortisone and hydrocortisone on plasma 17-OHCS concentrations in the dog have been studied. Δ¹-Hydrocortisone produced higher and more prolonged plasma 17-OHCS elevations than did equivalent doses of hydrocortisone administered orally. The half-life of Δ¹-hydrocortisone was found to be 1.37 times that of hydrocortisone. Intravenous administration of these steroids as the hemisuccinates gave lower but somewhat more prolonged elevations in plasma 17-OHCS concentrations than the corresponding free alcohols. It was concluded that Δ¹-hydrocortisone was metabolized more slowly than hydrocortisone and suggested that this may explain, at least in part, the enhanced potency of the Δ¹-steroid.

The preparation of PHE and their use in both tube and rapid plate HA and HAI procedures for the titration of influenza virus and viral antibodies are described. Such preserved cells have been stable, with respect to both resistance to lysis and sensitivity to hemagglutination by influenza virus, for a year. PHE showed good correlation in sensitivity with freshly washed human erythrocytes, and the rapid plate technic using PHE correlated well with tube HA and HAI procedures.

The radioactive rose bengal test, a very sensitive test of liver function, can be analyzed graphically to yield three quantities —uptake halftime, excretion halftime and fractional liver blood volume. The values in various liver diseases seem to be characteristic of the disease and to indicate whether cellular damage or obstruction predominates.

In order to study postictal recovery, groups of normal and Dilantin-treated mice were given a supramaximal electroshock and the durations of the hindleg components of the “first” seizure recorded. At various time intervals after the “first” seizure, a “second” supramaximal electroshock was given and the durations of the hindleg components of the “second” seizure recorded. This procedure was repeated, with longer time intervals between seizures, until the observed durations of the “second” seizure components were not significantly different from their “first” seizure values. In addition, the time (RT₅₀) required for 50% of animals to recover ability to exhibit a second tonicclonic seizure after a first was determined. The data obtained may be summarized as follows: The order in which the durations of the various components and the total duration of the “second” seizure returned to their “first” seizure values were the same in both groups: total seizure duration, terminal clonus, tonic extension, and tonic flexion. The total duration of a maximal seizure is relatively independent of marked changes in duration of the individual seizure components. Dilantin prolonged the recovery time of terminal clonus, tonic extension, and tonic flexion by approximately 300%, 150%, and 170%, respectively. In addition, Dilantin prolonged RT₅₀ by 325%. Evidence is presented which suggests that Dilantin acts to prolong the latent period before recovery of the tonic-extensor component starts, and that the recovery process per se, once initiated, is relatively unaffected by the drug.

Utilizing the tetrazolium technic, intracellular dehydrogenase activity has been localized in epiphyseal cartilage cells. The most favorabe substrates appear to be succinate. isocitrate, citrate, lactate and malate.

(1) The RNA content of 2484L treated C3H mammary adinocarcinomas was shown to be lower than that of untreated controls. (2) The nucleotide composition of both treated and control tumors appeared to be similar.

Anaerobic glycolysis is not increased in brain slices from rats injected with 4-8 mg of L-thyroxine, when compared to similar values from normal control rats.

Four strains of M. aureus were rendered resistant to erythromycin, carbomycin, oleandomycin, spiramycin and streptogramin by repeated subcultures in the presence of increasing concentrations of these antibiotics. Cross-resistance to about the same degree developed to all of these agents except to streptogramin; the latter produced homologous resistance at a slower rate than the others, but this was associated with a corresponding increase in cross-resistance to the other 4 antibiotics.

Intravenous reserpine in rabbits has been demonstrated by chemical means to release histamine from whole blood. 1. The minimal histamine levels were reached in one to 2 days with complete recovery to control values in 5 to 7 days. 2. Simultaneous serotonin determinations, revealed that the rate and degree of liberation, and the recovery of both histamine and serotonin were very similar. 3. Platelets, isolated 24 hours after reserpine, showed a marked reduction in both histamine and serotonin compared to control platelets.

The authors have been unable to find any report of results of paper electrophoresis of hemoglobins other than human. The present communication presents findings in 6 species of animals.

Methods. Electrophoresis was performed on “sandwich” type plate glass apparatus using Whatman 3MM paper in sheets 36 × 16 cm and 36 × 12 cm. Barbiturate buffer at pH 8.6 and M 0.025 was used in all preliminary experiments. Current was applied to 36 × 16 cm sheets at 300 volts and 5 milliamperes for 6 hours and to 36 × 12 cm sheets for 4 hours, constant temperature of 20°C was maintained. Under these conditions normal human hemoglobin (Hb. A) migrated approximately 5 cm during the run. In addition to electrophoresis alkali denaturation tests (1) and solubility tests (2) were carried out on all specimens (Table I). Rat hemoglobin was obtained from the pooled blood of adult Wistar rats killed at the end of experiments. Dog hemoglobin was from mongrel dogs used in experimental surgery. Rabbit hemoglobin was from albino rabbits used for pregnancy tests. Chicken hemoglobin was from pooled blood obtained from a poultry farm. Turtle hemoglobin was from several specimens of the common land turtle, Gopherus polyphemus. Alligator hemoglobin was from a single small specimen of Alligator missippiensis. Hemoglobin solutions were prepared in the usual manner and adjusted to approximately 5%, 3 lambda of the solution were applied to the paper.

Results. As shown in Fig. 1, dog and rabbit hemoglobins move as homogenous compounds at a rate slightly slower than fetal hemoglobin but faster than Hb. S while rat hemoglobin produces a trailing spot suggesting a mixture of hemoglobins; this we have been unable to resolve by varying the buffer or any other factors.

The toxic effects of 4-aminopyrazolo-(3,4-d)-pyrimidine (4-APP) have been studied in mice, rats, cats, and dogs. The agent was found to produce a fatty change in liver which was associated neither with necrosis nor with inflammation. Hepatic insufficiency was evidenced by hyperbilirubinemia, increases in one-stage prothrombin time and decreases in plasma fibrinogen. In rats fat nephrosis and depletion of hematopoietic elements in bone marrow were also seen. Hepatic insufficiency should be anticipated as a possible hazard in clinical trials with 4-APP.

Simultaneous electrophoretic and ultracentrifugal studies of the serum proteins of a multiple myeloma patient have conclusively demonstrated a considerable Bence-Jones proteinemia. The properties of the serum component correspond to those of one of the major urinary components.

The location and approximate concentration of gonadotropic activity in various fractions of human postmenopausal plasma has been determined. The plasma was fractionated by means of the cold ethanol method VI of Cohn and was assayed in immature female rats by estimating ovarian growth and secretory activity. Although traces of activity were found in Fractions IV-1, IV-4 and V, the bulk of the gonadotropically-active material was located in Fraction II + III. Subfractionation of Fraction II + III by successive precipitations with zinc and 20% ethanol freed the gonadotropins from the majority of the contaminating proteins.