Experimental Biology and Medicine

1. Preliminary experiments with the continuous production of monochromatic ultra-violet absorption images of living cells indicate that this may be accomplished by the flying spot television microscope technic.

Phenylpyruvic acid, phenyl-lactic acid and phenylacetic acid inhibit beef adrenal medulla DOPA decarboxylase. The implications of these data in phenylpyruvic oligophrenia are discussed.

The ability of a mixture of SK and human plasminogen to activate bovine plasminogen was shown to be dependent upon the concentrations of both components. The ‘activator' which was formed, may deteriorate on incubation even though the SK and human plasminogen when incubated separately were stable. The decrease was partially or completely restored by the addition of SK. The rate of disappearance of the ‘activator' was affected by the plasminogen preparation and by the initial SK concentration. The in-activation occurred at physiological pH as well as at pH 6.4. The in vitro and in vivo significance of the data were discussed.

Biochemical studies have been performed on a phenylketonuric infant from birth to the age of 34 days. Cord blood and newborn infant blood contained normal levels of phenylalanine, the serum phenylalanine rose to 62 mg % at 24 days, and decreased to 41% at 34 days. Phenylpyruvic acid was not excreted in detectable amounts until 34 days. o-Hydroxyphenylacetic acid was not present in excess at 5 days, but from 16 to 34 days was excreted at about one-fifth the rate usually observed in phenylketonuria. The minor abnormal metabolites indolelactic and p-hydroxyphenyllactic acids were not present at 5 days, but were present at 16 days.

Both glyoxal and β-ethoxy-α-ketobutyraldehyde (Kethoxal) were more potent virucidal agents than β-propiolactone (BPL). Glyoxal was less lytic for human erythrocytes than was Kethoxal or BPL. None of the chemicals showed significant toxicity for plasma proteins as measured by electrophoresis. These experiments indicate that glyoxal may be of potential value in the sterilization of human blood and plasma.

1. Plasma of normal adult female rats contains a factor capable of stimulating erythropoiesis in hypophysectomized and newborn rats, as judged by increased circulating red cell volume and hemoglobin. 2. This humoral erythropoietic factor is markedly increased during pregnancy. 3. Injections of plasma from non-pregnant and pregnant rats resulted in an increase in plasma and blood volumes of hypophysectomized and newborn recipients.

Rabbits injected with human serum “PTC” developed inhibitors to human PTC and also to human proconvertin. These two activities could be separated by differential adsorption.

Phenacyl bromide, p-chlorobromobenzene, or naphthalene tetrachloride inhibit growth of rats which ingested an 8% casein diet. The inhibition was alleviated by supplementary cystine, or by a 27% casein diet. Nitrogen balance remained positive in rats which ingested the 27% casein diet or the 8% casein diet which was supplemented with cystine. Intraperitoneal injection of naphthalene tetrachloride into adult rats on the high or low protein diet induced a negative nitrogen balance, accompanied by a decrease in the -SH and S-S content of the whole blood and in the protein content of practically all tissues. The implication of these observations is discussed in terms of possible interference of the hydrocarbons with the protein metabolism of the rat via direct binding of the hydrocarbons to the proteins through the -SH or S-S groups.

An alcohol precipitate obtained from heated bovine thrombin preparations promoted the early phases of the coagulation mechanism, but did not clot fibrinogen in the manner of the unheated preparation. These materials inhibited bovine thrombin at lower concentrations that human thrombin in the clotting of fibrinogen.

Derivatives of bovine fibrinolysin have been prepared which inhibited the enzyme from which they are derived, but did not inhibit human fibrinolytic activity.

1. Dietary fats of varying degrees of saturation were no different from each other with respect to their effect on aortic or hepatic cholesterol levels of cholesterolfed cockerels and only slightly with respect to serum levels. 2. When 0.1% Gallogen was added to the diet, serum and aortic levels were in proportion to the degree of saturation of the fat fed.

The tryptophan content of casein has been estimated using a rat growth assay method. Whether the results were calculated on the basis of the percentage of tryptophan in the diet or by using food consumption data to calculate the daily intake of tryptophan, essentially the same results were obtained. These animal assays indicate that casein (14.64% N) contains 1.25% tryptophan which is in good agreement with many reports based on microbioassays of alkaline and enzyme hydrolysates of this protein, but much lower than many commonly accepted figures obtained with chemical methods.

1) Dogs were depleted of body proteins by forced hemoglobin production superimposed on a low protein diet. Electrophoretic patterns indicated markedly decreased albumin values and A/G ratios in spite of relatively little change in total plasma proteins. In all instances there were marked increases in total α, β and γ globulins. 2) Anaphylactic reactions following immunization to horse serum were diminished or absent in the depleted animals as contrasted to controls.

Since the classic studies of Harvey(1) it has been generally accepted that diastolic filling of the heart resulted from venous return and auricular contraction. While the senior author(2) was studying the effects of anoxia and work on the glycogen content of the rat heart, it was necessary to make observations on the excised heart beating in saline. With each systole it was observed that fluid ejection from the aorta resulted in the jet propulsion of the heart in the solution and it was obvious that the heart must draw in fluid in each diastole to make possible subsequent systolic ejection; thus it was concluded that part of systolic energy was dissipated in diastole(2).

This paper presents the results of a study of pressure changes in intact and excised heart during systole and diastole.

Materials and methods. White rats weighing 300 to 450 g were anesthetized with pentobarbital 6 mg/100 g. The thorax was rapidly opened and the intact heart removed. A 20 gauge needle was inserted into the left ventricle and the needle connected to a three-way stop cock on a Statham strain gauge so that opening the stop cock recorded hydrostatic zero. The heart was placed in a pottery pie plate filled with saline. The three limb leads of the EKG were placed equidistant around the perimeter of the plate with the electrodes immersed in solution. Pressures and EKG were recorded with an Oscillograph. When desired the auricles were opened widely or excised with a pair of scissors. In vivo studies were performed on dogs anesthetized with pentobarbital. Artificial respiration was administered through an intratracheal tube and the chest was widely opened. Umbilical tape was placed around the superior vena cava and azygos veins.

1. Administration of Δ¹-cortisone and 9$aL-fluorohydrocortisone changes normal urinary excretion of steroid metabolites, indicating adrenal depression or altered steroid metabolism. 2. Only a small proportion of administered 9$aL-fluorohydrocortisone is excreted as $aL-ketolic metabolites; Δ¹-cortisone also is converted to a great extent to non-$aL-ketolic elements. 3. Elemental sulfur has been found to give a positive reaction with blue tetrazolium. This interferes with the quantitative determination of $aL-ketols in extracts of cat urines. 4. The $aL-ketols and sulfur can be separated by chromatography and each determined quantitatively by the blue tetrazolium reaction, using a modified method. 5. Administration of Δ¹E and 9$aL-fluorohydrocortisone causes an increase in urinary excretion of sulfur-containing compounds in cats, which leads us to conclude that these steroids alter sulfur metabolism.

(1) Intraprostatic deposits of 20-methylcholanthrene crystals induced three types of prostatic neoplasms in the ventral lobes of the Wistar rat's prostate gland: adenocarcinoma (2%), leiomyosarcomas (3%) and squamous cell carcinomas (30.3%). The squamous cell carcinomas and adenocarcinomas metastasized. (2) A detailed account of the distribution of invasion and/or metastases of these prostatic tumors to various organs are presented as well as an account on the growth characteristics of transplantable squamous cell carcinomas.

Rats were adapted to the force feeding of a moderate or high carbohydrate or high fat diet for several weeks before phlorhizinization. During the second day of phlorhizin injections, the animals were fed their respective diets, rendered protein free, and the urinary excretions of N.P.N. measured and compared to the outputs of starving animals. The feeding of either neutral fat or carbohydrate reduced N.P.N. losses. A similar sparing of endogenous protein occurred when animals, fed the high fat diet for 3 weeks, were given oleic acid, rather than mazola oil, on the second day of phlorhizinization.

1. The quantity of milk secured by litters of lactating rats treated with ergotamine 10 minutes before nursing was significantly less than that obtained by control offspring. Oxytocin administered before or after ergotamine restored normal milk letdown. 2. Topical application of ergotamine to living rat mammary tissue resulted in arteriole constriction but did not interfere with normal myoepithelial contraction induced by oxytocin. 3. Results indicate ergotamine does not inhibit milk let-down through vasoconstriction of mammary blood vessels thus prohibiting access of oxytocin to effector myoepithelium. Rather, a competition between ergotamine and oxytocin for the effector tissue or a neural block inhibiting release of oxytocin by posterior pituitary gland seem more probable.

Fermentations of glucose in the presence of carbon dioxide-C¹⁴ were carried out with strains of Actinomyces under aerobic and anaerobic conditions and with Lactobacillus bifidus. Fermentation products were isolated and degraded. It was found that succinic acid formed by these organisms had a high percentage of its radioactivity in the methylene carbons. When one strain of Actinomyces was grown in the presence of oxygen, 90% of the total activity of the succinic acid was found in the methylene carbons as compared to 62% when it was grown anaerobically.

A simple test for erythropoietin in deproteinized plasma extract is described. A total of 3 or 4 days is required for subcutaneous injection of the plasma extract and only one intravenous injection of Fe⁵⁹ is necessary. When concentrated plasma extracts are given to hypophy-sectomized rats, 6 to 9 fold differences in response between anemic and control groups are achieved. The deproteinized nature of the material enables cross-species examination of the erythropoietic factor.

1) Smooth brucellae were ingested by, and multiplied within, fibroblasts in tissue culture. Non-smooth variants and strain 19 neither multiplied nor survived long intracellularly unless the cells were massively infected. Brucellae remained viable intracellularly for over 30 days despite the presence of a bactericidal concentration of streptomycin in the tissue culture medium. Extensive cytopathogenicity was not evident except with Brucella suis which caused slow destruction of the fibroblasts. Under the same conditions of culture Salmonella typhosa multiplied rapidly intracellularly and caused complete cell destruction within several days. 2) Mixed infections of fibroblasts with smooth and non-smooth brucellae showed the intracellular environment to be highly selective for the smooth type. Mutation from I to S and M to S, followed by selection of the S type, occurred in several heavily inoculated tissue cultures. The addition of anti-S antiserum or DL-$aL alanine to the tissue culture medium did not affect the smooth-selective behavior of the intracellular environment.

Thirty-seven of 41 normal female rats, in which the ends of the tails were transposed into the abdominal cavity, showed a stimulation of hematopoiesis in the intra-abdominal vertebrae. Developing cells of all 3 types—erythroid, myeloid and megakaryocytes—were observed in these animals. In similarly treated hypophysectomized female rats only 1 of 16 animals showed any indication of response. The experiment provides another example of the depressed hematopoietic potential of the hypophysectomized rat.

1) Normal macrophages from the peritoneal cavity of the normal and shocked rabbit, and granulocytes from the irritated peritoneal cavity of the normal rabbit show no significant morphologic injury after immersion in normal rabbit plasma for one hour at 37°C. But after immersion in plasma from rabbits in hemorrhagic shock, a considerable percent of the cells show morphologic injury. A large number of granulocytes show changes similar to those characteristic of the L.E. cells. These changes are reversible if the shock plasma is replaced by normal plasma. 2) Shock plasma produces a severe depression of the phagocytic index; the percent of bacteria ingested is some 40% less than in normal plasma, and the percent of cells containing bacteria is some 30% less than in normal plasma. Shock plasma also reduces the bacteriostatic power of the phagocytes. The granulocyte in this respect appears to be damaged more severely than the macrophage. The extent of the injury produced by the plasma from rabbits in advanced shock appears to be no greater than that produced by the plasma of rabbits after two hours of shock, i.e. while they are still responsive to transfusion. The injurious property of plasma of the reversibly shocked rabbit persists, with undiminished potency, for at least four hours after transfusion. 3) It is concluded that a leucotoxin develops in the blood of the rabbit in hemorrhagic shock, and that this leucotoxin severely impairs the antibacterial potential of the animal.

Guinea pig liver homogenates reduce androstane-3, 17-dione to androsterone, epiandrosterone, androstan-17β-ol-3-one and androstane-3a, 17β-diol. The latter two compounds were observed only in trace amounts when addition of co-factors to the incubation mixture produced a marked increase in the amount of metabolites formed. Anaerobic conditions had no effect on the conversion produced by DPN. FDP and Na. The system which produced epiandrosterone exhibited an optimum pH of about 6.7. The formation of the other metabolites, however, was not affected by changes in pH. Approximately three-fourths of the ability to reduce androstane-3, 17-dione was in the supernatant fraction of homogenates centrifuged at 1500 x g.

1. The neuraminic acid (NA) content of 49 samples of human cerebrospinal fluid (CSF) was studied. In all instances the quantitative relationship of neuraminic acid to the cerebrospinal fluid protein was far in excess of that prevalent in serum. In 26 samples in which the total protein was 30 mg % or less, the neuraminic acid was 4-18 times in excess of that expected if it were derived from serum and bound to protein, as is the case in blood. Unlike its state in serum, most of the CSF neuraminic acid is in freely dialyzable form (60-80%). That moiety of CSF neuraminic acid which is derived from serum is in protein-bound, non-dialyzable form. 2. The origin of the dialyzable NA is unknown, but is not related to ingress of serum proteins into the CSF. The implications of these findings are discussed.

Incorporation of 20% fat into a corn-soybean oil meal diet increased ten to twenty-fold the vit. B₁₂ requirement of non-depleted chicks for optimal growth at 4 weeks of age. The requirement was between 0 and 5 γ per kg of diet (3% fat) and between 50 and 100 γ with the high fat diet. The high dietary fat did not deplete the chick of its vit. B₁₂ liver store or alter the absorption of dietary vit. B₁₂ and its subsequent storage in the liver.

1. Experiments with Fe⁵⁹ were carried out to test the hypothesis that hypoxia increases iron absorption. 2. Hypoxia was found to increase iron absorption by approximately one-third.

The details of a new method of cutting split thickness skin grafts from small animals are reported with a discussion of some of the advantages.

1. Determinations were carried out of the effect of extracts from developing chick leg muscle on the increase in area of coverslip cultures derived from chick leg muscle cells. 2. Maximal stimulation with the lowest extract concentration was obtained with extracts from muscle of the 19 day embryo and the 10 day chick. Extracts from the 12 day embryo and the one-year chick were less effective. 3. The activity of the muscle extracts was found to be unrelated to the rate of cell proliferation in chick muscle tissue.

1. Dogs cooled to 23°C 6 or 12 hours and then rewarmed, maintained sterility of blood during cooling and following rewarming, suggesting that the gastrointestinal tract and other bacterial foci maintained their integrity for at least 12 hours of hypothermia. 2. Hypothermic and normothermic animals intravenously inoculated with bacterial suspension were able to rid blood within 6 hours of 99% of the injected pathogens. Within 24 hours after injection of cultures of A. aerogenes and E. coli, normothermic animals cleared the blood completely of bacteria while dogs maintained hypothermic did not. However, within 12-24 hours after rewarming only 1 of 11 hypothermic animals had a bacteremia. 3. Inability to survive following prolonged hypothermia does not seem to be related to bacteremia.

The urinary excretion of 5-hydroxyindole acetic acid in patients with malignant hypertension and chronic glomerular nephritis is reported. Low levels were recorded for 3 patients exhibiting the malignant phase of hypertensive cardiovascular disease and in each patient with chronic glomerular nephritis. All patients exhibited extensive renal involvement and 5 of the 6 patients with malignant hypertension had severe arterial lesions. The 2 patients on whom determinations were made after removal of early carcinoid tumors did not show abnormal urinary 5-hydroxyindole acetic acid levels. Recovery studies show that proteinuria does not interfere with the determination of 5HIAA in the urine.

Aminoacetonitrile, with C¹⁴ in its cyanide group, was synthesized. When this compound was administered to weanling rats, the major portion of the radioactivity was eliminated in the urine, while a minor part appeared in respiratory CO₂. High initial C¹⁴ concentrations were observed in various body tissues. The proportion of this C¹⁴ due to unchanged nitrile decreased fairly rapidly with time. Labeling was found in tissue proteins, and to a lesser extent in liver lipids and glycogen. Also, in rats fed a diet containing isotopic aminoacetonitrile, radioactivity was detected in the sulfated polysaccharides of the enlarged epiphyseal plates of the bones. The presence of C¹⁴ in glycine and serine of liver protein and in urinary creatinine, allantoin, and hippuric acid, indicated an appreciable conversion of the cyanide radical into a carboxyl group. However, the major portion of the C¹⁴-containing metabolites was not identified.

Chronic dietary experiments in rats have revealed that months are necessary to evaluate the ultimate level of serum lipids resulting from sustained feeding of various levels of dietary protein. Quantitative changes in dietary protein can influence the levels of serum cholesterol and cholesterol/lipid phosphorus ratio. Reduction or elevation of dietary casein beyond a modest range (12-18%) will lead to ultimate elevation of serum cholesterol and phospholipid under conditions herein described. The insignificant change in cholesterol/lipid phosphorus ratio at low levels of dietary casein as opposed to its elevation at higher levels suggests that different mechanisms are operating in these two situations. Unusually high levels of serum cholesterol were achieved in old rats fed 40% casein. Coronary arterial atheromatous lesions developed in one-third of these animals, but in none of the other groups in the 6-month period.

1. Increasing x-irradiation of nursing female mouse caused increasing retardation of growth of suckling young. There is some effect even after 50 r exposure of the mother on the 3rd day post-parturition. There was pronounced effect at 200 r or greater. 2. There is no evidence of any toxic substance passing through the milk to the young. The effect seems to be one of simple starvation, probably through general debilitation of x-irradiated mother. 3. Newborn mice starved by an x-irradiated mother will recover rapidly if given to a normal lactating mother at 15 days postparturition, and by 30 days will be essentially like controls with respect to body weight. Recovery from initial starvation appears to be complete.

The prolactin and thyrotrophin content of various transplantable pituitary tumors was studied. 1. Prolactin was found in stilbestrol induced and stilbestrol dependent rat pituitary tumor and also in an autonomous mammotrophic mouse tumor in concentrations only 10% or less that found in bovine pituitaries. These tumors contained little or no thyrotrophin. 2. Two autonomous adrenotrophic tumors contained no detectable prolactin or thyrotrophin. 3. Two dependent thyrotrophic tumors contained no detectable prolactin while the thyrotrophin concentration was ten times that of bovine pituitaries. A five gram thyrotrophic tumor contains 5,000 times more thyrotrophin than one mouse pituitary. Blood levels of thyrotrophin were 2,000 times normal.

1. Phenaglycodol, a compound originally selected for clinical trial on the basis of its anticonvulsant and effective tranquilizing actions, has been shown to have a quieting effect on mice, cats and monkeys. 2. Its effects on blood pressure, respiration and electrocardiogram are minimal. 3. In studies on selected reflex arcs it has been shown to be a selective depressant of polysynaptic pathways at the spinal and supra-spinal level. 4. After doses of phenaglycodol causing sedation but not sleep in cats, the electroencephalogram shows a pattern of 9-12 c/sec. synchronous activity at both surface and deep electrodes.

Palmitic and oleic acids are the predominant non-esterified fatty acids of human plasma. Their concentrations fall proportionately after an injection of insulin.

Carbutamide, a sulfonylurea compound capable of lowering blood sugar, did not have an insulin-like action in increasing glycogen deposition of isolated rat hemi-diaphragm. Furthermore, this drug did not produce any enhancement of the insulin effect. In fact, when higher concentrations were used there was a significant inhibition of the insulin effect. This inhibition is somewhat non-specific since it was also observed when sulfadiazine was substituted for Carbutamide. It is suggested that this inhibition, the inhibition of insulinase, and the in vitro reduction of the epinephrine and glucagon response which have been attributed to the sulfonylurea compounds might all be due to a non-specific protein poisoning action of these drugs in higher concentrations.

Strips were prepared from aortae of rats and their reactivity to epinephrine tested by means of a lever system that recorded shortening of the smooth muscle. A 2% salt diet increased the sensitivity of the vessel to 1-epinephrine in both sexes. Male aortae in all instances had greater sensitivity to epinephrine than those of females under the same conditions. Castration markedly increased the sensitivity to epinephrine in females, especially when the castrates were placed on the salt diet. In contrast, male aortae were not particularly changed by castration.

The mechanism of the hypoglycemic action of carbutamide in normal dogs was studied by means of pancreatic-femoral and mesenteric-femoral cross-circulation experiments. The results of these acute experiments suggest that: 1) carbutamide hypoglycemia is accompanied by secretion of insulin; 2) carbutamide does not cause degranulation or other significant changes in appearance of pancreatic A and B cells; 3) clearance of carbutamide from blood is relatively slow, as reported by others(21); 4) some dogs are refractory to the hypoglycemic action of carbutamide in doses of 50 mg/kg I.V. The results are consistent with the hypothesis that insulin secretion is one of the results of carbutamide action. The continued administration of carbutamide in animals and men with reduced pancreatic reserve may stimulate the islets of Langerhans to regeneration or to exhaustion. Although some evidence suggests that islet growth does occur (10,12), the second alternative has not been ruled out and should be investigated before the sulfonylureas can be recommended for the prolonged treatment of diabetes mellitus.

In the rabbit, plasma phospholipide synthesis continued at a reduced rate in absence of liver, intestine and spleen. Removal of kidney further reduced plasma phospholipide synthesis. The rate of synthesis of kidney, lung or muscle phospholipides from phosphate precursors does not appear to be altered by evisceration.

Primidone (Mysoline) is in part converted to phenobarbital by dog and by man. Concentrations of phenobarbital found in plasma of patients receiving therapeutic doses of primidone are high enough to exert a significant antiepileptic effect.

A hemagglutination test is described, wherein the sera of individuals diagnosed as having contracted viral hepatitis agglutinate the red blood cells of the rhesus monkey in dilutions of 1:8 or greater, as contrasted with the sera of a majority of apparently healthy blood donors and of jaundiced patients in whom a diagnosis of viral hepatitis was excluded. The observation, although made empirically and requiring confirmation, appears to offer promise and is being investigated at greater lengths.

Three natural and 2 synthetic estrogens were compared for hypotensive activity in metacorticoid hypertensive rats. The most active compound was 16-epiestriol 3-methyl ether.

It has been shown that high speed vibration of cells of P. pestis in the presence of “ballottini” is accompanied by disintegration of the bacillary forms with concurrent release of antigenic material. At least one antigen was present in the lysed material which could not be demonstrated when cells were not lysed. 2. P. pestis and P. pseudotuberculosis appear to share 2 free “envelope” antigens as well as a single somatic antigen. In addition, it has been shown that P. pestis possesses a minimum of 1 “envelope” and 3 somatic antigens which are not shared with P. pseudotuberculosis. No antigens which could be considered specific for P. pseudotuberculosis were revealed.

Noludar and pentobarbital alone and in combination with morphine were compared as to their respiratory minute volume depressant effects in rabbits. No statistically significant difference could be found between Noludar alone and in combination with 0.603 mg/kg of morphine. Pentobarbital, on the other hand, was significantly more depressant in combination with morphine at low pentobarbital doses but not at anesthetic levels.

Inorganic phosphorus content of blood and liver and hepatic glucose-6-phosphatase activity are increased in scorbutic guinea pigs. Glucose-6-phosphate, fructose-6-phosphate and fructose-1,6-diphosphate contents of liver are decreased in scorbutic guinea pigs. Fructokinase activity of brain remains unchanged in scorbutic guinea pigs. Considerable increase in weight of livers of scorbutic guinea pigs has been observed.

A novel method for the sensitive and specific microestimation of steroids has been described. This method depends upon the selective oxidation or reduction of respectively hydroxy- and ketosteroids by highly purified hydroxysteroid dehydrogenases of bacterial origin. The method permits the estimation of 3a-hydroxysteroids, 3β-(and 17β-) hydroxysteroids, 3-ketosteroids and 17-ketosteroids.

Molybdenum, iron and calcium ions have been shown to be specifically required when Azotobacter vinelandii O is fixing atmospheric nitrogen. Ammonium ion eliminates the need for molybdenum and calcium and has a sparing action on the requirement for iron. High levels of iron stimulate the knallgas reaction of A. vinelandii O fixing N₂ in the presence of H₂, but molybdenum in the range tested had no effect on this reaction. Low concentrations of calcium ions in a nitrogen-fixing culture result in an extended lag phase of growth. Decrease in the concentration of phosphate in the presence of low levels of calcium brings about a further increase in the lag, but this increase is not observed if the calcium supply is adequate.

1. Uricolysis occurs in whole blood as well as in plasma incubated with white or red blood cells. The rate of uricolysis is not dependent on the hyperuricemia in normals. There is marked defect in uricolysis in leukemics under the same experimental conditions. 2. Hyperuricemia in leukemia, may be in part due to diminution in the rate of uricolysis.

No evidence of a positive anti-globulin test was found in dogs following injection of acetylphenylhydrazine. This is in contradiction to the results of others.

It is shown that Russell's viper venom (Stypven) does not correct the prolonged one-stage prothrombin time of patient with a congenital hemorrhagic diathesis previously thought to be deficient in factor VII. The factor deficient in this patient which is referred to as the Stuart factor, is readily distinguishable from both the Prower factor and factor VII since the prolonged prothrombin times of plasma deficient in either of these two factors are completely corrected by Stypven. The finding that Stypven does not correct the clotting defect of Stuart factor deficient plasma implies that certain modified one-stage “specific” assay methods for prothrombin using Stypven as a source of factor VII are not, in fact, specific, and measure both prothrombin and Stuart factor.

Gey's strain HeLa cells were cultured in Earle T-60 flasks for almost a year in media composed of 0.25 g. yeast and 0.25 g. lactalbumin hydrolysate/1 Earle solution with 10% human or horse serum. Respiration was determined manometrically and lactate formation chemically. In preliminary experiments, 0.25% Difco trypsin (1:250) when used to separate cells, depressed respiration in some instances. Respiration of suspensions of HeLa was constant for an hour but decreased slightly after first hour. Metabolism of two lines of HeLa cells was compared. Respiration and glycolysis of cells grown in media containing homologous human serum was higher than that of cells grown in media containing horse serum. Human HeLa cells had a metabolism like that reported for tumor tissue in that respiration was low (Qo2 = 6.0) and aerobic glycolysis moderate (QO2 LA = 12.0). They differed from tumor tissue in that anerobic glycolysis was low (QN 2 LA = 5.5) and p-phenylenediamine stimulated respiration 338% (Qo2 = 26.3). There was no Pasteur Effect and Fermentation Excess was negative.

Cortisone acetate was given daily at 4 dose levels, for 50 days, to young female Long-Evans rats, initially 38 days of age and weighing approximately 100 g each. The upper 2 dose levels were in the range giving nearly complete cessation of growth. In the upper 2 dose levels, there appeared to be a slight reduction in growth inhibiting action after the first 10 days, but all 4 groups still had a marked depression of growth during the 40th to 50th day of treatment. For comparison, anterior pituitary growth hormone and stanolone were given to plateaued adult female rats of the same strain. These hormones, at dose levels capable of giving a major growth response during the first 20 days of treatment, gave little response after the first 20 days.

The previously demonstrated reduction of serum albumin and elevation of gamma globulin and mucoprotein in experimental guinea pig tuberculosis is associated with a slight elevation of total serum hexose which is to the greatest extent accounted for by a slight rise of the carbohydrate containing alpha globulin. No evidence was obtained that the carbohydrate content of this small increment differs from that of the normal alpha globulin. In contrast the marked rise of gamma globulin is associated with an only very small increase of gamma globulin associated carbohydrates. This suggests that the excess gamma globulin in experimental tuberculosis differs from the normal gamma globulin by a very low carbohydrate content and suggests a different nature and possibly a different origin of this increment. The serum protein-bound lipids are not altered in experimental tuberculosis.

Disturbance in energy metabolism, as indicated by increased basal oxygen consumption and growth retardation, is prevented when rats receiving a fat-free diet are given daily supplements of methyl linoleate (100 mg) or cottonseed oil (200 mg). However, similar supplementation with a saturated fatty acid (methyl palmitate, 100 mg) in no way modifies the development of the fat-deficiency syndrome.

Improved procedures for preparation of tissues for chemical analyses have been presented. These procedures have definite advantages over all previous procedures; they are saving in both time and effort; they permit a uniform routine for all types of tissues; they produce homogenous materials that insure adequate sampling, and they enable numbers of tissues to be taken from the same experimental animal and handled satisfactorily.

Two sexually isolated varieties were found among 155 clones of Stylonychia putrina collected in Southern California plus 5 clones produced in the laboratory. In Variety I, at least 15 mating types have been found; in Variety II, 11 mating types.

The changes induced in concentration of rhamnose, mannose, fucose, glucose and galactose in the kidney of rabbits treated with various steroids were studied using a semi-quantitative chromatographic method. The findings are discussed in relation to the pathogenesis of diabetic glomerulosclerosis.

A new colorimetric procedure is substituted for microtitration in the measurement of ammonia by the Conway method. The ammonia is oxidized by addition of hypobromite, and the excess hypobromite determined by its power to decolorize phenosafranin. The range of the method as described is 0.05 to 0.5 micromole. The procedure has been applied to brain tissue and to blood, and to the measurement of glutamine. It has also been used for the determination of non-protein nitrogen after Koch-McMeekin digestion, the microdiffusion being omitted.

(1) Nodal rhythm, ventricular extrasystoles, flutter and fibrillation were precipitated by norepinephrine in cats, under pentobarbital anesthesia, treated with harman methosulfate. Migration of the T-wave into the R-wave at onset of ventricular fibrillation was similar to that described before epinephrine induced arrhythmias. (2) The effects of norepinephrine on cardiac excitability were confirmed using isolated papillary muscles in which norepinephrine had a greater potency than epinephrine in eliciting automatic contractile activity.

Rabbits were subjected to x-irradiation in dosages of 500, 750, and 1,000 r. Serum glutamic oxaloacetic transaminase estimations were accomplished at 3, 6, and 24 hours post-irradiation. A significant increase in G-O-T activity could be identified at 6 hours in the rabbit group subjected to 500 r, and as early as 3 hours post-irradiation in the 750 r and 1,000 r animal groups. For the latter animals, the noted increase at 6 hours served to segregate 75% of the group on an individual basis. In rabbits subjected to 625 r of x-irradiation no relation was found between the serum G-O-T elevation and survival time.

1. Pathogenesis of severe arteriosclerosis and disseminated myocardial necrosis, induced in the albino rat by standard renal injury, was investigated. These cardiovascular lesions are due to autointoxication with parathyroid hormone, and this hormone apparently contains an important hypertensive factor. 2. Evidence was presented that excess production of parathyroid hormone, as a consequence of renal dysfunctions, is mediated through the adrenal cortex and that mineralocorticoids are essential for direct or indirect activation of the parathyroid gland. To some degree, androgens can enhance the mineralocorticoid effect, whereas estrogens seem to inhibit emergence of muscular necrosis. The incidence of cardiovascular injury in intact and adrenalectomized rats was found to be substantially higher in male than in female animals. 3. It was shown further that, in the absence of parathyroid glands, mineralocorticoids in excessive dosage fail to produce cardiovascular necrosis, whereas excess parathyroid hormone assures emergence of typical lesions in complete absence of mineralocorticoids. In fact, by providing abundant supplies of parathyroid extract, cardiovascular necrosis was induced without renal injury and even in absence of kidneys. 4. Implications of these findings upon the concept of “metastatic calcification” and upon pathogenesis and therapy of renogenic cardiovascular necrosis and hypertension in human beings are discussed. It is suggested that subtotal parathyroidectomy be tried in preference to adrenalectomy in attempts to arrest rapidly progressive sequellae of malignant hypertension.

Iodide and chloride clearances were determined in anesthetized and unanesthetized dogs. The control gastric iodide clearance was 10-50 times greater than the chloride clearance. The iodide clearance could be 95% suppressed by anions known to block the thyroid iodide trap but chloride clearance was practically unchanged. In the presence of the blocking agent, gastric iodide and chloride clearances were similar. These data suggest at least 2 mechanisms for gastric secretion of iodide: (a) a specific iodide secretory system and (b) a non-specific mechanism in common with chloride secretion which continues to function after blockade of (a).

Evidence has been presented that: (1) Frequency of abortive parthenogenesis in the Broad Breasted Bronze turkey is as high as 80% when based on unincubated eggs studied histo-cytologically. (2) There are varietal differences which affect the frequency of occurrence of this type of development. Possibly, environmental differences also affect the frequency of abortive parthenogenesis.

Secretion of corticosterone into the adrenal venous effluent of the rat was markedly influenced during the first 10 minutes of collection by the nature of the anesthetic employed for cannulation procedure. Ether anesthesia resulted in a significant elevation of corticosterone concentration and corticosterone flow rate during this period as compared with the values observed when Nembutal was used. This difference was eradicated during subsequent collection periods of 10 to 25 minutes and 25 to 85 minutes following cannulation. The relationship of these findings to the possible activation by ether and inhibition by Nembutal of the pituitary-adrenocortical system is discussed.

1. Tissue culture method can be successfully used to convert Histoplasma capsulatum to the tissue phase. 2. Six of 13 isolations did not convert to the tissue phase in absence of HeLa cells; with all isolates conversion was more complete and rapid in presence of cells. 3. The yeast phase can be propagated and maintained in tissue culture mediums without HeLa cells in 100 ml centrifuge bottles. 4. Rotation of cultures is necessary to assure maximum growth of yeast cells in tissue culture medium. 5. The technic can be successfully used to convert other diphasic systemic fungi, Blastomyces dermatitidis, Sporotrichum schenckii, and Coccidioides immitis to their tissue phase. 6. Candida albicans and Cryptococcus neoformans can be propagated in tissue culture medium without HeLa cells.

1. An extract of Rauwolfia serpentina was administered daily by subcutaneous injection to 20 rats. The daily dosage was equivalent to 1 mg/kilo body weight and was continued from 21st to the 65th to 70th days of life. At autopsy body weights and weights of pituitary gland, thyroid, ovaries, and adrenals were obtained. 2. Application of analysis of covariance leads to the conclusion that at this dose level administration of Rauwolfia serpentina to young rats resulted in (1) a diminution of weight of thyroid by 40%, at the 1% level; (2) a similar reduction in weight of pituitary gland by 8%; (3) no significant change in either ovarian or adrenal weights, after making allowance for body weight differences.