Experimental Biology and Medicine

Deficiencies of hemophilioid factors are not necessarily genetic in origin. Serum activities of plasma thromboplastin antecedent (PTA) and of plasma thromboplastin component (PTC) were depressed in 20 patients with a variety of hepatocellular disorders. PTA and PTC activities were also reduced in several patients who developed a deficiency of vit K subsequent to inability to adsorb fats. PTA and PTC were depressed in a number of patients following administration of 4-hydroxy coumarin compound. In these latter 2 groups, PTA and PTC activities rapidly increased toward normal following administration of vit K analogue. Significant deficiency of antihemophilic factor (AHF) was not found in any of the above conditions.

Methyl ethyl ketone has been isolated from normal human urine and identified. Its possible origin was briefly discussed.

Liver extract and crystalline vit. B₁₂ were administered in equivalent amounts to 2 groups of normal subjects, and total serum vit. B₁₂ levels, bound vit. B₁₂ levels, and urinary excretion of vit. B₁₂ compared in the 2 groups. Concentration of free vit. B₁₂ was higher in the group receiving crystalline vitamin. Over the 24-hour period, percentage of bound vitamin serum was greater in the group receiving liver extract. Urinary excretion of vit. B₁₂ was significantly lower in the group receiving liver extract.

A homogenate of a biopsy specimen of liver obtained from a patient with phenylpyruvic oligophrenia did not catalyze detectable conversion of phenylalanine to tyrosine, under conditions whereby liver homogenates obtained from normal individuals were active in catalyzing such conversion. Addition of a fraction prepared from rat liver (which exhibited relatively low activity alone) to the inactive phenylketonuric liver homogenate resulted in striking activation of the conversion of phenylalanine to tyrosine.

Evidence is presented to show the presence of the stable Fraction II of phenylalanine hydroxylase in the liver of a patient with phenylpyruvic oligophrenia. The biochemical significance of this finding is discussed.

G. tetragena, strains 6007 and 10875, failed to acquire resistance to penicillin after repeated transfers in or on media containing it, nor did the procedure induce visible variation or reversion of the usual dissociant forms. Growth on streptomycin-agar increased the tolerance of each variant to 5 to 30 times the original amount of streptomycin which may be of clinical significance. There was no evidence that variant forms better fitted than the original one to grow under new adverse circumstances were either induced or purposefully developed. The change seemed to be a gradual adaptive one.

Clipping of restrained rats resulted in a much lesser additional body temperature fall than a similar treatment of nonrestrained animals. This is interpreted as supporting the hypothesis that the less erected hair of the restrained animal results in a greater rate of heat loss than the fully erected hair of the nonrestrained animal.

1. Adrenalectomy produced no significant change in weight of rat's pituitary gland. Cytologically, beta basophils decreased in number progressively while acidophils were not altered in number or appearance. The relative per cent of delta basophils was unaffected, but these cells were hypertrophied and exhibited some vacuolation 56 and 100 days after adrenalectomy. The silver-staining capacity of delta basophils, as revealed with protargol, was markedly reduced. 2. Gonadotropic and thyrotropic hormone contents of pituitary were decreased after adrenalectomy, while the corticotropin content was elevated 2- to 3-fold. No difference in growth hormone content was found when whole pituitaries were analyzed by the epiphyseal plate test. 3. Correlations between pituitary cytology and hormone content of the gland suggest that the delta basophil is the source of corticotropin.

1. Concentration of free amino acids from 6 tissues of normal and vit. E-deficient rabbits has been determined by Dowex-50 chromatography. 2. There was a general increase in free amino acids of muscle, heart, spleen, and liver of dystrophic animals. Only glycine and serine were markedly reduced. 3. Kidney and especially brain showed a general decrease in free amino acids. 4. Concentration of glycine was more consistently reduced in various tissues than other amino acids.

The androgenic and anabolic effects of testosterone, testosterone propionate, methyl testosterone, methyl androstenediol, androstanolone and 17-ethyl-19-nortestosterone (Nilevar) were compared. The greatest androgenic responses were obtained with testosterone propionate while the most marked anabolic effects were observed with either testosterone propionate or 17-ethyl-19-nortestosterone. Compared to testosterone propionate, methyl androstenediol and androstanolone showed a small increase in the anabolic: androgenic potency ratio but a much more marked increase was observed with 17-ethyl-19-nortestosterone.

1. Some of our results confirm previous findings: hematocrits of rats castrated as adults are lower than those of intact animals; testosterone propionate returned hematocrits to normal after castration, but did not cause elevation in hematocrits of intact or castrated animals above normal. Estradiol benzoate did not significantly lower hematocrits of intact and castrate rats. 2. Other findings are new: hematocrits of neonatally castrated adult rats were not significantly different from those of intact adults of the same age, and thus differed from those of animals castrated as adults and examined 14 days later. Aqueous testosterone returned the hematocrits of castrates to normal. There was a trend in both intact and castrates for sesame oil to increase the hematocrit. Because sesame oil is widely used as a supposedly neutral vehicle for hormones, the possibility that it has an effect upon the hematocrit warrants further investigation.

Activity of the enzyme system concerned with synthesis of the peptidic link in p-aminohippuric acid was studied simultaneously with rate of uptake of isotopically labelled glycine and lysine into the protein of rat liver slices. The synthetase system increased by 40% in the hyperthyroid state. Incorporation of labelled amino acids, on the other hand, was uninfluenced in the same situation.

Thyroid I¹³¹ uptake by restrained rats was higher at one hour, and lower at 4 hours, than that of nonrestrained control animals. The lower the body temperature of the restrained animals at 4 hours, the lower the I¹³¹ uptake by the thyroid. Restrained hypothermic rats had a significantly greater thyroid uptake of I¹³¹ than equally hypothermic nonrestrained animals. It is concluded that hypothermia of restrained animals can not be ascribed to depressed basal metabolism.

From an outbreak of 11 cases of aseptic meningitis near Cambridge, England, during the autumn of 1955, echo-9 virus was isolated from 4 patients. In addition, 7 cases and two family contacts showed rising titres of neutralizing antibody to this virus. From 2 subsequent outbreaks of febrile headache in this region during the summer and autumn of 1956, echo-9 virus was isolated from 6 patients and rising titres of neutralizing antibody were detected in sera from 8 patients. These data, plus the recent echo-9 isolations from spinal fluids as well as from the alimentary tract of aseptic meningitis cases in other outbreaks, strongly suggest that echo-9 virus has caused severe and mild cases of this syndrome. English strains isolated in 1955 and 1956 were found to produce myositis and paralysis in infant mice indistinguishable from that produced by group A Coxsackie viruses.

Quantitated inocula of 14 tissue culture cell lines which are remarkably similar in vitro were implanted into cheek pouches of unconditioned Syrian hamsters. All cell lines grew in the cheek pouch when 1 × 10⁶ cells were implanted, but only those cell lines derived from neoplastic tissue produced tumors when inocula contained 1 × 10⁴ cells. Other experiments indicated that this difference in the growth potential of cell lines deriving from neoplastic tissue also may be delineated in hamsters conditioned with cortisone acetate by implantation with 1 × 10³ or fewer cells.

A new intestinal antipropulsive test in intact insulin-treated rat is described. The insulin test provides certain advantages over the commonly-used charcoal meal test for screening of synthetic antispasmodic or antipropulsive agents. The use of both tests permits determination of the effect of the agent on both normal and hyperactive states of intestinal muscle and provides information relative to the mechanism of action of the agent. The specificity and some limitations of the test are discussed.

Glomerular filtration rate and renal plasma flow were determined by inulin and para-amino hippuric acid clearance tests in 19 normal subjects, in 14 cholera patients and in 6 patients suffering from acute gastroenteritis. In patients suffering from cholera and acute gastroenteritis, glomerular filtration rate and renal plasma flow were greatly diminished to the same degree, when these patients had apparently normal urination after transfusion of saline. The diminished clearance values might be due to renal anoxia causing temporary diminution in blood flow through the cortex of the kidney and not due to any specific toxins of the disease.

1) Thiocarbanilide, in a 1 to 1 ratio or less, effectively inhibited the anticoccidial activity of nicarbazin when given parenterally as well as orally. 2) Thiocarbanilide also antagonized the effect of nicarbazin on the depigmentation of egg shell color. 3) In both instances, the antagonistic effect of thiocarbanilide was correlated with decreased plasma concentrations of dinitrocarbanilide. 4) It is suggested that thiocarbanilide may interfere with the activity of nicarbazin through competition in absorption.

Pouches from control and test rats fed BAPN were removed after 6, 12, and 18 days. Extraneous material such as collections of leukocytes, fibrin, and areas of hemorrhage were removed before the pouches were extracted in 4% perchloric acid, ethanol, and ether. Grossly the test pouches were poorly formed, and microscopically they showed decrease in fibroblastic maturation and collagenic fiber formation with relative increase in homogeneous eosinophilic Schiff positive material. Fat free protein residue was analyzed for hexosamine and hydroxy-proline concentration. More nitrogenous material was consistently obtained from test pouches during perchloric acid extraction. Hexosamine concentration in different pouches varied from 0.82 to 1.17% of dry fat free protein. Significant alterations with age or variations from control values were not observed. In control pouches the hydroxy-proline concentration gradually increased to approximately 4% in 18 days. Hydroxyproline concentrations in the test pouches were significantly less than in controls of corresponding age.

The instrument to be described was developed to fill the need for an infusion pump with which rate of delivery could be continuously varied over a wide range during injection. The present pump has met this need and has been employed for 9 months in experiments on small animals in which rate of injection has been critical.

Details of construction and mode of operation of this pump will be apparent from Fig. 1 and 2. The motor (Bodine Electric Co., Chicago, 1/50 H.P., 1725 rpm, with gear reduction to 11.5 rpm) turns the first drive cone (A). The 4 identical drive cones are knurled cast aluminum. The conical portion is 4′′ long and the large and small diameters are 3′′ and .95′′ respectively. Cone A drives cone B by means of rubber ring (1/4′′ by 18′′ circumference) which provides contact between the 2 cones. The point of contact of the ring with the cones is adjustable by means of yoke K, rod L and setscrew M (Fig. 1). It can be seen that when the largest portion of cone A is driving B, B rotates 3.16 times as rapidly as A; conversely, when the smallest portion of A is driving B, A rotates at 3.16 times the speed of B. Thus, a 10-fold range of speed is achieved by the first pair of cones. Cone C is coupled to B and both turn at the same speed. Cone D, however, is driven by C through a mechanism identical to that linking cones A and B. Range of speed for the 2 pairs of drive cones is therefore 100-fold. To prevent slippage of the cones on the rubber rings it is necessary to have considerable tension on the 2 pairs of drive cones.

Lactogenic hormone (LTH) alone or together with testosterone propionate (TP) produced a significant increment in the weight of the seminal vesicle. When given with TP, LTH produced an increase in the amount of glandular tissue in the ventral prostate. Growth hormone (STH) alone or together with TP did not produce any consistently significant increase in the weight of the accessories. When TP, LTH and STH were administered simultaneously to the same test animals, the weight response of all accessories was significantly greater than that to androgen alone. The data are interpreted as indicating that a synergism among the 3 hormones may operate to stimulate sex accessory growth in the male Sprague-Dawley rat.

1. The influence of time and dosage upon sodium excretion by rats was investigated after cortisone and hydrocortisone administration. 2. Results of time-response studies show that large doses cause initially weak and transient retention of sodium and subsequently strong excretion of the ion. Smaller doses reduce progressively duration of retention phase and hasten onset of secondary diuresis. 3. Two factors appear to influence the response of sodium excretion. During a fixed time-interval of urine collection, increasing doses cause excretion, no change and retention of sodium. Similar changes in excretion of the ion are seen with a given dose when collection interval is progressively shortened.

1. A lengthening of saline-re-calcification time beyond normal occurred in plasma deficient in any one of the following: platelets, AHF, PTC, PTA, Hageman factor, proaccelerin, or proconvertin. It also occurred in certain acquired multiple factor deficiencies, and in heparinized plasma. 2. Brain thromboplastin recalcification times were normal in platelet, AHF, PTC, PTA and Hageman factor deficiencies, but prolonged in proconvertin or proaccelerin deficiency or heparinized plasma. 3. Plasma thromboplastin recalcification times were normal in these same deficiencies and, in addition, in proconvertin or proaccelerin deficiency. This latter activity seemed due to incorporation of (pro) convertin and (pro) accelerin into the plasma thromboplastin complex, rather than to any free proconvertin and proaccelerin in solution. 4. Formed plasma thromboplastin is inactive in absence of optimal calcium concentration.

1. Only one of 10 dogs submitted to 15 to 30 minutes of potassium citrate produced asystole was a long-term survival (90% mortality). 2. Ten dogs were submitted to 30 minutes of retroperfusion of the coronary sinus, and 4 died from cardiac causes, a 40% mortality. 3. Ten additional dogs were subjected to combination of 30 minutes retroperfusion and potassium citrate provoked asystole. Two of the 10 died from cardiac complications, mortality 20%. 4. Certain aspects of cardiac metabolism in the retroperfused heart with and without potassium citrate induced asystole have been studied in 10 dogs. There appears to be an additionally reduced degree of metabolism under these latter circumstances.

Aliquots of supernates prepared from stools of volunteers with induced afebrile (Marcy) and febrile (FS) nonbacterial gastroenteritis and capable of inducing human disease were administered by stomach tube to cats. No recognizable disease was produced.

The effects of ultrasonic treatment on human brain thromboplastic suspensions have been presented. Although causing a loss in clotting potency, ultrasonic treatment does not affect the toxicity of intravenous tissue thromboplastin nor has it destroyed the TPase-labile component. The results are discussed and possible implications presented.

A modified method of CAM inoculation is described which greatly reduces nonspecific lesions and increases the accuracy of titrations.

1. Concentration of tissue activator of plasminogen in the rat uterus has been estimated in different stages of the estrous cycle. 2. An increase in concentration was recorded during late estrus, whereas castration had no influence upon the concentration. 3. These results are in accordance with those found for human endometrium and myometrium.

A homogenate of urethan-grown S. nitrificans exhibited a marked increase in oxygen uptake when supplied with many esters. Carbamates generally exerted a lesser though significant stimulation of gas consumption. Growth on urethan therefore produced cell material containing esterase(s) characterized by broad spectrum activity. But whether carbamates are hydrolyzed by these or specific enzymes is not yet known. Acetylcholine showed the least activity of all substrates tested, although choline caused an appreciable increase in oxygen uptake.

1. 2-C¹⁴ labeled acetate was injected into the milk cistern of the right front quarter of a lactating cow. This quarter was subsequently milked separately from the other 3 quarters and radioactivities of milk fat constituents from each were determined. 2. The data confirm that there is a delay of several hours between synthesis and secretion of milk fat. 3. A surprisingly small amount of the injected acetate diffused into the other 3 quarters or into the rest of the body. This can be accounted for by its rapid utilization for milk synthesis near the site of injection. 4. Fatty acids of all chain lengths and glycerol seem to be synthesized from acetate in the mammary gland itself. The glycerol synthesis, however, is on a smaller scale.

Purified bovine prothrombin preparations may lose the property of clotting fibrinogen but retain esterase activity. It is likely that a single molecule possesses the 2 activities. When biothrombin has lost clotting activity and only esterase activity remains, it is called esterase thrombin.

1. The effects of exposure of albino rats to 2 hours of low, high and ambient environmental oxygen tensions upon intestinal iron absorption were studied. 2. Total serum irons were increased 116%, 84.8% and 42.4% at environmental oxygen tensions of approximately 54, 125, and 1390 mm Hg respectively.

The information presented shows that the HA test can be utilized for determining adenovirus antibody response. The test may be carried out with heated adenovirus antigen and yields results which relate to a current adenovirus infection. Hemagglutination antibody titrations can be performed at room temperature and readings may be made up to 18 hours after the initial 2 hour incubation without change in titration end-points. Although the HA test does not achieve type specificity, this test appears to be more specific than the CF test. Huebner et al. (17) have reported 74%–90% heterotypic crossing in the CF test. The HA test reported here deals only with types 4 and 7 adenovirus infections and by extending these studies to include other adenovirus type infections it may be possible to utilize this test more specifically to determine infections to other types.

Based on preliminary data the HA test appears to be superior to the CF test in regard to determining infections in persons having CF antibodies in their acute serum due to previous adenovirus infections. The HA test and CF test show close correlation with respect to antibody titer rises and time of antibody appearance so that the HA test appears to be comparable to the CF test in its diagnostic value. Studies are in progress comparing HA, neutralization, and CF antibody titer responses in Naval recruits who have received the adenovirus vaccine.

Application of this HA technic to the immunology of other viruses should be considered. The ease of performing this test may provide a simple laboratory tool for determining serological responses to virus infections.

1. Progestational activity of a series of 17-substituted 19-nortestosterone derivatives was investigated using both intrauterine and systemic assays in rabbits. Several of the substances studied possessed a high order of progestational activity. This activity was not related to either androgenic or anabolic potency of these compounds. Of the compounds studied 17-butenyl appeared to be the most potent, possessing about 10 times and 2 1/2 times the activity of progesterone in intrauterine and systemic assays, respectively. 2. Progesterone-like activity of these 19-nortestosterones was further demonstrated by the ability of some members of the series to maintain pregnancy in ovariectomized rabbits.

Plasma, brain, and muscle electrolytes were determined in mice given DCA and aldosterone. DCA increased the concentration ratio of K (K_c/K_e) without affecting the ratio of Na (Na_e/Na_c) in brain; it increased the concentration ratio of K moderately in muscle and decreased the concentration ratio of Na. Aldosterone increased concentration ratios of Na and K in both brain and muscle. An explanation of this difference in the effect of the two steroids on skeletal muscle electrolyte is proposed.

Two wetting agents, Deceresol-OT and Duponol W.A., when added to suspensions of Sh. dysenteriae, Sh. paradysenteriae, Salm. typhosa, or Sh. ambigua, consistently enhanced pathogenicity of the organisms for mice inoculated intraperitoneally.

The d and l forms of 5-hydroxy-tryptophan were prepared. Only the l form was found to be acted on by 5HTP decarboxylase, to increase brain serotonin levels in vivo, and to be pharmacologically active.

Mature male “wild-strain mice,” isolated from weaning, were placed in groups of 6 for 4 hours a day for 10 days. The social or dominance rank of each mouse was determined. Adrenal and body weights were obtained for each mouse. A significant relationship was found between adrenal weight and social rank. It is concluded that high-ranking mice have smaller adrenals than do low-ranking mice.

The data demonstrate that amplitudes of R, S and RS of the ECG of normal hens are approximately half those of normal males. The effects of estrogen on the ECG of adult males were studied. The voltage of these waves of estrogenized males is decreased about 30 to 35%.

The effects of acetylcholine on pulmonary vascular bed of intact dogs were studied in an experiment in which the drug could be excluded from the systemic circulation. While larger doses (usually greater than 0.1 mg) injected into the pulmonary artery were followed by increased pulmonary vascular resistance, smaller doses lowered pulmonary vascular resistance. Exclusion of extrapulmonary factors indicated active pulmonary vasoconstriction and vasodilatation. The larger doses also caused bronchomotor activity. However, the lack of intratracheal pressure changes with pulmonary vasodilating doses of acetylcholine or vasoconstricting doses of norepinephrine, and the absence of pulmonary hypertension with nebulization of acetylcholine indicated that pulmonary vascular resistance alterations were not dependent upon changes in bronchomotor tone.

(1) Weights of turkey poults fed a purified type diet were increased significantly upon addition of distillers dried solubles, distillers dried solubles ash, molybdenum (0.0254 ppm) or a mixture of distillers dried solubles, dried whey, fish solubles, an antibiotic fermentation residue, and forage juice. Although differences of as much as 2% in bone ash were observed, statistical significance was not obtained. (2) Both liver and intestinal xanthine dehydrogenase activities were significantly higher in supplemented groups as compared to control birds. A significant (P<0.05) correlation was determined between liver molybdenum content and level of enzyme activity. (3) Under the conditions of this study a portion of growth promoting activity of distillers dried solubles ash would appear to be explained on the basis of the molybdenum content.

An agent producing a cytopathogenic effect has been isolated in bovine embryo tissue culture cells from upper respiratory tissues of cattle having the disease of infectious bovine rhinotracheitis. Lack of growth in bacteriological culture media, resistance to antibiotics such as penicillin and streptomycin, and ability to pass through fine porosity filters demonstrates that this agent is a virus. Reproduction of a disease in experimental cattle identical to that which occurs naturally, protection tests in cattle using tissue culture virus and infectious material from naturally occurring cases, as well as neutralization tests with serum obtained from naturally and experimentally infected cattle, show that this virus is the etiological agent of infectious bovine rhinotracheitis. Serum neutralization tests also indicate that viruses isolated from different parts of the country are closely related antigenically.

Using C¹⁴ glucose to label the body glucose pool, it was found that growth hormone treatment for 4–5 days in the hypophysectomized dog, resulted in increased rate of glucose release by the liver and plasma glucose uptake by the tissues. The plasma glucose concentrations were also increased. The significance of these findings is discussed.

1. Injections of approximately 3.7 to 60 I.U. of prolactin daily for 5 to 10 days, beginning on the 16th day of gestation, prolonged pregnancy in rats. The largest doses and longer treatment were most effective in preventing parturition, resulting in death of most of the young when pregnancy was extended beyond 23 days. 2. Prolactin failed to prolong pregnancy in rats ovariectomized on the 19th or 20th day of pregnancy, while injections of progesterone into intact rats extended pregnancy. These results as well as histological examination of corpora lutea, suggest that prolactin acts by extending functional activity of corpora lutea of pregnancy. 3. Lactation was usually initiated in prolactin-injected rats by the 21st day of gestation, independent of the occurrence of parturition. However, milk secretion was not as marked in rats which failed to be delivered of their young as in parturient rats.

Reversal of hyaluronidase effects on concentration of negatively charged colloids in the dermis of normal and cortisone treated rabbits was studied by serial measurement of liquid junction potentials. In normal animals, hyaluronidase lowers negative charge density in the tissues; recovery is half completed in about 4 hours. In animals treated with cortisone initial charge density is lower than in normal animals. After hyaluronidase is injected into the dermis, the lowest levels are reached. Half recovery, however, occurs in 2 1/2 hours. Serial electrometric measurements following hyaluronidase injection are suggested as a means of studying hormone and drug effects, and disease states in connective tissue.

Serum albumin, serum γ globulin, crystalline glucagon and crystalline insulin were observed to bind to diaphragm in vitro and to be eluted slowly on continuous washing with buffered solutions. A lack of reproducibility of binding of insulin at low concentrations is attributable in part to variable adsorption to glassware. A definite saturation of binding sites on diaphragm was not observed at the highest concentrations employed. Competitive inhibition was observed in the presence of heterologous proteins. Binding capacity was not altered by treatment of diaphragms with formaldehyde. Binding of insulin by isolated rat diaphragm in vitro is not demonstrably of biologic significance but is attributable to nonspecific surface adsorption of proteins.

Less than 10 organisms of an unadapted strain of Streptococcus pyogenes, type 18, caused fulminant infection in BALB, BALB/c, AKR, DBA/2, C58 and C57 Black inbred mice. Pen bred Swiss albino mice were significantly more resistant to infection. Inbred BALB and C58 mice that survived the original inoculum underwent latent or chronic infection; organisms were recoverable from untreated mice for at least 3 to 4 months. Data were obtained which revealed that susceptibility of mice to infection depended on strain of serotype of the organism tested as well as the genetic constitution, age and sex of the host. These studies demonstrated that a spectrum of diseases could be produced by employing a single strain of Streptococcus pyogenes, type 18, and different strains of inbred mice. The application of these findings to study of streptococcal infections and their late sequelae was discussed.

Trypan blue injected into developing rat embryos (10 days to birth) fails to stain the central nervous system. This impermeability in the developing central nervous system exists at the time when blood vessels invade the brain. It cannot be assumed, however, that the mechanisms which prevent protein-bound dyes from penetrating the cerebral blood vessels are the same for all other substances.

The pharmacological activity and fluorescence characteristics of extracts of an appendiceal carcinoid tumor could be accounted for entirely by its serotonin content. The amount of serotonin in the carcinoid tissue was about 1000 times that of normal tissue, and its enzymatic ability to form serotonin from 5-hydroxytryptophane was about 35 times that of normal human appendiceal tissue.

1. In growing rats, hypophysectomy has been shown to significantly increase the level of kidney lysozyme specific activity (SA) up to 2-fold. Other tissues, particularly the spleen, show constant SA regardless of hormonal condition of the animal. 2. Thyroid has a marked depressing effect upon kidney lysozyme SA when given to either untreated or hypophysectomized growing animals.

1. A strain of a filterable mouse leukemia agent has been developed from a spontaneous Ak leukemia through several successive cell-free inoculations of newborn C3H mice. 2. This agent induced acute, generalized leukemia within 2 1/2 to 3 1/2 months in 40 to 100% of the inoculated newborn C3H mice (Bittner substrain). 3. Newborn mice had to be inoculated; adult C3H mice were with few exceptions resistant to inoculation of the cell-free agent. 4. The leukemic agent could be preserved at −70°C from 46 days to 5 1/2 months, with no apparent loss of infectivity.

1. The chemotherapeutic properties of 2 newly discovered sulfonamide antidiabetic agents have been studied in infected mice and compared to a commonly used triple sulfa mixture. Data demonstrating that 1-(p-methylphenylsulfonyl)-3-butylurea is devoid of any antibacterial activity and that 1-(p-aminophenylsulfonyl)-3-butylurea is active as an antibacterial agent have been presented. Lack of antibacterial activity of 1-(p-methylphenylsulfonyl)-3-butylurea can be attributed to absence of the para-amino group. This para-amino group is present in each of the ingredients in the triple sulfa mixture as well as other sulfanilamide-like compounds. 2. It has been demonstrated that failure of 1 - (p-methylphenylsulfonyl)-3 - butylurea to protect infected mice was due to a true lack of antibacterial activity, and death of test animals was not attributed to toxicity. 3. Large daily doses of 1-(p-methylphenylsulfonyl)-3-butylurea have been shown not to interfere with therapeutic activity of tetracycline in infected mice.

1. The present studies have shown that, in addition, significant loss of kidney function can result from exposure to this material. Such kidney debilitation could be of considerable consequence if it occurred together with severe liver damage. The time of occurrence of maximal kidney effects correlates well with time of death after NNDMA. as well as with that of rise of blood NPN in rabbits and in dogs. 2. No histologically demonstrable changes in functional kidney cells were evident in these animals nor in those examined by other investigators (3,4). However, other investigators found local vascular congestions in liver, lungs and intestine. The present experiments indicate this to be true in kidneys which show significant loss of function. The suggestion(3) that an outstanding property of NNDMA is its affinity for vasculatures of liver, lung and intestine may also apply to vasculature of kidney.

Although the thyroid gland influences both the status of the thymus and serum cholinesterase activity, thymectomy alone has no influence on serum cholinesterase activity.

No change in concentration of folic acid was found in eggs during incubation for 3 weeks.

A strain of rabbit fibroblasts designated strain RM3–73 requires folic acid, nicotinamide, pantothenic acid, pyridoxal, riboflavin, and thiamine for continuous proliferation in a medium containing 2% (v/v) dialyzed horse serum. The concentration of each vitamin which permits optimal proliferation was determined by propagating the cells serially in media containing graded concentrations of the compound. A group of eight amino acids which are not required for serial propagation in the presence of pyridoxal replaced the requirement for this vitamin under experimental conditions employed.

Coxsackie B2 virus was repeatedly isolated from a serum specimen obtained 5 days prior to the onset of clinical meningitis. The virus was also demonstrated in the stool specimen during the prodromal phase and in convalescence. The convalescent serum contained specific antibodies which were not present either in the virus-bearing serum or in sera collected before the onset of this illness.

Administration of estrogen to normal male rats increased the radioactivity of blood serum following Ca⁴⁵ injection, whereas bone remained unchanged. Parathyroid extract depressed radioactivity of both bone and blood serum but when estrogen was given in addition radioactivity was increased in both. Parathyroidectomy depressed the radioactivity of bones and serum but estrogen again increased the radioactivity of bones slightly and serum significantly. These data indicate that estrogen plays a role in calcium metabolism independent of the parathyroid hormone in aiding retention of calcium.

Estrogen, in an amount estimated to equal that secreted in late pregnancy, was injected daily into lactating rats for 4 days along with Ca⁴⁵. On the 5th day, radioactivity of blood serum, bones and young was compared to a similar group of lactating rats not treated with estrogen. Average gain in weight of the litters of the 2 groups was similar. However, radioactivity in serum and bones of treated group was significantly higher than in the controls. The radioactivity in litters of the 2 groups was similar. These data indicate that estrogen plays a role in the retention of calcium by the animal body.

(1) In the normal rat most of the Ca⁴⁵ is bound to albumin and α-globulin. (2) After parathyroidectomy the Ca⁴⁵ decreases in the albumin and increases in the α-globulin fraction. (3) In animals treated with estrogen most of the Ca⁴⁵ was present in the β and gamma-globulin components. (4) In the parathyroidectomized-estrogen-treated animals highest amounts of Ca⁴⁵ were found in either albumin, α- or β-globulin fractions.

A microbiological method for biochemical screening of compounds of potential interest in the chemotherapy of leukemia, using mutants of B. subtilis resistant to amethopterin and E. coli mutants resistant to purine-antagonists, has been described. Greater growth suppression by a test compound in the resistant strain, when contrasted to the parent strain, has been sought (collateral sensitivity). This phenomenon of collateral sensitivity has been observed for several chemical analogs of the pyrimidine moiety of thiamine. Possible significance of these findings for studies in intermediary metabolism and for cancer chemotherapy has been discussed.

Isolation and cultivation in tissue culture of a cytopathogenic agent recovered from tissues of cattle with mucosal disease from 2 separated herds is reported. A homologous tissue culture system was used with bovine embryonic kidney cells and newborn, antibody-free calf serum. The virus was inactivated by temperatures above 50°C for 15 minutes, would pass an 03 Selas filter and had an 11th passage titer of 10^-5. In sheep, the virus stimulated formation of neutralizing antibodies. A preliminary neutralization study indicated that antibodies are common in animals from herds having no previous history of mucosal disease.

Our experiments indicate that L. acidophilus R-26 could not use either 3′, 5′-TDP, TDP, TTP, D-CDP, or D-CTP for growth, although the compounds were fully utilized after hydrolysis. It was concluded that the organism could not be used directly to detect deoxynucleoside polyphosphates in tissue extracts.

The presence of heparin in normal blood is demonstrated by the “carrier” technic, using radioactive sulfate as precursor. It appears to be found in blood in a combined form so that preliminary decomposition of the complex is required before it can be isolated.

Solutions of 16–32% urea and of 10% sodium salicylate were used to separate I¹³¹ labeled rabbit anti-rat organ antibodies from insoluble rat organ residues acting as antigens. Following removal of these eluting agents, the antibody retained its specificity for rat organs as measured by in vivo and in vitro technics. Measured by in vivo technics, rabbit gamma globulin was resistant to the altering or denaturing effect of these reagents.

1. Constant and reproducible curves of blood radioactivity levels are found in normal humans and dogs after an oleic acid test. 2. Following pancreatectomy and induced pancreatitis in dogs, the curves obtained with oleic acid test were within normal range while those obtained with triolein were depressed.

1. The role of gonadal, adrenal, and hypophyseal hormones on growth and somatotropic actions of a transplantable, functional mammotropic pituitary tumor (MtT) was studied in rats. 2. Although this tumor is autonomous (in that it grows in normal rats of both sexes) it is highly responsive to estrogen, the rate of tumor growth and body growth of the host being roughly proportional to the quantity of estrogen administered. Hormones of mammotropes stimulate body growth and growth of many organs. Somatotropic and mammotropic effects are marked in hypophysectomized rats bearing mammotropic tumors; thus they are independent of other pituitary factors.

1. Fraction studies with plasma from a dog treated with novobiocin showed that novobiocin is extensively bound to plasma albumin. 2. In vitro experiments with crystalline bovine albumin showed that the binding is a reversible process.

A method for evaluation of milk “let-down” in mature lactating rats on 14th day postpartum is described. Amount of milk obtained by litters of 6 young after 30 minutes nursing expressed as percent litter body weight is used as criterion of response. Results obtained follow a normal distribution pattern with a mean value of 3.8 ± .19. The results of this study may serve as a basis for evaluating data from experimental alteration of milk “let-down” in lactating rats.

1. The delayed thrombin generation in samples of horse plasma was made normal by 1) addition of normal human plasma, or 2) addition of reabsorbed heated human serum, or 3) addition of washed human or horse platelets, or 4) freezing of the horse plasma with its normal content of platelets. 2. These results indicate that the clotting defect of the horse plasma investigated is caused by lack of a factor similar to the Hageman factor. 3. Blood platelets, after washing or freezing, apparently substitute for the effect of the lacking plasma factor. 4. The coagulation defects in horse plasma apparently reflect various types of hemophilioid deficiencies.

1. 17-α-hydroxyprogesterone-17-n-caproate (HPC) is an active progestational compound possessing prolonged activity, in confirmation of Junkmann(3). 2. Modification of the oily vehicle has resulted in an earlier peak response to HPC in virgin female rabbits, without affecting duration of action. 3. It has been verified that 17-α-hydroxyprogesterone is an extremely weak progestational agent.