Experimental Biology and Medicine

Administration of cortisone prior to and subsequent to sensitization of the virginal female guinea pig with known quantities of antibody, inhibited the response of the smooth muscle of the small bowel to antigen, but failed to alter the sensitization of uterine smooth muscle.

Cortisone fails to obliterate the spreading reaction caused by hyaluronidase. This substance seems to inhibit at times somewhat the increased capillary permeability induced by 500 to 900 γ of hyaluronidase. An alkaline exudate or leukotaxine induces an increase in the local capillary permeability. Thus, as shown (1,2), the materials are inhibited by adrenal cortical extract or by cortisone. When, however, the exudate is acid in reaction, the local increased capillary permeability is not repressed by either cortisone or by adrenal cortical extract. This is shown to be referable to another factor liberated in the later stages of an acute inflammatory reaction. This substance is concerned with the mechanism of increased capillary permeability in inflammation. This factor is termed “exudin.” Consequently the initial increase in capillary permeability in the development of inflammation is referable to leukotaxine. With the progress of the acute inflammation the pH of the exudate becomes acid and the mechanism of sustained increased capillary permeability is referable to exudin. The degree of acidity does not seem to be the cause of exudin formation. Exudin seems rather to be liberated about concomitantly with the development of an acid reaction in the inflamed area. The nitrogen content of exudin is about 10%. The amino nitrogen values before and after hydrolysis suggest that one is dealing possibly with a dipeptide to which there may be a prosthetic group attached. Further studies, however, will perhaps throw more light on this point. Exudin, tested for the presence of histamine, fails to contract the strip of guinea pig intestine. When extracted for histamine by the method of Bar-soum and Gaddum, there is no evidence that exudin contains any histamine. Its behavior toward cortisone and its indiffusibility indicate that leukotaxine and histamine are dissimilar in nature to exudin.

An accentuation of hyaluroni-dase activity has been observed in skin which had been treated locally for 5 months with desoxycorticosterone, 11-dehydro-l 7-hydroxy-corticosterone and progesterone. Only the adrenocortical steroids produced a characteristic alteration in the slow phase of spread. These alterations are interpreted as representing possible alterations in connective tissue ground substance, specifically related to steroids of adrenocortical origin.

1. The phosphorus content reaches a maximum per mg of uterine tissue in the immature untreated hypophysectomized animal. 2. The phosphorus content per mg uterine tissue is lowest in the adult and adolescent group in both the untreated and estradiol treated rats. 3. The phosphorus content per mg uterine tissue of estradiol treated animals is less than in the untreated animals. 4. Estradiol increases uterine weight and presumably stimulates phosphorus turnover but it does not increase the phosphorus content of the uterus. 5. The importance of estrogenic and pituitary hormones with respect to phosphorus metabolism in the uterus of the rat has been emphasized.

Bilateral, symmetrical, electrolytic lesions in the hypothalamus between the suprachiasmatic and caudal ventromedian nuclei prevented the goitrogenic response of the rat to thiouracil feeding. The iodide-concentrating capacity of the thyroid, however, increased to approximately the same extent as that of intact rats similarly treated. It appears that hypothalamic lesions of this type impair the secretion of thyrotrophin by the pituitary. It is not known whether this interference with thyrotrophin secretion is qualitative or quantitative, nor whether hypothalamic control is exerted by neural or hormonal mechanisms. The available evidence points to the secretion of a hypothalamic hormone, however.

(1) A liter of a 10% combined fat emulsion was injected subcutaneously into hyaluronidase treated areas in each of 22 patients. (2) Each of 8 of the latter received the combined fat emulsion both by the intravenous and the subcutaneous routes. (3) Six patients in this series received 3 consecutive hypodermoclyses of the emulsion. (4) The general clinical reactions and the changes in blood lipids, urinary lipids and oxygen consumption were studied. (5) The clinical effectiveness of this type of administration for parenteral fat was established.

1. Tissue slices from various human cancer and control tissues and mouse breast cancer were incubated in TTC solution with and without the enzyme inhibitors: fluoride, malonate and azide and the ^g of TTC reduced/mg of acetone dried tissues determined. 2. Both control and tumor tissues exhibited a linear relationship between the endogenous and fluoride inhibited TTC reduction. The tumors were characterized by a decreased fluoride inhibition as compared to homologous controls. Malonate inhibition of tumor tissue also tended to be minimal. The findings were discussed in terms of the metabolism of tumor tissue.

The administration of ACTH to fasted rats has no appreciable effect on LGGase, GGGase and cobalt-activated GGase activity of crude extracts of liver, muscle, lung, spleen, kidney or skin.

Carotene solubilized in water by the use of Tween 40 will support growth in vit. A-deficient rats when as little as 1.6 γ is injected intramuscularly daily. For maximum growth, approximately 4 to 6 times as much carotene is required parenterally as orally. Aqueous carotene is also well utilized when given subcutaneously. Carotene dissolved in cottonseed oil and administered parenterally is essentially not utilizable.

(1) The lens and auditory sac showed defective development or were completely missing in 42% of 150 chick embryos infected with Newcastle disease virus at 45 hours of incubation. In 150 control embryos, inoculated with normal allantoic fluid, no similar defects were found. (2) In subsequent experiments the specific defects were obtained in 90% of the N.D.V. infected embryos. Virus samples of such known teratogenicity if stored in hermetically sealed ampules at—50° C retained their teratogenic properties for at least 4 weeks.

(1) The distribution of iodo-acetate and of potassium iodide, both labelled with I¹³¹, has been determined in rats three hours after intraperitoneal injection. (2) The thyroid gland does not show a high selective uptake of iodoacetate. (3) Uptake of iodo-acetate by tissues other than the thyroid is about twice that of iodide. There is no evidence of selective absorption by any tissue studied.

The growth response of normal and castrate hyperthyroid rats to supplementation with vit. B₁₂ and liver concentrate powder has been investigated. Both supplements were effective in partially counteracting the growth depression induced by feeding toxic levels of iodinated casein to normal and castrate rats of both sexes. Vit. B₁₂ was less effective than 1:20 liver powder for normal males and oophorectomized females. There were little difference in response to these supplements in normal females and castrate males. A sex difference in the growth response of the hyperthyroid rat to a factor present in liver concentrate and not identical with vit. B₁₂ is suggested.

Indirect evidence is presented to show that a portion of infused dextran (“M-acrodex”) is broken down to dextrose in the phlorizinized starved dog.

(1) The relative effects of 16 dinitrophenols in inhibiting the oxygen uptake mechanism of spores of Myrothecium verrucaria were studied. (2) The alkyl-, cyclo-alkyl-, or aryl- groups substituted ortho to the hydroxyl group in the 2,4-DNP molecule significantly increased the poisoning effect on this oxidative mechanism as compared to 2,4-DNP. (3) Effective concentrations of all active compounds manifested their full effect within an hour following contact with the spores. (4) The inhibitory effect as caused by all the alkyl substituted 2,4- and 2,6-DNPs averaged 85-90%. (S) Concentrationwise, the 6-n-hexyl-2,4-DNP compound proved the most potent, while dinitrosaricylic, picramic, and picric acids were significantly ineffective.

The hemoglobin from sickle cell anemic individuals does not differ significantly in primary amide content from normal hemoglobin. Normal human hemoglobin was found to contain 26 primary amide groups per mole.

(1) Weanling rats of the Sprague-Dawley (SD) strain have a lower choline requirement for protection against renal hemorrhage and a higher requirement for maintenance of normal liver fat than was observed in rats of the Alabama Experiment Station (AES) strain. (2) Forty-two-day-old rats of either strain grew rapidly and failed to develop hemorrhagic kidneys when they were fed the choline-dencient basal diet supplemented with only vit. B₁₂ and folacin. However, liver fat of the SD strain rats was 33% (dry basis) as compared to 62% for the AES strain rats. (3) The addition of vit. B₁₂ and folacin to the basal diet supplemented with suboptimal choline chloride markedly suppressed the accumulation of fat in the liver, reduced the incidence of renal damage and increased the choline content of the livers of the SD strain of rats. Liver fat values averaging 12.8% (dry basis) were obtained with a 0.20% choline chloride supplement in the presence of vit. B₁₂ and folacin as compared to a requirement of 0.40% choline chloride in the absence of vit. B₁₂ and folacin.

The X-ray sensitivity of E. colt is reduced if the cells are irradiated in the presence of certain chemical agents. BAL, ethanol, and sodium hydrosulfite increase the dose required to kill a given number of cells in suspensions at 2°C by a factor of 3-4. Other sulfhydryl compounds, glycols, and low-molecular-weight alcohols also increase the survival of E. coli suspensions.

Thyroid fragments implanted into the spleen of thyroideo tomized rats are functional and capable of maintaining normal growth and plasma protein-bound iodine concentrations. No morphologic evidence of hyperactivity of the implants could be observed. It therefore appears that the liver does not destroy and excrete the thyroid hormone to a significant extent, when the hormone is present in physiologic quantities.

Thonzylamine caused a slight reduction in the extent of tuberculosis in experimentally infected guinea pigs and caused a slight but significant increase in the length of survival. It did not supplement the effects of streptomycin.

The intravenous or local administration of relatively crude bull testis extract is capable of causing increased vascular permeability in rats. Purification of the hyaluronidase component of this extract results in loss of its capacity to induce increased vascular permeability. The purified hyaluronidase appears to retain its spreading activity.

The administration of testosterone propionate in doses of 10, 25, or 50 mg daily for a period of 10 days to 13 patients with diabetes mellitus failed to alter significantly the responsiveness to insulin in all but one. It is suggested that the presence of the side-chain attached to carbon atom 17 in the steroid molecule may be essential for this activity.

Ethionine induces necrosis of the distal portion of the proximal convolutions in the rat kidney. On a normal diet renal involvement is seen more frequently and to a greater extent in male animals. A protein depletion diet increases very markedly the susceptibility of the kidney to the nephrotoxic action of ethionine in male and female rats. Methionine inhibits the renal toxicity of ethionine while glycine and alanine are without effect.

In the enlarged adrenals of renal hypertensive rats there Is a relative increase in medullary size, whereas in the enlarged adrenals of rats treated with ACTH there is a relative decrease in medullary size. Large numbers of PAS positive intracytoplasmic bodies are found in the cortical fascicular zone in the adrenals of rats with severe chronic renal hypertension, but were not produced in comparable numbers with ACTH. It is concluded that factors in addition to ACTH are concerned in the growth of the adrenal in renal hypertension.

1. Acute hepatic artery ligation in the rabbit resulted in 100% mortality in this experiment. 2. Massive penicillin therapy following hepatic artery ligation in rabbits resulted in a 66% survival rate. 3. Ascites produced by thoracic caval constriction in the rabbit is not prevented or appreciably affected by previous hepatic artery ligation.

The role of vit. B₁₂ and folacin in the biosynthesis of choline and methionine has been studied and the following conclusions are warranted: 1. In the presence of adequate methyls (from choline), vit. Bi2 alone was as effective for methionine synthesis as when combined with folacin. When fed alone, folacin was ineffective. 2. In the presence of a limited supply of methyl groups, both folacin and vit. Bi2 were essential for the normal development of weanling rats. 3. Vit. Bi2 and folacin were essential for the synthesis of choline and of methionine from aminoethanol, homocystine, and a limited supply of betaine. 4. As a methyl donor for the synthesis of methionine from homocystine, betaine was at least as effective as choline, mole per mole. 5. Under the experimental conditions employed, the rate of synthesis of the methyl group or utilization of methanol for the methy-lation of homocystine to methionine and aminoethanol to choline was not fast enough to support normal development of weanling rats.

Exposure to −12°C for 2 hours significantly increased both the free and total acetylcholine concentration of rat brain.

1. Three strains of a filterable virus closely related to West Nile have been isolated from the blood of children living on the outskirts of Cairo, Egypt. 2. The 3 strains of virus appear to be immunologically identical. 3. In addition to a strong relationship and apparent identity with West Nile virus, the Egyptian virus crosses, although to a lesser degree, with both Japanese B and St. Louis encephalitis viruses. This was found by both neutralization and complement fixation tests. 4. The virus produces a marked encephalomyelitis in monkeys inoculated intracerebrally. Lesions are found throughout the spinal cord and brain including the cerebellum. 5. Chimpanzees develop a silent infection following intracutaneous infection of the virus. This is characterized by a viremia lasting 3 days followed by the appearance of both neutralizing and complement fixing antibodies. 6. The Egyptian virus was not neutralized by human gamma globulin collected in the United States, nor by 24 different hyperimmune sera, each containing antibodies to an antigenically distinct virus type. 7. Infection with this virus has been widespread in the local Egyptian population in 1950, with more than 70% of the inhabitants aged 4 years and over having both neutralizing and complement fixing antibodies.

The oxygen consumption of liver and spontaneous mammary tumor slices from female mice was initially slightly increased and then progressively decreased by increasing concentrations of added testosterone.

1. Labelled colchicine was detected only in some organs of normal and tumor-bearing mice. 2. The uptake of colchicine in sarcoma 180 is as high as the uptake of liver. 3. Spleen is one of the organs having the highest concentration of colchicine in normal mice. 4. It was found that there was no colchicine in the spleen of tumor-bearing mice.

In a study of an homologous series of aliphatic quaternary ammonium and piperidinium compounds in mice, it was found that an increase in the chain length of 2 alkyl groups from 2 to 4 carbons results in an increase in the acute toxicity. Compounds with branched chains are relatively less toxic than compounds with straight chains. Piperidinium derivatives have the same range of toxicity as their aliphatic analogues. Administration of an anesthetic dose of Nembutal (66 mg/kg), 15 minutes prior to an injection of the quaternary ammonium derivative, reduces the toxicity of all but 4 compounds. These 4 compounds have methyl branches on 2 of the alpha carbons and at least one of the other substituents on the nitrogen is an ethyl group.

In a series of 57 sera collected from poliomyelitis patients during the acute or subacute stage of their illness, 9 (16%) gave strongly positive reactions, 5 (9%) gave questionable reactions and 43 (75%) gave negative reactions when tested for the presence of hemagglutination-inhibitory substances against Col-SK virus. All of 12 sera from patients convalescing from Jap B encephalitis gave negative reactions. In a control group of 80 sera from healthy individuals 2 (2.5%) gave positive reactions, 4 (5%) gave questionable reactions, and 74 (92.5%) gave negative reactions. Of 14 positive or questionable poliomyelitis sera, 5 contained significant neutralizing antibody levels against Col-SK virus as determined by mouse test. The significance of these findings is discussed.

A polysaccharide yielding osazone resembling that of Arabinose with a melting point of 166°-168°C has been isolated from the supernatant of the Haffkine plague vaccine as well as from the specific soluble protein and, to a less extent, from the bacterial debris of both virulent and avirulent protective plague strains. It is absent in non-protective avirulent plague and pseudotuberculosis organisms. The protective substance of the plague bacillus is probably a polysaccharide-protein complex.

(1) The serum iron and hemoglobin values were determined for 275 healthy women ranging in age from 17 to 86 years. Mean values for all women were 116 ± 33 μg% and 13.2 ± 0.9 g%, respectively. (2) Analysis of variance indicated a difference of serum iron levels among different age groups which was significant at the 1% level. The regression coefficient of serum iron on age was —.343, indicating a decrease of serum iron with advancing age. This decrease was not accompanied by a decrease in hemoglobin values, nor was there a correlation between serum iron and whether the menopause had been reached or passed. (3) There is no apparent nutritional or medical significance to the decrease of serum iron levels with age in healthy women.

Certain problems of cardiodynamics can be studied profitably by the use of an improved isolated mammalian heart preparation. The mode of relaxation of the ventricle, for example, may hold further information concerning the control of cardiac output, for it is increasingly apparent that the remarkable property of the heart is not only that it can pump so much, but also that it can fill so efficiently. A study of the relaxation process of the ventricle is difficult, however, when large volumes of blood are being pumped by the heart.

In order to study the relaxation process adequately an isolated heart preparation should possess the following attributes: (a) generate ventricular pressure pulses of normal contour, (b) possess easy control of atrial and ventricular systolic and diastolic pressures, (c) pump no blood, i.e., have a fixed ventricular volume, and (d) not deteriorate over a period of many hours. The preparation to be described possesses all these characteristics to a satisfactory extent.

The preparation. Heyman and Kochman ( 1 ) apparently introduced the idea of using a donor dog to supply the blood and pressure head for the perfusion of the coronary arteries of the isolated heart. Although their method has not been used extensively it has an advantage over the use of mechanical perfusion systems, namely, that the presence of the intact lungs, liver and kidney of the donor dog can maintain a more normal blood chemistry.

Fig. 1 shows the schema currently used. Blood from the femoral artery of the donor dog enters and ascending aorta via a cannula (A) introduced through a common carotid artery. The aorta is tightly ligatured at the arch (B), as are all head vessels (C) and the left subclavian artery (D).

(1) Tromexan in amounts of .005 to .2 mg partitions from 1 ml of acidified plasma into 5 ml of 2,2,4-trimethylpentane of the extent of 97.2 ± 3.5%. (2) The optical density of such solutions varies in accordance with Beer's law. (3) On this basis a procedure for estimating plasma and serum concentrations of Tromexan by ultraviolet spectrophotometry was evolved.

The intravenous administration of sodium lactate causes a statistically significant increase in Tm_PAH in man. The average maximum increase amounted to 30% of the preinjection levels, and the effect was observed in each of 8 subjects tested twice. Neither the observed increases in pH₈ and (BHCO₃) nor the changes in renal blood flow appear to bear a causal relationship to the increases in Tm_PAH produced.

The technical assistance of Mrs. Elsie Beard, Miss Margaret McCollum, Mr. Arthur Dinan, and Mr. W. Irving Jones is gratefully acknowledged.

(1) An extensive segmental gastric resection in which 74% of the acid secreting portion of the stomach by weight is excised, leaving 10% of the entire stomach as the upper fundic pouch anastomosed to the antrum will protect against the histamine-in-beeswax provoked ulcer in the dog. (2) A segmental resection in which 25% of the stomach by weight is left as the fundic pouch will not protect against the histamine-in-beeswax provoked ulcer, whether or not a complemental vagotomy is done.

The creatine phosphate content of the white breast muscle of the fowl was found to be over twice as great as that of the red leg muscle. In the “flying” bird, the pigeon, the red breast muscle and the leg muscle contained approximately the same levels of this high energy phosphate.

The effectiveness of 2,3-dimer-captopropanol (British anti-lewisite) in increasing the elimination of injected P³² and Sr⁹⁰ in mice was investigated. It is concluded that BAL is essentially without effect although, in the case of P³², the rate of uptake in bone appears to be reduced during the first hour. No such differences could be demonstrated after 24 hours.

By the use of 3 criteria, namely: the presence of circulating antibodies in the serum; the mode of development of the skin reactions to intradermal injections of antigen, and the passive transfer of anaphylactic sensitivity to guinea pigs; it has been demonstrated that the sensitivity induced in rabbits to foreign proteins while incorporated in Freund's adjuvant is of the usual anaphylactic or foreign protein type.

The animals simultaneously develop a tuberculin sensitivity due to the M. tuberculosis organisms injected but this in no way alters the quality of the sensitivity developed to the foreign proteins. The persistence of the skin reactions to the higher concentrations of the serum antigens appears to be a reaction to the marked necrosis of tissue that occurs in these highly sensitized animals to the concentrated antigens rather than to any fundamental qualitative difference in the type of reactivity induced in the animals.

(1) Thymic weight loss in 14 hours offers an accurate quantitative endpoint by means of which to measure the action of cortisone when administered by different routes. (2) Under conditions of our experiment, cortisone was much more effective when administered by subcutaneous route as compared with the oral or intrasplenic route. The subcutaneous:oral ratio is about 1:15. (3) Sensitivity of adrenalectomized mice to the thymolytic action of cortisone did not differ from that of intact animals. (4) Administration of small doses of cortisone, which caused thymic involution when given by subcutaneous route, failed to do so when given by intrasplenic or oral route. This suggests that inactivation by the liver, previously found in in vitro experiments, also plays a role in vivo. If the dose administered through the portal routes is large, sufficient amounts escape inactivation to produce a maximal effect upon the thymus.

In 9 of 10 patients under treatment with ACTH or cortisone, the phagocytic activity of the neutrophilic leukocyte decreased. In the remaining case an increase was observed which was shown to be due to the development of specific antibody during treatment.

Electrophoretic, ultracentrifugal and immunochemical analyses of a protein extracted in buffered saline from tissues injured by the application of a tight tourniquet to one hind leg of the dogs indicate that large quantities of serum albumin enter the injured tissue. However, little or none of the α- and β-globulins enters the tissue. The dye, T-1824, injected intravenously before release of the tourniquet appears in the tissue of the injured leg only.

The proposed method of assay consists of incubating cellular suspensions derived from the rabbit thmus with adrenal cortex preparations at 37°C for 24 and 44 hours. The number of viable cells in the incubated and treated suspensions were determined by the method of unstained cell counts. The cytocidal unit was defined as that amount of reagent which killed 50% of the cells as compared to a control, incubated suspension. The cytocidal unit was found to be 0.025 γ for 17-hydroxycorticosterone and 0.1 γ for corticosterone and cortisone.

Small doses of the adrenal cortex preparations had a delayed cytocidal effect whereas large doses of adrenal cortex extract and corticosterone had an immediate effect.

The data show that the resistance of alien strain (CF₁) mice to the progressive growth of lymphosarcoma 6C3H-ED (originating in C3H mice) is greatly impaired by intensive treatment with cortisone acetate. Seventeen of the 23 CF₁ mice which survived treatment with cortisone died of progressive growth of lymphosarcoma implanted from C3H mice, whereas only 2 of 37 untreated mice developed progressive tumors.

It should be pointed out that the course of cortisone dosage used to bring about the effects described were excessive, actually constituting a lethal concentration for some of the CF₁ mice. The nature of the change in resistance shown by these experiments has not been established, but it might be inferred that it is associated with the well known depression of normal lymphoid tissue and immune responses which large doses of cortisone induce.

Addendum. After the above manuscript was submitted for publication, a paper by E. L. Howes (Yale Jour. Biol. And Med. 1951, v23, 454) came to our attention. This paper described the progressive growth of adenocarcinoma EO771 or 775 when transplanted in Rockland Swiss mice treated with cortisone.

The rate of protein absorption by normal mice has been measured using radioiodinated human serum albumin as a trace protein. No change in this rate could be shown after whole body X-irradiation of the mice although a change in tone and motility of the gastrointestinal tract was noted. So absorption of unhydrolyzed protein could be demonstrated.

The inhibitions of the cholinesterases of mouse brain, bovine erythrocytes and horse serum produced by N-allylnormorphine were, measured and compared to the inhibitions produced by morphine. N-allylnormorphine is at least as potent an inhibitor as morphine. The implications of this finding for the theory of morphine action are discussed.

Electron micrographs of typical inclusion (Negri) bodies from the hippocampus of a mouse inoculated with rabies virus are shown together with smaller inclusions that may be early forms of the larger bodies. No particles have been recognized that could be identified with elementary bodies of this virus.

The major portion of the acetate-replacing factor activity of rat liver homogenates has been demonstrated to be associated with the mitochondria.

1. Normal rats respond to the intraperitoneal injection of renin with an intense, transient proteinuria. ACTH administration results in a 60 to 65% increase in proteinuria in the hour following renin administration over and above that shown by non-ACTH treated controls. 2. The normal, spontaneous proteinuria of the male rat is significantly increased by ACTH administration.

When mice were infected by the intranasal route with large doses of influenza virus and immune serum was introduced into the lungs 1 hour later, the animals died even though neither the amount of infectious virus in the lungs at the time the serum was given nor the amount subsequently produced was as great as the corresponding amounts found in the lungs of mice that had been infected with small doses of virus and that were protected when immune serum was given 24 hours later. The findings suggest that a large amount of influenza virus introduced directly into the mouse lung may, by itself, be sufficient to kill the animal without any subsequent multiplication of the agent.

The administration of DCA to normotensive rats following a figure-of-eight “renal tie” and uninephrectomy has been found to induce consistently a marked hypertension. Similar treatment of both intact and uninephrectomized rats drinking physiologic saline was ineffective in this respect. Hypertension was readily induced in the “renal tie” animals despite the absence of excess sodium chloride and their older age.

1. Observations on the effects of activation of the adrenal cortex by ACTH on the B-glucuronidase activity of plasma, leukocytes, and urine are reported. 2. ACTH administration evokes a marked rise in plasma and white cell glucuronidase activity. Plasma glucuronidase activity correlates well with the total corticosteroid level elicited by the ACTH in that it increases during administration and decreases following administration although the day-to-day parallel is variable. 3. Urinary glucuronidase activity fails to indicate significant correlations either with plasma and white cell glucuronidase or with the corticosteroids.

A method is presented for the ultramicro estimation of serum calcium and magnesium with ethylene-diamine-tetraacetate in the presence of eriochrome black T, at a pH of 10.0 to 10.5. The serum (0.02 to 0.05 ml) is titrated directly and the titration is completed in less than one minute; thus the speed of this determination compares favorably with that of the flame photometer. For rapid and approximate calcium value, the mean magnesium concentration of human serum (1.64 mE/L) is subtracted from the total calcium plus magnesium thus determined. For precise calcium values, magnesium is determined on an aliquot of serum and the value subtracted from the calcium plus magnesium values. For precise magnesium values, calcium is determined on an aliquot of serum and the value subtracted from the calcium plus magnesium values.

Rapid intravenous administration of procaine amide was found effective in reverting ventricular tachycardia, produced in dogs by large doses of cardiac glycosides, to normal sinus rhythm. In most instances, the reversion was temporary and additional injections of procaine amide were necessary in order to maintain the normal rhythm. Such additional injections, repeated several times in these experiments, did not produce electrocardiographic signs of cumulative toxicity.

Although procaine amide effectively reverted ventricular tachycardia in the dog experiments, the same agent failed to increase the lethal dose as determined by limited “cat unit” studies. A similar relation was noted by Emerson in studies on procaine(6) and by Stanbury and Farah(7) in studies on magnesium.

The development of slow idioventricular rhythms and cardiac arrest in several experiments following procaine amide administration demonstrates a danger of drug termination of digitalis-induced ventricular tachycardia. As Gold(8) has emphasized, complete A-V heart block concomitantly produced by the digitalis may be masked in the electrocardiogram by the ventricular tachycardia and the abolition of the latter arrythmia by quinidine or other drugs may result in ventricular arrest.

The administration of large amounts of folic acid inhibited the increase of the urinary coproporphyrin in rabbits given Rose Bengal and exposed to ultraviolet light. The administration of small amounts of vit. B₁₂ had the same effect. The combination of folic acid and vit. B₁₂ had no advantage over either substance given separately. The administration of nicotinamide in large amount produced a significant but only partial inhibition.

In view of the fact that nicotinamide, folic acid, and vit. B₁₂ are all intimately related to cellular metabolism, it seems highly likely that the marked increases of urinary coproporphyrin under these experimental conditions are likewise so related, but whether to the metabolism of the bone marrow, or some other system of cells is not clear. Further studies of these relationships are in progress.

A study has been made of blood and liver glutathione after 500 r total body x-irradiation. The data show first, that irradiation generally causes a decrease in blood glutathione; and second, in some cases, a decrease in liver glutathione.

Several characteristic crystal patterns have been observed microscopically in dry smears of pure gastric mucus from canine Heidenhain pouches. Although their chemical composition has not been established, there is reason to believe that they are neither pepsinogen, pepsin, nor one of the mucins.

Under the conditions of these experiments, 11-desoxycortisone (25 mg) lowered the tooth pain threshold, but did not alter the wrist pain threshold, and cortisone (25 mg) did not alter the tooth or wrist pain thresholds. An alteration of the pathophysiologic state initiating pain is suggested as an explanation for the pain relief afforded by cortisone.

(1) Pure aqueous humor from beef eyes will support survival but only limited growth of chick fibroblasts. (2) A mixture of aqueous humor and 20% desiccated embryo extract results in extensive proliferation of chick fibroblasts which is superior to that obtained in media prepared from desiccated horse serum, embryo extract and saline. (3) The possible explanations and significance of these findings have been discussed.

(1)Three crystalline antibiotics were fed to 100 male and female rats to determine whether they could counteract the retardation of growth induced by feeding thyrotoxic amounts of Protamone. On a ration adequate in vit. B12, penicillin or neomycin but not streptomycin were completely effective in overcoming the growth-inhibition, while on a ration deficient in vit. B12, all 3 antibiotics were only partially effective. On the deficient ration, the antibiotics reduced the survival rate of the hyperthyroid rats. (2) At least part, if not all, of the effectiveness of the antibiotics in counteracting growth retardation could be attributed to the increase in food consumption and to an increased efficiency in converting food into body weight gains. The antibiotics did not alter the 74% increase in basal oxygen consumption induced by the Protamone, nor did they change the action of the latter on adrenal and spleen weight. Thymus atrophy was partially or completely prevented by the antibiotics.

Unilateral renal diseases resembling hydronephrosis, pyonephrosis, or pyelonephritis were induced in rats by partial ureteral constriction and intravenous injection of E. coli. Arterial hypertension developed when chronic unilateral renal infection was produced.

It has been found that with the bismuth cathode tube, a 50 uC dose can be used efficiently, and that a tube-to-patient distance of 17 cm will cover a sufficiently large area to make variations in gland size unimportant to the accuracy of the test.

The growth stimulatory and inhibitory action of several analogues of phenyllactic acid on L. casei were studied. DL-mandelic acid was found to be a potent inhibitor of phenyllactic acid utilization by this organism. The action was not competitive, except over a narrow range of inhibitor to nutrilite. Mandelic acid did not interfere with the functioning of phenylalanine.

Administration of alloxan to white rats causes an increase in the DNA and the histone of liver nuclei. Direct measurements indicate no change in cytoplasmic RNA, but, indirect evidence suggests an increase in nuclear RNA.

(1) Changes in circulating leukocytes which occur after anaphylaxis have been compared with those after pyrogen injection. Following both of these reactions, there is immediate leukopenia followed by polymorphonuclear leukocytosis. (2) Two types of response, although superficially very similar, are distinguishable by direct eosinophil counts which reveal uniform rises with anaphylaxis and uniform falls with the pyrogenic reaction. (3) The mechanism of these hematologic changes is not touched upon in the present study. (4) It is concluded that the leukocyte changes after anaphylaxis are not due to pyrogen contamination.

Serologically reactive substances (“soluble antigens”) which were readily detectable by precipitation and by complement fixation with Cryptococcus antiserum were found in the spinal fluid, blood serum? and urine of a patient with cryptococcal meningitis. Absorption of the antiserum with solutions of purified Cryptococcus polysaccharide removed its capacity to react with the fluids of the patient. The reactive substances in the patient's fluids were relatively heat stable and it is probable that they represent capsular polysaccharides of the fungus with which the patient was infected.

Serological reactions of the Quellung type were obtained with the capsules of Cryptococcus cells from the centrifuged sediment of the spinal fluid.

A method has been described of treating cats with an estrogen plus progesterone to assure uteri in vivo highly responsive for hours to oxytocics. Modifications of this method applied to juvenile and castrated cats indicated that either type of hormone would establish some brief uterine responsiveness but that both together were required to give a maximal and prolonged sensitivity to drugs. The two hormones in the cat are synergistic, or at least mutually complementary, and not antagonistic in respect to their effects upon the sensitivity of the cat's uterus in vivo to drugs.

Further details are given concerning the movements of proteins on filter paper using M/10 solutions of sucrose solution and sodium potassium tartrate as developing solvents in the first and second dimension, respectively. It is shown that mixtures of proteins, such as serum albumin and cytochrome c, may be separated from each other on paper, since the former moves in the first dimension, whilst the latter does not and both proteins move in the second dimension. The amount of cytochrome c moving in the second dimension is proportional to the amount applied to the paper. The chromatographic patterns of purified proteins e.g. crystalline human serum albumin, α, β, and γ globulins, and fibrinogen differ from each other in a characteristic manner. The differences are more marked when the proteins are mixed with surface active agents with which they form complexes; and these complexes move at different rates on filter paper. Complexes of surface active agents with albumin move with greater ease along the first dimension than those formed with globulins; the latter tend to remain at the point of origin. Almost all the protein complexes, however, move in the second dimension. The pattern given with a blood plasma is a composite of the patterns of the individual protein fractions, the basal fractions being apparently due to globulin. The patterns of chromatograms of pathological cases (multiple myeloma, liver cirrhosis, cancer of the stomach, lupus erythematosis) differ significantly from the normal, and are often characterized by the presence of heavy basal fractions due probably to increase in the globulin content of the plasma.

The chromatograms of dried snake venoms (cobra, Russel viper, Agkistrodon piscivorus) show various distinctive features.

1. CFW and dba mice were pretreated with small (125 and 250 mg./kg) doses of cysteine (pH 1.0 or 6.5) before irradiation with lethal doses of x-rays (600 r). It was found that 10 to 14 days after irradiation the percentage of surviving animals was consistently higher in cysteine pretreated group than in irradiated not pretreated controls. These results illustrate “protective” effect of cysteine against radiation. 2. In new mice prepared with high doses (more than 800 mg/kg) of cysteine the x-ray treatment was followed immediately by death or agony. In mice treated by one of these factors alone, the condition was affected only after several days. Thus, the rapid lethal effect of combined treatment with cysteine (high doses) and x-rays is interpreted as potentiation of one factor by the other. 3. This effect of high doses of cysteine was absent in x-ray treated mice possessing copious peritoneal exudate due to the growth of free S-37 cells in the peritoneal fluid. This failure of cysteine is attributed to inactivation (oxidation) of cysteine by some factor of the peritoneal fluid. In these mice with exudate pretreated with small doses of cysteine, the decrease in the cell number (in the exudate) induced by irradiation was followed by a steady increase, while in irradiated controls the decrease continued until their death. 4. Pretreatment with small doses of cysteine (intravenously) had unfavorable effect on survival of mice injected (i.p.) with radioactive colloidal gold. 5. An explanation of the above phenomenon as manifestation of cysteine penetration from blood stream into tissues after increase of blood vessels permeability by high doses of irradiation is offered.

The use of N-acetyl 4-amino antipyrine (NAAP) in the measurement of body water has been explored. The substance is uniformly distributed in the various tissues in close proportion to the water content, its metabolism is negligible and it is excreted slowly. The volumes of distribution of anti-pyrine and NAAP were compared in normal human subjects and the values found to agree well. NAAP has certain advantages over antipyrine: it is negligibly bound to plasma proteins; the value for body water may be calculated from a single plasma sample and a urine sample; its colorimetric estimation obviates the need for an ultraviolet spectrophotometer.

Pulsations in the esophagus at the level of the left atrium and pressure pulses within the left atrium were simultaneously recorded in normal dogs and in the same animals following the production of mitral insufficiency.

An identifiable and consistent pattern in the esophageal tracings was observed in the normal animals. Following the production of mitral insufficiency, characteristic alterations of the esophageal pulse wave contour were observed. These very closely paralleled the pressure tracings obtained simultaneously within the left atrium itself.

It was also possible to detect a fall of the regurgitation pressure which resulted from hemorrhage or a rise which was produced by aortic compression. A ratio of approximately 3 to 1 was observed between simultaneously recorded atrial and esophageal pulse pressures in the dogs with mitral insufficiency.

It is suggested that the contour of the pulse waves of the esophagus at the level of the left atrium, recorded with this technic, closely reflects that of the pressure pulses of the left atrium and that, within limits, a pressure relationship of a quantitative nature exists.

1. The effects of large doses of total body radiation upon the blood pressure, blood volume, and hematocrit of normal adrenalectomized and spinal adult rats have been studied. 2. From the data presented it appears that the cause of the immediate hypotension following such irradiation is depression of the nervous system.

(1) The African toad (Xenopus laevis) responds to the injection of pineal extract by a contraction of melanophores, in a manner similar to that previously described for amphibian tadpoles, prior to metamorphosis. This response is suggested as the basis for a simple assay of mammalian pineal body. (2) Preliminary chemical studies suggest that the melanophore contracting substance is a relatively stable, low molecular weight compound.‡

(1) Water excretion by hydrated rats has been studied to ascertain the alterations after ligation of the inferior vena cava at 3 levels in relationship to the renal veins. In addition, the effects of unilateral nephrectomy, hypophysectomy and thoracic lymph duct ligation, and various other control procedures have been observed. (2) The antidiuretic effect of inferior vena cava ligation in the hydrated rat is shown to be independent of increased renal venous pressure, and is also shown to occur in the absence of the pituitary gland. (3) The critical importance of the lymphatic pathway in maintaining an effective circulating blood volume in the presence of acute venous obstruction is demonstrated by showing the lethal effects of hydration in animals with combined caval and thoracic duct ligation, procedures which are separately tolerated by the rat.

The “nervous syndrome” condition produced in rats after 10 weeks or longer on a low protein, niacin-free regimen has been shown to respond to thiamine supplementation. Complete protection from the convulsions was affected when sulfasuxidine was added to the ration. Analyses of the rations for thiamine showed that as much as 70% of the vitamin was destroyed in the non-sulfasuxidine ration, while very little destruction occurred in the ration containing the drug. The addition of 0.2% dl-phenylalanine did not increase the thiamine requirement.

Cortisone in the doses used in these experiments was not effective in reducing the extent of muscle or skin necrosis in rabbits after 2 standard local cold injuries.

American torula yeast, when used as the sole protein source in Vit. E free diets, induced dietary necrotic liver degeneration in rats in the same manner as reported for European yeasts. Other American yeasts did not produce the defect. With 30% torula yeast in the diet the incidence of liver degeneration in 15 consecutive experiments was 100%, the average survival time was 45.5 days.

L-Cystine reduced the incidence of the defect considerably; however, even 1.0% did not give full protection when supplementation started at the 25th day. Five mg % vit. E prevented the defect completely.

Necrosis of the liver developed very suddenly after an experimental period of 22 to S3 days and was accompanied by kidney lesions, forestomach ulcers, acute lung edema and hemorrhages in the lungs, in the lymphatic tissues and occasionally in the intestine.

Acknowledgment is made to Mr. Louis E. Lowman and Mr. William Beane for technical assistance.

The addition of ethyl alcohol to liver and kidney slices from adult male rats produces a stimulation in oxygen consumption followed by a progressive inhibition as the concentration of the ethyl alcohol is increased.

The synthesis of vit. A and closely related compounds constitutes a field of investigation engaged in by numerous organic chemists. Once a new product has been prepared, its chemical structure elucidated and its spectrophotometry characteristics established, there remains the question of its biologic potency, for which no rapid methods of determination are available. The biologic assay procedure as given by USP XIV, including a depletion and assay period, requires a minimum of 46 days or a maximum of 73 days for completion. (U.S. Pharmacopoea XIV, p. 790, 1950).

This report offers a method for identifying as well as roughly evaluating the degree of biologic potency in terms of vit. A activity which is—(1) more rapid; (2) more specific; (3) requires no preliminary depletion period or use of specialized diets. This method depends upon the response of the skeletal structures of the young growing animal to excessive intake of vit. A. This response is specific in the sense that no other vitamin or any compound not closely related structurally to vitamin A gives a similar picture. The syndrome of hypervitaminosis A, with its emphasis upon changes in skeletal structures, has been repeatedly reaffirmed ever since its recognition by Collazo and his associates. Its characterization as an acceleration of normal growth sequences in long bones, first suggested by Wolbach and Bessey ( 1 ) was fully described by Wolbach( 2 ). This response is readily recognized histologically in suitably prepared material when high dosage levels are maintained even for short periods of time. If one uses a weanling rat (the test animal that is most convenient and responds with great uniformity at high dosage levels) a tentative diagnosis can be made within 5-6 days, to be confirmed by histological findings in another 10 days, if the material under investigation has a potency in the range of vitamin A.

A fraction isolated from adrenal venous blood of cows has been demonstrated to have androgenic activity and to contain a compound giving the reaction for 17-ketosteroids. Its exact chemical structure has not been established due to the small quantity of the substance available for analysis.

Phosphorus uptake and oxygen consumption by particles from rat liver, with a-ketoglutarate as substrate, are almost completely inhibited by 0.0007 N Ca⁺⁺ and completely inhibited by 0.0014 N Ca⁺⁺; this effect may be a specific one upon phosphorus uptake or transfer, as the oxygen uptake with pyruvate and hexose diphosphate as substrate is not markedly impaired at the same Ca⁺⁺ concentrations. Citrate used in concentrations equivalent to the calcium had no protective effect against the Ca⁺⁺ inhibition, but larger ratios of citrate to calcium afforded partial or complete protection. Attempts to obtain oxidative phosphorylation with washed particles obtained from Walker 256 rat tumor homogenates, using a-ketoglutarate as substrate, were unsuccessful. Wide variations in concentration of magnesium appeared to exert no effect on the phosphorus uptake and oxygen consumption of washed rat liver particles.

When homogenates, particles, and supernates of rat liver, rat brain, and the Walker rat tumor 256 were subjected to the action of calcium with hexose diphosphate and pyruvate as substrate, the maximal inhibiting effect on the phosphorus uptake of homogenates and particles was noted with liver, an intermediate effect was obtained with brain, and a minimal effect was found with tumor. In all cases the phosphorus uptake, lactate production, and oxygen uptake of supernates were unaffected by calcium chloride at concentrations up to 0.0028 N.

Mice injected in a foreleg with pertussis vaccine alone or combined with precipitated diphtheria-tetanus toxoids were frequently paralyzed in the injected foreleg following infection with mouse encephalomyelitis virus. The frequency of paralysis in the inoculated extremity was 7 or more times greater than among the untreated controls.

Precipitated diphtheria-tetanus toxoid had less effect and typhoid-paratyphoid vaccine had none.

The injection of vaccines did not increase the frequency of paralysis, irrespective of location, but pertussis vaccine consistently shortened the period between the introduction of virus and the onset of paralysis.

The average urinary excretion of vit. B₁₂ in 14 subjects on normal diets was found to be 31 my per day. Similar values were obtained in 28 subjects maintained for 3 days on the following diets: milk, alkaline ash? low protein, and high protein. A somewhat higher value was found in 8 subjects on high purine diets, and the increase over normal was shown to be due to interfering compounds. On the third day of fasting, 6 subjects were found to excrete an average of 62 my vit. B₁₂ per day.

The average CF excretion by the subjects on the above diets accounts for only part of the total FA compounds excreted, and appears to be correlated with the total FA values. Differences in vitamin B₁₂ excretion resulting from dietary changes are not similar to those observed in CF and FA excretion.

1. The observed rise in sodium and the decrease in potassium in the allantois fluid of chick embryos treated with cortisone are the reverse of the reported changes in the urine of human subjects receiving this steroid, and resemble those seen in adrenal insufficiency. 2. The increased chloride and sugar levels in the fluid of the allantois following therapy with the higher dosages of cortisone resemble directionally the renal effects of cortisone in man.

Extracts of Rain-lily bulbs have been tested for their effect on the growth of tumor and embryo of tumor-bearing eggs. The results obtained with aminopterin in the same tests are included for comparison. Acid aqueous and acetone extracts of the plant bulb inhibited tumor growth 81% and 94%, respectively, without affecting the embryo. Aminopterin inhibited tumor growth 87% and embryo growth 19%.

Thin sections of Molluscum contagiosum lesions from man have been examined with the electron microscope. Cells were seen in the prickle cell layer of the epidermis which contained only a very few cytoplasmic elementary bodies. These are considered to be early infected cells. Cells were seen with varying numbers of elementary bodies scattered through the cytoplasm while others were entirely filled with elementary bodies forming a mature inclusion. The elementary bodies in the early infected cells differed somewhat in shape from most of the elementary bodies seen in the larger inclusions. The mature inclusion bodies were divided into locules by septa. The locules contained mature elementary bodies. The elementary bodies appear to form from the material composing the matrix of the septa by a process of segmentation and condensation. The elementary bodies themselves were sectioned and appeared in some instances to have a formed cortex with a less dense interior.

Using p-nitrophenyl sulfate as substrate no sulfatase activity could be demonstrated in costal cartilage at varying stages of differentiation or in bone providing no red blood cells or marrow elements were present.

At one hour after injection of 100,μc of C¹⁴-formate into mice with advanced Ak-4 leukemia, it has been shown by means of blood smear autoradiograms that about 90% of the leukemic cells are highly active as compared to about 5% of the more mature lymphoid elements. No active polymorphonuclears, erythrocytes, or thrombocytes were observed within one hour.

Total body chloride was measured in normal man by the dilution of bromide and it has been estimated that nearly half the total body chloride is intracellular.

p- (2,4-Diamino-s-triazinyl−6) -aminophenyl p'-aminophenyl sulfone has been synthesized and shown to inhibit the growth of M. tuberculosis and E. coli in vitro at a concentration of 1.2 × 10^-4 moles per liter, the inhibition being reversed by p-aminobenzoic acid.

1. Pyridoxine deficiency was produced in weanling male and in young adult female mice fed (a) a purified diet containing 9 other crystalline B vitamins, and (b) a semi-purified diet containing a low level of liver powder. 2. Acrodynia appeared among both the weanling and young adult mice fed these 2 diets. 3. As little as 0.166 mg of added crystalline pyridoxine-HCl per kg of the purified diet protected the mice from acrodynia and resulted in a good growth response by the weanling mice.

(1) Digestion and destruction of the living esophageal mucosa in cats and dogs can be accomplished by bile, pancreatic juice, and mixtures of the two. (2) Bleeding from esophageal erosions attending the complete diversion of bile and pancreatic juice into the esophagus in the dog causes a severe secondary anemia. (3) The development of the esophagitis and anemia in the dog is not altered by total excision of the acid secreting portion of the stomach.

(1) The daily inhalation of cigarette smoke failed to produce peptic ulcers in dogs. (2) Cigarette smoke abets the ulcerogenic effect of histamine in beeswax; this is believed to be a nicotine effect. (3) Cigarette smoke does not stimulate gastric secretion in dogs.

In 112 autologous and homologous anterior chamber grafts of ovarian tissue in castrate and non-castrate males, and spayed and non-spayed females, the percentage of homologous takes was highest in non-spayed females, next highest in castrate males and spayed females. The lowest percentage of such takes was in normal males. The incidence in the different categories is not considered to be statistically significant with the possible exception of the non-spayed group of females. The ages of the donor grafts were not significant in relation to growth of fragments at the ages investigated. There was no apparent influence of the tissue transplants upon the weight gain of the recipient rabbits.

Both stable and unstable emulsions of fat have been administered intravenously to rats. The stable emulsions caused a moderate increase in total lipids of the spleen, liver, and lungs; whereas, a very marked increase was observed when unstable emulsions were given. Much of the increase due to the latter type of emulsion was caused by large extracellular fat globules. The removal of the excess fat in the tissues has been studied and found to be rapid in the case of stable emulsions, and slower but definite in the case of the unstable emulsions.