Experimental Biology and Medicine

Small amounts of antibody and excess antigen sensitize guinea pig skin so that challenge with intravenous antigen and dye elicits passive cutaneous anaphylaxis. The significance of the biological reactivity of antigen-antibody complexes is briefly discussed.

Measurement of antibodies in acute and convalescent serums from college students hospitalized because of acute, nonbacterial respiratory infections revealed 5 cases of influenza-like illness and 6 cases of viral pneumonia with 4-fold or greater increases in reactivity with Sendai virus measurable by hemagglutination-inhibition and neutralization in ovo. Thirteen of 110 patients hospitalized with the diagnosis of infectious mononucleosis were found to develop increases in serum reactivity with Sendai virus during their illnesses. Efforts to isolate a virus from throat washings of 17 of these patients were unsuccessful. The possible role of Sendai virus in the etiology of these illnesses and the significance of the antibody responses were discussed.

In vitro studies on mast cells isolated from rat peritoneal fluid revealed: Metachromatic granules were partially removed by testicular hyaluronidase, but when rats were pretreated systemically with Escherichia coli endotoxin, 2 mg iv., this effect of hyaluronidase was much more extensive. E. coli endotoxin, 0.5%, did not influence the mast cells in vitro. Metachromatic staining of the cells was prevented, and if previously stained the color was abolished, by compound 48/80 or protamine sulfate. These agents and serotonin, 0.5%, neither degranulated nor disrupted the cells, but they produced cell clumping. Heparin, over 0.005% caused cell disintegration, whereas 0.5% hyaluronic acid or histamine did not appear to affect the cells adversely. Hyaluronic acid seemed to protect them to some degree against degranulation.

Employing the dog's hind-leg which permits a controlled study of bleeding time, it has been determined that refrigeration of blood for 10 minutes at 1-2°C causes it to lose its normal hemostatic properties, not recoverable by heating to 37°C. Furthermore violent aeration of blood in air, by liberating CO₂ and driving the pH to the alkaline side, also renders blood non-hemostatic, although by returning the liberated CO₂ to blood, by shaking in atmospheric air containing 9% CO₂, hemostatic action may be recovered. The mechanism of these phenomena, their application as routine methods in the laboratory, and their implication in technics of blood transfusion are discussed.

(1) The extent of biochemical alteration in experimental bowel obstruction shock was related directly and consistently to actual rectal temperatures developed, in both normothermic control rats and those made poikilothermic by chlorpromazine. (2) The value of hypothermia in protecting rats from bowel shock has been demonstrated. (3) Chlorpromazine was beneficial in shock only when its use led to enhancement of hypothermia. Its use at a higher environmental temperature facilitated development of hyperthermia, and aggravated the biochemical syndrome of bowel shock. Chlorpromazine appears to have no specific protective action against shock. (4) Possible mechanisms are discussed.

A DLSH reaction (statistically significant decrease in liver NPSH) was secured in mice by administration of very large amounts of insulin, epinephrine or nor-epinephrine, or tourniquet traumatization. Trauma was as effective in inducing a DLSH reaction in alloxan diabetic or adrenal demedullated mice as in normal mice; hence the trauma induced DLSH reaction was not attributable to endogenous insulin or adrenal medullary hormones. The trauma induced DLSH reaction was practically abolished by either adrenalectomy or administration of the ganglion blocking drug chlorisondamine dimethochloride. This indicates that both adrenal cortical and sympathetic nervous system activities are required for the reaction.

Feeding of 3/4 or 1/2 lb trichloroethylene-extracted soybean oil meal/day/100 lb to young calves for 10 days induced aplastic anemia and caused death 14-25 days after feeding of the toxic material had ceased. DL-batyl alcohol injected by various routes after feeding of the toxic agent had ceased, but before appearance of clinical symptoms, failed to prevent development of aplastic anemia.

(1) Feeding of cobaltous chloride to cholesterol-fed chickens reduced the incidence and severity of their aortic lesions and caused blood cholesterol concentrations to be lower than in controls.

The amounts of cholesterol in β-lipoprotein were similarly reduced and ratios were increased. There was no consistent effect on lipid phosphorus concentrations. Cobalt in the diet did not seem to influence activity of the thyroid. When given either in the diet or intravenously, cobalt did not change the structure of the pancreatic α-cells. (2) Subcutaneous injection of cobaltous chloride generally increased the incidence and severity of aortic lesions and concentration of cholesterol in blood. These findings are similar to those of Caren and Carbo in the rabbit(8). (3) The influence of cobalt in the diet on lipid pattern and aortic lesions was opposite to that of parenterally administered cobalt. It did not produce other effects indicative of previously reported specific systemic actions. Siperstein, Nichols, and Chaikoff(1) concluded that blood-cholesterol-lowering effect of ferric chloride given in the diet was due to precipitation of bile acids, resulting in decreased cholesterol absorption. It is concluded here that the effect of cobalt in the diet of cholesterol-fed chickens similarly is due to a local action in the intestinal tract.

Absorption of triglyceride labeled with I¹³¹ in the rat bearing Walker Carcinoma 256 has been studied. The amount of fat recovered from thoracic duct lymph of the tumor rat is significantly less than that recovered from lymph of normal and pair-fed control rats.

(1) One or 2 doses of 7 mg/kg of TC or 2.5 mg/kg of MC destroyed the litters of rats when given after 10th day of gestation period. (2) Placentas survived the fetuses. (3) Maternal response to dosage consisted in depression of myeloid tissues of bone marrow, peripheral WBC and prothrombin time. (4) Four consecutive litter destructions in utero with TC were well tolerated by a group of rats, did not impair their subsequent fertility or reproduction, nor cause abnormalities in subsequent offspring. (5) The same doses of TC or MC given prior to fertilization, or implantation were less effective in litter destruction.

Histochemical studies were conducted on effects of avitaminosis C on the reproductive tract of the male guinea pig, and an attempt was made to differentiate these changes from those which result from inanition in the presence of an adequate dietary intake of ascorbic acid. Results indicate that scurvy may cause an inability to utilize other essential dietary substances resulting in more severe cachexia than that noted in inanition controls. These studies suggest that changes in the cytochemistry of the Golgi apparatus are involved in spermatogenic arrest. Furthermore, as noted in this investigation, the relation of scurvy and/or inanition to reproductive physiology is indicated by altered carbohydrate metabolism in testes and seminal vesicles. These changes were more marked in scorbutic animals than in inanition controls.

1) Sputum specimens from patients with acute and chronic pulmonary diseases were examined cytologically by the Papanicolaou technic. Certain abnormalities in exfoliated ciliated epithelial cells (ciliocytophthoria) were observed in specimens from a high proportion of individuals with viral respiratory infections; in contrast, ciliocytophthoria was seen only rarely in sputum preparations from patients with acute or chronic pulmonary disease of non-viral etiology. 2) Sputum cytology may prove valuable for prompt differential diagnosis between viral and bacterial respiratory infections.

1. When the entire facial nerve of the alligator was transected intracranially, there were chromatolytic changes in the facial nucleus, but none in either superior olive. 2. When both the facial and auditory nerves were sectioned, chromatolytic changes were particularly noticeable in the contralateral superior olive. This indicates that the olivocochlear bundle which Rasmussen has described in the cat is also present in the alligator.

Human serums and serums from several species of animals have the capacity to kill and disintegrate T. vaginalis with great rapidity. This activity of serum is destroyed by exposure of the serum to 56°C for 30 minutes.

1) The whole carcass retention of fluoride by rats receiving a fluoride drinking water containing a combination of Ca⁺⁺, Mg⁺⁺, Fe⁺⁺, and PO₄— ions was significantly less than in rats drinking a chemically pure fluoride water containing no additional inorganic ions. 2) In contrast, studies of the individual ions at various concentrations show only small fluctuations in the proportion retained. No significant influence on retention was noted when any of the ions were studied singly. 3) The decrease in retention seen in rats ingesting the water containing a combination of ions suggests that the influence of the ions may be additive or synergistic. This may be important since most natural waters contain a group of several inorganic constituents. More attention should be placed on inorganic composition of the communal water supplies before fortifying them with fluoride. It is possible that if the combination of major inorganic constituents of natural waters exceeds a yet-to-be determined concentration, optimal fluoride concentration would need to differ from the now accepted level of 1 ppm in order to produce maximum anticariogenic benefits.

1. Banana ingestion (12 g/kg/24 hr) by normal adults produced an average 2-fold increase in urinary excretion of 5-OH indoles. This was not of such magnitude as to suggest an erroneous diagnosis of malignant carcinoid. 2. Collection of 24-hour urine specimens and use of a quantitative test are advocated over collection of random urine specimens. Positive screening tests should be checked by a quantitative method.

A rise in antibodies to types 1-4 of adenoviruses during illness can be detected in human serum by employing hemagglutination technics. Although the original method described does not measure an increase in antibodies directed against a single type of virus, type-specific antibodies can be titrated if serum is mixed with a pool of heterologous viruses before tanned rbc treated with homologous virus are added. This modification measured type-specific antibodies solely in 4 of the 8 sera tested. The antigen adsorbed to tannic acid-treated rbc is probably not the infectious viral particle and is separable from it by high speed centrifugation.

1) An investigation was made of changes in virulence of L. pomona for hamsters following serial transfers in a fluid medium. L. pomona strain W, which did not initially kill hamsters, later produced a lethal effect. Both strain W and O of L. pomona decreased in virulence (increase in LD₅₀ values) during first 4 medium passages. After 4 to 8 passages in media an increase in virulence was observed. Titrations of subsequent medium passages (12 to 24) of strain W indicated a gradual loss of virulence. 2) In vitro hemolysin production could not be correlated with virulence. An explanation for virulence alterations is proposed and the possible significance of hemolytic activity of leptospirae is discussed.

The fate of thiamine, labeled with sulfur-35 in the thiazole moiety, has been studied in rabbits after oral and parenteral administration of doses in the physiological range. Analyses of the urinary and fecal sulfur fractions for 4 to six 24-hour periods following administration of radiovitamin, gave average total recovery of radiosulfur of 77% after administration by stomach tube, 86% after intramuscular injection, and 54% after intravenous injection. When oral dose of radiovitamin was followed 6 hours later by intramuscular injection of a 20-fold larger dose of non-radioactive thiamine, an additional 17% of radiosulfur was recovered in urine. In all cases, the neutral sulfur fraction of urine contained more than 50% of the recovered radiosulfur, and the greater portion of this was excreted in the first 24-hour period following administration of radiovitamin. Isolation of main metabolites following oral administration indicates that unchanged thiamine-S³⁵ and thiazole S³⁵ moiety together account for approximately 95% of radiosulfur of the neutral sulfur fraction in the first 24-hour period, and are excreted in a ratio of 2 : 1, respectively.

Six strains of influenza A virus were isolated in New York during the 1957 pandemic. Close antigenic relationship was found among these strains and 2 Far East isolates. and slight antigenic relationship to older prototype strains was also demonstrated. One of the local isolates, RI/5/57, was found to be a very sensitive indicator of the presence of hemagglutination-inhibiting and neutralizing antibodies against the 1957 strains of influenza A virus in human serum. Antibodies to the RI/5/57 strain were demonstrated in patients of various ages obtained from 1940 to 1957. The sensitivity of the 6 New York strains to non-specific inhibitors of hemagglutination varied from apparent insensitivity in 3 strains to extreme sensitivity in 2.

1. Dietary restriction of copper in adult rats was followed by prompt decrease in serum copper and a somewhat slower and less extensive depletion of liver copper. 2. Feeding of diets high in copper was followed by increase in concentration of copper in both serum and liver. The increase occurred more rapidly in serum than in liver during the first 20 days. Thereafter, rate of increase was greater in liver than in serum. 3. It is concluded that concentration of copper in serum and liver is markedly influenced by dietary intake of copper and that there is a close correlation between serum and liver copper concentrations.

1. Administration of highly unsaturated fats to the rat effected a greater intestinal absorption of cholesterol than similar administration of saturated fats. 2. Following continued ingestion of highly unsaturated fat, there was evidence that the fall in plasma cholesterol observed was accompanied by a marked increase in excretion of both cholesterol and bile acid in the bile. 3. The possible relationship of reduction in plasma cholesterol to the observed change in bile cholesterol and bile acid excretion is discussed.

1. C¹⁴-labeled lysine administered intraperitoneally into the rat is incorporated within hours into the saline-soluble and -insoluble non-collagenous protein and collagen of biopsy-connective tissue. Radioactivity is confined to lysine in all fractions of connective tissue except for a portion which is converted to hydroxylysine in the collagen. 2. Following appearance of radioactivity in all fractions of connective tissue, decay occurs rapidly in saline-insoluble non-collagenous protein and saline-soluble protein and slowly in the collagen fraction. Thirty days after injection of labeled lysine approximately 10% of maximum activity remained in the saline protein fractions while 30% remained in collagen. 3. Incorporation of labeled lysine into the non-collagenous protein of connective tissue is not influenced by tissue age while its incorporation into the collagen appears to decrease with tissue age. 4. Specific activity of labeled hydroxylysine from a 300-day old tissue of a female rat was approximately twice that of a comparably aged tissue from a male rat.

1. Lysine-2-C¹⁴ becomes incorporated and hydroxylated in connective tissue incubated in Krebs-Henseleit medium for 12 hours. 2. Specific activity of collagenous and non-collagenous protein in the tissue studied in vitro was greater than that in tissue studied in vivo. 3. Hydroxylation of labeled lysine appears to occur more effectively in connective tissue of rats given labeled lysine intraperitoneally than in the connective tissue incubated with lysine-2-C^14.

The folic acid and Vit. B₁₂ content of CSF and B₁₂ in serum was studied in more than 300 neurological cases. The PGA levels in CSF in a number of cases were elevated; high values were scattered among a variety of neurological conditions, but were high in four-fifths of cases with MS. The titre of B₁₂ in CSF and serum of MS patients was elevated in one-third and reached the upper limit of the normal range in another third of the cases. We assume that increased levels of these vitamins in CSF are not merely an indication of increased permeability of the blood-brain barrier, but of etiological significance for MS.

No significant relationship has been found between the diphasic response in the benzol-induced leukopenia in rabbits and the presence or absence of adrenal glands.

A method and an apparatus for routine electroejaculation of guinea pigs were developed, the semen coagulum or “vaginal plug ”examined, and the coagulation process described. A series of proteolytic enzymes were tried for their liquefying activity and 0.1% chymotrypsin was found to result in consistent liquefaction and good sperm motility. Mean ejaculate volume was 0.5 ml and mean motility at 39°C was 51%. There was no evidence of gross morphological or histological changes in the reproductive system of guinea pigs electroejaculated at biweekly intervals.

This report describes a simple apparatus for continuous intravenous infusion of fluids in dogs, employing a standard assembly of reservoir bottle and flexible connecting tubing that leads to the dog's harness and a plastic catheter dwelling in an external jugular vein. Suspension of the apparatus by a swivel above the dog's cage permits the dog almost complete freedom of movement during long periods of continuous infusions.

1. The data demonstrate that both penicillin and aureomycin supplementation increased food intakes of rats subsisting on diets deficient in either or both thiamine and riboflavin, but neither antibiotic influenced food consumption when adequate thiamine and adequate riboflavin was administered. In general, growth of test animals paralleled food consumption, with indications that antibiotic supplementation improved food utilization, particularly when thiamine intake was sub-optimum. 2. The data further indicate that the amount of thiamine accounted for was increased by feeding rats diets containing either penicillin or aureomycin, or a combination of the 2 antibiotics. However, the data show that thiamine absorption was improved in the presence of antibiotics, particularly, penicillin, as indicated by lower thiamine concentration in ceca and feces, and by higher thiamine concentration in liver and urine. 3. While the data do not exclude the possibility of riboflavin synthesis in the animal body, they do indicate that if synthesis occurred it was not influenced by the presence or absence of the antibiotics under investigation. There was evidence of increased riboflavin absorption in the presence of antibiotics as indicated by lower concentrations of vitamin in ceca and feces and a higher concentration in liver.

A liquid scintillation technic for counting total plasma or serum activity of C¹⁴ and H³ is described. The serum is complexed by the quaternary ammonium base, Hyamine and dissolved in phosphor-containing toluene with the addition of ethanol. Correction for quenching is effected by recounting an added internal standard after determination of the observed count. The method was shown to be rapid and accurate for various volumes of serum.

1. Peripheral blood ammonia concentrations were elevated following convulsions due to electroshock therapy in patients, convulsions in dogs due to metrazol, and active voluntary exercise in human subjects. 2. The possibility that increase in circulating blood ammonia may be a factor in hyperpnea of muscular exercise is discussed.

The necessity of implanting an occlusion bag to secure a base line at zero flow has been eliminated. This has been accomplished by placing the pick-up electrodes in the magnetic field to eliminate the transformer emf induced in the input lead circuit. As the sleeve is moved parallel to flow axis a point is found at which the a.c. voltage between electrodes vanishes regardless of whether the magnet is on or off. Thus, the baseline is obtained by merely switching off the magnet. The circuit has been improved to a point where the animal can be handled without interference with the recording of blood flow.

The male AxC rat exhibits a genetically-determined anomalous development of the genitourinary apparatus including absence of one kidney, ureter, vas deferens, and seminal vesicle associated with testicular hypoplasia on the same side. Detailed histochemical and histological analysis of the postnatal development of the defective testis reveals that this testis can not be differentiated from the normal testis until puberty, at which time it exhibits a total failure of tubular maturation and spermatogenic function. Interstitial cells, however, continue to develop normally and exhibit normal endocrine function and removal of the normal testis does not lead to atrophy of the prostate or seminal vesicle. Surgical extirpation of the vas deferens during the prepuberal period does not lead to any alteration in subsequent development of the normal testis. It may be concluded that this instance of testicular hypoplasia represents a genetically determined defect with delayed somatic manifestation.

1) Soluble BSA anti-BSA complexes formed in regions of extreme antigen excess are less anaphylactigenic for the mouse than similar complexes formed in regions of low antigen excess. 2) The hypothesis is advanced that anaphylactigenic activity of these complexes for the mouse is dependent upon their capacity to react with complement.

1. No interarterial intercoronary anastomoses between any branch of right and left coronary artery could be demonstrated at autopsy in normal dogs with a vinyl plastic injection mass. 2. Following surgical production of arteriovenous fistula (10-12 mm long) between main pulmonary artery and left atrium, interarterial intercoronary anastomoses between right and left coronary arteries were demonstrated at postmortem examination in 5 of 7 dogs examined at intervals of 2 days to 14 weeks following surgery. These anastomotic communications were arterial in size.

After treatment of human synovial fluid with hyaluronidase, the hyaluronate and proteins can be separated by paper electrophoresis. The electrophoretic distribution of the protein fractions of normal synovial fluid differs from that of the protein fractions of both normal serum and osteoarthritic synovial fluid.

HeLa adapted measles virus (Edmonston strain) was propagated in suckling mice through 24 serial intracerebral passages. The virus produced a lethal infection in most animals 7 to 13 days following inoculation. Identity of mouse propagated agent was established by serum neutralization tests in tissue culture and in suckling mice. Further identification was established by protection of suckling mice born of mothers immunized with tissue culture measles virus. Fluid phase from tissue cultures inoculated with late mouse passage materials did not contain infectious material either for HeLa cells or for suckling mice. Infections, however, were readily induced when cells were employed as inoculum.

(1) Individual infectious particles were isolated from the mycelial phase of H. capsulatum using Chambers micromanipulator. The particles consisted of germinated and non-germinated tuberculated macroconidia, germinated and non-germinated microconidia and non-branching mycelial fragments. Each particle was placed into a sterile capillary tube inserted into a 21 gauge sterile inoculating needle. Aseptic technics were followed, in addition, streptomycin and penicillin were added to the collecting and inoculating fluids. Microscopical examination of each capillary tube was made prior to and immediately after inoculation procedure to ascertain the disposition of the particle. Fertile Leghorn eggs and the white Swiss mouse were the experimental animals. (2) Infection resulted from all particles with one exception, mycelial fragments, in the mouse. The total percentage of infectivity in the 182 inoculated mice was 5%. Of the 571 inoculated chick embryos approximately 8% became infected. The most infectious particle was the mycelial fragment. (3) Inoculation of single particles from the mycelial phase of H. capsulatum proved that the minimal infection inoculum can be units other than the macroconidium. In each instance the infectivity rate was relatively small but no physical protection was given the inoculated particle.

Two newly established cell strains capable of continuous cultivation, one derived from adult bovine kidney (MDBK) and one from adult ovine kidney (MDOK), are discussed. It is suggested that the strains may be useful for those interested in veterinary virology in particular and virus-host relationships in general.

The fate of Bacillus megatherium bacteriophage intraperitoneally inoculated into mice has been investigated. Active bacteriophage particles were recovered from spleen and liver at a time when none were recovered from blood, lung, kidney, and brain. The relationship of this occurrence to possible phagocytosis of bacteriophage particles is discussed. Introduction of bacteriophage suspensions of high titer into the gastro-intestinal tract by feeding and gastric lavages led to an irregular but consistent recovery of active phage particles from the blood circulation. Conversely, phage particles inoculated intraperitoneally were recovered in samples of urine and feces.

When rats are kept on a “Low Potassium Diet ”that contains only near minimal maintenance levels of Mg, there develop cardiac necroses which can be prevented both by KCl and by MgCl₂ administration. On the other hand, Na₂HPO₄ (unlike equivalent amounts of NaCl) rapidly provokes development of severe cardiac necroses, nephrocalcinosis and muscular cramps before this diet, in itself, produces any obvious morbid changes. The particularly severe K and/or Mg-deficiency syndrome induced by Na₂ HPO₄- supplements, in animals on this diet, can be prevented by either KCl or MgCl_2. These observations highlight the importance of PO₄- and Mg-ions in the development of the syndrome usually ascribed to K-deficiency.

1) An adenovirus vaccine evaluation study was carried out at San Diego Naval Training Center. Recruit population was studied. Kidney cell tissue culture vaccine containing adenovirus types 3, 4 and 7 was used, inactivated with formalin. Half of the men in 39 companies were given vaccine while the other half were given placebo. Observations were made from March 11 to July 3, 1957. Adenovirus isolations and complement fixing antibody response in acute and convalescent serums were employed as indices of infection. Types 4 and 7 adenovirus infections were prevalent in the control population. 2) During 3 through 9 week post-vaccination period, weekly admission rate/1,000 was 22.3 from the vaccine group, and 26.2 from the control group, a 15% difference. However, on the basis of adenovirus isolation the weekly admission rate was 3.4 from the vaccinated compared to 10 from the control, a difference of 66%. When 4-fold or greater complement fixing antibody rises to adenovirus antigen were employed as evidence of infection, the weekly admission rate was 3.4 from the vaccinated group and 12 from the control group, a difference of 72% in favor of the vaccine. On the basis of above findings, adenovirus vaccine significantly reduced number of adenovirus infections in the vaccinated group compared to the control group. 3) In non-study companies from Mar. 5 to May 5 adenovirus infections accounted for 80% of Dispensary admissions. Forty % of admissions from the control group of the study companies were due to either types 3, 4 or 7 adenovirus while only 15% from the vaccine group were due to these agents.

1. An explosive outbreak of Asian influenza beginning the week ending June 15, occurred at San Diego Naval Training Center. The peak admission rate of 192.2 from the recruit population occurred the following week. After the first 3 weeks the rates subsided but continued to be 2 to 3 times the pre-epidemic levels through the first 2 weeks in November. Serological and virus isolation studies showed that approximately 75% of admissions from onset of epidemic through November were due to Asian A influenza infections. A few were due to influenza B infections. Admissions due to adenoviruses occurred also in increasing numbers during Sept., Oct. and Nov. On the basis of serological studies essentially all admissions to the Dispensary from onset of epidemic to Nov. 16 were due either to influenza or adenovirus infections. 2) The general lack of immunity to Asian influenza virus in the recruit population is reflected in the high attack rate during the first 3 weeks of the outbreak and high percentage of individuals showing an antibody response to Asian A antigen in the 3-survey companies bled before and 4 weeks after the onset of the epidemic.

In 5 subjects receiving clinically effective oral doses of Nilevar, there was no alteration in adrenocortical function as measured by plasma 17-OHCS levels before and after intravenous ACTH.

1. Plasma copper was determined in 29 patients with sprue in relapse, 27 patients with sprue in remission, and 6 patients with megaloblastic anemia of pregnancy. 2. Mean plasma copper concentration (± 1 S.D.) in patients with sprue in relapse was 87 ± 33 μg%; in treated patients, 114 ± 33; and in patients with megaloblastic anemia of pregnancy, 159 (105 − 242). 3. It is suggested that hypocupremia in sprue may frequently be due primarily to an inability to synthesize the protein portion of ceruloplasmin. In a few patients malabsorption of copper may be a contributing factor.

Two experiments are reported in which regional blood flow under controlled perfusion pressure was being measured by direct means along with systemic blood pressure and in which Traube-Hering waves were seen to occur. Rhythmic changes in the regional blood flow were observed which were synchronous with but opposite in direction to the changes in systemic blood pressure. This agrees with the conventional view that changes in blood pressure in this situation are secondary to changes in peripheral resistance.

Rabbits were treated with dicumarol and dicumarol plus ACTH and prothrombin times and bleeding times determined. A slight increase in bleeding time was observed after 3 days on dicumarol, in 50% of the rabbits. Half the rabbits died in the next 24-72 hr from generalized non-specific hemorrhage. This was not related to the previous increase in bleeding time. Bleeding did recur and continue from the cut ear but hemorrhage was much more extensive than this. It is suggested that determination of bleeding time is a marked stress procedure in rabbits.

A method for purification of erythropoietin from the filtrate of acidified, boiled plasma prepared from phenylhydrazine anemic rabbits utilizing DEAE-cellulose ionexchange columns has been described. The active erythropoietic factor thus prepared has been partially characterized and shown to be a low molecular weight, acidic glycoprotein.

(1) Doses of 0.05 mc of Y⁹⁰Cl³ solution and of CrP³²O⁴ colloidal suspension in 0.05 or 0.1 ml were injected into ventral surface of tails in mice. At intervals of 1 minute to 3 days after injection, tails were serially amputated and preserved until 3rd day when every mouse was sacrificed; their organs (liver, lung, spleen, kidney), tails and heart blood (0.3 ml) were emulsified, dried and tested for radioactivity content with a Geiger Tube Detector. (2) Results showed only a narrow range of radioactivity for each organ specimen calculated as a product of the average blood radioactivity in the same group of mice (amputated at the same time). (3) The calculated and tabulated data show that a significant proportion of CrP³²O⁴ was translocated into the liver from the tail; for Y⁹⁰ the initial storage in the tail was very high, but some translocation occurred, presumably by excretion. (4) The pattern of Y⁹⁰ and CrP³²O₄ distribution in the body after injection into the tail was comparable to delayed results of intravenous injection; this resemblance was interpreted by high vascularization of the tail. (5) It was concluded that the method of serial tail amputation may be useful for outlining the dynamics of isotopes spread in bodies of experimental animals.

(1) Ethyl alcohol does not influence cardiac output of the rat in doses of 5 ml of 20% by volume solution in 200-250 g rats. (2) Blood flow values to the major organs calculated from the fractional uptake of Rb⁸⁶ or iodoantipyrine (I¹³¹) show the following: cutaneous blood flow (considering the skin as a whole) hepatic arterial inflow, portal flow other than splenic, and bronchial blood flow are unaffected by alcohol; coronary blood flow is increased 31%; renal blood flow is reduced 32%; cerebral blood flow is 90% greater in the alcohol treated than in the pentobarbitalized animal.

1) Mice were capable of being protected against a number of isolates of C. neoformans by passive immunization with antiserum. This protection was limited to the immunization period and was type-specific. 2) In vivo and in vitro phagocytosis of Cryptococcus cells has been demonstrated. When large and small capsule variants were studied separately, the former were practically resistant and the latter relatively susceptible to ingestion by mouse PMN. 3) The probable mechanism for the protection achieved has been discussed and a possible use for antiserum has been proposed.

Experimental mumps virus infections in guinea pigs are characterized in vivo by a positive delayed hypersensitive skin test reaction to viral antigen and in vitro by a cytotoxic effect of mumps virus antigen on cells from tissues cultivated from mumps-infected animals. Cortisone protects against this in vitro cytotoxicity of mumps virus.

Follicle-stimulating activity was detected in blood plasma of adult female castrate rats as early as 7 days post-operative by injection of plasma into immature hypophysectomized female rats for a 4-day period. Follicle-stimulating activity continued to increase slowly but progressively as the postoperative interval increased to a 4 months' period. With this bioassay method no direct evidence for circulating interstitial cell-stimulating activity was found at the highest dosage level tested.

Serum cholesterol level was determined in 5 series of rats receiving either different fats in their diets or different fats plus fluoride to determine whether the use of fluoride is associated with an increased serum cholesterol level. After administration of 2 mg of fluoride daily throughout a 10-week period there was no significant change in serum cholesterol level of rats regardless of their dietary fat source or the presence of fluoride. Comparison of body weight gains of the various groups fails to suggest that any of the fats used resulted in increased fluoride toxicity.

After intraperitoneal injection of a mixture of Ca⁴⁵ and Sr⁸⁹ in the rat, the Sr/Ca Observed Ratio in parts of a long bone, the femur, was obtained 15 minutes to 48 hours after injection. A maximal Sr content appeared in bone within 30 minutes, with the shaft showing discrimination against Ca; during the next 48 hours the Observed Ratios fell but remained consistently higher in shaft portions of the bone. Data on blood Sr/Ca ratios are also presented.

Adrenal function was studied in 6 diabetic patients with hyperglycemia controlled by DBI (N B' phenethylformadinylimourea) or by DBI and insulin. DBI administration did not significantly affect baseline plasma hydrocortisone levels or response to ACTH. These results appear to eliminate adrenal steroid suppression as a possible mode of action of this biguanide.

A procedure is described which utilizes a combination of filter paper and ion exchange chromatography to separate the urinary metabolic products of adrenaline. After intravenous infusion of dl-adrenaline-2-C¹⁴ in 6 human subjects, 73 ± 5% of infused radioactivity was recovered in 24 hour urine. Radioactivity which appeared in urine was distributed among various metabolic products as follows: adrenaline, 4%, 3-methyl-O-adrenaline, 5%, 3-methyl-O-adrenaline conjugate, 42 ± 7%; 3-methoxy-4-hydroxymandelic acid, 27 ± 3%; 3, 4-dihydroxymandelic acid, 12 ± 4%; 2 unidentified fractions, 4%.

The effects on plasma cholesterol level of 4 fats each supplied at 3 levels in a high and a low protein diet were studied in the cholesterol-fed chicken. On the low protein diet a marked hypercholesterolemia was observed which was not affected by type of fat studied or level of supplementation. On the high protein diet, plasma cholesterol levels were essentially normal regardless of source and level of dietary fat. Since the fats used represented a wide range in the product EFA x TSFA, a lack of relationship between this product and plasma cholesterol level in the growing chicken is indicated.

(1) A method for rapidly determining the mobility equilibrium of a compound is described. The procedure provides an analytical rather than the conventional trial and error method for determining paper electrophoretic characteristics of the components in a mixture. (2) Some factors affecting ion movement and mobility equilibria in open-type paper electrophoresis units are discussed. (3) An application of the method in separating urea and several indole derivatives is presented.

Electrophoretic separation of fetal and maternal plasma proteins in goats and sheep revealed that the fetal plasma lacks γ-globulins and exhibits an additional peak in the area of α₁-globulins. This peak probably corresponds to fetuin. Quantitative conditions of fetal plasma proteins seem to depend largely on age of the fetus. In goats, the percentage of total plasma proteins and of plasma albumin increases; percentage of fetuin decreases with the progressing age of the fetus. The remaining plasma proteins do not seem to be affected. In fetal sheep the percentages of total plasma proteins, plasma albumin and probably of β-globulins increase; levels of fetuin and α₁-globulins decrease with age. The percentage of α₂-globulins remains unaffected.

In both bovine kidney and HeLa cultures, extensive focal nuclear degeneration followed inoculation with a HeLa-adapted IBR virus strain. The pattern of nuclear and cytoplasmic degeneration was similar for the native host (PBK) and the HeLa cell. In cells of both types, nuclear inclusions were Feulgen-negative, and morphologic nuclear changes were accompanied by a reduction of DNA.

Newborn mice of the ZBC strain injected intravenously with living spleen cells taken from donors of the same strain in which tolerance to Ce tissues was previously induced, became tolerant to this strain and accepted homologous skin grafts from Ce donors. This would indicate that acquired tolerance can be transferred to an isologous individual at birth by the intravenous injection of spleen cells taken from a tolerant donor.

1) Chronic treatment with posterior pituitary preparations containing ADH induces significant enlargement of tibia cartilage width in normal male and female rats; oxytocic hormone is ineffective in normal animals. 2) In hypophysectomized animals posterior pituitary hormones do not produce modification of the epiphyseal cartilages.

A mean increase of 393 mg in daily excretion of creatine and an increase of 0.77 g in daily excretion of total creatinine chromogen has been observed in 6 subjects after oral administration of 30 mg 17α-ethyl-19-nortestosterone daily for 6 weeks.

1) Injections of 1-25 μg of DNP into chicken eggs, prior to incubation resulted in embryonic mortality after 3 days incubation, but did not produce lens changes. 2) Injections of 25-200 μg of DNP after 5 days incubation did not produce lenticular changes. 3) Cataracts were produced in embryos and hatched chicks from eggs injected with 200-500 μg of DNP after 8 days incubation. The size of cataract was roughly the same in hatched chicks as in embryos from the 500 μg group. 4) Injection of 750 μg of DNP after 15 days incubation produced an opacity evident after 20 hours. However, the opacity apparently regressed since the eyes of chicks hatched from this group appeared to be normal.

When rats are swimming at 19 or 28°C, swimming time is markedly reduced by chlorpromazine (10 mg/kg) treatment. This difference is not due to differences in body temperature but is probably caused by sedation and muscular paralysis caused by this drug. At 9°C the detrimental effect of bath temperature on swimming time is so marked that chlorpromazine fails to display any action whereas at 32°C the stress is so small as to allow the animal to perform as well even when treated with chlorpromazine.

Organic phosphate insecticides. OMPA, EPN, malathion, chlorothion and phostex, which are rapidly metabolized in liver, markedly increased hexobarbital sleeping time in mice, while BFP and TEPP did not. Malathion and chlorothion did not alter toxicity of hexobarbital. TEPP hastened onset of action of barbital while chlorothion did not. Certain anticholinesterases which are rapidly metabolized by liver may compete for enzymes which are responsible for destruction of hexobarbital.

1. In Vit. B₁₂ deficient rat there is a decrease in serum levels of albumin and α₁-globulin. In PGA deficiency these fractions are affected to a lesser extent but there is, in addition, a significant drop in levels of β- and γ-globulins. Reductions in serum levels of albumin, α₁-globulin and γ-globulin in combined deficiency of Vit. B₁₂ and PGA are greater than those in single deficiencies of either vitamin. 2. In Vit. B₁₂ deficiency, an increase in proportion of β-globulin is attended by increase in relative concentration of free Vit. B₁₂ in serum. It is suggested that there may occur a preferential synthesis of β-globulin in the deficient state whereby mobilization of the available vitamin may be effected. 3. The observed alterations in serum level of γ-globulin are similar to the known effects of corresponding vitamin deficiencies on antibody synthesizing ability of the animal.

Chemical analysis of rat skin was independent of the area from which the sample was taken. Increases in hexosamine and decreases in hydroxyproline concentration found in skin necrotized with croton oil was paralleled to a lesser degree in uninjured skin. This systemic response was also found as an increase in circulating concentration of hydroxyproline in blood. Despite the statistical significance of these changes, the markedly increasing ratio of hexosamine to hydroxyproline in necrotic tissue with time was not paralleled significantly by the control.

Extracts of human rabbit tissues inhibited heparin-induced lipemia clearing activity of serum. Extracts of spleen, kidney and liver were strongly inhibitory; those of other organs exercised slight or moderate inhibition. The tissues contained inhibitory components which appear to be associated with proteins since they are nondialyzable, heat-labile and inactivated by trypsin.

1. It has been demonstrated that chlorpromazine, perphenazine, proclorperazine, promazine, thiopropazate, promethazine and reserpine are effective antagonists of mescaline-induced mortality in CF₁ mice. 2. No significant protection was found with use of sodium phenobarbital, meprobamate, benactyzine, atropine, hydroxyzine, ethoxybutamoxane, SY-21, diphenylhydantoin, trimethadione, lysergic acid diethylamide, serotonin, amphetamine, morphine, pyrilamine, diphenhydramine, iproniazid, pilocarpine, and arecoline. 3. It is suggested that the protective effects of the ataractics against mescalineinduced mortality are centrally mediated.

Ultrathin sections of tumorous spleen and liver from chickens of the so-called line 15 with erythroblastosis have been examined in the electron microscope. In cells of these organs spherical or spheroid structures were observed, composed of a central dense core of about 260 A, surrounded by a less dense material and limited by an outer membrane, varying in diameter from 570 Å to 760 Å. The structure of these particles resembles that of particles described in other tumors of chickens and in other tumors of known viral origin. No particles of similar character were observed in cells of organs from control healthy chickens.

Tumorous spleens and livers from chickens with granuloblastosis were examined in ultrathin sections. In the cytoplasm of the affected cells changes were observed similar to those seen in erythroblastosis, another member of the fowl leukosis complex. Particles were observed in the cytoplasm of cells of similar size and structure to that of particles in erythroblastosis. Average total diameter of the particles was found to be 670 Å and that of the central dense core was 260 Å. No particles of similar size or structure were observed in ultrathin sections of organs from young healthy chickens of similar breed.

Adult capons were treated with prolactin (LTH), follicle stimulating hormone (FSH), luteinizing hormone (LH), singly or in combination with a constant progesterone (Pg) dosage. In addition, Pg was administered alone. Pg was effective in activating the castrate feather papilla, these findings confirming earlier responses obtained in both intact sexes of fowl by others. FSH and LH were both ineffective in stimulating capon feather regeneration; in combination with Pg, the reaction was similar to that toward Pg alone. LTH uniformly caused molt of the capon feathering whether given alone, in combination with Pg, or alone as a highly purified preparation. The positive performance of LTH in the male castrate feathering described here, together with the observed lack of response toward FSH and LH in this fowl, suggest that prolactin may be the immediately effective agent in stimulating the feather papilla to production of a new plumage.

After intravenous administration of heparin-S³⁵ to dogs, it is rapidly cleared from the bloodstream. The highest radioactivity is found in liver and lung. However, no measurable radioactivity is detected in any organ after 48 hours.

Continuous administration by vein of 300 mg of corticosterone daily for 48 hours to 2 normal men caused sharp rise in urinary aldosterone, retention of sodium, and depression of urinary 17-hydroxycorticosteroids the first day. On the second day aldosterone values began to fall, while sodium retention persisted and 17-hydroxysteroid output was negligible. During the recovery period aldosterone excretion remained below baseline levels for 2 or 3 days, whereas both profuse sodium diuresis and normal 17-hydroxycorticoid excretion appeared promptly on the first day. Essentially similar but less intense effects were exhibited by 1 patient who received 200 mg of corticosterone daily for 2 days. The data suggest that normal man is capable of performing 18-oxygenation of corticosterone to aldosterone. Should further study establish that this conversion occurs within the adrenal cortex, the tentative thesis will be fortified that corticosterone is a physiological precursor in the biosynthesis of aldosterone.