Experimental Biology and Medicine

Analytical ultracentrifugation of the sera of patients with manic-depressive psychoses demonstrated different mean concentrations of protein fractions than were found in patients with schizophrenia. Values were similar for the comparative patient samples within 2 different hospital populations. Sera from manic-depressive and schizophrenic patients could be differentiated by their 19S levels.

A polysaccharide has been isolated from heat-treated or sonically disrupted cells of S. aureus. The polysaccharide is readily attacked by bovine testicular but not by bacterial hyaluronidase. Under the influence of subinhibitory concentrations of penicillin G and nafcillin, the production of the polysaccharide is greatly increased. The possible influence of penicillin on a fundamental pathway or enzyme system involved in the breakdown of the polysaccharide is discussed.

1) Progesterone, 17 α-hydroxy-progesterone and 20 α-hydroxy-pregn-4-en-3-one were measurable in ovarian plasma or ovarian tissue following gonadotropic stimulation in 2 members of the Genus Lyssodes, M. mulatta and M. irus. 2) The presence of these progestins was not dependent upon the occurrence of ruptured follicles or recently formed corpora lutea. 3) Shortly after intravenous administration of human chorionic gonadotropin, the synthesis and release of these steroids were measurably increased.

A method for tissue culture of Hydra cells is described. The removal of parasitic microsporadia from the Hydra cells and addition of Hydra bud extract to the culture medium are essential for a successful growth in vitro. Under the proposed procedures, the cultivated cells apparently have the features only of interstitial cells. Mitotic division in some of the cultured cells has been demonstrated.

Uteri of pregnant hamsters, and of pseudo-pregnant hamsters in which decidualization has been induced, reveal areas of opacity when viewed by oblique light after clearing by the benzyl-benzoate process. These opacities have been shown to be associated with decidua(1–3), and the presence of glycogen in them has been suggested(2).

During gestation, the first of these areas which appear opaque after clearing develop coincidentally with decidua and mark implantation or decidualization regions prior to visible external swelling. They increase in mass, distending the uterus as localized swellings, late on the sixth day of gestation. They reach a peak in terms of mass per conceptual swelling on the seventh day of development, and begin to regress from the antimesometrial area on the eighth day, i.e., at 7 days and 12 hours.

This paper reports results of chemical analyses of these tissues in comparison to the remainder of the uterus.

Materials and methods. At the relative peak of development of the areas in question (sixth and seventh days), it is possible to make a transverse slit in the uterus at the swollen gestational sites and, with fine forceps, peal from the endometrium the denser mass of the embryo and its surrounding decidua. Examination of material separated by the above technique and then cleared, showed that the peeled out region comprised almost all the opaque material (Fig. 1 and 2). Such “peeled out regions”subsequently are referred to as “opacities,”whether or not they have been subjected to the clearing process and thus actually appear opaque. Samples of the remaining portions of the uterus, from which all mesometrium was removed, were analyzed for comparison.

One series of determinations was carried out on material which had been cleared prior to the separation of the opaque regions.

Mice treated with a RES-stimulating agent (restim) were less susceptible than control mice to subsequent infection with opsonized S. typhimurium cells. A similar enhancement of resistance was observed when restim-treated mice were challenged with bacterial cells exposed to EDTA rather than antibody. Preliminary results indicate an increase in lysozyme in the blood of restim-treated mice.

Weanling male rats were maintained on the U.S.P. rachitogenic diet, with and without chlortetracycline supplementation, for a period of 35 days. Representative animals were sacrificed at intervals and the femurs removed, extracted and ashed under comparable conditions. The resulting data show that incorporation of 1 g chlortetracycline per kg in the rachitogenic diet slowed the rate of mineral depletion from the bones (femurs) as measured by the conventional bone-ash procedure.

Experiments on rats, calciphylactically sensitized with dihydrotachysterol (DHT), indicate that challenge with an aluminum dextran chelate (Al-Dex) prior to sensitization causes selective calcium deposition in the vagi, with gastric dilatation and respiratory difficulties. If, under otherwise comparable circumstances, the rats are challenged with Al-Dex after DHT sensitization, there is intense calcification of the analphylactoid shock organs (lips, paws, esophagus, mediastinum and joints), but not of the vagi. The observations show that, by varying the “critical period“between sensitization and challenge, a neurotropic calciphylaxis limited to the vagi, or a calcifying anaphylactoid reaction can be produced with considerable selectivity, at will.

Three experiments have been conducted to evaluate the site of action and effectiveness of the antioxidant ethoxyquin on egg yolk pigmentation. Results of the first experiment conducted during a period of high ambient temperatures indicated an average improvement of 53.8% in yolk pigmentation due to the feeding of 125 ppm of the antioxidant in addition to dehydrated alfalfa meal in the diet of laying hens. In the absence of ethoxyquin, dietary loss of xanthophylls amounted to 24% and loss of betacarotene was 32%.

The second experiment conducted during cooler weather resulted in an improvement of 21% in yolk pigmentation due to ethoxyquin feeding. Dietary losses of carotenoids in the second study were negligible.

The result of the third experiment indicated approximately 10% of the xanthophylls were destroyed in vivo. This in vivo loss of xanthophyll pigments was prevented by either the feeding of 125 ppm ethoxyquin in the diet or administration of the antioxidant by capsule (50 mg, 3 times daily) or intramuscular injection (50 mg, 3 times daily).

1. The specific precipitates prepared with A substance from 6 out of 8 high-titer anti-A antisera showed the presence of β _2A-globulin. This was demonstrated with 3 different antisera specific for this component, utilizing eluates prepared by excess A substance and also by acid extraction. 2. Antisera made against eluates of specific precipitates contained antibodies specific for normal β _2A-globulins. 3. All 3 classes of immunoglobulins were encountered in the specific precipitates of the majority of the sera. Antibodies corresponding in antigenic type to ordinary 7S γ-globulins, made up the bulk of the precipitates studied.

The isoelectric optimum of 6 types of human adenoviruses in undialyzed monkey kidney harvest fluids has been established as pH 3.15 ± 0.15. Live and formalin-inactivated viruses were concentrated efficiently by elution of isoelectric precipitates from Celite columns. These observations appear to have several practical applications, as in the preparation of more potent vaccines.

Production of hemagglutinin by dengue virus types 1, 2, 3 and 4 has been demonstrated in the fluid phase of HeLa cells infected either with unadapted mouse brain virus or with high passage level HeLa cell adapted virus.

Cell cultures inoculated with high passage levels of HeLa cell adapted virus of all 4 types showed the presence of hemagglutinin in infected fluids at the time of the first appearance of the cytopathic effect. High titers of hemagglutinin correlated with high TCD₅₀ titers.

Adaptation to in vitro conditions favored the production of hemagglutinin in cell cultures inoculated with dengue types 1, 3 and 4. Under the experimental conditions, type 2 unadapted mouse brain virus yielded higher concentrations of hemagglutinin than did type 2 high passage level HeLa cell adapted virus.

The effect of the engulfment of crystalline and non-crystalline silica dusts and anthracite on oxygen uptake, glycolysis and amino acid incorporation into protein of rat peritoneal macrophages was studied. A marked inhibition of these processes occurs in presence of silica dusts, tridymite being the most effective. The effects on oxygen consumption and glycolysis but not on amino acid incorporation almost disappear when results are correlated to the number of viable cells. Anthracite slightly stimulates macrophage metabolism in anaerobic conditions. It is suggested that protein synthesis is the primary site at which silica particles exert their toxic action.

Feeding of lithocholic acid to chicks and hens caused a rise in their serum cholesterol in work of Edwards(1) and Edwards et al. (2). When this compound was fed to mice or rats it caused a depression in serum cholesterol in the work of Beher et al.(3), Howe et al.(4) and Portman and Bruno (5). Recently Howe and Bosshardt(6) demonstrated that in the presence of a mixture of succinyl-sulfathiazole and oxytetracycline in the diet of mice, lithocholic acid no longer had a serum cholesterol-depressing effect while hyodeoxycholic acid was still effective. They interpreted these findings to mean that lithocholic acid was active by virtue of conversion to an unidentified compound under the influence of the intestinal bacteria. The investigation reported here was carried out to determine if the cholesterol elevating action of lithocholic acid in the chick might also be due to some other compound produced from lithocholic acid in the chicken's gastrointestinal tract.

Experimental. Four plastic canopy germ-free isolators (American Sterilizer Co.) were used for the experiment. The isolators were sterilized by spraying with 30% beta-propiolactone solution; each air filter (Fiber-glass 50) was sterilized with 1 ml methyl bromide; water bottles, cages and other equipment were autoclaved; feed was sterilized with ethylene oxide.

Eggs of Single Comb White Leghorns were obtained from a local hatchery after 18 days of incubation. The eggs were washed in detergent and passed through 2% HgCl₂ solution into the isolators. Twelve eggs were placed in each isolator. After hatching, 6 chicks were placed in each of 2 wire cages in each isolator. The basal ration was fed to the chicks in one cage and the lithocholic ration to the chicks in the other cage.

Cultures were taken daily for 3 days and each 3 days thereafter.

A transmissible agent with biological and physical properties similar to those of viruses in the myxovirus group was recovered from the pooled tissues of 2 white-footed mice (Peromyscus leucopus) trapped in Virginia in March, 1962. The recovered agent produced degenerative changes in a variety of primary and continuous cell cultures, induced fatal infections in suckling mice when inoculated intracerebrally and hamsters when inoculated intracerebrally or intraperitoneally. Five of 8 white-footed mouse serum pools contained hemadsorption inhibiting antibody directed against the new agent in titers ranging from 1:20 to 1:160 but sera obtained from laboratory mice and from wild animals other than Peromyscus did not. A number of human beings were found to have hemadsorption inhibiting antibody against the new virus in their sera. This observation is taken as evidence that these individuals had experienced infection with the new agent or one closely related to it.

Three immunologically unrelated enterovirus-like agents, antigenically unrelated to any of the recognized human prototypes were isolated from stool specimens of patients suffering from bacillary dysentery in Mexico City in 1957. The viruses were not pathogenic for suckling mice nor for cultured cells of human origin. Indirect evidence based on antibody studies would indicate that 2 of these viruses may belong to the human enterovirus group. The relationship of these agents to the clinical disease observed remains undetermined.

(1) The effect of supplementation of diet with high levels of KCl and NaCl on Cs¹³⁷ retention in rats was studied; the level of added alkali metal ion in each case was 5%. (2) Both Na⁺ and K⁺ increased the rate of excretion of Cs¹³⁷ but also considerably enhanced water consumption. (3) The response was probably due to the diuretic action of the salts rather than to specific ion antagonism. (4) The exposure dose to the animal from Cs¹³⁷ was reduced by a factor of about 0.7 by high salt (Na⁺ or K⁺) intake.

When 1 μg estradiol benzoate and 3 mg progesterone were administered to ovariectomized mice for 19 days, mammary gland development as measured by DNA was only 57.6% of that observed in mice on day 18 of pregnancy. Addition of thyroxine at level of 3 μg/100 g BW increased the DNA by 12.5%. Hydrocortisone acetate at a level of 100 μg/day increased DNA by 7% while the combination of the two hormones increased DNA by 17%. Lactogenic and growth hormones were ineffective as synergists. The highest level of DNA was obtained in ovari- and adrenalectomized animals given 100 μg Prednisone and varied EB and P, an increase to 11.67 mg/100 g BW or 19.9%.

The activities of 2 mitochondrial respiratory enzymes were determined in the liver of normal rats and of rats treated with cortisone and/or anabolic steroid Methandrostenolone. The activities of malic dehydrogenase and DPN-cytochrome-c-reductase were decreased in cortisone treated rats. This depression was reversible and interruption of treatment resulted in normalization of activities of both enzymes. Simultaneous treatment of rats by cortisone and the anabolizer did not, however, prevent this depression. The activities of both enzymes were increased in rats treated with Methandrostenolone alone.

Chick embryo fibroblasts infected in vitro with RSV exhibit striking alterations in their metabolism which include a decrease in oxygen consumption, increase in glucose utilization and lactic acid production, and an increase in release of lactic dehydrogenase into the culture fluids under aerobic conditions. These changes are associated with the increased aerobic glycolytic activity of these cells which is characteristic of Rous sarcoma and other neoplastic cells.

In all cases, an increase in heart rate resulted from intravenous infusion of fluid into dogs with chronic cardiac denervation, the increments ranging from 4 to 31 beats per minute. In those cases in which it was measured, right atrial pressure increased; in individual experiments, the increase in pressure was proportional to the volume infused. In the experiments with dextran, the maximal increase in right atrial pressure preceded the maximal increase in heart rate by intervals ranging from 4 to 54 minutes. Part of this wide variation was explained by the fact that the peak of the increase in rate often was approached so slowly that it was difficult to locate it precisely in relation to the time axis. In the experiments with saline, there was a clearly defined lag of 2 1/2 minutes in the example shown in Fig. 1, but in the others the small degree of cardiac acceleration made it impossible to decide if a lag was present. Similarly, a lag between the maximal increase in venous pressure and the maximal increase in heart rate has been reported by Coleridge and Linden(5) in a study of the Bainbridge reflex in intact anesthetized dogs.

When exercised, dogs with cardiac denervation achieve a maximal heart rate in 90 to 100 seconds. An increase in the transmural atrial pressure, due to an increase in depth of respiration and in return of blood to the heart, might be expected to develop early in exercise and to be sustained throughout the period of work. In the experiments shown in Fig. 1 and 2, the maximal transmural atrial pressures were achieved in 15 seconds and 15 minutes, respectively, but the corresponding maxima in heart rates did not occur until 2 1/2 and 45 minutes later, at which times the transmural pressures had declined considerably from their maximal values.

Corticosterone in plasma and urine was determined by spectrofluorometry in sulfuric acid and by direct spectrophotometry in ethanol. Plasma levels and urinary excretion of corticosterone were elevated in rats with subcutaneous lymphomas. The high plasma concentration of corticosterone in these rats may help to explain the reduction in splenic, thymic and lymphoid tissue of lymphoma-bearing rats and certain interactions of exogenous steroids, quantity of lymphoma tissue and tumor-necrotizing effect of K. pneumoniae polysaccharide.

Acetyl-β-methylcholine is a racemic compound. Acetyl-D-β-methylcholine is hydrolyzed by acetylcholinesterase; the Lisomer is not. Acetyl-L-β-methylcholine is an inhibitor of the hydrolysis of the Disomer and of acetylcholine by acetylcholinesterase. Therefore the generally available acetyl-β-methylcholine is composed of half substrate and half inhibitor. Two other structurally related compounds are also cholinesterase inhibitors when tested under conditions comparable to those obtaining in the use of acetyl-DL-β-methylcholine.

The average total amount of acid phosphatase produced by 46 strains of S. aureus in a sub-group of phage group I lysed by phage 80, was found to be 25% greater than the average of 19.1 units for 30 strains in the sub-group not lysed by phage 80. In turn, both of these phage group I sub-groups produced more enzyme than the 69 phage group III strains, for which the mean was 12.0 units. There were no significant differences in the amount of acid phosphatase produced by strains in phage group I without an 80 in their pattern or in phage group III when the strains within each of these 2 groups were segregated according to either penicillin-resistance or mercuric-ion resistance. The percentage of strains resistant to mercuric ions was much greater among penicillin-resistant strains than in penicillin-sensitive strains.

The pteridine compound triamterene was studied in 11 dogs using the stop flow method. These data support the hypothesis that the site of action of triamterene is the distal tubule of the kidney and that triamterene produces its natriuretic effect by reducing Na reabsorption in this area. No evidence was obtained for a proximal renal tubular effect.

Ten synthetic peptides, structurally related to bradykinin or to kallidin, were compared with bradykinin for (a) stimulant activity on the excised rat's uterus, (b) i.v. vasodepressor activity in the anesthetized rat, (c) increased capillary permeability effects in rabbit's skin, and (d) coronary dilator activity in the excised cat's heart.

The decreasing order of activity of these peptides was as follows: (1) bradykinin and 6-glycine bradykinin, (2) 7-glycine kallidin, (3) 7-glycine bradykinin, (4) 9-citrulline bradykinin; 1-citrulline bradykinin; bisnitrobradykinin, (5) arginylheptaglycylarginine; 1,9-biscitrulline bradykinin; fully protected bradykinin; and retrobradykinin. None of the bradykinin analogues manifested anti-bradykinin activity on the excised rat's uterus or in rabbit's skin.

The mammary glands of intact virgins and ovariectomized mice after 33 days had DNA levels essentially equal. Only after 150 days of involution was a significant decrease in DNA observed. Following the weaning of normal 20 day lactating mice, the DNA level was reduced from 21.35 mg/100 g to 9.95 mg after 5 days, a decrease of 53.4%. At 20 days of involution, DNA had decreased to 6.07 mg/100 g, a total decrease of 71.6%. This level of DNA is comparable to the virgin condition. Hydrocortisone acetate injected daily at 100 μg/day for 5 days or 250 μg/day for 10 days was effective in retarding involution.

Daily feed consumption was determined in 34 lactating rats of the Sprague-Dawley-Rolfsmeyer strain. Feed consumption increased at a decreasing rate until it was 2.5 to 3 times normal at day 20 of lactation. Standard deviations, calculated separately for each day, increased as the means increased. Total feed consumption varied from 9.6 ± 3.7 g (Mean and S.D.) on day 1 to 38.7 ± 8.1 g on day 20 of lactation. Feed consumption per 100 g body weight was 3.5 ± 1.4 g, on day 1 and 14.1 ± 3.1 g on day 20, then returned to normal by 9 days after weaning the pups.

The experiments discussed here indicate that a potential difference, requiring the expenditure of metabolic energy, is maintained across the chorioallantoic membrane, the inner surface being negative. Its value is somewhat smaller than the potential observed across most other biologically active membranes (1), and it is unlikely that these small electric forces would act directly as organizing agents in a tissue graft on the membrane. Nevertheless the potential difference may be an expression of an underlying metabolic asymmetry, maintained for example by a gradient of CO₂ or O₂ across the membrane, which could be related more relevantly to the observed polar reconstitution of the organs in the experiments of Weiss and Taylor.

Adenovirus stocks prepared by serial passage of the virus in rhesus monkey kidney cells were found to be contaminated with the simian papovavirus SV40. The infectivity titer and number of particles of SV40 in each of 4 virus stocks examined were higher than those of adenovirus. The virus stocks had an average adenovirus titer of 10^3.6 TCD₅₀/ml and an adenovirus particle count of about 10^7.6 particles/ml. The same stocks had an average SV40 titer of 10^6.0 TCD₅₀/ml and an SV40 particle count of about 10^9.0 particles/ml. Antigenicity of the adenovirus and SV40 components was tested by inoculation of rabbits and guinea pigs, using the untreated stock, and stocks heated in water and in MgCl₂. When the virus stock was heated at 50°C for 30 minutes in water, the adenovirus component lost all of its infectivity and a large part of its antigenicity, while the SV40 component retained both infectivity and antigenicity. When the virus stock was heated in M MgCl₂ at 50°C for 30 minutes, the adenovirus component again lost all of its infectivity but it retained all of its antigenicity. However the SV40 component was reduced both in infectivity and in antigenicity.

Methods have been described for quantitative plaque assay of herpes simplex virus either in bottle culture or in Leighton tubes. The overlay employed was devoid of agar, peptones and serum, which are viewed as undesirable constituents. Use of agar overlay results in production of plaques from only a small fraction of the potential plaque-forming population. The inhibition of plaque formation by agar can be reversed by addition of protamine sulfate at a level of approximately 800 μg/ml. Possible uncertainties associated with the use of mixtures of agar and protamine can be avoided by substitution of methyl cellulose with which a further 10-fold increase in sensitivity of the assay was obtained.

1. The biological half-life for in vivo screened I¹³¹ labeled rat fibrinogen injected intravenously in normal rats was found to be 1.5 days. After 6–7 days the apparent biological half-life changed to 2.7 days. 2. The apparent whole-body turnover of I¹³¹ fibrinogen after whole-body X-irradiation was the same for both irradiated and control rats. However, the data on plasma I¹³¹ fibrinogen indicate that fibrinogen leaves the vascular space at an enhanced rate after whole-body irradiation.

1. Serum cholesterol and phospholipid determinations were made in 4 groups of subjects: 79 normal men without evident disease (“well controls,”WC), 80 men suffering from a variety of chronic disorders but without evidence of coronary artery or cerebrovascular disease (“sick controls,”SC), 223 men who had recovered from a frank myocardial infarction (MI), and 150 men who had recovered from a frank cerebral thrombosis (CT). In each group the cholesterol-phospholipid relationship appeared linear on a log-log scale and linear log-log regressions were calculated for each of the 4 groups. 2. In the 2 control groups, WC and SC, showing no evidence of coronary or cerebral atherosclerosis, the quantitative relationship between cholesterol and phospholipids was the same, confirming reports of other workers. Also, age did not affect this relationship. 3. In the 2 atherosclerosis groups, a substantial and highly significant abnormality in the quantitative relationship between cholesterol and phospholipids was observed (both groups showing the same abnormality). 4. There is, therefore, a disturbance in the normal balance between these 2 kinds of serum lipid in men with atherosclerosis. This imbalance was present over the entire range of observed cholesterol levels and is thought to represent some important and consistent lipid abnormality characteristic of atherosclerosis.

1. A total of 223 men recovered from myocardial infarction were allocated to treatment groups and serum cholesterol and phospholipids determined (I) before and (II) following 6 months of uninterrupted treatment with No estrogen, Premarin, Lynoral or Anvene. 2. The abnormality in interlipid relationships characteristic of men with clinical atherosclerosis was unaltered in the subjects receiving Premarin, but this abnormality was abolished by treatment with Lynoral. 3. There is no necessary correlation between the feminizing, lipid-altering and outcome-altering effects of estrogen therapy in men with myocardial infarction.

Lipid analysis of chick aorta intima and HeLa cell tissue cultures confirms histological studies indicating that total lipids and cholesterol are significantly reduced per unit volume of cells when very small amounts of calf aorta extracts or the mucopolysaccharide, chondroitin sulfate A are added to the cultures. The lipid clearing effect noted in cells is often associated with stimulation of cellular growth and with a process of lipid dilution by cellular multiplication. However, in the case of one aortic extract, lipid clearing took place without significant stimulation of cellular growth. The latter suggests a specific cellular clearing effect as distinguished from lipid dilution by cell division and growth.

Two studies were conducted in an attempt to learn more concerning the effect of bilateral nephrectomy and previous exposure to fluoride upon rate of fluoride absorption in the rat. The first study was conducted in weanling animals having no previous exposure to fluoride and indicated that bilateral nephrectomy significantly decreased the rate of fluoride absorption by 41.9% during the first 2 hours after fluoride administration and by 41.7% 4 hours thereafter. In the second study, this effect was substantiated using animals exposed to 2 different levels of fluoride. Previous exposure to 125 mg of fluoride decreased the rate of fluoride absorption by 21.9% and when the animals were in addition nephrectomized, a further decrease of 13.7% was noted. After exposure to 250 mg of fluoride, the rate of fluoride absorption was decreased by a similar value of 21.9%. Nephrectomy further decreased the rate of fluoride absorption and the combination of previous exposure and nephrectomy decreased the rate of fluoride absorption by more than 65%. These data suggest that when kidney function becomes impaired the body responds by decreasing the rate of fluoride absorption.

A technic is described for labeling rat lymphocytes, collected from intestinal lymph, with radiochromium without impairing their motility. After intravenous administration, these labeled cells disappeared rapidly from the circulating blood, reappeared to a limited extent within 6 hours, then continued to disappear at a very slow rate (about 1.1% per hour). When the rat's intestinal lymphatic trunk was cannulated, 4 to 7% of the labeled lymphocytes appeared in the lymph within 4 days, with maximal lymphatic radioactivity occurring between the 24th and 36th hours.

Change in the pH of the incubation medium has been shown to have opposite effects upon influx rates of cations and of anions into mitochondrial substance. Alkalinization was associated with increase in influx of Na⁺ and K⁺; and conversely, acidification resulted in more rapid influx of Br^- and Cl^-. Increased sensitivity to pH change was observed in the presence of the buffer ion imidazole. At pH 7 imidazole carries a positive charge and inhibition to influx of Na⁺ was observed. The inhibition was absent at pH 8.5 with conversion to the uncharged free base.

Approximately one thousand Sprague-Dawley white rats were subjected to the stress of a celiotomy at various intervals (0 to 30 days) before inoculation with Walker 256 tumor cells. The low resistance noted when cells were injected at the time of stress (as measured by a take of 90%) increased gradually from day to day reaching the control value (60%) by the eighth day, but continued to show an increased resistance for several more days, with a take of only 41% on the 18th day. The resistance decreased rather sharply to a take of 79% on the 20th day, coming back to the control level of 60% within the next few days.

1. Chromatographic and radio-autographic evidence is presented showing that progressive degradative changes occur in L-ascorbic acid dissolved in water and kept at 25°C for a 72-hour period. 2. When a human subject received 20 μc of freshly dissolved L-ascorbic-1-C¹⁴ acid solution, little or no C¹⁴ appears in his respiratory CO₂. 3. Men who were given similar samples of L-ascorbic-1-C¹⁴ acid aged for 36 and 72 hours, respectively, excreted 30.6% of the ingested C¹⁴ as respiratory CO₂.

A series of injections of papain protease, in doses of 0.2 or 0.3 mg, was administered into the proximal tibial metaphysis of 89 immature rabbits and the effect on the epiphyseal cartilage plates and on longitudinal growth observed for periods up to 4 months. Twenty-six of 63 animals followed 1 to 4 months had a relative elongation of the injected tibia of 3 to 6 mm. Significant retardation and moderate damage to the epiphyseal cartilage plates of the injected tibia occurred in 3 animals on the higher dosage schedule. Histological studies of the tibial epiphyses a few days after one or two injections of papain disclosed transverse splitting through portions of the hypertrophic zone and a marked increase in thickness of the cartilage plates.

1. A simplified method for isolation of kidney and liver cell membranes is reported. 2. Aldosterone is weakly bound to kidney subcellular fractions as demonstrated by the techniques of equilibrium fractionation and equilibrium dialysis. 3. The liver and kidney cell membranes studied were found to either take up the steroids according to their lipid solubility or to bind them in accordance with the polarity rule for steroid-protein interaction. 4. Spirolactone failed to displace aldosterone from the kidney cell membranes.

These studies indicate that a virus particle, bacteriophage φX 174, incapable of infecting mammalian cells, can cross the placenta and gain access to the foetal circulation. The possibility of in vitro contamination of foetal blood samples during cardiac puncture of the foetus can be excluded since: a) the washing procedure of the foetus was shown to be effective after deliberate contamination of the foetus's skin; b) after injection of phage into pregnant guinea pigs, separate bleedings of 0.5 ml and 0.1 ml from the foetal heart usually showed a 5-fold difference in amount of φX in the 2 samples; this difference was not observed in deliberately contaminated foetuses from pregnant guinea pigs not injected with φX. It is unlikely that endotoxin contamination of phage preparations was responsible for the transplacental passage of virus because of virtual absence of biologically detectable endotoxin in the preparations of phage employed and the failure of an injection of endotoxin with phage to increase the amount of phage found in the foetal circulation. There was no apparent difference in phage transfer between foetuses of markedly different weights, 56–120 g.

The route of transplacental passage of phage is as yet uncertain. The failure to find a gradient in concentration from amniotic fluid to foetal blood (the latter sometimes had a higher phage concentration) suggests that phage is not transferred by this route. Previous studies of placental transfer of large proteins indicate that the splanchnopleure is the principal transfer organ in guinea pigs(7).

The question arises as to whether the placenta is an effective barrier in preventing intrauterine viral infections. The following findings are relevant to this question: a) in the guinea pig, the difference between maternal and foetal blood virus concentrations was approximately 105⁵–10⁶ and transfer usually was not demonstrated when maternal φX concentration was below 10⁷/ml; b) the φX particle is an unusually small virus (approximately 250 angstroms in diameter).

Cellulose acetate discs or agar are useful supporting media for titration or identification of antigen or antibody. The cellulose acetate fluorescent spot (CAFS) method is simple to perform and read and requires only a few lambda of antiserum or antigen; the fluorescent impregnated agar (FIA) method is essentially the same as the fluorescent antibody technique applied to tissue. In the titration of ABSA, AHGG and AHS both fluorescent methods were more sensitive than the ring precipitin test, when the indirect staining was used. In addition the methods permitted selection of proper fixatives for a given antigen and determination of the proper concentration of labelled antisera for demonstration of tissue antigen.

Ten daily doses of boiled normal human urine equivalent to 0.5, 1.0, 2.0, 4.0, 6.0, and 12.0 ml per 100 g body weight were administered by gastric tube to normal rats. Doses equal to 1% of the rats“weights evoked thrombocytosis. Larger doses induced progressively greater responses. The throm-bocytosis-promoting activity of normal urine was comparable to that in similar amounts of normal plasma. Urine from a polycythemic subject appeared to contain increased activity. These studies are in accord with the thesis that thrombopoiesis is subject to humoral regulatory control.

The oral effectiveness of 4 progesterone-like compounds to stimulate mammary gland growth (DNA) of ovariectomized rats is reported. Norlutin and Norlutate plus E.B. were ineffective in stimulating increasing DNA in comparison with E.B. alone. Lutoral and Provera plus E.B. at the 1 mg level or above stimulated comparable mammary gland growth to that observed at end of pregnancy or after 1 μg E.B. + 3 mg P. The injection of 1 or 2 μg E.B. plus 3 or 6 mg Norlutin Enanthate was also effective in stimulating DNA values comparable to normal pregnancy. The losses in body weight of most groups during the 19-day feeding period were apparently due to the unpalatable nature of feed containing these compounds.

Clone NCTC 1469 tissue culture line of C3H mouse liver, maintained in chicken serum-containing medium, was found to provide sensitive assay systems, based on production of plaques and cytopathic changes, for laboratory adapted and recently isolated strains of mouse hepatitis viruses, as well as for some naturally occurring strains in mouse feces.

The incorporation of C¹⁴-labeled hexoses into glycogen compartments of avian liver and glycogen body was studied in young chicks which had been fasted 20 hours and tube-fed a test gruel at 1/2 and at 1–1 1/2 hours prior to isotopic administration. The results indicate that in contrast to rapid liver incorporation of the hexose label, the avian glycogen body was not active as far as glycogenesis was concerned for at least 24 hours after isotope injection. Only a trace of the galactose label was found in the glycogen body 8 hours later, no activity from glucose or fructose-C¹⁴ being found at any time in this structure throughout the 24-hour post-injection observation period.

Difluoromalonic acid and its di-amide inhibit the oxidation of a number of organic acids by a strain of Pseudomonas aeruginosa although no inhibition could be demonstrated in sonicates of the cells. The di-amides of monofluoromalonic, tetrafluoro-succinic and hexafluoroglutaric acids are essentially inactive.

Several chemicals were tested and found to be in vitro inhibitors of chicken serum alkaline phosphatase. Sodium borate inhibited serum phosphatase more in a line bred for high serum level than in a random bred control line. Subsequent studies with this inhibitor on tissues from the two lines showed that intestinal phosphatase activity was depressed significantly more than that in bone, liver, and kidney. These studies indicate that the major portion, if not all, of the increased serum phosphatase of the high line originates in the intestine.

Leghorn cockerels were fed for 20 months on diets containing either no supplement, whole egg powder, egg oil equivalent to that contained by egg powder or these diets in combination with corn sterols, the latter added in amounts equivalent to that supplied by 5% corn oil. An additional group was given egg powder for 12 weeks and then placed on the control ration for the remainder of the experimental period. The inclusion of corn oil sterol retarded the atherogenesis that was accentuated by the addition of egg powder or egg oil. Total fat deposition in the abdominal section of the aorta was reduced in the groups fed the sterols. The brief exposure to egg powder did not leave lasting effects on those animals that were changed over to the control diet. Egg oil appeared to be similar in its atherogenicity to egg powder. Sterol excretion patterns suggested that the beneficial effect of the corn sterols was not due to interference with cholesterol absorption alone.

Oxygen uptake by planktonic cyprids of Balanus amphitrite niveus was elevated by addition of 0.5, 1, and 5 mg of copper ion per liter of sea water. Concentrations from 10 to 500 mg/1 progressively depress oxygen absorption. Analysis of individual patterns relates increased oxygen consumption to increased motor activity and energy cost of active transport.

Two successive experiments, each lasting 8 weeks, were conducted with a total of 100 weanling male rats. Animals actively immunized with crystalline jackbean urease and given either 2.2 or 22.0 I.U of Vit. A daily following a 4-week depletion period grew faster and were more efficient in feed utilization. Both immunizing with urease and increasing the dietary Vit. A level reduced ammonia production and ureolytic activity in the gastrointestinal tract. The evidence indicates that depression of ammonia production and urease activity in the gastrointestinal tract is beneficial to animals fed limited levels of Vit. A.

Various levels of EDTA and zinc were added to the poult diet containing isolated soybean protein. One hundred mg of EDTA/kg were equivalent to approximately 8 mg/kg zinc in supplementing the basal diet. When 7 or 10 mg of zinc/kg were added to the diet, 100 mg of EDTA/kg were equivalent to approximately 19 mg of zinc/kg. The data indicate that EDTA improves the effectiveness of added zinc as well as increasing the utilization of zinc already present in the basal diet. Perosis increased as the zinc level of the diet increased to approximately 30 mg/kg and decreased as zinc was added above this level.

When 95 strains of Ps. aeruginosa were assayed for lysogenicity on a series of indicator strains, all bacteria were found to be lysogenic. From the patterns of lysogenicity obtained with respect to the indicators we conclude that the majority of the 95 strains tested were lysogenic for more than one bacteriophage. When the patterns of lysogenicity obtained in this study were compared with patterns of phage sensitivity with respect to 12 bacteriophages, it appeared that the lysogenicity patterns showed more utility with respect to grouping strains of this organism. Approximately 30% of the strains tested were found to be untypable with respect to the bacteriophages used, whereas all strains could be typed on the basis of lysogenicity patterns. Finally, the resistance of a given strain of Ps. aeruginosa to a series of bacteriophages appeared to be related to the degree of lysogenicity exhibited on a series of indicator bacteria.

Inactivated influenza viruses administered to mice intranasally or by aerosol induced transient heterologous immunity to infective influenza viruses. This immunity was reflected by reduction of pulmonary viral titers and by a reduced occurrence of gross lesions. This effect is considered to be an example of viral interference in that it required time to develop and persisted for at least 3 days; and because protective activity of inactivated viruses in mice could be correlated with interfering activity in the chick embryo. Interference modified but did not completely inhibit infection, and strain specific immunity of lessened degree developed to the challenge virus following the modified infection.

Ethionine administration resulted in rapid and essentially complete inhibition of liver urocanase activity in the rat, while the activities of histidase and histidinepyruvate transaminase were relatively unaffected. Rats receiving ethionine in their diet, after showing a severe weight loss during the first week, resumed a rate of growth parallel to that of the control animals; there was, however, no restoration of urocanase activity during the period of adaptation to dietary ethionine. Thus, the straight deamination pathway for the degradation of histidine is not essential for the growth of the rat.

Administration of nethalide, an adrenotropic beta receptor antagonist, to dogs anesthetized with cyclopropane, trichloroethylene, or halothane was found to decrease markedly the sensitivity of the myocardium to epinephrine induced arrhythmias. From 4–15 times the minimum dose of epinephrine causing multiple ventricular ectopic beats during the control period was required to produce this arrhythmia 15 minutes after intravenous nethalide. Ventricular fibrillation was not observed with even large doses of epinephrine after nethalide but did occur during the control period in 2 of 3 dogs receiving halothane. Some possible problems that might be encountered with the use of this agent as an adjunct to anesthesia are discussed.

Weekly implantation of BroadBreasted Bronze turkeys with 24 mg of DES increased the incidence of aortic ruptures. Poults fed a 16% protein diet developed more aortic ruptures (58%) than turkeys fed a 28% protein diet (29%).

Serum from the agouti and paca, rodents closely related to the guinea pig, exerted an inhibitory effect in vivo on the 6C3HED mouse lymphoma equal to that of guinea pig serum. Agouti, paca, and guinea pig sera have a measurable level of L-asparaginase activity; whereas human and nutria sera, which are inactive against the 6C3HED lymphoma, do not. The results support the hypothesis that this enzyme is the active principle responsible for inhibition of this tumor.

When a rabbit antiserum to human macroglobulin was reacted with human sera by immunoelectrophoresis, several distinct macroglobulin precipitin lines were revealed. Internal to the major beta-2M line all sera showed a second, and some a third, line. A fourth macroglobulin line appeared opposite the serum source in most sera examined. The major macroglobulin line was forked at its anodal end in some sera. The major macroglobulin line had a single curvature when produced by the rabbit antiserum used in most of these experiments, but had a double curvature when produced by a horse antiserum. These findings could be due to multiple antigenic sites on macroglobulins of a single molecular species, or to antigenically distinct macroglobulin molecules.

In rats, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (p-CF₃O-CCP) was more potent than 2,4-dinitrophenol (DNP) in causing marked oxidative and expiratory effects but was without apparent effect in mice at an equivalent dose. Toxicity of p-CF₃O-CCP in mice prevented further observations since increasing the dose proved lethal. DNP was active in both mice and rats.

Non-particulate cellular cytoplasmic constituents from human liver were isolated by physicochemical procedures. These substances were reactive as antigens in complement fixation tests against sera from patients with lupus erythematosus and other diseases. Results indicated that the soluble fraction constituents were not extracted from the cell nuclei. Fractionation of the soluble fraction by isoelectric precipitation at low ionic strength produced 3 active precipitates each of which reacted differently in complement fixation tests with sera from patients with lupus erythematosus. Enzymatic studies suggested that the substance active as antigens were RNA-protein and protein.

A peptide material has been extracted from rat urine which decreases blood pressure of normal rats, raises blood pressure of rats with sympathetic system blocked, and causes contraction of uterine muscle. The activity of the extract is greatly increased in rats treated several weeks with DCA, but there is no clear relationship between the increase in the active substance and the onset of the hypertension. In renal hypertension pressor activity of the urine extract was not increased, even after hypertension had developed. It is highly probable that 2 or more peptides are involved. Anephrotensin, bradykinin and or kallidin may be the active agents, or the activity may be due to some as yet undescribed substance or substances. The possibility that angiotensin or vasopressin is the active component has been ruled out.

The rapid removal of 10% and 25% blood volume in anesthetized dogs (sodium pentobarbital) produced a marked rise in peripheral 17-OHCS when reported as γ/100 ml or γ/plasma volume, whereas the expansion of vascular volumes by 10% and 25% with dextran caused only slight increases in steroid levels. In the 10% hemorrhagic group, 17-OHCS increased without a significant decrease in mean arterial pressure. In animals which were hemorrhaged 5% of their blood volume every 10 minutes until 35% had been removed, 17-OHCS progressively increased and became significant only after 25% hemorrhage. The alterations in peripheral plasma 17-OHCS which occur during the expansion or reduction of vascular volumes are better described when these values are corrected for circulating plasma volume.

According to Lewis(1), the nonapeptide bradykinin is an important mediator of the acute inflammatory reaction. It is, therefore, of interest that orally given proteases were recently shown to antagonize 3 pharmacologic actions of bradykinin: vasodilatation, vascular permeability and smooth muscle contraction(2). The objects of the present investigation were: first, to assess the action of orally given streptokinase against the fourth pharmacologic action of bradykinin, namely, pain production; and second, to appraise the anti-inflammatory action of orally given streptokinase in a controlled animal and clinical experiment.

Methods. 1. Analgesia study. The cantharidin blister technic of Armstrong et al. (3) was slightly modified. This study comprised 30 apparently healthy students and technicians. A band-aid containing 2 drops of tincture cantharidin was applied to the skin of the flexor surface of the forearm the evening before the day of the experiment. When the band-aid was removed the next morning, the area showed a blister with a diameter of about 1.5 to 2 cm. Blisters remained relatively unchanged for 24–48 hours. Intact blisters were opened and the separated epidermis cut away exposing the blister base, which was bathed in Ringer's solution warmed to 37°C. Before the application of bradykinin, the bathing solution was removed and the area dried with filter paper. 0.2 ml bradykinin (0.1 μg) was applied to the dry blister base and the following information recorded:

1. Length of time for pain to develop.

2. Duration of pain.

3. Intensity of pain. This factor was evaluated by each subject on a 1 to 4 + basis.

Ten control subjects received placebo tablets; 10 treated subjects received tablets containing streptokinase 10,000 units; 10 treated subjects received aspirin, 5 grains. Control and experimental groups received one tablet every 15 minutes for one hour following pretreatment control determinations.

Monkeys pretreated with stabilized colloidal thorium dioxide (Thorotrast) are more sensitive to the emetic action of intragastrically administered staphylococcal enterotoxin. The amounts required to elicit vomiting in 50% of the animals tested was reduced to about one-twentieth of that required for untreated monkeys when Thorotrast was given 18 hours before the enterotoxin challenge. The degree of sensitization was the same for both of the 2 different antigenic types of enterotoxin tested. Fresh blood was observed in the vomitus when the interval between Thorotrast and enterotoxin was reduced to 4 hours.

Three cases of clinical renal transplantation have been performed, using donor organs of different blood type than that in the recipient patient. Graft function occurred in all 3 cases, being most retarded in the kidney subjected to the longest ischemia. The kidneys provided by living donors have essentially normal function 9 and 5 weeks after transplantation.

Sera from farmer's lung patients and non-farmer's lung control individuals were reacted with trichloroacetic acid extracts of moldy hays. All the sera from patients with acute symptoms and two-thirds of the total which included recovered farmer's lung patients formed specific precipitin lines with the moldy hay extracts. No precipitin lines were obtained with sera from healthy farmers who had no history of farmer's lung or with extracts of good hay. Evidence is presented to show that farmer's lung is associated with the presence of specific antibodies in the patients“sera against antigens found in moldy hays and that these antigens are the cause of farmer's lung.

When administered locally via the conjunctival sac, 5-iodo-2′-deoxyuridine delays the onset of symptoms of experimentally produced pseudorabies keratitis in rabbits. The incubation period was approximately 20 hours longer in treated animals than in controls.

When laying hens were fed a diet containing 0.25% histamine, egg production stopped. When this diet was preceded by a diet containing 0.1% histamine, egg production was not affected. The adaptation of hens to histamine in their diets is discussed. The gross pathology of hens fed a diet containing 0.25% histamine is described.

Assay of lysozyme activity in homogenates of kidney of irradiation chimeras has shown that a marked increase in activity is associated with the presence of grafted homologous bone marrow. This increase began about 3 days after injection of the marrow cells, reached its peak 20–40 days and returned to normal 100–200 days after treatment. A significant increase in weight of kidney, maximal at 9 days after treatment, was observed in the animals given homologous bone-marrow cells.

Prolactin activity appeared in the blood of normal young women during the first 5 and last 10 days of a 28-day menstrual cycle, whereas no activity was demonstrable during days 6 to 18 of the cycle. Blood prolactin activity appeared to be present in the latter part of the cycle of a normal woman receiving the gonadotropin inhibitor, Enovid. No blood prolactin activity was found in a normal male subject during a 31-day period. These observations suggest that prolactin acts in opposition to the other 2 pituitary gonadotropins, FSH and LH, in the normal human menstrual cycle.

Ovulation was delayed in 20 of 63 hens following stimulation of a ventromedially limited region of the preoptic brain. The effect was produced following electrical stimulation with stainless steel or platinum electrodes, or by placement of electrodes without passage of current. In accord with our previous findings in the anterior median eminence, the delays in ovulation are believed to originate in the delay in or blockade of the release of ovulation-inducing hormone.

Feeding a diet containing 0.3% of thiouracil (TU) by dry weight for 18 months to a subline of C3H mice produced a nearly 100% incidence of hepatoma in both sexes. Controls given no TU had definite but significantly lower incidence of this tumor. When the inbred strain of mice TM was used, no hepatomas were observed in experimental nor in control animals. Apparently genetic susceptibility must be present for development of hepatomas and for their increased incidence in mice treated with TU.

Pertussis-sensitized mice were tested for their reactivity to endotoxin as expressed by lethality and induction of hemorrhagic necrosis in implanted tumors. It was found that, whereas pertussis treatment increased the susceptibility of animals to the lethal effect of endotoxin, transplanted tumors became more resistant to its necrotizing effects. Induction of tumor necrosis by an anaphylactic reaction was not affected by pertussis treatment. Tumor-bearing mice injected with pertussis vaccine did not become more susceptible to the lethal effects of endotoxin than did animals not bearing tumors.

1. Administration of 2-amino-4-arsenosophenol hydrochloride to Eperythrozoon-infected mice bearing transplantable tumors resulted in a lowering of plasma LDH early in the course of tumor development. 2. Splenectomy of Eperythrozoon-infected mice resulted in a marked rise in plasma LDH. 3. One week or more after tumor implantation, plasma LDH was elevated in all mice, with or without the presence of demonstrable Eperythrozoon in the blood. 4. It is suggested that Eperythrozoon infection contributes to the early rise in plasma LDH in mice inoculated with transplantable tumors.