The biology of affective disorders.

The biology of affective disorders. The clinical features, genetic data, psychopharmacological studies, hormonal abnormalities, and biochemical observations all serve to define major depressive illness as an inherited neurochemical disorder affecting the hypothalamus, and probably involving monoamine pathways. While the precise nature of the defect and its mode of transmission remain obscure, the rapid development of this field in less than two decades permits optimism that major depressive illness illness will be among the first "functional" psychiatric disorders to have its chemical pathology elucidated.


Modified procedure for the recovery of naturally accumulated poliovirus from oysters. Methods were compared for their ability to recover poliovirus from oysters (Crassostrea gigas) which had been allowed to accumulate virus via normal filtration activities. Clarification procedures included glycine-NaCl and polyelectrolyte extraction methods followed by a variety of acid precipitation concentration methods. Polyelectrolyte flocculation followed by a beef extract-supplemented acid precipitation carried out at pH 3.5 yielded the most efficient recoveries. Direct assay of homogenates was found to be an unreliable method for determining the initial virus concentration in "naturally infected" oysters.


Viral heat resistance and infectious ribonucleic acid. High-titer suspensions of poliovirus 1 and coxsackievirus B-2 were shown to contain a heat-resistant fraction when heated for 65 min at temperature ranging from 56 to 70 degrees C. The addition of ribonuclease to the heated suspensions eliminated plaque production in the cell cultures, indicating that the resistant fraction was infectious ribonucleic acid that had been liberated from ruptured viruses during the heating process.


Adsorption of enteroviruses to soil cores and their subsequent elution by artificial rainwater. The adsorption and elution of a variety of human enteroviruses in a highly permeable, sandy soil was studied by using cores (43 by 125 mm) collected from an operating recharge basin on Long Island. Viruses studied included field and reference strains of polioviruses types 1 and 3 and reference strains of coxsackie virus B3 and echovirus types 1 and 6. Viruses suspended in treated sewage effluent were allowed to percolate through soil cores, and the filtrate was assayed for unadsorbed viruses. To determine the likelihood of desorption and mobilization, soil-bound viruses were subjected to a rinse with either treated sewage effluent or simulated rainwater which reflected the anion, cation, and pH characteristics of a typical northeastern United States rainfall. The results demonstrated that all polioviruses tested, including both reference and field strains, adsorbed extremely well to cores. Adsorption was somewhat reduced when clean, unconditioned soils were used. Soil-bound poliovirus strain LSc was not significantly mobilized by flooding columns with either a sewage effluent or rainwater rinse. One virus was mobilized by both types of rinses. The amount of viruses mobilized by rainwater rinses ranged from 24 to 66%. Variable adsorption-elution results were observed with other enteroviruses. Two guanidine-resistant mutants of poliovirus LSc demonstrated a soil adsorption-elution profile different from that of the parent strain. The data support the conclusion that soil adsorption-elution behavior is strain dependent and that poliovirus, particularly strain LSc, represents an inappropriate model.


Procedure for the recovery of airborne human enteric viruses during spray irrigation of treated wastewater. Because of the relatively low number of indigenous enteric viruses recovered from secondary wastewater effluents, their presence in air (aerosols) as a result of wastewater spray irrigation requires extensive sampling. Methodology to allow the recovery of indigenous enteroviruses from aerosols generated at an operational wastewater irrigation site was tested under both laboratory and field conditions.


Structural changes associated with poliovirus inactivation in soil. The loss of infectivity of poliovirus in moist and dried soils was a result of irreversible damage to the virus particles. The damage included (i) dissociation of viral genomes and capsids and (ii) degradation of viral ribonucleic acid (RNA) in the soil environment. Under drying conditions, capsid components could not be recovered from the soils. Further studies in sterile soils indicated that, under moist conditions, the viral RNA was probably damaged before dissociation from the capsid. However, in sterile, dried soil, RNA genomes were recovered largely intact from the soil. These results suggest that polioviruses are inactivated by different mechanisms in moist and drying soils.


In vivo effect of the Syndets Idet 5L and Swanic 6L ON ATPase activity in the teleost, Channa punctatus. Channa punctatus were exposed to various sub-lethal concentrations of Idet 5L and Swanic 6L for 25 and 50 days. Brain and gill homogenates were analyzed for enzyme activity from treated and untreated fish. Enzyme inhibitions were highest in oligomycin-insensitive Mg++ ATPase, with pronounced effects (over 65% inhibition) after 50 days exposure of fish to 7.5 mg/L concentration of Swanic 6L. Fish exposed to lowest concentrations of syndets showed an insignificant activation of Na+-K+ and oligomycin-sensitive Mg++ ATPases in the gill.


NADH-dependent reduction of D-proline in Clostridium sticklandii. Reconstitution from three fractions containing NADH dehydrogenase, D-proline reductase, and a third protein factor. The enzyme system from Clostridium sticklandii catalyzing the NADH-dependent reduction of D-proline was co-purified by chromatography on DEAE-cellulose at pH 8.2 and ammonium sulfate fractionation, and resolved into fractions containing three different protein components, NADH dehydrogenase, D-proline reductase and a third protein factor, by chromatography on DEAE-cellulose at pH 7.0. Upon recombination of the fractions containing the three different protein components, the NADH-dependent reduction of D-proline was successfully reconstituted. The NADH dehydrogenase fractions oxidized NADH in the presence of artificial electron acceptors, and were inhibited by p-hydroxymercuriphenylsulfonate (50% at 80 nM). They contained 3--4 different enzyme bands as revealed by polyacrylamide-gel electropherograms stained with the NADH-dependent reduction of 2,3,5-triphenyltetrazolium chloride. D-Proline reduction was also coupled to a leuco-methylene blue-generating system containing D-glucose and glucose-oxidase (EC 1.1.3.4). Circumstantial evidence indicated that, among the clostridial proteins, only D-proline reductase and the third protein factor were needed for this reaction.


The effects of ultraviolet and ionizing radiation on herpesviruses, SV40 and adenoviruses in relation to the small-plaque effect. The small-plaque effect occurs with a wide range of herpesviruses following irradiation with ultraviolet light. The 37 per cent survival (D37) values, or dose required for one lethal hit (e-1), for herpes simplex, pseudorabies and pigeon herpesviruses in different cells indicate a broad spectrum of host-cell repair capacity. Other DNA-containing viruses such as SV40 and adenoviruses, which also replicate in the cell nucleus, show the small-plaque effect. Ionizing irradiation of herpes simplex virus type 1 (HSV-1) showed but little reduction in plaque-size.


Factors affecting the growth and titration by immunofluorescence of simian foamy virus. This paper presents some observations concerned with the growth of simian foamy virus and some modifications which should be introduced to the fluorescence assay of foamy virus. The modified procedure is the most sensitive method described for the titration of foamy virus. Examination of the optimal conditions for the growth and titration by fluorescence assay of simian foamy virus showed that the virus was particularly sensitive to changes in virus and cell concentration. At the low cell concentrations employed previously a "saturation-type" response was obtained with high titre virus and virus adsorption efficiency was decreased as input virus was diluted. Maximum virus production was obtained with high cell concentrations at input multiplicities of 5 and 10. At high multiplicities of infection more than 90 per cent of the cells adsorbed virus but only 45 per cent became infected, this appeared to be related to cell DNA synthesis.


Experimental infection of mink with influenza A viruses. Brief report. Mink were found to be susceptible to the intranasal inoculation of human, swine, equine and avian influenza A viruses. The viruses were recovered until the 7th post inoculation (p.i.) day from the respiratory tract. The inoculated mink showed antibody response against these viruses. Contact infection in mink with A/Kumamoto/22/77 (H3N2) was possible.


Avian pox virus. An ultrastructural study on a cherrug falcon. Brief report. The ultrastructure and maturation of avian pox particles is described in the rare case of a naturally infected hawk (Falco cherrug). As in other cells infected by fowlpox virus two types of inclusion bodies are encountered in the cytoplasm: firstly assemblies of fluffy filamentous material apparently giving rise to immature virions. They are thought to present virus factories (inclusion body B). Secondly mature virus particles budded into extensive groupings (Bollinger bodies, inclusion body A) which display a fine structure identical to other strains of fowlpox virus.


Serum lipoprotein lipids after gemfibrozil treatment. The changes of serum lipoprotein lipids including the phospholipids of twelve hyperlipoproteinemic patients were studied after a four weeks treatment with 1200 mg/day gemfibrozil. There was a decrease of all VLDL lipids as well as of LDL and HDL triglycerides and an increase of HDL cholesterol. The phospholipids were influenced in different directions, particularly their concentration in the HDL was constant. The LDL/HDL lipid ratios remained elevated after gemfibrozil. To evaluate the effect of a hypolipidemic drug in more detail, the analysis of the complete lipoprotein lipid moiety and the ratios within and between the lipoproteins would be helpful.


Effects of oxandrolone on plasma lipoproteins in patients with type IIa, IIb and IV hyperlipoproteinemia: occurrence of hypo-high density lipoproteinemia. The effects of oxandrolone on plasma lipids and lipoprotein composition in patients with type IIa, IIb and IV hyperlipoproteinemia were investigated. Oxandrolone (4 mg/day, for 28 days) induced a significant reduction in plasma and VLDL triglyceride. Reciprocal elevation of LDL cholesterol was observed in 8 of 10 type IV patients and in 2 of 5 type IIb patients. In patients with type IV hyperlipoproteinemia, significant reduction of plasma apolipoproteinB and VLDL apolipoprotein B and inverse LDL apolipoprotein B increment was observed after treatment with oxandrolone. HDL cholesterol was decreased. Hypo-high density lipoproteinemia defined as HDL cholesterol concentration below 40 mg/100 ml occurred in 7 of 10 type IV, 3 of 6 type IIb and 1 of 4 type IIa patients.


Dextran sulfate precipitable (light) plasma lipoproteins in genetically active and passive rats fed two types of diet. Genetically active and passive female rats were fed a purified sucrose/fat diet or a stock diet from the age of 11 to 27 weeks. The age-related increase in plasma total cholesterol concentration was much more pronounced in active rats fed the purified diet than in active rats fed the stock diet. No significant alteration with age took place in the plasma cholesterol level of passive rats fed the stock or the purified diet. The increase in body weight was somewhat greater in active and passive rats fed the purified diet than in those fed stock diet. The lipoprotein distribution at the age of 27 weeks was estimated using both polyacrylamide gel electrophoresis and dextran sulfate/Ca++ precipitation. As compared with the stock diet, the purified diet effected a decrease in the relative amount of prebeta and beta lipoproteins in both active and passive rats. The major effect was, however, a pronounced increase in the relative amount of dextran sulfate precipitable "light prealpha" lipoproteins with no effect on "heavy prealpha" or alpha lipoprotein percentages. Alpha lipoprotein percentages were slightly higher in active than in passive rats. The results confirm our earlier observation that there might be a dissociation between the dietary effect on the plasma total cholesterol level and on the lipoprotein distribution. The data also suggest that the hyperlipemic sucrose/fat diet causes a shift from the prebeta and beta lipoproteins to the "light prealpha" fractions.


Effect of intravenous injection of CDP-choline, S-adenosyl-methionine and citiolone in subjects with hyperlipemia. The effect of some drugs (CDP-choline, S-adenosyl-methionine and acetyl-homocysteinethiolactone) stimulating a number of liver enzymatic activities was studied in hyperlipemic patients to explore the role of liver function in hyperlipidemia. Recently a close connection between liver impairment and hyperlipemia has been observed. Twenty-eight, 13 and 10 hyperlipemic subjects received i.v. doses respectively, of CDP-choline (300 mg), S-adenosyl-methionine (SAMe) (45 mg) and citiolone (300 mg) in 100 ml saline solution during a 30 minute period. Plasma lipid fractions were evaluated basally and at the end of the injection. CDP-choline caused a significant decrease in plasma total lipids (p less than 0.001), cholesterol (p less than 0.01) and triglycerides (p less than 0.01) as well as a slight increase in beta - LP (p less than 0.05). SAMe injection was followed by a significant fall in plasma total lipids (p less than 0.05), cholesterol (p less than 0.001) and beta - LP+pre-beta LP/alpha - LP ratio (p less than 0.05), while alpha -LP increased significantly (p less than 0.05). Citiolone caused only a decrease in plasma cholesterol values (p less than 0.001). The data suggest that the hypolipidemic effect of the three compounds is due to a common mode of action, i.e., the stimulation of hepatic phospholipid biosynthesis, though in a different way.


High density lipoprotein cholesterol and myocardial infarction or sudden coronary death: a prospective case-control study in middle-aged men of the Oslo study. The high density lipoprotein (HDL) cholesterol concentrations of frozen specimen obtained in 1972-73 are reported from 93 men aged 40-49 years who later developed coronary heart disease (CHD), and for 186 controls. Mean HDL cholesterol of CHD-patients was 7.9% lower than that of controls matched for smoking habits and serum concentrations of triglycerides and total cholesterol (p 0.05 for 82 men who had myocardial infarction, n.s. difference for 11 with sudden coronary death), and 10.2% lower (p 0.05) than that of controls who were not matched for the parameters mentioned. The present prospective study confirms that HDL cholesterol is inversely associated with the risk of developing CHD in middle-aged men.


5'-Nucleotidase: solubilization, radiochemical analysis, and electrophoresis. 5'-Nucleotidase (5'-NT, E.C. 3.1.3.5) of cultured human and rodent cells was rendered soluble using the zwitterionic detergent Zwittergent 314. Optimal activity of 5'-NT was obtained when sonicated cells were incubated in solutions containing 0.75% (w/v) Zwittergent. A method was developed for the determination of the activity of 5'-NT in which the unutilized substrate, [14C]-AMP, was precipitated with lanthanum chloride and the soluble [14C]-adenosine was measured by scintillation counting. 5'-NT isozymes were separated using agarose gel electrophoresis and isoelectric focusing in polyacrylamide gel. The zones of enzyme activity were established by precipitation of unutilized [14C]-AMP with LaCl3, removal of soluble [14C)-adenosine by washing gels in water, and autoradiography. The zones of 5'-NT appeared as clear zones on darkened X-ray film. When analyzed by agarose gel electrophoresis, fibroblasts derived from human skin and rat liver produced a single zone of 5'-NT activity. The 5'-NT isozyme of rat cells migrated faster than that of human cells and was easy to distinguish. The presence of detergent in the sample and in the gel enhanced enzymatic activity and improved the separation of the isozymes. Isoelectric focusing resolved 5'-NT of human fibroblasts into two molecular forms, one of which focused in the region of pH 6 and the other at pH5.


Expression of galactose-1-p-uridyltransferase in Chinese hamster x human galactosemia somatic cell hybrids. Lymphocytes from a patient with classic galactosemia (GALT deficiency) were hybridized with a Chinese hamster cell line.  Electrophoretic valuation of GALT in 31 independently derived interspecific hybrid clones failed to demonstrate expression of the human GALT gene even when human chromosome 9 was present. Possible mechanisms for this lack of expression are presented.


Effects of inhibitory ligands on the aerobic carbon monoxide complex of cytochrome c oxidase. 1. In the presence of both CO and O2, ox heart cytochrome c oxidase forms a 607 nm-peak intermediate distinct from both the cytochrome a2+a3 2+CO and the cytochrome a3+a3 2+CO ('mixed-valence') CO complexes. 2. This aerobic CO compound is stable towards ferricyanide addition, but decomposed on treatment with ferric cytochrome a2 ligands such as formate, cyanide and azide. 3. Addition of formate or cyanves rise to a complex with alpha-peak at 598 nm, not identical with any azide complex of the free enzyme, but possibly a cytochrome a3 2+NO complex produced by oxidative attack of partially reduced O2 on the azide. 4. The results support the idea that although the initial reaction of oxygen is with cytochrome a3 2+, the next step is not an oxidation of the ferrous cytochrome a3, but a transfer of O2 to a neighbouring group, such as Cu+, to give Cu2+O2- or similar complexes. 5. The aerobic CO complex is then identified as a3+a3 2+COCu2+O2-; a similar compound ('Compound C') is formed by photolysis of a3+a3 2+CO (the 'mixed-valence' CO complex) in the presence of oxygen at low temperatures.


Partial purification and characterization of deoxyguanosine kinase from pig skin. Deoxyguanosine kinase, which catalyses the phosphorylation of deoxyguanosine to form deoxyguanosine 5'-monophosphate, was purified 1024-fold from extracts to newborn-pig skin. This activity requires the presence of a bivalent cation and a nucleoside triphosphate, which functions as a phosphate donor, ATP being twice as effective as CTP or GTP and 4 times as effective as UTP. The enzyme appears to have a molecular weight of 58500 as determined by Sephadex-column chromatography. Optimal enzymic activity was observed at pH 8.0; however, the enzyme remained active over a broad pH range of 5.5-9.0. Several deoxyribonucleoside and ribonucleoside monophosphates and triphosphates were tested as effectors of catalytic activity. Effective inhibitors were dGMP [Ki(app.) = 7.6 x 10(-5) M] and dGTP [Ki(app.) = 2.1 x 10(-5) M]. Both of these inhibitors acted in a competitive manner. A Km(app.) of 3.2 x 10(-7) M was measured for deoxyguanosine and a Km(app.) of 3.3 mM was determined for MgATP. Of the four major deoxynucleosides tested, this catalytic activity appears to phosphorylate only deoxyguanosine; thus the enzyme is a specific deoxyguanosine kinase.


Purification of pig synovial collagenase to high specific activity. 1. Pig synovium in tissue culture secretes a specific collagenase in a latent form. 2. The latent enzyme was concentrated by (NH4)2SO4 precipitation and activated with 4-aminophenylmercuric acetate, and the active enzyme was purified by chromatography on Ultrogel AcA44, DEAE-cellulose, heparin-Sepharose and a zinc-chelate medium to a specific activity of 53 400 units/mg. of protein. 3. The enzyme was shown to be essentially homogeneous by polyacrylamide-gel electrophoresis. 4. The purified collagenase digested collagen to give the characteristic three-quarter and one-quarter pieces.


Evidence for the occurrence of an ecto-(adenosine triphosphatase) in rat epididymal spermatozoa. Intact spermatozoa from rat cauda epididymis possess a Mg2+-dependent ATPase activity that hydrolyses externally added [gamma-32P]ATP. The ATPase reaction was linear with time for approx. 6 min and there was no detectable uptake of ATP by these cells. The ATPase activity of the whole spermatozoa was not due to leakage of the intracellular enzymic activity, contamination of the broken cells or any possible cell damage during incubation and isolation of spermatozoa. The activity of the enzyme was strongly inhibited (approx. 85%) by p-chloromercuribenzenesulphonic acid (50 microM) or the diazonium salt of sulphanilic acid (50 microM), which are believed not to enter the cells, whereas ouabain (0.5 mM), NaF (10 mM), NaN3 (2.5 mM) and oligomycin (5 microM) had no appreciable effect on the activity of the spermatozoal APTase. There was little loss of ATPase activity from the cells when washed with 0.5 mM-EDTA and an iso-osmotic or hyperosmotic medium. These data are consistent with the view that the observed ATPase activity is located on the external surface of spermatozoa. The sperm ecto-ATPase activity is resistant to the action of proteinases (50 micrograms/ml), namely trypsin, chymotrypsin and Pronase. Studies with various unlabelled phosphate esters indicate that the sperm ecto-ATPase is not a non-specific phosphatase and it has high degree of substrate specificity for ATP.


Characteristics of the rat liver microsomal enzyme system converting cholecalciferol into 25-hydroxycholecalciferol. Evidence for the participation of cytochrome p-450. Properties of the rat hepatic cholecalciferol 25-hydroxylase have been studied. An assay system has been developed in which 25-hydroxycholecalciferol production is linear for at least 2h in both homogenates and microsomal fraction. Furthermore, the initial reaction velocity is linearly related to the amount of liver tissue or microsomal fraction. This enzyme system also metabolizes an analogue of cholecalciferol, namely dihydrotachysterol 3, into 25-hydroxydihydrotachysterol 3. The 25-hydroxylase is in the microsomal fraction and not in mitochondria. It has a Km of 44 nM for cholecalciferol and 360 nM for dihydrotachysterol 3. Its activity is not altered by dietary concentrations of calcium and phosphorus. Vitamin D-deficient rats have higher activities of the hepatic 25-hydroxylase than those receiving 25 ng of cholecalciferol daily. The 25-hydroxylase is inhibited by metyrapone. An atmosphere of CO/O2 (9:1, v/v) inhibits the reaction by 87%. This inhibition is partially reversed by white light. Additionally, cholecalciferol and 25-hydroxycholecalciferol competitively inhibit aminopyrine demethylase. These results support the idea that the cholecalciferol 25-hydroxylase is a cytochrome P-450-dependent mono-oxygenase.


The reaction of cytochrome oxidase with oxygen in the fission yeast Schizosaccharomyces pombe 972h-. Studies at subzero temperatures and measurement of apparent oxygen affinity. 1. Cytochrome alpha 3 in whole-cell suspensions of the fission yeast Schizosaccharomyces pombe reacted in the reduced form with CO to give a photodissociable CO complex with absorption maxima at 429, 543 and 591 nm in CO-liganded reduced-minus-reduced difference spectra. 2. Other CO-bound haemoproteins, cytochromes P-420 and P-450, were not photodissociated under the conditions employed. 3. Measurements of the rates of reassociation of CO with cytochrome alpha 3 after flash photolysis over the temperature range from -101 to -109 degrees C gave a value for Eact. of 28.6 kJ/mol. 4. Between -94 and -106 degrees C, O2 reacted with cytochrome oxidase in intact cells to give an oxygenated intermediate (compound A). 5. At -70 degrees C compound A was converted into a second spectrally distinct intermediate (compound B). 6. Electron transport, indicated by the oxidation of cytochromes alpha + alpha 3 and cytochrome c, did not occur until the temperature was raised to -50 degrees C. 7. At room temperature cytochfome oxidase was oxidized to 50% of its steady-state concentration by 0.35 microM-O2.


Turnover of adenosine 3':5'-cyclic monophosphate in chicken erythrocytes. Turnover of cyclic AMP was studied in intact chicken erythrocytes. Production of cyclic AMP was stimulated by adrenaline and then blocked by propranolol. The decline in the cyclic AMP concentration under these conditions is solely due to its intracellular degradation, whereas efflux of the nucleotide, although existing in these cells, does not contribute significantly to the change in its concentration. Intracellular degradation of cyclic AMP follows a first-order kinetics with a half-life of about 6 min. Similar half-lives were obtained at widely different adrenaline concentrations or when the ration of propranolol to adrenaline was varied by 25-fold. Theoretical equations were applied to calculate the rates of cyclic AMP synthesis and degradation in the intact cells under different experimental conditions. Maximal adrenaline concentrations raise the rate of cyclic AMP synthesis and its steady-state concentration by about 10-fold. The addition of caffeine causes a further 33% increase in intracellular concentration of the nucleotide, which is in good agreement with the theoretical increase computed from its slowed-down degradation.


Macrophage-fibroblast interactions in collagenase production and cartilage degradation. Rabbit bone-marrow macrophages and fibroblasts were cultured, independently or together, with pieces of 35S-labelled cartilage or at the surface of dried [14C]collagen gels. Each type of cell, cultivated alone, rapidly degraded the proteoglycan of cartilage, but only the fibroblasts degraded collagen. The co-culture of both types of cell had no consistent effect on the rate of proteoglycan degradation, but it stimulated the rate of collagen degradation. In parallel, the accumulation of collagenase in the culture fluid was enhanced but not that of neutral proteinase. Coinditioned media from macrophage cultures added to cultures of fibroblasts had the same effect as the living macrophages in stimulating the production of collagenase. Their action was itself enhanced when the macrophages had been activated by concanavalin A-stimulated spleen-cell factors. These data suggest that fibroblasts may act as effector cells in producing collagenase and degrading collagen in response to soluble factors released by macrophages under the control of lymphocyte factors.


Ultrastructural and virological aspects of Langat virus-induced SSPE in suckling hamsters. The morphology of subacute sclerosing panencephalitis (SSPE) lesions in hamsters following i.p. inoculation of Langat virus was studied by light microscopy and electron microscopy. The lesions' temporal relationship to virus presence and antibody production was studied by immunofluorescence, virus isolation from brains and brain cell cultures, and by antibody assay of serum and spinal fluid. All 10-day-old hamsters infected with Langat virus developed SSPE lesions, most prominently in the cerebellum, without any overt signs. The lesions appeared 10 days after infection, progressed in severity until Day 21 and persisted unaltered at least 3 months. They consisted of neuronal degeneration, calcification, and intracellular lipid accumulation. Virus was isolated from Days 3 to 15 and disappeared on Day 21. Demonstration by cerebellar immunofluorescence of viral antigen was observed 10 days after inoculation. Antibody appeared in the serum on Day 6, rose to very high titres by Day 21, and thereafter declined. Cell-associated virus was not demonstrable in negative brain-cell cultures. These findings suggest that SSPE in hamsters, associated with Langat virus infection, is biologically different from measles SSPE. Purkinje-cell lipid accumulation and mineralization sparing mitochondria were manifestations of the response of neurons to cell injury at the threshold of irreversibility.


Allosteric oscillatory enzymes: influence of the number of protomers on metabolic periodicities. The study of a concerted allosteric model for an enzyme activated by the reaction product shows that this system can generate sustained metabolic oscillations regardless of the number of protomers constituting the enzyme. The analysis extends the results previously obtained in a dimeric model for the phosphofructokinase reaction which produces glycolytic periodicities. When the substrate and product concentrations evolve on comparable time scales, the amplitude of oscillations significantly drops as the number of enzyme subunits evolves from 2 to 8. The width of the domain of substrate injection rates which produce oscillations and the periodic variation in enzyme activity also depend on the number of protomers and on the time scale structure of the system. Theoretical predictions are compared with the experiments on glycolytic oscillations in yeast and muscle, and with the structural characteristics of phosphofructokinase. The results are also discussed in relation with the mechanism of cyclic AMP oscillations in the slime mold Dictyostelium discoideum.


The influence of adenosine on intermediary metabolism of isolated hepatocytes. The effect of adenosine was tested on the energetic metabolism of fed rat liver cells after isolation. The cells were incubated in a buffered saline medium with glucose (5 mM) and adenosine (1 mM) for 30 minutes at 37 degrees C. This increased the concentration of the adenylic nucleotides ATP (+57 per cent, ADP (+39 per cent). Cyclic AMP was increased (+50 per cent) and the intracellular inorganic phosphate decreased (-22 per cent). These changes were accompaned by a decrease of glycogenolysis, glucose consumption and lactate production. Measurement of glycolytic intermediates showed decreased concentrations of fructose 1,6-bis-phosphate and 3-phosphoglycerate proportional to the increase in ATP concentration. The near-equilibrium of the glyceraldehyde 3-phosphate dehydrogenase-phosphoglycerate kinase system was not modified by adenosine. The decrease of the NAD+/NADH ratio along with the increase of the ATP/ADP X PO4 ratio explains the decrease of 3-phosphoglycerate. The decrease in glucose consumption can be explained by the cross over at the phosphofructokinase stage with the decrease of fructose 1,6-bisphosphate. The major part of adenosine was deaminated as indicated by an increase in the production of ammonia and urea. The effects of inosine, or adenosine along with an inhibitor of adenosine deaminase (pentostatin) suggest that adenosine acts on the glucose consumption through adenylic nucleotides. However the increase of the adenylic nucleotide level cannot totally explain the other metabolic changes: decrease of the NAD+/NADH cytoplasmic ratio, constancy of this ratio in mitochondria, decrease of gluconeogenesis from lactate. A direct action of adenosine can therefore be expected.


[Preparation and seric form of rabbit C reactive protein]. The preparation of rabbit C-reactive protein (CRP) involves a single step affinity chromatography. This preparation takes advantage of the calcium-dependent affinity of CRP for an agarose gel bearing 2-aminoethanol dihydrogen-phosphate as a ligand. A prior chromatography on agarose gel without the ligand allows the uptake of the serum amyloid P-component (SAP). The CRP prepared according to this method is able to form precipitating complexes in agarose with rabbit lipoproteins. The specificity of these interactions is studied. It is demonstrated that CRP-High Density Lipoproteins (HLD) association produces a second precipitation arc when the pure CRP is revealed by a specific antiserum in agarose. Moreover, CRP in the serum is shown to be in the bound form only, and the binding involves Low Density Lipoproteins (LDL) exclusively.


Conformational effects of coenzyme binding to porcine lactic dehydrogenase. Changes induced on addition of the coenzyme, NADH or NAD+, to porcine lactic dehydrogenase isoenzymes H4 and M4 have been studied by hydrodynamic and spectroscopic methods. As shown by ultracentrifugal analysis, the native subunit structure remains unchanged on holoenzyme formation; an approximately 5% increase of the sedimentation coefficient, parallelled by a slight decrease of the partial specific volume (less than 1%) indicate a significant change in the native tertiary and/or quaternary structure of the enzymes, corroborating earlier calorimetric data (Hinz and Jaenicke, 1975). The binding constant for the enzyme from skeletal muscle (M4) and NADH are found to be in agreement with KD-values obtained from equilibrium dialysis, as well as spectroscopic and thermal titration experiments (8 muM). Far UV circular dichroism measurements do not show significant changes on ligand binding, indicating unchanged helicity or compensatory conformational effects. In the near UV, ligand binding is reflected by an extrinsic Cotton effect around 340 nm; in the range of aromatic absorption no changes are detectable. The experimental results suggest that there are gross structural changes on coenzyme binding to lactic dehydrogenase which do not affect the intrinsic spectral properties normally applied to analyze transconformation reactions in protein molecules.


[Prostaglandins, cyclic nucleotides and heart adaptation to acute and chronic pressure overload]. The content of prostaglandins (PGs) and cyclic nucleotides was determined in the rat heart at different time following coarctation of the abdominal aorta. The animals were divided into three groups according to the rate and degree of hypertrophy. The rats with the best adaptability (the highest level of hypertrophy) showed the highest myocardial PG content. An increase in myocardial weight in response to the overload correlated with an enhancement in PGE/PGF2 ratio. The relationship between the myocardial levels of PGs and cyclic nucleotides during the period of adaptation was also observed. Prostaglandins are suggested to play an important role in the heart adaptation to pressure overloads.


[Differences in the structure of adrenalin and ACTH receptors in adipose cells of rats]. The treatment of adipose tissue of the rat epidiymis with trypsin decreased the lipolytic action of ACTH by 95%, and the lipolytic action of adrenalin by 50%. Supplementation of the incubation sample with lysolecithin suspension led to a complete loss of the tissue sensitivity to ACTH and a 40% decrease in the sensitivity to adrenalin. A conclusion is made about a different structure of adrenalin and ACTH-receptors in the plasma membrane of adipose cells.


[Circadian rhythm of changes in phospholipids and activity of non-specific phosphomonoesterases in rat liver]. The circadian rhythm of the content of total and individual phospholipids and the activity of alkaline and acid phosphatases was revealed in the liver of 130 white rats by the biochemical and histochemical methods. The concentration of phospholipids and enzymatic activity rise during the day time (9 a.m. - 15 p.m.). The results obtained suggest a good agreement between the structural and metabolic features of the liver during its maximal functional activity.


The cyclic adenosine monophosphate phosphodiesterases of myoblasts, fibroblasts, and their somatic cell hybrids. Three forms of cAMP phosphodiesterases are found in mouse L cells (fibroblasts) and rat skeletal myoblasts. The myoblast enzymes can be resolved by chromatography on DEAE-cellulose and the fibroblast enzymes by chromatography on DEAE-Biogel. The myoblast enzymes are "high affinity" cAMP specific forms and have different molecular weights, while all L-cell enzymes have an apparent molecular weight of 450,000. Only one of the L-cell enzymes is able to hydrolyze both cyclic guanosine monophosphate (cGMP) and cAMP. Hydrolysis of the latter is stimulated by micromolar amounts of cGMP. The myoblast x L cell hybrids possess at least five phosphodiesterases, three of which can be identified as being of myoblast or fibroblast origin. One of the fibroblast enzymes appears to be modified in hybrids. The entire phosphodiesterase regulatory system of the myoblasts is active in the hybrids.


Enzymatic degradation and partial biosynthetic reconstitution of microsomal and mitochondrial membranes. Guinea pig liver microsomal and mitochondrial membranes were degraded with phospholipase C and D followed by partial biosynthetic reconstitution. Activities of phosphatidylinositol synthetase in microsomal membranes and NADPH-cytochrome c reductase were almost completely lost after phospholipase C and D treatment; almost complete restoration of the original activity was achieved after biosynthesis of phosphatidylcholine in degraded microsomes, but was not reparable after biosynthesis of cytidinediphosphodiglycerides (CDP-diglycerides). The mitochondrial biosynthesis of polyglycerophosphatides was completely retained after degradation of these membranes with phospholipase C, but after similar treatment with phospholipase D, only about one-quarter of the original activity remained, the relative composition of polyglycerophosphatides being significantly different. The activity of NADPH-cytochrome c reductase of microsomes represented about 76% of the original activity after phospholipase C treatment, but only approximately 1% after treatment with phospholipase D. Although this activity could not be restored with CDP-diglyceride synthesis, it was restored to about 75% of the original activity after the biosynthesis of phosphatidylcholine in these fragments. These and additional experimental findings are discussed in terms of the relation between structural organization of lipids and proteins and enzymatic activities of membrane-bound phospholipid-synthesizing enzymes in microsomal and mitochondrial membranes isolated from guinea pig liver.


Changes induced by cold adaptation in the brown adipose tissue from several species of rodents, with special reference to the mitochondrial components. (1) The effects of cold adaptation upon the brown adipose tissue have been studied in rats, hamsters, mice, and guinea pigs. (2) Striking effects were found for total tissue as well as at the mitochondrial level, e.g., increases in protein and phospholipid contents, changes in phospholipid fatty acid composition (a decrease in the percentage of palmitic and palmitoleic acids and an increase in stearic and linoleic acids), and a change in the mitochondrial polypeptide composition (a marked increase in a 32000 molecular weight polypeptide, except for hamsters). (3) In situations where animals exhibit a greatly enhanced capacity for nonshivering thermogenesis (cold adaptation for rats, mice, and guinea pigs, birth for guinea pigs, and hibernation ability for hamsters, dormice, and garden dormice), brown fat mitochondria are characterized by the occurrence of large amounts of the 32000 molecular weight polypeptide characteristic of these mitochondria.


In vivo preparation of [32P]cAMP and thin-layer chromatography of cAMP-related compounds. A simple and rapid method for preparing [32P]adenosine 3'5'-cyclic monophosphate (cAMP) is described. A culture of an Escherichia coli mutant which excretes cAMP about 150 times faster than does a wild-type strain was incubated overnight with [32P]orthophosphate of high specific activity (e.g., 4000 Ci/mol (1 Ci = 37 GBq). The [32P]cAMP which accumulated extracellularly was then purified to 99.9% radiochemical purity in less than 4 h by adsorption to charcoal and alumina column chromatography. A two-dimensional chromatography system using a PEI-cellulose plate is also described which should prove useful for studying cAMP metabolism with 32P- or 3H-labeled cAMP or ATP.


The properties of purified low molecular weight phosphoprotein phosphatase from rabbit heart. A simplified procedure for the purification of low molecular weight phosphoprotein phosphatase acting on muscle phosphorylase a has been described from rabbit heart. The enzyme was purified to homogeneity by acid precipitation, ethanol treatment, and chromatography on Sephadex G-75 and Sepharose-histone. The purified enzyme showed a single band when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; the molecular weight calculated by this method was 34 000. The S20, W value and Stokes radius for the enzyme was 3.35 and 24.0 A(1 A = 0.1 nm), respectively. Using these two values, a molecular weight of 35 000 was calculated. Purified enzyme showed a wide substrate specificity and catalyzed the dephosphorylation of phosphorylase a, glycogen synthase D, phosphorylated histone, and phosphorylated casein. Kinetic studies revealed the lowest Km with glycogen synthase D and maximum Vmax for the reaction with phosphorylase a.


Components and mechanism of action of ATP-driven proton pumps. We have studied the composition of ATP-driven proton pumps from bovine heart mitochondria and have reconstituted the oligomycin-sensitive ATPase complex from its individual components. The complex contains 9 to 10 subunits of which 5 are assembled in the soluble F1 protein, 2 are required for the attachment of F1 to the membrane and 2 form the proton channel within the membrane. With the help of information obtained from studies of the chloroplast and the bacterial proton pumps, we can tentatively assign a function to each of the subunits of the pump. The position of F1 outside of the membrane seen in electron micrographs of negatively stained preparations, does not appear to be an artifact. Evidence from immunological studies, chemical derivatizations as well as further electron microscopy (positive staining and freeze-etching), support this statement. We describe in this paper a 28 000-dalton polypeptide which has been isolated from the mitochondria membrane and is required for the reconstitution of oligomycin-sensitive ATPase and 32Pi-ATP exchange activity. We propose a mechanism of action of the proton pump in which the key energy-yielding reaction is the binding of Mg2+ to the protein. The function of the proton gradient is to displace Mg2+ from this site to permit cyclic repetition of the binding process. Essential for this scheme is the cyclic opening and closing of the proton channel. We have outlined our present approaches to test this hypothesis.


Selective spin-labeling of the ribosomal proteins of 70S ribosomes from Escherichia coli. We have used a series of N-(1-oxyl-2,2,5,5-tetramethyl-3-pyrrolidinyl) maleimide spin labels of different length to label, covalently and selectively, the most reactive sulfhydryl groups of 70S ribosomal proteins of Escherichia coli. Under short periods of labeling (1--2 min), less than two spin labels per ribosome are incorporated and were shown to be distributed mainly on five ribosomal proteins in the following order: S18 greater than S21, L27 greater than S17, and S12. With a long period of labeling (3 h) up to 13 spin labels are attached to the ribosome, and protein S1 is the most labeled. The shape of the electron paramagnetic resonance (epr) signal shows two components with a predominance for the strongly immobilized orientation, and the percentage of these components in each spectra has been evaluated. When the distance between the nitroxide group and the maleimide-attaching group exceeds 6 A (1 A = 0.1 nm) the strongly immobilized orientation disappears. The effect of magnesium ions on these selectively spinlabeled ribosomes shows that the dissociation into subunits does not affect the epr signal, but more spin labels are incorporated into the subunits if labeling is performed under conditions of dissociation.


Inhibition of rat and human adipocyte adenylate cyclase in the antilipolytic action of insulin, clofibrate, and nicotinic acid. Clofibrate (Atromid-S), nicotinic acid, and insulin are known to be potent hypolipidemic and antilipolytic agents. The present study was undertaken to define the mechanism of action of this latter effect on isolated rat and human fat cells. Sodium clofibrate (0.42 mM), nicotinic acid (0.42 mM), and insulin (100 microU/mL) were shown to inhibit norepinephrine-stimulated lipolysis in rat and human adipose cells and this inhibition was associated with a reduction in intracellular 3',5'-cyclic AMP levels. A similar cyclic AMP lowering effect was demonstrated with insulin in the presence of procaine-HCL, which uncouples the adenylate cyclase system from lipolysis. This insulin effect was attributed to inhibition of adenylate cyclase. A direct and significant inhibition of adenylate cyclase in membrane fractions obtained from isolated human adipocytes was demonstrated for all three antilipolytic agents. The common membrane site of action of these agents whereby adenylate cyclase activity is depressed, thus decreasing cyclic AMP production and free fatty acid (FFA) mobilization from adipose stores, implies a central role for the adenylate cyclase system. These findings are consistent with the view that the hypotriglyceridemic effects of clofibrate, nicotinic acid, and insulin may be partly explained by deprivation of FFA substrate for hepatic very low density lipoprotein synthesis.


Effect of carbon sources on the rates of cyclic AMP synthesis, excretion, and degradation, and the ability to produce beta-galactosidase in Escherichia coli. We have determined the rates of adenosine 3',5'-cyclic monophosphate (cAMP) synthesis, excretion, and degradation, and the cAMP pool size in Escherichia coli grown on various carbon sources. We have found that the cAMP pool size increases in approximate proportion to increases in the cAMP synthetic rate. Although the combined rate of excretion and degradation of cAMP is in approximate proportion to the cAMP pool size, no such regular relationship is seen between the cAMP pool size and either the excretion rate or the degradation rate. Using a method which we have developed for determining the cellular efficiency of enzyme production (termed 'cellular' rate), we have reexamined the relationship between cAMP pool size and the rate of beta-galactosidase production. Although there exists an overall trend of increasing rate of beta-galactosidase production with increasing cAMP pool size, large variations in the rates of beta-galactosidase production are seen even under culture conditions which yield similar cAMP pool sizes. This suggests that the intracellular level of cAMP cannot be the unique regulator of beta-galactosidase production.


Horseradish peroxidase. XXXIV. Oxidation of compound II to I by periodate and inorganic anion radicals. The one-electron oxidation of horseradish peroxidase compound II to compound I by sodium periodate was observed. The bimolecular rate constant for the NaIO4--compound II interaction is equal to 9.5 +/- 1 x 10(-3) M-1s-1 at room temperature. Irradiation, using ultraviolet light, of the solution containing compound II and persulfate in the presence of bicarbonate, chloride, or bromide, leads ot the fast accumulation of compound I due to the oxidative action of SO4, CO3, Cl2, and Br2 anion radicals, which are products of the photolysis.


Polioviruses and other enteroviruses in urban sewage from Laval (Canada): presence of non-vaccinal strains of poliovirus. During a 13-month period (August 1, 1977-August 31, 1978) 55 samples of 2-4 L of raw sewage from an urban residential area were collected and studied for the presence of human enteroviruses. Viruses were recovered from 47 (90.5%) of the samples. Of the 201 viral isolates propagated, 180 (89.5%) were identified by the serum neutralization test as poliovirus types 1,2, or 3; 16 (8%) were identified as coxsackie B viruses; and 5 (2.5%) could not be identified by the methods used. While all polioviruses, types 2 and 3 isolates, were vaccine-like (rct-/d-) or intermediate strains, 14 poliovires, type 1 out of 55 selected isolates, were found to be non-vaccine-like (rct+/d+), 8 were rct-/d-), and 33 were intermediate strains. Out of nine samples submitted to the serodifferentiation (McBride) test, six poliovires, type 1 rct+/d+ and three rct+/d-, were related to wild strains.


Effect of divalent cations on the inhibition of alkylsulfatase induction and the generation of ATP in Pseudomonas aeruginosa after exposure to exogenous uridine 5'-triphosphate. In the absence of added Mg2+, alkylsulfatase induction in resting cells of Pseudomonas aeruginosa was inhibited 17% by exogenous 0.05 mM UTP. Under these conditions, the cells converted UTP to ATP and rapid degradation of these nucleotides did not occur. In the presence of 0.73 mM Mg2+, 0.05 mM UTP repressed the synthesis of the enzyme by 71%. Under these conditions, the cells rapidly degraded both ATP derived from UTP as well as residual UTP. In the presence of Mg2+ and 0.1 mM UTP, full repression of alkylsulfatase formation occurred whereas Mg2+-depleted cell suspensions were still capable of synthesizing 47% of the enzyme under these conditions compared with control levels. The inhibition of alkylsulfatase induction was highly specific for UTP. Some inhibition was observed with exogenous uracil, uridine, and pyrophosophate but only at concentrations greater than 1.0 mM. Exogenous UMP and UDP (2mM) had no effect.


Quantitative determination of cytomegalovirus IgG antibody by enzyme-linked immunosorbent assay (ELISA). Quantitative determination of cytomegalovirus (CMV) human IgG antibody by ELISA performed in polystyrene trays was carried out and compared with results obtained by the routine complement fixation (CF) test. The antibody titres of all the sera determined by ELISA were higher than those detected by the CF test. Of the 27 sera negative (less than 1:8) by the CF test, 22 of them were also negative (less than 1:50) by ELISA. The other five CF negative sera were found to have low levels of antibody by ELISA. ELISA is a sensitive and specific assay for CMV antibody. The CMV antigen adsorbed readily to the microtitre plates and the test was simple to perform. The results of ELISA could be obtained in 5 to 6h. A description of the test procedure is given including the method of CMV antigen attachment to the plates. The possibility of using paired sera at a single dilution in the ELISA test to detect seroconversion is discussed.


ICMSF methods studies. XII. Comparative study for the enumeration of Clostridium perfringens in feces. As the second phase of an international comparative study for the enumeration of Clostridium perfringens, four methods were compared for "total" and spore counts of C. perfringens in fecal specimens: the SFP (Shahidi-Ferguson perfringens) agar (A), TSC (tryptose-sulfite-cycloserine) agar (B), SC (sulfite-cycloserine) agar (C), and neomycin blood agar (D) methods. In both the total and spore count procedures, the confirmed C. perfringens counts in method D were lower than in methods A, B, and C. Little differences among methods were found in the percentages of presumptive colonies confirmed as C. perfringens. The nonspecific counts in methods A and D were generally greater than in B and C, but nonspecific microorganisms did not interfere in the enumeration of C. perfringens spores by any of the four methods. In overall performance, methods B and C were superior to A and D. The mean C. perfringens spore count was only 0.17 log lower than the mean total count. Spore counts alone are, therefore, adequate in investigations of C. perfringens outbreaks.


Protective effect of polymyxin B sulfate in experimental enterobacterial infection in mice. The mouse model of intraperitoneal enterobacterial sepsis was used to evaluate the anti-endotoxic effect of polymyxin B sulfate. Single or multiple therapeutic doses of polymyxin, administered either before or after lethal challenge with Serratia marcescens, produced statistically and clinically significant protective effects.


[A case of giant fibroadenoma of the breast in adolescence]. Starting from a personal observation of a giant fibroadenoma of the breast in a girl aged 13, the authors review the possible causes of massive swelling of the breast in adolescence. Next, they analyze the salient anatomoclinical aspects of giant fibroadenoma, benign phylloid cystosarcoma, and virginal mammary hypertrophy. The conclude that the surgeon should do everything possible to arrive at the histological diagnosis of fibroadenoma intraoperative--that is, extemporaneously--in order to avoid treating as a malignancy a possible benign phylloid cystosarcoma.


Morphometric method for evaluating the neurovirulence of live poliomyelitis vaccine. A method for measuring morphological changes caused by residual neurovirulence in live poliomyelitis vaccine is described. The entire area of 16 standard parts of the central nervous system is measured directly under the microscope using a morphometric eyepiece grid. The method can be used to supplement the conventional system of scoring morphological lesions.


[Bone maturity and physical performance]. We have examined the relation between physical performance (as measured with the CAHPER tests), bone age (Greulich and Pyl's wrist evaluation) and dental age (Garn's method). Our study included 164 boys and 149 girls aged from 7 to 11. In spite of interesting significant simple correlation between most of the CAHPER tests and bone age measurements for the pre-pubescent period, the latter remains of negligible value in predicting physical performance in the studied children when variables like chronological age, body height and body weight are already included in the prediction equation.


Effect of age on angiotensin II receptors from rat brain. 1. Angiotensin II receptor binding was studied in specific regions of rat brain at different ages from birth to 14 weeks. 2. The number of specific angiotensin II receptors increased in all regions during the first 2 weeks of life and then decreased to adult levels. Peak numbers of receptors were up to 10 times the adult numbers. 3. The midbrain and thalamus-hypothalamus had maximum numbers of angiotensin II receptors at 2 weeks of age, whereas the rest of the brain regions had maximum number at 1 week. 4. Saralasin-infusion experiments suggested that circulating angiotensin-related peptides could reach brain angiotensin II receptors in 2 week old rats, but not in 6 week old rats. 5. It is postulated that the centrally mediated actions of circulating angiotensin II may be particularly important in the newborn.


Hormonal changes with long-term converting-enzyme inhibition by captopril in essential hypertension. 1. Captopril was shown to be as effective as hydrochlorothiazide in lowering the blood pressure in patients with moderately severe essential hypertension. 2. With the combination of captopril and hydrochlorothiazide satisfactory control of blood pressure was maintained over 8 months. 3. Inhibition of angiotensin converting enzyme by captopril in man was associated with falls in plasma angiotensin II and urinary aldosterone and rises in angiotensin I and plasma renin. 4. No change in venous concentrations of bradykinin could be demonstrated during therapy. 5. Captopril attenuated the hyperaldosteronism and hypokalaemia associated with diuretic therapy.


Effects of an angiotensin converting enzyme inhibitor on psychosocial hypertension in mice. 1. Renin-associated, chronic psychosocial hypertension of 150-160 mmHg develops in groups of mice interacting socially in complex population cages. 2. The blood pressures of 16 males in a cage were measured and an intraperitoneal injection of the angiotensin coverting enzyme inhibitor captopril (SQ 14,225) was given. Three hours later blood pressures were measured again. 3. During the first 3 weeks of psychosocial hypertension SQ 14,225 was without effect. But at 1 month and subsequently up to 7 months, SQ 14,225 reduced blood pressure to the normal range of 120-130 mmHg. 4. Plasma renin activities were not related to the extent of blood pressure reduction by SQ 14,225. Hence other factors in addition to the renin-angiotensin mechanism play a part in maintaining chronic psychosocial hypertension.


A novel orally active converting-enzyme inhibitor YS 980: effects on blood pressure in spontaneously hypertensive rats. 1. The effects of long-term treatment with the angiotensin I converting-enzyme inhibitor YS 980 were examined in stroke-prone spontaneously hypertensive (sp-SH) rats. Development of hypertension was markedly blunted in the YS 980-treated animals. 2. Effective converting-enzyme inhibition was confirmed by significant increases in plasma angiotensin I (ANG I) and plasma renin concentration, inhibition of the pressor responses to intravenous ANG I and potentiation of the depressor responses to intravenous bradykinin. 3. Urinary free aldosterone excretion was decreased but no changes in urinary sodium and potassium excretion were observed. 4. The pressor responses to intravenous leucine-enkephalin were reduced. 5. The pressor responses to injection of ANG I and bradykinin into the lateral brain ventricle were unaltered. 6. We conclude that the antihypertensive action of YS 980 in sp-SH rats cannot be explained by the inhibition of the plasma renin-angiotensin system alone. Effects on other peptide systems must be considered.


Plasma noradrenaline correlates with alpha-adrenoreceptor-mediated vasoconstriction and blood pressure in patients with essential hypertension. 1. The relationships between plasma noradrenaline concentration at rest and blood pressure, as well as increase in forearm blood flow in response to a brachial artery infusion of the alpha-adrenoreceptor-blocking agent phentolamine, were investigated in hypertensive and normotensive subjects of similar age. 2. In 44 hypertensive patients plasma noradrenaline correlated with systolic, diastolic and mean blood pressures, but no difference in the mean plasma noradrenaline concentration was found. 3. In 11 patients and 14 normotensive subjects alpha-adrenoreceptor blockade resulted in a similar increase in forearm blood flow. Only in the patients, however, was this increase related to plasma noradrenaline and blood pressure. 4. In patients with established essential hypertension plasma noradrenaline can be considered to be a marker of alpha-adrenoreceptor-mediated vasoconstriction, which, in part, determines the height of the blood pressure.


Sympatho-adrenal and cardiovascular response to mental stress and orthostatic provocation in latent hypertension. 1. Seven latent hypertensive patients and seven matched controls were subjected to standardized mental stress and orthostatic provocation. 2. Mental stress increased blood pressure by approximately 25%, heart rate by 25 beats/min, plasma glycerol by 50% and plasma cyclic AMP by 25% in both groups. Plasma glucose and renin activity were unchanged. Plasma noradrenaline and adrenaline were essentially unchanged during stress. 3. There was an insignificant tendency towards higher noradrenaline levels in latent hypertensive subjects and two of these subjects displayed an exaggerated noradrenaline response to standing. 4. Our results indicate that the physiological responses to mental stress are caused by selective neuronal activation, rather than by generalized sympatho-adrenal activation. Latent hypertension does not seem to be associated with adrenergic hyperactivity or receptor supersensitivity, except possibly in individual cases.


Adrenocorticotropin inhibition of mineralocorticoid hormone production. 1. Adrenocorticotropin (ACTH)-induced steroidogenesis, obtained by continuous administration of ACTH for 3 days, produces in man (a) sustained elevations of plasma deoxycorticosterone and cortisol concentrations, (b) transient elevations of plasma aldosterone and 18-hydroxycorticosterone concentrations that return to near-control values, and (c) brisk initial increases in plasma 18-hydroxydeoxycorticosterone and corticosterone concentrations that fall to 20-68% of peak values 30 h thereafter. 2. Dexamethasone (8 mg/day, orally) treatment for 2 days in man permits a greater postural increase in plasma aldosterone and 18-hydroxycortisone concentrations. A dampening effect of ACTH is suggested. 3. An ACTH-initiated inhibition of 11 beta- and 18-hydroxylation is proposed to be operative in regulation of mineralocorticoid hormones.


Reactive hyper-reninaemia to angiotensin blockade identifies renovascular hypertension. 1. Saralasin and converting enzyme inhibitors SQ 20881 and captopril induced increases in plasma renin activity to greater than 14 ng h-1 ml-1 in 43 out of 44 patients with untreated renovascular hypertension when studied in the seated position and on normal sodium intake. This degree of response was absent in patients with normal-renin essential hypertension and present in only three out of 26 with high-renin essential hypertension. 2. Reductions of greater than approximately 9% in diastolic pressure in response to these three drugs occurred regularly in renovascular hypertension (95%) but also frequently in high-renin (65%) and normal-renin (26%) essential hypertension. 3. Prior sodium depletion abolished the specificity of the renin and depressor responses to angiotensin blockage for renovascular hypertension. 4. Some patients with bilateral renovascular and all with malignant hypertension also exhibited these responses to angiotensin blockade that are characteristic of unilateral renovascular hypertension.


Haemodynamics of ACTH-induced hypertension in sheep. 1. Administration of adrenocorticotropic hormone (ACTH) to sheep produced increases in mean arterial pressure within 24 h associated with an increase in cardiac output and cardiac rate. Both cardiac output and blood pressure remained elevated over the 5 days of ACTH treatment. 2. Administration of ACTH during beta-adrenoreceptor blockade resulted in an increase in blood pressure without changes in cardiac output at 24 h. 3. Administration of a combined steroid infusion over 5 days produced increases in cardiac output identical with the effects of ACTH but with a substantially smaller effect on blood pressure. 4. These data suggest that the observed changes in cardiac output produced by ACTH treatment may be associated with high blood concentrations of adrenocortical steroids rather than being necessary for the development of the hypertension.


Comparative beta-adrenoreceptor-blocking effects and pharmacokinetics or propranolol and pindolol in hypertensive Africans. 1. The beta-adrenoreceptor-blocking effects of pindolol were compared with those of propranolol and a placebo in a double-blind cross-over trial involving nine hypertensive African patients. 2. Heart rate, systolic blood pressure and diastolic blood pressure were measured at rest and immediately after exercise before and at intervals up to 6 h after oral administration of the drugs. In addition, plasma pindolol and propranolol concentrations were determined at the same intervals. 3. Pindolol diminished systolic blood pressure at rest and after exercise and antagonized exercise-induced tachycardia, but had no effect on resting heart rate. Propranolol diminished systolic blood pressure predominantly after exercise and reduced both resting and exercise heart rate. Both drugs had no effect on diastolic pressure. 4. The mean plasma concentration reached a peak at 2 h for each drug and this coincide with the interval at which maximal beta-adrenoreceptor-blocking effect was observed.


Influence of selective and non-selective beta-adrenoreceptor blockade on the haemodynamic effect of adrenaline during combined antihypertensive drug therapy. 1. Haemodynamic effects of adrenaline were studied in 27 hypertensive patients, succesively during treatment with propranol and metoprolol. In 12 patients beta-adrenoreceptor blockade was combined with diuretics and in 15 patients the blockade was combined with vasodilators. 2. During propranolol adrenaline caused a marked pressor effect: there was a considerable rise in systolic as well as in diastolic blood pressure and a marked fall in heart rate. During metoprolol there was only a slight rise in blood pressure and an increase in heart rate. 3. Forearm blood flow was decreased by adrenaline during propranolol and was increased during metoprolol. Calculated vascular resistance showed opposite changes. 4. Results were essentially the same when beta-adrenoreceptor blockade was combined with diuretics or with vasodilators and did not differ from previous results obtained in patients treated by blockade alone. 5. If adrenaline infusion can be considered as a model for acute stress, our results seem to favour a selective beta 1-adrenoreceptor blocking agent over a non-selective one, even when the blocker is combined with a diuretic or a vasodilator.


Long-term effect of sotalol on plasma lipids. 1. We studied the effect on plasma lipids of sotalol given orally over a 12 month period to patients with essentially hypertension. 2. Plasma free fatty acid concentration was lower than initially at 1, 3, 6 and 12 months. The difference was significant (P less than 0.01) at 1 and 3 months. 3. Plasma cholesterol (VLDL + LDL-cholesterol) increased during treatment. Plasma total cholesterol increased from 5.49 +/- SD 0.94 mmol/l at the beginning to 6.37 +/- 1.10 mmol/l at 12 months (P less than 0.01). 4. HDL-cholesterol concentration and the ratio of HDL-cholesterol to total cholesterol decreased significantly. The ratios were 0.28 and 0.18 at the beginning and at 12 months respectively (P less than 0.001). 5. Plasma triglycerides increased simultaneously from 1.14 +/- 0.31 to 1.89 +/- 0.99 mmol/l (P less than 0.01).


Effect of captopril on renal vascular tone in patients with essential hypertension. 1. The effect of acute inhibition of angiotensin-converting enzyme by captopril (50 mg) on renal haemodynamics and function was assessed in nine patients with essential hypertension on unrestricted sodium intake (n = 8) or low sodium diet (n = 1). 2. Captopril induced a rapid and significant decrease in arterial pressure, which was maximal within 60 min. 3. Effective renal plasma flow (ERPF) increased, glomerular filtration rate (GFR) did not change and filtration fraction (FF) decreased after captopril. No change in sodium excretion and a decrease in urinary potassium occurred. 4. In the patient on low sodium diet, captopril induced striking increases in GFR and ERPF (64 and 106% respectively). 5. The logarithm of baseline plasma renin activity was positvely correlated with the change in ERPF and negatively correlated with changes in FF and renal resistance. 6. The results indicate that in patients with essential hypertension angiotensin participates actively in the maintenance of renal vascular tone at the efferent arteriolar level. A possible influence of kinins remains to be defined.


[Possible pathogenetic factors of gallstone formation in populations with a high standard of living (author's transl)]. The present work points out the variety of conditions favouring gallstone formation.--Recent research suggests that fibre-depleted, high-fat foods may lead to cholesterol gallstone formation in man. On the other side civilized diets contribute to the carcinogenesis of large bowel.--Experimental observations have indicated that hypovitaminosis C may favour pathogenesis of these diseases in a different way.


Insulin release and eflux of [32P]Phosphate from islets of rat Langerhans treated with iodoacetic acid and the anomers of D-glucose. To clarify the insulin-releasing mechanism, we studied insulin release and the efflux of [32P]phosphate by glucose at 0.1 mM/min of gradient level or at 16.7 mM, and other metabolism in islets of rat Langerhans. When treated with 1 mM iodoacetic acid (IAA) plus the anomers of D-glucose at 2.8 mM for 6 min at 37 degrees C, islets elicited insulin at half the control rate under the step-wise stimulation by glucose and at the same rate as the control under the slow-rise stimulation by glucose. Using islets treated with IAA plus the alpha anomer at 16.7 mM, the step-wise stimulation secreted insulin at half a rate of the control and the slow-rise stimulation at the rate lower than the control, which was not significantly different from the control rate. Treatment with IAA plus the beta anomer at 16.7 mM inhibited insulin release under both types of stimulations by glucose. The step-wise stimulation caused the same rapid efflux of [32P]phosphate from IAA-treated islets  as from the control islets, except for islets treated with IAA plus the beta anomer at 16.7 mM. The rate of glucose utilization in islets was inhibited by all IAA-treatments to the same extent, being merely half the control rate. Treatments with IAA plus the anomers at 16.7 mM significantly reduced the formation of [3H]-cAMP and the activity of protein phosphokinase in islets, while in the presence of the anomers at 2.8 mM IAA produced no significant effect. Neither IAA-treatments altered the uptake of 45Ca and the ATP content in islets. The uptake of [14C]IAA was significantly enhanced by the presence of the beta anomer at 16.7 mM to two times the control level. On the basis of these results, we suggested that the B cell might contain both glucoreceptors and rate-sensors of glucose controlling insulin release and the former might be less sensitive to IAA as compared with the latter.


The effects of dopamine on renal excretion of sodium, phosphate and cyclic AMP in thyroparathyroidectomized dogs. With dopamine (0.5 microgram/kg/min) infusion into the renal artery of thyroparathyroidectomized dogs, urine output and inorganic phosphate excretion increased significantly (p less than 0.05), but the increase in sodium excretion was low and not statistically significant. However, natriuresis and phosphaturia due to the infusion of dopamine were accelerated more markedly by the pretreatment with phenoxybenzamine. Dopamine was infused into the renal artery indoses too small to affect renal hemodynamics (0.02-0.05 microgram/kg/min) after the treatment with phenoxybenzamine and alprenolol with the result that phosphate and sodium excretion increased significantly (p less than 0.05). The excretion rate of cAMP did not change. This suggests that the effect of dopamine on sodium and phosphate excretion is directly influenced by alpha adrenergic activity in the kidney. The mechanism of natriuresis and phosphaturia by dopamine is, however, independent of changes in parathyroid hormone and the adenyl cyclase-cAMP system.


Glucagon secretion during the development of insulin-secreting tumors induced by streptozotocin and nicotinamide. Serial oral glucose tolerance tests in rats treated with streptozotocin and nicotinamide showed that blood glucose levels after glucose loading were suppressed significantly 7 months after treatment as compared to those of earlier stages. Post-glucose plasma insulin levels were significantly elevated at the 9th to 12th month and concomitantly fasting plasma glucagon levels rose significantly. At that time pancreatic islet cell tumors were demonstrated in all of the rats in this experiment. Post-glucose plasma glucagon levels, however, did not show remarkable changes throughout the observation. In spite of hyperinsulinemia, post-glucose plasma glucagon levels of tumor-bearing rats were significantly lower than those of body weight adjusted controls. It is inferred from the study that secretory activity of pancreatic A-cells of tumor-bearing rats is restrained by excess insulin released from islet cell tumors.


Stimulation of pituitary-adrenocortical system by ginseng saponin. Effects of preparations of saponin mixture and isolated ginsenosides, extracted from the root of Panax ginseng, on plasma corticotropin (ACTH) and corticosterone concentrations in rats were determined by the radioimmunoassay and competitive protein binding method. When ginseng saponin mixture was administered to rats intraperitoneally, plasma ACTH and corticosterone increased significantly 30, 60 and 90 min after the treatment. The kinetic pattern of the increase in plasma ACTH was almost parallel to that in plasma corticosterone. Isolated ginsenoside, protopanaxadiol or protopanaxatriol glycoside, also increased plasma corticosterone. The ginseng-induced increase in plasma corticosterone was suppressed by pretreatment with dexamethasone. Thus the ginseng saponin was found to act on the hypothalamus and/or hypophysis primarily, and stimulated ACTH secretion which resulted in increased synthesis of corticosterone in the adrenal cortex.


Some properties of hog thyroidal membrane-bound adenosine tri-phosphatase: proteolytic activation of Mg-dependent activity. The ATPase preparations from the hog thyroid was preincubated with various amounts of trypsin. The activity of Mg-ATPase was consistently elevated. On the contrary, the Na, K-ATPase activity decreased with increasing amounts of trypsin. The effects were similar to those which were observed in the enzyme preparations treated with basis polyamino acids as previously reported. This phenomenon seemed to be specific in the preparations from the thyroid. The Mg-dependent activity was increased after pretreatment with trypsin or poly-L-lysine (PLL) when CTP, ITP and UTP were used as substrate. Thus the substrate specificity of Mg-ATPase was low. The enzyme-kinetics using ATP as substrate showed that the increase in activity was due to an increase in Vmax and not to a change in Km. The activity of Mg-ATPase was increased even after 30 min of preincubation with trypsin, while the Na, K-ATPase activity was almost diminished. These results suggest that the activity of Mg-ATPase in the preparation from the thyroid is specifically changed by the modification of the molecular environment of the enzyme with trypsin or basic polyamino acids.


Suppressive effect of pentagastrin on pituitary-adrenocortical secretion. The effect of pentagastrin on the pituitary-adrenocortical secretion was examined in male rats. In the morning the intraperitoneal injection of this peptide produced a slight, but significant, decrease in the plasma corticosterone levels, but it had no effect on the evening rise due to circadian periodicity and the stress-induced elevation of the plasma corticosterone level. Following intracerebroventricular administration of pentagastrin, plasma corticosterone tended to decrease and in in vitro incubation of rat pituitary tissue the addition of pentagastrin elicited a suppressive effect on the ACTH release from the tissue into the medium. It was suggested that gastrin-like peptide might control the secretion of ACTH.


Clinical trials in cancer research.

Clinical trials in cancer research. This is a review paper which gives a discussion of various aspects of clinical trials in cancer research. Since the conduct of the first randomized controlled clinical trial in cancer patients in the mid-1950's, substantial progress has been made in the utilization of the clinical trial technique for the evaluation of therapeutic efficiacy. The important elements of a protocol are given with some discussion of items to be considered in designing a protocol. The types of clinical trial (phase I, II, III) are defined, and the place of each phase of study in the context of the search for new treatments is delineated. A comprehensive discussion is given of the elements in the comparative clinical trial (phase III), including objectives, consierations in planning (comparability of treatment groups stratification of patients, feasibility and size of study, and prospective versus retrospective studies). Brief descriptions are given of designs for comparative clinical trials and a trial in oat cell lung carcinoma is discussed in some detail. Finally, some comments and references are given concerning the analysis of clinical trials.


An outbreak of foal perinatal mortality due to equid herpesvirus type 1: pathological observations. Twenty-nine cases of EHV1 infection occurred on a property, mainly in full term foals. Some foals were stillborn, some were born alive but weak and soon died and others were healthy at birth, became ill and died within 3 days of birth. Apart from voluminous, oedematous and atelectic lungs there were no gross lesions. Microscopically the lungs showed oedema, pneumonitis and bronchiolitis with intranuclear inclusions and, in many of the foals that survived over 6 hours, there was also hyaline membrane formation. Microscopic lesions were also seen in the liver, adrenal, thymus and spleen of some of these foals.


Involvement of glucocorticoids in the development of the secondary palate. Genetic differences between various inbred strains of mice in the levels of glucocorticoid receptors embryonic in maxillary mesenchyme cells appear to be reflected in the magnitude of the responses to steroids in these cells. High levels of glucocorticoids cause significant growth inhibition in maxillary mesenchyme cells with subsequent alterations in the production of extracellular matrix components. The presence of higher levels of cytoplasmic glucocorticoid receptor proteins may be one factor which could predispose those strains such as A/J to a greater inhibition of craniofacial growth in vivo by glucocorticoids and therefore increase the frequency of cleft palate production. Furthermore, women with infertility treated with glucocorticoids to support pregnancy give birth to infants with a marked decrease in birth weight [98]. Pharmacologic doses of glucocorticoids can also cause a dramatic reduction in the growth of a number of fetal tissues in mice and humans. In fact, there is evidence that glucocorticoids may be a causative factor in the production of cleft palate in primates [52]. The nature of the molecular elements which determine the biochemical and physiologic responses to glucocorticoids in the palate still remains largely unknown. Although in the mouse there is some evidence to suggest that the major histocompatibility locus (H-2) might be involved, the level(s) at which this control is exerted is unknown. It is possible that this locus may regulate in some manner the level of glucocorticoid receptors and the response to glucocorticoids in the secondary palate. Moreover, there is evidence to suggest that other genes distinct from, but closely linked to the H-2 locus may be important in determining both the strain-dependent differences in susceptibility to glucocorticoid-induced cleft palate and the intracellular levels of cyclic AMP in the secondary palate. It is also apparent that glucocorticoids in conjunction with other hormones or growth factors such as epidermal growth factor and agents which regulate cyclic nucleotide metabolism are essential for the normal development of the secondary palate. Excesses or deficiencies in either the level of these growth regulators and/or in their receptors in specific fetal tissues at defined periods in development are likely to lead to certain fetal malformations. Definition and integration of the genetic, biochemical, and endocrine factors which are involved in the control of cellular growth as influenced by alterations in the composition of cell surface and extracellular matrix components should provide some insights into the events associated with normal palatogenesis.


The ultrastructure of renomedullary interstitial cells in short duration hypercalcemia induced by vitamin D3. Under short duration hypercalcemia induced by pharmacological doses of vitamin D3 significant ultrastructural changes were observed in the renomedullary interstitial cells of rats. The most striking alteration was the degranulation of these cells accompanied with the increase in volume of the rough and smooth-surfaced endoplasmic reticulum, enlargement of the Golgi apparatus and occurence of osmiophilic inclusions probably of lipid nature in mitochondria. The ultrastructural changes can be regarded as an expression of the increase of a synthetic and secretory activity of the renomedullary interstitial cells and they may be associated with an enhanced production of prostaglandins or other lipid hormonal substances than prostaglandins under condition of hypercalcemia.


[Effects of diethyldithiocarbamate on ejaculatory inhibition and change in monoamine levels of accessory glands (author's transl)]. Effects of diethyldithiocarbamate(DDC) on erection and ejaculation were investigated and monoamine levels in central nervous system, gonad and accessory glands were determined in dogs. Intraperitoneal administration of 50, 75 and 100 mg/kg of DDC suppressed ejaculation 1 hour after administration, although penial erection did occur. This suppression of ejaculation was recovered to some extent 3 hours after DDC and completely recovered 24 hours after DDC. One hour after the administration of 100 mg/kg of DDC, noradrenaline (NA) level in the caudate and serotonin (5-HT) level in the posterior hypothalamus decreased significantly, as compared with the control. However, there were no significant changes in the regions of anterior hypothalamus and hippocampus which are known to play an important role in sexual behavior. In the epididymis, NA and adrenaline levels decreased significantly, though 5-HT levels increased, as compared with those of the control. In the prostate and posterior urethra, NA levels decreased significantly, as compared with the control. The seminal emission induced by hypogastric nerve stimulation induced a gradual rise in the posterior urethral pressure. When the pressure reached the maximum level, rhythmic alterations of the pressure considered to be a phenomenon associated with ejaculation occurred. DDC (100 mg/kg) abolished both seminal emission and rhythmic alterations of the posterior urethral pressure. These findings indicate that the sperm transport may be inhibited by decrease of NA level in accessory glands.


[Effect of thiol compounds on experimental liver damage (I). Preventive effect of tiopronin (2-mercaptopropionylglycine) on liver damage induced by carbon tetrachloride (author's transl)]. Effects of oral administration of tiopronin (2-mercaptopropionylglycine) on acute liver damage induced in rats by intraperitoneal administration of carbon tetrachloride (CCl4) were investigated. Tiopronin suppressed increase in serum transaminase activity, accumulation of liver triglyceride and decrease of liver glucose-6-phosphatase activity induced by CCl4. CCl4 induced a significant decrease of nonprotein thiol (NPSH) in the liver 24 hr after administration, but this decrease did not result in an increase of nonprotein disulfide in the liver. Tiopronin suppressed the decrease in NPSH induced by CCl4, but did not influence NPSH in normal rats 24 hr after administration. In addition to these biochemical findings, it was also noted that tiopronin prevented necrosis and decrease of glycogen in liver, as determined histologically. Other compounds such as cystamine, cysteine and glutathione proved to have a preventive effect on CCl4-induced liver damage as in the case of tiopronin. It was revealed that such preventive effect correlated to some extent with NPSH content in liver as well as serum transaminase activity and histological findings.


[Neuropharmacological studies on tolperisone hydrochloride (author's transl)]. Neuropharmacological properties of tolperisone hydrochloride (2,4'-dimethyl-3-piperidinopropiophenone hydrochloride) were investigated in mice, rats and cats. Tolperisone inhibited the spontaneous movement and methamphetamine-induced hyperactivity in mice and the ED50 was approx. 50 mg/kg, s.c. At this dose, tolperisone did not prolong the pentobarbital-induced sleeping time. Tolperisone inhibited convulsions induced by pentylenetetrazol, nicotine and maximum electric shock, but did not affect convulsions induced by strychnine and picrotoxin. Tolperisone induced muscle relaxation in mice and rats in several pharmacological tests, but did not affect neuro-muscular transmission. Tolperisone did not affect conditioned avoidance response in rats and methamphetamine-induced rotational behaviour in nigro-lesioned rats. Tolperisone reduced decerebrated rigidity in cats with i.v. administration of 5 approximately 10 mg/kg and intraduodenal administration of 50 approximately 100 mg/kg. Tolperisone elicited a slight drowsy pattern in the spontaneous EEG of cats at 5 approximately 10 mg/kg, i.v., and inhibited the EEG arousal response and pressor response to stimulation of mesencephalic reticular formation or posterior hypothalamic area. These results suggest that inhibition of the activity in the gamma pathway descending from the mesencephalic reticular formation may be involved in the mechanism of muscle relaxant action of tolperisone.


[Effectiveness of a new "paramunity" inducer (PIND-AVI) for human beings and animals]. Attenuated and by treatment with gamma rays inactivated avian Pox viruses (PIND-AVI) stimulate in man and animals selectively the T-lymphocytes. In this way PIND-AVI helps to repair a reduced reactivity of lymphocytes damaged by cytostatic chemotherapy: The lymphocyte stimulation by phytohemagglutinin could be normalized. In immunodeficient patients PIND-AVI showed very good effects in the treatment of herpes Virus infections.


New cell line. Epithelial cell line and subline established from premalignant mouse mammary tissue. A cell line and subline with epithelial characteristics were established from mouse mammary hyperplastic alveolar nodules (HAN). The cells do not grow in suspension cultures in vitro or form tumors in vivo. The cells do produce significant amounts of C-type and A-type virus and low amounts of plasminogen activator.


Induction of alkaline phosphatase activity in HeLa cells by sodium butyrate. The induction of HeLa cell alkaline phosphatase activity by sodium butyrate could be inhibited by the coadministration of caffeine or theophylline. The inhibitions were dose dependent, and at any given concentration the potency was theophylline greater than caffeine. Although the induction by sodium butyrate was more sensitive to the inhibition by the xanthines than was that produced by 5-iodo-2'-deoxyuridine, the magnitudes of the increases in cyclic AMP concentrations after treatment with the xanthines were similar in the inhibition of both types of induction. The induction of alkaline phosphatase activity by sodium butyrate also produced a shift in the thermostability pattern of the enzyme, with a proportionately greater increase in the heat-labile, rather than heat-stable, form of the activity.


Ultrastructural and autoradiographic observations of hamster cheek pouch epithelium grown in vitro. Epithelial outgrowths from hamster cheek pouch explants were cultured for varying periods of time up to 22 days. Growth of the epithelial sheets was monitored, employing colcemid for demonstrating mitotic activity and tritiated thymidine for DNA synthesis. Mitoses and thymidine uptake were observed among epithelial outgrowths at a considerable distance form the original explant. The epithelial nature of the growing cell sheets was confirmed, employing electron microscopic techniques. The cells exhibited the presence of tonofilaments, desmosomes, ribosomes, Golgi, mitochondria, and rough endoplasmic reticulum. The cultured explants were treated with cyclic nucleotides in order to investigate their modulatory effects on epithelial cell differentiation. Dibutyryl cAMP induced marked mitotic inhibition (46.3%) in our assay, which was increased to 57% with the addition of theophylline. Dibutyryl cGMP showed only a mild (5%) stimulatory effect on mitotic activity. Dibutyryl cAMP enhanced keratinization in the epithelial cell outgrowths with the biogenesis of keratohyalin granules, whereas dibutyryl cGMP did not produce any observable alterations.


Improved basal medium for Y-1 mouse adrenal cortex tumor cells in culture. I. Dependence of growth and steroid response on calcium ion concentration. An improved basal medium is presented that required only minimal supplementation with dialyzed fetal bovine serum or bovine serum albumin and fetuin to be comparable to Ham's F-10, which requires 15% horse serum (HS) and 2.5% fetal bovine serum (FBS) for the growth and function of Y-1, mouse adrenal cortex tumor, cells. Cell monolayers maintained for up to 2 weeks without any protein supplementation have retained their steroid response to ACTH. The medium differs from Ham's F-10 in its buffer composition and higher calcium-ion concentration. This medium should be a useful adjunct to studies pertaining to steroid and lipid intermediary metabolism, the retention of a specialized physiological function in a chemically defined medium, and the mechanism of hormonal response.


Concurrent replication of a papovavirus and a C-type virus in the CCL 33 porcine cell line. A papovavirus, CCL 33 PV, isolated from a porcine cell line, CCL 33 (GT), was characterized. Based on a comparison of four isoenzyme systems, CCL 33 (GT) and CCL 33 (ATCC), obtained directly from the American Type Culture Collection, appeared to be the same. In addition to the previously characterized C-type virus of CCL 33 cultures, CCL 33 (GT) produced CCL 33 PV in high quantity, but CCL 33 (ATCC) produces a papovalike virus, presumably the same as CCL 33 PV, in barely detectable amounts. Serological results showed that CCL 33 PV is apparently identical to a papovavirus (SPV) isolated by Newman and Smith after inoculation of CCL 33 with concentrated porcine trypsin. These studies extend the characterization of this papovavirus, demonstrating that CCL 33 PV is weakly hemagglutinating after neuraminidase treatment, has a high affinity for a component of fetal bovine serum and is highly infectious in appropriate porcine cell systems rather than very defective as reported previously. Moreover, it was concluded from the data that CCL 33 PV is probably indigenous to the CCL 33 porcine cell line.


Serological studies on the antigenic relationship between herpes simplex virus and varicella-zoster virus. If crossreacting antibodies between varicella-zoster virus (VZV) and herpes simplex virus (HSV) exist, one would expect more positive reactions with VZV in a group of HSV positive patients than in a group of HSV negative patients. This statement can only apply to a group of individuals where positive and negative reactions with respect to HSV and VZV are evenly distributed. Such a distribution can only be found among children. Therefore, the relationship between HSV-1 and VZV was the only one which was considered in this investigation, since the incidence of HSV-2 antibodies in children is very rare. The sera from 197 children were examined using the neutralization test (NT), the complement fixation test (CFT) and the indirect immunofluorescent assay (IFT) and could be classified as either HSV positive (80) or HSV negative (117). The children's ages were similar in both groups. Approximately the same proportion of VZV positive sera was found in both groups when examined using IFT (53% in the HSV positive and 47% in the HSV negative group). However, when the CFT was applied the proportion of VZV positive sera in the two groups differed markedly (22% of the HSV positive sera and 38% of the HSV negative sera). These findings suggest that crossreactivity observed between HSV and VZV in acute HSV and VZV infections is evidently not dependent on crossreacting antibodies but is apparently confined to the cellular level of the immune response.


Generation of virus specific cytotoxic T cells in vitro. III. Spleen cells stimulated by viral antigens generate alloreactive cytotoxic T cells. Cytotoxic T lymphocytes (CTL) from DBA/2 strain mice primed with Sendai virus (SV) in vivo were activated by secondary stimulation of spleen cells with viral antigens in vitro and analyzed for their target antigen specificity. These effector cells lysed syngeneic Sendai virus infected target cells, marginally a variety of non-infected targets and had a strong cytotoxic effect on H-2b targets. Studies on the antigenic requirements revealed that all SV preparations which generated specific CTL also induced the alloreactive populations. Similar results were found in the response to Newcastle disease virus (NDV) and some influenza A viruses; all these viruses were mitogenic for lymphocytes. Experiments on the cellular requirements indicated that virus specific and alloreactive cells can be separated by their requirements for help and for restimulation. By competition experiments both activities could be attributed to clearly separable T cell subpopulations. The induction mechanism of alloreactive T cells by viral antigens is discussed.


Generation of effector cells from T cell subsets. I. Similar requirements for Lyt T cell subpopulations in the generation of alloreactive and H-2 restricted killer cells. Lyt T cell subsets involved in the generation of H-2 restricted and alloreactive cytotoxic effector cells were analysed using anti Lyt antisera. Our data show that Lyt 1, 2, 3+ T cells are required for the induction of primary and secondary H-2 restricted and TNP-specific killer cells. In contrast, primary and secondary H-2 restricted and virus-specific T effector cells were obtained from selected Lyt 2, 3+ T cell populations and were not dependent on the presence of Lyt 1, 2, 3+ T cells. Allogeneic responses to selected K, I, or D region differences were obtained only in the presence of Lyt 1, 2, 3+ T cells; yet alloreactive killer cells were effectively generated from selected Lyt 2, 3+ T cell popluations deprived of Lyt 1, 2, 3+ T cells, when responder and stimulator cells differed at either K + D, K + I, I and D regionre is no qualitative difference between alloreactive and H-2 restricted cytotoxic responses in their requirements for particular Lyt T cell subsets. The findings indicate that the number of different antigenic determinants rather than their association with MHC self determinants is critical for the requirement of Lyt 1, 2, 3+ T cells during the sensitization phase.


The role of angiotensin in the control of blood pressure during sodium depletion. Sodium depletion was induced in dogs to raise plasma renin activity (PRA) from 1.11 to 26.48 ng/ml/hr. Little overall change in blood pressure (BP) occurred, but cardiac output (CO) and central venous pressure fell, while total peripheral resistance and heart rate (HR) increased. A nonapeptide converting enzyme inhibitor (CEI) produced a fall in BP which was linearly related to log. PRA; the intercept with PRA was at 1.05 ng/ml/hr, close to the average value for dogs on a normal diet. The fall in BP with this agent was not accompanied by an increase in HR or CO. When Sar1-Ala8 angiotensin II was used to antagonize the action of angiotensin, the fall in BP was also linearly related to log. PRA. However, for a given level of PRA this fall in BP was less than that achieved with CEI and the intercept of BP fall with PRA was 2.6 ng/ml/hr. Again with this agent there was little change in HR or CO as BP was reduced. Thus, both antagonists lowered peripheral resistance without exciting the homeostatic reflexes indicating that, as PRA rose above the normal resting level, the angiotensin generated had both a direct and indirect effect in maintaining BP.


A serological survey of St Lucia.

A serological survey of St Lucia. A serological survey of a random sample of 541 of the population of St Lucia was undertaken. The prevalence of antibodies to dengue, herpes virus, VZ, rotavirus, rubella and syphilis is described and compared with other communities.


Angiotensin and sodium balance: their role in chronic two-kidney Goldblatt hypertension. The purpose of this study was twofold: 1) to determine whether the failure of rats with chronic renovascular hypertension to respond to the angiotensin II antagonist (AIIA) with a decrease in mean blood pressure (BP) was dur to the agonistic effect of the antagonist; and, 2) if this was not the case, to examine whether a positive sodium balance impaired the reversal of the hypertension, after unclamping, in the rats that did not respond to angiotensin inhibitors. For this purpose, rats with chronic, two-kidney Goldblatt hypertension (one renal artery clamped and contralateral untouched) were tested for their BP response to the AIIA (1-Sar-8-Ala-angiotensin II) and to the converting enzyme inhibitor (CEI) SQ20,881, which is devoid of agonistic effect. Approximately 50% of the rats responded to both inhibitors either with no change or with a decrease in BP of less than 20 mm Hg (nonresponders). The other 50% had a decrease in BP of 20 mm Hg or greater (responders). The decrease in BP produced by the AIIA and the CEI correlated significantly (r = 0.76). Nonresponders to both inhibitors were unclamped or sham unclamped. A positive sodium balance was produced before surgery by injecting either 400 or 1000 microEq of sodium and was maintained for 12 hours. Direct BP significantly decreased 12 hours after surgery in the unclamped rats despite a continuous positive sodium balance. In the sham unclamped rats, BP did not change. These data indicate that the failure to respond to the AIIA is not due to the agonistic effect of this peptide. Furthermore, these data suggest that a positive sodium balance is not a major pathogenetic factor in maintaining the high BP in the nonresponder rats, since a positive sodium balance failed to maintain the hypertension after unclamping.


Role of renin-angiotensin system in chronic renal hypertensive rats. The role of renin-angiotensin system has been examined in the maintenance of hypertension in acute and chronic two-kidney (36 weeks) and chronic one-kidney (12 weeks) Goldblatt hypertensive rats using three inhibitors of this system. The inhibitors used were URI-73A, a synthetic analog of lysophosphatidylethanolamine, which inhibits renin both in vivo and in vitro, SQ14,225, a potent converting enzyme inhibitor, and [Sar1, Thr8] angiotensin II, an angiotensin II antagonist. When the inhibitors were administered in acute (high renin) hypertensive rats, they all lowered blood pressure significantly. However, in the chronic (low renin) hypertensive phase, both renin and converting enzyme inhibitors lowered blood pressure, whereas, Sar1, Thr8 failed to lower blood pressure. The renin inhibitor lowered plasma renin activity (PRA), and SQ14,225 and [Sar1, Thr8] Ang II increased PRA. Further studies on water and electrolyte balance with one-kidney model hypertensive and uninephrectomized control rats showed no change in plasma volume. However, there was increased 24-hour urinary output and increased sodium excretion. This study indicates that in chronic renal hypertensive rats, blood pressure reduction is possible by either renin on converting enzyme inhibitor, but not by angiotensin antagonists. Since volume did not change either during the development or reversal of hypertension, volume did not appear to play a major role in the maintenance of hypertension.