A direct radioimmunoassay for plasma renin in mice and its evaluation.

The pioneering work of Cohen, S., Taylor J.M., Murakami, K., Michelakis, A.M. and Inagami, T. ((1972) Biochemistry 11 4286-4293) with isolation of submaxillary renin and the first direct radioimmunoassay for renin subsequently described by Michelakis A.M. Yoshida H., Menzie, J., Murakami, K. and Inagami, T ((1974) Endocrinology 94, 1101-1105) was reinvestigated and confirmed. In addition a detailed evaluation of the assay is described by characterization of the immunoreactivity of the labelled renin and demonstration of immunological identity between plasma renin and purified renin standards. Assay and equilibrium conditions were characterized and optimized and the association constant for the antibodies determined to be of the order of 10(11)1/mol. The validity of the direct assay for renin was further investigated by a comparison with another radioimmunoassay for renin; the antibody trapping method, which measures the enzymatic activity of renin in plasma. The correlation coefficient is high (0.91) but does not exclude that the direct assay measures a hypothetical prorenin in plasma as well.


Direct measurement of high molecular weight forms of renin in plasma.

Gel filtration of normal mouse serum revealed the presence of high molecular-weight forms of renin, in addition to the well-recognized 40 000 dalton form. The distribution of renin was measured by using both the antibody trapping method, which measures renin's enzymatic reactivity, and by a direct radio-immunoassay, which detects the renin molecule by its antigenic properties. A well characterized and distinct peak corresponding to a molecular weight of 200 000 or greater was always present. A less distinct peak was seen between 70 000 and 140 000 daltons. The well separated and symmetrical peak at 40 000-45 000 daltons was secreted from both kidneys and submaxillary glands and disappeared almost completely after combined nephrectomy and sialoadenectomy in contrast to the forms of higher molecular weight. The higher the molecular weight the lower was the specific enzymatic reactivity found to be. The 200 000-dalton peak was mainly detectable by the direct assay, although it was demonstrated to contain enzymatic reactivity to a small extent. The 40 000-dalton peak had a specific enzymatic reactivity and molecular weight identical to that of pure submaxillary renin.


Quantitative polyacrylamide gel electrophoresis and specific activities of human somatotropin and its derivatives.

A preparation of human pituitary somatotropin examined in quantitative polyacrylamide gel electrophoresis is conformationally compact. The derived molecular weight by quantitative electrophoresis is consistant with the known mass of the hormone and value obtained by other methods. A plasmin-modified somatotropin is more compact and shows full activity in immunoassay, and in lymphocyte binding and somatotropic assays, with enhanced lactogenic activity. The N-terminal 134-amino acid fragment and a hendekakaihekaton fragment exist in non-monomeric forms. The N-terminal fragment has immunologic and biologic activity, with greatest activity in the in vivo somatotropic assay. The hendekakaihekaton fragment exhibited only marginal activity. All preparations showed heterogeneity of charge in quantitative electrophoresis with discreet charge isomerism recognizable for the native and plasmin-modified preparations.


Primary structures of alpha-crystallin A chains of elephant, whale, hyrax and rhinoceros.

As part of a study of the evolutionary development of the eye lens protein alpha-crystallin the 173-residue A chain of this protein has been studied in elephant, whale, hyrax and rhinoceros. The primary structures were inferred mainly from amino acid compositions of peptides obtained by enzymic digestions and CNBr cleavage. The positions of substitutions, as compared to the known bovine A chain, were confirmed by Edman degradation. In accordance with the previously observed slow rate of evolution of the A chain only a small number of substitutions was found among these species. Elephant and hyrax share a number of unique substitutions, strongly indicating a common ancestry of these two species within the mammalian class.


Rat liver alpha-tocopherol binding protein.

1. The properties of rat liver cytoplasmic alpha-tocopherol binding protein have been studied. 2. The binding protein sedimented in the 3 S region of sucrose density gradients, and gel filtration indicated an approximate molecular weight of 30 500. 3. Of the tissues examined by the present assay, binding was detectable only in the liver. 4. Optimal binding was achieved by incubation at 26 degrees C for 4 h and was independent of pH between 7.4 and 9.0. 5. Pronase completely abolished binding. The binding protein was, however, almost completely resistant to trypsin, and unaffected by RNAase, DNAase, triacylglycerol lipase, and phospholipase C. 6. A variety of tocopherol analogues and other lipid-soluble compounds were tested for their ability to compete for binding. Only alpha-tocopherol and to a lesser extent alpha-tocotrienol and gamma-tocopherol exhibited competition. alpha-Tocopherol acetate, alpha-tocopherol quinone and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid had no effect on binding. 7. Tocopherol binding was reversible, and the tocopherol was not metabolized during incubation.


Studies on the flavins in rat liver mitochondrial outer membranes.

The incorporation of radioactivity derived from [2-14C] riboflavin into the flavins of rat liver mitochondrial outer membranes was studied. These membranes were found to contain about 0.6 nmol of non-covalently bound flavins per mg protein; the majority is in the form of FAD (73%) and FMN (24%). The membranes also contain about 1.5 nmol per mg of covalently bound flavins. After labeling, radioactive flavins appeared in the non-covalently bound flavins for about 4 h. Most of this radioactivity was in FAC (77%). Neither the rate nor extent of this labeling was affected by cycloheximide (1 mg/kg) administered 30 min prior to the radioactive riboflavin. With the covalently bound flavins, radioactivity was incorporated into the coenzymes for at least 18 h, but the rate of incorporation was much slower. After cycloheximide, radioactive flavins continued to appear in covalently bound flavins for about 2 h, but then stopped. Labeling of both types of flavins after [14C] riboflavin was considerably slower than the incorporation of [3H] leucine into outer membrane proteins. These results suggest that with flavoproteins from the mitochondrial outer membranes, the incorporation of flavins occurs after synthesis of the various apoenzymes is complete.


Iron removal from transferrin. An experimental study.

We have studied the facilitation of iron transfer from transferrin to desferrioxamine by various anions. Most of the anions which can substitute for HCO-3 in the ternary complex of transferrin - Fe - HCO3 do not facilitate iron transfer; anions which do facilitate iron transfer do not necessarily form stable ternary complexes. Combinations of anions effective in transfer have a less-than-additive effect, suggesting a common reaction pathway. We suggest that the transfer of iron from transferrin to desferrioxamine involves a substitution step and a subsequent chelation step, and that the efficiency of the overall reaction is a function of both these attributes of the anion.


Formation and characteristics of hepatic dexamethasone-receptor complexes of different molecular weight.

The dexamethasone-binding receptor protein in rat liver cytosol has a Stokes radius of 61 A and a sedimentation coefficient of 4.0 S. In contrast, cell nuclei labelled with [3H]dexamethasone in vivo or in vitro (reconstitution experiments with [3H]dexamethasone-labelled cytosol and isolated unlabelled nuclei) contain a high-salt-extractable dexamethasone-receptor complex with a Stokes radius of 30-36 A and a sedimentation  coefficient of 3.2 S. Exposure of liver homogenate or 1000 X g homogenate supernatant to low ionic strength during preparation of cytosol resulted in conversion of the 61 A to a 36 A complex very similar to the intranuclear form of dexamethasone receptor. 61 leads to 36 A complex-converting activity was present in both the 100 X g-10 000 X g sediment of liver homogenate, from which it could be extracted by hypotonic media, and in the liver cell nuclei, from which it could be extracted by hypertonic media. Mild digestion of the 61 A dexamethasone-receptor complex with trypsin also gave rise to a complex with a Stokes radius of 36 A. Reconstitution experiments with isolated liver cell nuclei indicated that both the 61 A and 36 A dexamethasone-receptor complexes were taken up by the nuclei; reextraction of the nuclei incubated with the 61 A complex revealed that this form had been converted to the 30-36 A complex. Further digestion of the 61 and 36 A [3H]dexamethasone-receptor complexes with hypotonic extract of the 1000 X g-10 000 X g sediment of liver homogenate or with trypsin resulted in formation of a third complex with Stokes radius of 19 A and a sedimentation coefficient of 2.5 S. The approximate molecular weights of the 61, 36 and 19 A dexamethasone-receptor complexes were calculated as 102 000, 46 000 and 19 000, respectively, and the frictional ratios of the molecules as 1.84, 1.38 and 1.00, respectively. It is concluded that the nuclear 30-36 A dexamethasone-receptor complex is formed from the cytosol 61 A complex by proteolytic digestion and that this latter protein contains at least two sites with a relatively high sensitivity to protelytic cleavage.


Regulation of extracellular protease production in Candida lipolytica.

Production of extracellular protease by Candida lipolytica NRRL Y-1094 was derepressed upon transfer to carbon-, nitrogen- or sulphur-free medium but not upon transfer to phosphorus-free medium. The protease activities produced under the three nutrient limitations had alkaline pH optima and similar substrate and inhibitor specificities. Any one of the following three conditions was found to be sufficient for derepression of extracellular protease: (a) "poor" carbon source, (b) cysteine intracellular pool below 0.5 micronmol/g dry weight cells and (c) ammonia intracellular pool below 10 micronmol/g dry weight cells. Thus, extracellular protease production in C. lipolytica was subject to at least three different regulatory controls, carbon, sulphur and nitrogen repression. Intracellular cysteine and ammonia appeared to be the metabolic signals for sulphur and nitrogen repression, respectively. Anabolic glutamate dehydrogenase did not act as a regulatory protein mediating nitrogen repression. Exogenous protein had an inductive effect on extracellular protease production.


The partition of glycosaminoglycan-quaternary ammonium complexes, II. The effects of polymer molecular weight and sulfation.

Previous results have shown the possibility for obtaining high-resolution separations of glycosaminoglycans by partition in butanol/aqueous two-phase systems containing quaternary ammonium salts. In this paper, the effects on partition behavior of both polymer molecular weight and sulfation were examined. Two series of fractionated chondroitin sulfate polymers were isolated in which the molecular weight and sulfation varied systematically. In the molecular weight series the six samples, spanned the range from 3200 +/- 300 to 19 700 +/- 500 and each sample carried 0.8 sulfate groups per uronic acid residue. In the sulfation series, each sample had an essentially constant molecular weight of 13 000, but the sulfation varied from 0.58 to 0.88 sulfate groups per uronic acid. The C50 of each of these samples was determined in the 1-butanol/aqueous NaCl phase system containing 1% hexadecylpyridinium chloride. In the series wherein the molecular weight varied, the C50 increased with molecular weight up to 12 000 where a limiting value was reached. In the series wherein the sulfation varied, a linear relationship was found between the C50 and the square of the number of anionic substituents per disaccharide. These results show that fractionation by partition techniques will be sensitive to the anionic nature of the polymer, but for the common connective tissue glycosaminoglycan, there will be no fractionation according to molecular weight.


Subcellular localization of enterokinase (enteropeptidase EC 3.4.21.9) in rat small intestine.

The subcellular localization of enterokinase is controversial. In this study, enterokinase was extracted from a soluble fraction and a brush border fraction of rat small intestine by differential centrifugation. The soluble fraction contained 41% of the initial enterokinase activity while the brush border fraction contained only 4.6% of the initial activity. In contrast, alkaline phosphatase monitored as a brush border marker, yielded 26.3% in the brush border fraction and only 6% in the soluble fraction. Further separation of the soluble fraction on a Sepharose 4B column revealed three peaks of enterokinase activity. One small peak (3%) of a bound enzyme (Mr, 2 - 10(6)) and two larger peaks of free enzyme (Mr, 3 - 10(5) and 9 -10). In contrast, alkaline phosphatase major fraction was in a high molecular weight peak of bound enzyme. When the brush border fraction was chromatographed only a single peak of bound enterokinase and alkaline phosphatase were found. In the lower part of the small intestine, no brush border-bound enterokinase was found, while the peak of alkaline phosphatase was the same as in the upper intestine. These data suggest that enterokinase activity in the rat intestine is mainly in a free form localized in the mucin and soluble fraction and to a negligible extent in the brush border.


Rapidly metabolized glycoproteins in a neuroblastoma cell line.

The metabolism of neuroblastoma cell glycoproteins was examined using L-E13H]fucose. Incubation of monolayer cultures with [3H]fucose resulted in a rapid uptake of the radioactive precursor and its incorporation into acid-insoluble macromolecules. Less than 3% of the [3H]fucose that was isolated from neuroblastoma cells by trichloroacetic acid precipitation was associated with glycolipids. The metabolism of fucosylated macromolecules was studied in cells which were labelled to a steady state, and then reincubated under conditions which limited reutilization of the radioactive precursor (40 mM unlabelled fucose). During reincubation of the cells, we observed a rapid metabolism (27% by 2 h) of the prelabelled macromolecules which stabilized within a cell generation time to give an overall rate of turnover of 9%. This rapid loss of radioactivity from the cells was not due to exocytosis since less than 4% of the [3H]-fucose was lost into the media as macromolecules during a 5 h reincubation period. The presence of 40 mM fucose in the media did not affect cell growth until after 24 h of incubation or cellular protein synthesis until after 15 h of incubation. When the metabolism of neuroblastoma cell glycoproteins was measured in the presence of 1.8 - 10(-4) M cycloheximide, there appeared to be a less rapid decrease in cell-associated specific activity, and an increased reutilization of [3H]fucose. Although the major proportion of the radioactivity remained as e13H]fucose, extensive incubation of neuroblastoma cells with this radioactive precursor led to increased amounts of tritium associated with other cellular components; However, a rapid rate of glycoprotein metabolism could also be demonstrated with cells incubated with [14C]fucose. This eliminated the possibility that the above results were restricted to the tritiated precursor and merely a reflection of hydrogen-tritium exchange.


Metabolism of naturally occurring propenylbenzene derivatives. III. Allylbenzene, propenyl benzene, and related metabolic products.

The acidic and neutral urinary metabolites of allylbenzene, propenylbenzene, 1'-hydroxyallylbenzene, and cynnamyl alcohol were identified and related through a common metabolic scheme. The rat metabolizes allylbenzene to 1'-hydroxyallylbenzene which can rearrange to yield cinnamyl alcohol whick is further metabolized. This mechanism is proposed to account for the appearance of "propenyl type" metabolites from allylbenzene compounds. Propenylbenzene is oxidized to cynnamyl alcohol. Both 1'-hydroxyallylbenzene and cunnamyl alcohol are excreted unchanged in the neutral extract when given to rats. Allylbenzene and 1'-hydroxyallylbenzene yield basic ninhydrin-positive metabolites. Allylbenzene is first oxidized on the benzylic carbon to form 1'-hydroxyallylbenzene, which is further oxidized to form phenyl vinyl ketone, which condenses with the secondary amines piperidine and dimethylamine to form tertiary aminopropiophenones (Mannich bases). Analogous compounds, cinnamyl alcohol and propenylbenzene, do not yield Mannich base metabolites. This proposed metabolic scheme is consistent with the chemical mechanism operative in the synthesis of Mannich base from allylbenzene via chromic acid oxidation followed by amine addition.


Effect of the protein synthesis inhibitor cycloheximide on RNA synthesis in radish seedlings.

The effect of cycloheximide on rRNA and poly (A) (+) RNA synthesis is studied in radish seedlings. When used at a concentration of 50 microng/ml cycloheximide selectively blocks rRNA synthesis without altering poly (A) (+) RNA synthesis. Processing of the 2.3 X 10(6) dalton pre-rRNA is severely impaired. This observation should help in studies of mRNA metabolism in plants. When the concentration of cycloheximide is lowered, it is possible to completely block protein synthesis without preventing RNA synthesis. This implies that the effect of cycloheximide on RNA synthesis is probably more complex than usually assumed.


Differentiation in vivo and in vitro of pyruvate kinase isozymes in rat muscle.

The M type isozymes of Pyruvate-kinase have been studied by isoelectrofocusing in thin layer acrylamide ampholine gel, during the ontogeny of rat muscle in vivo and during the differentiation in vitro of myoblasts of a line established by Yaffe. In both cases, multiple subbands have been seen; the most acid (pHi 5.2) was the predominant band in myoblasts and in fetal muscle at the 15th day. Several more cathodic bands appear sequentially in vitro as in vivo, one of them corresponding to the "M2" or "K" band (predominant in kidney). The most cathodic band M1 (pHi 7.3), characteristic of the adult muscle, appears at the 9th day of culture in vitro in multinucleated myotubes and at the 20th day of fetal life in vivo. Kinetic results confirm these electrofocusing results, showing in fetal muscle and in myoblasts a sigmoid saturation curve of pyruvate kinase activity with phosphoenolpyruvate as substrate. This allosteric kinetic is progressively replaced by a Michaëlian kinetics in vitro as in vivo. Consequently, the studies in vitro may serve as a model for myogenesis in vivo, and may contribute to the understanding of the significance of the multiple forms of pyruvate-kinase during this myogenesis.


Bépridil a new effector of oxidative phosphorylations.

Bépridil action on oxidative phosphorylations depends on the nature of the oxidized substrate. Thus it seemed to act as an uncoupler on the FAD-linked substrates as we have shown with succinate. But this property could not be applied to the NAD-linked substrates. Thus, in the presence of Bépridil, ADP would have opposite effects on the electron flow across the first site of oxidative phosphorylations; from one side the direct electron flow was decreased while from the other side, the reverse one was increased. Confirming this difference in its action, Bépridil did not affect the oxidative phosphorylation property of the cytochrome oxidase as could be deduced from the invariability of TMPD + ascorbate oxidation whether Bépridil was added or not. Moreover the effect of Bépridil on the P/O ratio was a dramatic demonstration of its selectivity. As a matter of fact this ratio was increased to a value near 5 for the NAD-linked substrate while it was decreased to near 0.4 with the FAD-linked substrate.


[Presence of a pathway for biosynthesis of UDP-glucose in the Golgi apparatus of rat hepatocytes].

Glucokinase, phosphoglucomutase and glucose-1-phosphate uridylyltransferase are the three enzymes involved in a microsomic pathway for the synthesis of UDP glucose. Evidence is given, in this paper, for the localization of these three enzymes in a Golgi-rich fraction of rat liver. This fraction is prepared, from smooth microsomes, by the means of a discontinuous four-step sucrose gradient. Three of the lighter fractions (d = 1.08-1.13) are enriched in the Golgi markers (galactosyltransferase, sialytransferase and thiamin pyrophosphatase), especially the one with density 1.13. The three enzymes we are interested in are enriched in the two upper hands (d 1.08-1.11), which display an activity for the biosynthesis of UDP-glucose from glucose equivalent to the one obtained in a crude microsomic preparation, and which are not contaminated by other subcellular components.


Lipids of the yeast Hansenula anomala.

An analysis of the free lipids of Hansenula anomala was performed. The main fatty acids obtained by saponification of whole cell crude lipids were palmitic, C18:1, C18:2 and C18:3 acids. In mitochondrial lipids the tri-unsaturated acid was present as traces. Fatty acid composition of each class of lipids was also determined. Phosphatidylcholine and phosphatidylethanolamine were the main phospholipids; phosphatidylserine, phosphatidylinositol and cardiolipin were also characterized. The most abundant sterols were ergosterol and lanosterol. An acetate of a 24-ethyl cholesterol was also isolated. Two glycolipids, a galactosyl diglyceride and a glucosyl ceramide were identified; concerning the galactosyl diglyceride, the content of C18:3 acid was higher than in other lipid classes. In the glucosyl ceramide, the main fatty acid was alpha-hydroxy C18:0 acid; C16:0, C18:1, C18:2 and C18:3 acids were present too. The long chain base was shown to be C18-phytosphingosine (4-hydroxy C18 sphinganine). Some similarities and differences with Saccharomyces cerevisiae are discussed.


Origin of the acetyl moiety of acetylcholine synthesized in rat striatal synaptosomes.

The subcellular localization of the AcCoA compartment supplying the cytoplasmic choline acetyltransferase (ChAc, EC 2.3.1.6) was investigated using a purified preparation of rat striatal synaptosomes (B fraction). It was first demonstrated that the SRA of the [14C]ACh synthesized during a 10 min incubation period was equal to the SRA of the [2-14C] and the [3-14C]pyruvate added to the isolated nerve terminal suspension. The experimental results can be summarised as follows: (i) No modification in the amount of [14C]ACh synthesized from [2-14C]pyruvatetion in the amount of [14C]ACh synthesized from [2-13C]pyruvate could be detected after the addition of high concentrations of either carnitine, acetylcarnitine or acetyl phosphate to the synaptosomal suspension. (ii) Under experimental conditions in which the amount of [1,5-14C]citrate taken up by passive diffusion into the cholinergic nerve endings would allow detection of the possible formation of the labelled ester, no [14C]ACh could be recovered. (iii) The SRA's of the individual carbon atoms of the Krebs cycle intermediary compounds when the cycle is fed with [2-14C] and [3-14C]pyruvate were calculated as a function of the STA's of each of these two precursors (a and a' respectively), of the number of 14CO2 dpm produced in the Krebs cycle from each of these two labelled compounds (D2 and D3 respectively), and as the function of the rate y of exchanges of molecules between the tricarboxylic acid cycle and other metabolic compartments. The experimental value obtained from a 10 min incubation, after the nerve endings had reached a steady metabolic activity, indicate that if the acetyl moiety of ACh was derived from some Krebs cycle intermediary compounds, its SRA could never exceed 55 per cent that of the [2-14C]pyruvate from which it is produced, (iv) No correlation could be found between the rate of [14C]ACh formation and changes in the Krebs cycle activity induced by sodium cyanide, 2-4 dinitrophenol and Ca2+ free medium. (v) The lack of significant [14C]ACh synthesis from [1-14C]acetate in striatal synaptosomes is consistent with the failure of fluoroacetate to modify the amounts of 14CO2 as well as of [14C]ACh formed from [2-14C]pyruvate. These results were interpreted as a confirmation of the presence of a low AcCoA synthetase activity in the nerve terminals. To reconcile all these data, it is proposed that pyruvate is transformed into AcCoA outside the mitochondria by the action of some cytoplasmic pyruvate dehydrogenase-like enzyme.


[A comparative study of the role of creatine phosphokinase isoenzymes in energy metabolism of skeletal and heart muscle].

It has been shown that the contents of mitochondria and mitochondrial isoenzyme of creatine phosphokinase are almost identical in skeletal and heart muscles. In mitochondria from both types of muscle creatine phosphokinase is functionally coupled to ATP-ADP translocase. This kind of coupling ensures complete conversion of mitochondrial ATP energy into the energy of creatine phosphate and effective control of oxidative phosphorylation by the creatine phosphokinase reaction. It has also been shown that all isoenzymes of creatine phosphokinase from heart and skeletal muscle have very similar kinetic properties. Significant differences have been found to exist between isoenzyme patterns of these muscles and also in distribution of different isoenzymes in the cells. In skeletal muscle cells creatine phosphokinase is present mainly as cytosolic MM isoenzyme; about 6% of total cellular activity is localised also in mitochondria. Due to high activity of cytosolic isoenzyme the total activity of creatine phosphokinase is about three times higher in skeletal muscle than in cardiac muscle. It has been also shown that phosphoenolpyruvate and glucose-6-phosphate do not have any inhibitory effect on creatine phosphokinase.


[Substrate inhibition as a cause of oscillations in an open irreversible enzymic reaction S1 + S2 in the presence of E(R,T) leads to S1' + S2'. A mathematical model].

A mathematical model of an open irreversible reaction S1 + S2 (formula: see text) catalysed by an olygomeric enzyme E(R, T) has been analysed. It is assumed that the enzyme undergoes the concerted conformational transitions R in equilibrium T in conformity with the theory of Monod, Wyman and Changeux, and one of the substrates (S2) produces inhibition of the enzyme, thus shifting the equilibrium between the two enzyme forms in the direction of T formation. A simple graphical explanation is given to the hysteresis of the input characteristic approximately v ([S1]) (approximately v is the reaction rate at d[S2]/dt=O) which gives rise to self--oscillations. The hysteresis occurs both in the case of allosteric and isosteric substrate inhibition.


[The phospholipid composition of organs and tissues of rabbit].

The phospholipid compositions of 11 organs and tissues of rabbit (brain, heart, lung, liver, muscles, kidney, spleen, bone marrow, pancreas, adrenula, and blood plasma) were comparatively studied by micro-thin-layer chromatography. Total lipid extracts of liver and brain are the most abundant in phospholipid (61,8% and 56,3%, respectively) and that of bone marrow and spleen are the poorest (1,6% and 7,1%, respectively). All the organs and tissues, except blood plasma, contain phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidylinositol, sphingomyelin and diphosphatidylglicerol; the majority of organs contain phosphatidic acid. Lyso-phosphatidyl-choline was detected in blood plasma, bone marrow, adrenula and liver. Phosphatidylglicerol was found only in the lungs. Brain has the lowest content of phosphatidyl choline (34,3%) and the highest contents of phosphatidyl ethanolamine (36,2%) and phosphatidyl serine (12,6%), whereas the heart is the richest in DPG (about 11%). Diphosphatidyl glycerol is the most abundant in the heart (approximately 11%) and phosphatidyl inositol-- in the adrenula (8,1%). Plasma is found to have a high content of choline-containing lipids; however, phosphatidyl serine and phosphatidic acid are absent altogether. Some of the organs and tissues have a similar phospholipid composition; namely brain and kidney, spleen and bone marrow, adrenula and liver.


[Structural studies on D-glyceraldehyde-3-phosphate dehydrogenase from rat skeletal muscle].

The amino acid composition of glyceraldehyde-3-phosphate dehydrogenase isolated from rat skeletal muscle was determined. Tryptic peptide maps of the carboxymethylated protein were compared with those of the corresponding derivative of the rabbit muscle dehydrogenase. Evidence from the amino acid analysis, N-terminal amino acid and the position of the peptides in two-dimensional chromatography-electrophoresis suggests a high degree of homology between the two enzymes. However, the rat muscle glyceraldehyde-3-phosphate dehydrogenase is characterised by a markedly lower histidine content. Some differences are also revealed in the peptide maps of the rat and rabbit dehydrogenases. Four cysteine residues per subunit of rat apoenzyme may be carboxymethylated in the absence of denaturing agents (pH 8,4 75 min). Two of them are modified at pH8,0 within 30 min and are both found in the peptide, which has been demonstrated to contain the most reactive cysteine residue. This suggests cysteine residue N 153 to be the second in order of reactivity towards iodacetate. No difference in the accessibility of SH groups to iodacetic acid has been found between apoenzyme samples incubated in 0,15 M NaCl at 20 degrees and 4 degrees respectively. This indicates that enzyme inactivation caused by dissociation which occurs at low temperature brings about no measurable alterations in the SH group reactivity as compared to that observed at 20 degrees.


[Studies of oxygen uptake (Mehler reactions) in chloroplasts in the presence of silico-molybdic acid].

Silico-molybdic acid (SiMo) alone or in combination with diurone is known to provide for the functioning of various sites of electron transport within photosystem II (PS-II). Using SiMo, the ability of PS-II to competitive reduction of O2 with SiMo in the presence of some activators of O2 uptake (e. g. catalase"ethanol, malonate, oxalate and glyoxalate) was shown. Oxygen uptake (Mehler reaction) with those compounds may be accomplished against the diurone block. The Mehler reactions studied occur in PS-II at the site, uncoupled with phosphorylation, up to the primary acceptor Q, The Mehler reactions with FMN and ferredoxin confirm the fact that PS-II functioning in the presence of SiMo is possible only after its reduction. Photoreduction of NADP is inhibited irrespective of whether the oxidized or reduced form of SiMo is added.


[Benzoyl-DL-arginine-p-nitroanilidase from buckwheat seeds. Properties and substrate specificity].

A proteolytic enzyme, hydrolyzing N-benzoyl-D,L-arginine-p-nitro-anilide (BAPAase), has been isolated from the buckwheat seeds (Fagopyrum esculentum). The enzyme was purified 400-fold and was homogeneous according to isoelectrofocusing and disc electrophoresis in polyacrylamide gel. The molecular weight of the BAPAase was determined to be 65000 by gel-chromatography and 70000 by polyacrylamide gel electrophoresis. The sedimentation coefficient of the BAPAase was found to be 4.3 S, the isoelectric point--pH 4.5. The enzyme split peptide, esteric and amide bonds formed by carboxyl groups of lysine and arginine in synthetic substrates. The enzyme did not hydrolyse fibrinogen, did not activate chimotrypsinogen, weakly hydrolyzed histones and casein and strongly--protamine. The BAPAase did not hydrolyse albumins and globulins from the buckwheat seeds, and weakly hydrolyzed glutelins. The study of the products of the hydrolysis of salmine and sturine by BAPAase showed that the enzyme split internal peptide bonds in these substrates, and, thus, it is an endopeptidase.


[Effect of dissociating agents on Chlorella glutamate dehydrogenases].

Effect of urea beta-mercaptoethanol and guanidine--hydrochloride on two Chlorella glutamate dehydrogenases (GDH's) have been studied. Both GDH's are inactivated irreversibly by 2-3 M guanidine hydrochloride. Urea above 4 M rapidly inactivates only NH+4-induced NADP--GDH. Constitutive NAD(P)--GDH is stable in 8M urea solution at room temperature for a long time. beta-Mercaptoethanol does not effect significantly the stability of constitutive GDH in 8m urea. Urea above 1M being in reaction mixture inhibits constitutive GDH in a competitive manner to L-glutamate and uncompetitively regards to alpha-oxoglutarate. Taking this into account, one may conclude that L-glutamate and alpha-oxoglutarate seems to bind to different groups on the enzyme molecule.


Tissue-binding factor in schizophrenic sera: a clinical and genetic study.

The hypothesis that pathologic immune mechanisms, characterized by production of brain autoantibodies, operate in schizophrenia, was the basis for this study. Binding of serum globulin substance by human brain septal region obtained at autopsy was measured by radioimmunofixation assay in 27 schizophrenic probands, 28 first-degree relatives, 12 patients with primary affective disorder (depression), and 117 normal controls. Schizophrenic individuals tended to have higher levels of brain-serum affinity than controls. Age and sex did not appear to affect results. Within families, elevation of serum-binding activity showed intra sib-pair resemblance, distinguished healthy relatives from probands and ill relatives and relatives of probands with positive sera from relatives of probands with negative serum activity. Serum activity distinguished well relatives from normal controls and was independent of clinical state. This suggests that brain-serum affinity may be compatible with characteristics of a genetic marker of vulnerability to schizophrenia. Within sib-pairs, concordance rates for elevated serum activity and for subtype diagnosis, mode, and age of illness onset were positively related. This finding supports clinico-genetic disposition in a subgroup of schizophrenic persons. To determine distribution patterns of antigenic components, selected schizophrenic and normal sera were tested against human liver and mouse brain, thymus, and liver. Wide tissue cross-reactivity was observed in schizophrenic, but not in normal sera, a finding consistent with overlap of serological reactions affecting specific tissues in autoimmune processes. The assay employed in the present study and investigation of inheritance of brain-serum affinity have not previously been reported.


Evoked potentials of schizophrenics in several sensory modalities.

This is the initial report of results obtained by a comprehensive approach to investigation of evoked potential (EP) characteristics of psychiatric patients. EP to left and right medion nerve, visual (checkerboard flash), and auditory (click) stimuli were obtained for 14 monopolar scalp and one EOG derivation in 32 schizophrenics and 16 nonpatients, matched for age and sex. Results showed amplitude attenuation of EP events 100 msec poststimulus or later in the patients' EPs of all modalities and in most lead locations. Attenuation of later EP events thus appears to be a general characteristic of schizophrenia. Comparisons of chronic paranoid and undifferentiated with other patient subtypes revealed a relatively specific correlate of the chronic subgroup, increased negativity of a negative somatosensory peak occurring 60 msec poststimulus (N60) at contralateral central leads.


ECT versus chlorpromazine in mania.

The efficacy of ECT in the treatment of depression is well recognized. In mania its efficacy is less well acknowledged. This report compares ECT, chlorpromazine, and no active treatment in mania. Twenty-eight control patients diagnosed as manic were selected from consecutive admissions to the University of Iowa Psychiatric Hospital in the period 1935-1941 when there was no active treatment given. Twenty-eight patients treated with ECT were selected from the period 1945-1949, and 28 chlorpromazine-treated patients were selected from the period 1958-1964. Symptoms sufficient to fulfill a research diagnosis of mania had to be documented in the record for all patients. Results compare symptomatology, duration of hospital admission, discharge condition, discharge category, social recovery, and follow-up. Both ECT and chlorpromazine were superior to no active treatment for outcome measures considered. Ten chlorpromazine-treated patients did not respond satisfactorily to the chlorpromazine treatment but recovered with ECT treatment.


Effect of marijuana and placebo-marijuana smoking on psychological state and on psychophysiological cardiovascular functioning in anginal patients.

Ten male anginal patients with angiographically documented coronary artery disease, in a randomized double-blind crossover study, smoked one marijuana cigarette (containing 18 mg of delta-9-THC) on one morning and one placebo marijuana cigarette (containing 0.05 mg of delta-9-THC) on a successive morning. Significant increases occurred in average cognitive and intellectual impairment scores, derived from the objective content analysis of 5 min of speech, 30 mins after smoking the marijuana cigarette as compared to the placebo marijuana cigarette, and these scores decreased to near presmoking levels 60 min after smoking. No significant average changes occurred in anxiety or three hostility scale scores following smoking marijuana. Sizable individual differences were noted in the psychological responses to marijuana smoking due, presumably, to personality differences and/or differences in THC pharmacokinetics. Significant psychocardiovascular hemodynamic correlations, as measured by echocardiography, were observed during placebo-marijuana smoking between hostility inward scores and systolic blood pressure and ejection fraction, overt hostility outward scores and diastolic blood pressure, as well as between anxiety scores and stroke volume and left ventricular end-diastolic dimension and left ventricular diastolic volume. These significant psychophysiologic correlations were all eliminated during marijuana smoking. In view of associated findings that marijuana smoking decreased myocardial oxygen delivery, decreased exercise time until the onset of anginal pain, and increased myocardial oxygen demand in anginal patients, the use of marijuana by such patients is clearly inadvisable.


Can callosal speed of transmission be inferred from verbal reaction times?

Filbey and Gazzaniga (1969) found that verbal reaction times were shorter to right than to left visual field stimuli. They interpreted this reaction time difference (30 to 40 msec) to reflect callosal transmission time, i.e., the delay required for information received in the right hemisphere to be acted upon by the verbal left hemisphere. We have performed four verbal reaction time experiments with normal subjects, utilizing differing hemifield stimulus presentations and task requirements. Stimuli were: small lights (light-emitting diodes); checkerboard pattern briefly flashed; small circles; consonant-vowel-consonant triads, either meaningful or nonsense. Contrary to Filbey and Gazzaniga's observations, we found no difference between verbal reaction times to left and right half-field presentations, or a significantly shorter reaction time with left-field presentations, depending upon experimental conditions. Faster reaction times with left-field stimuli were found in left-handed as well as right-handed subjects. Our data indicate that it may be premature to infer callosal speed of transmission from verbal reaction times to half-field stimuli. The paradoxical finding of faster verbal reactions to right hemisphere visual inputs does not appear to be related to handedness, and it occurs with meaningful stimuli; this finding remains unexplained.


Concentrations of adenine nucleotides in erythrocytes of patients with gout.

Gout has long been known as a disease of the high social class or of those with achieved social status. Brooks and Mueller (1966) studied the relationships between behavior and serum uric acid levels among university professors. They indicated that personal characteristics of drive, achievement, and leadership are positively associated with the uric acid levels in plasma. Hansen and Dimitrakondi (1974) reported a marked decrease in ATP levels in psychotic depression and significant correlation between ATP concentration and mood scores. We estimated the adenine nucleotides in erythrocytes of gouty patients.


Control of the pilocarpine release rate through hydrogels by plasma treatment.

The rate of release of an aqueous solution of pilocarpine hydrochloride sequestered in hydrogel-type materials can be reduced by plasma treatment of the polymer surface. Two plasma techniques were used. The first involves exposure of the hydrogel to the effects of a glow discharge sustained in argon, a process known as CASING. (Crosslinking by Active Species of Inert Gases). The second technique involves the deposition of a thin film by plasma polymerization of organic gases. The gases used in this study were ethane, ethylene and tetrafluoroethylene. Hydrogels were prepared by photopolymerization of 2-hydroxymethyl methacrylate (HEMA), and copolymerization of HEMA with methyl acrylate. The CASING treatment was found to be least effective. The most successful method was the plasma polymerization of tetrafluoroethylene, which yielded an order of magnitude reduction in the flux rate of pilocarpine with a film thickness of 0.25micronm. Polymerization conditions bringing about a crack-resistant film were critical in obtaining the best results.


Studies on copolymers of collagen and a synthetic polymer. First report--experimental study on biocompatibility of laminar copolymers of collagen and a synthetic polymer.

To develop biomaterials for semiparmanent substitution, we are studying copolymers of collagen and a synthetic polymer. In this study, in order to test tissue compatibility, the laminar copolymers were prepared from bovine collagen treated with proctase to remove telopeptides and synthetic polymers such as polyethylene, polyvinylalcohol, silicon gum etc., by the OKAMURA-HINO metthod, namely polymerization achieved by the introduction of cross-lincages, applying plasma discharge and gamma-irradiation. These copolymers were implanted into dorsal subcutaneous tissue in rabbits and removed after 1, 2, 3, 4 and 6 weeks to be examined by light microscope and electron microscope. One or two weeks after implantation, we observed that a large number of fibroblasts gathered on the surface of the grafted collagen. After 3 or 4 weeks, bridges of collagen fibrils were observed between the copolymers and rabbit tissue. These copolymers were firmly adhered to the tissues and separable only with difficulty. These results showed that the copolymers maintain the capacity for extreme strong bonding for at least 6 weeks and possess high tissue compatibility. From that we concluded copolymers are useful as a biomedical material.


Biogenesis of the purple membrane of Halobacterium halobium.

A protein closely resembling the purple membrane protein pre-exists in the cell membrane of H. halobium prior to the appearance of functional bacteriorhodopsin. It is associated with a differentiated membranous structure which has been isolated on a sucrose gradient and appears to be a precursor of the purple membrane. The identity of the precursor protein as a form of the purple membrane protein was established in different ways: (1) The cell proteins were labelled in vivo with 14C-proline during dark aerobic growth, the label was "chased", and the cells transferred to the illuminated near-anaerobic conditions under which purple membrane is optimally synthesised (induction conditions). Cell lysates were fractionated on sucrose gradients at different times after induction. Label first found in the precursor fraction appeared within 24 h in the purple membrane fraction. (2) SDS-urea-acrylamide gel electrophoresis of the purple membrane protein and the precursor showed only one protein band whose migration coincided with that of the purple membrane band. (3) The amino-acid analysis of the purified precursor was very similar to that of the purple membrane. The absorption spectrum of the precursor showed little of the characteristic absorption of bacteriorhodopsin at 570 nm. A major band appears at 412 nm, the exact nature of which is not known. The difference spectrum (reduced versus oxidised) of a purified fraction showed only traces of cytochrome. Thin-layer chromatography of an acetone-soluble lipid extract indicated the presence of retinal and beta-carotene. Cells grown in the presence of nicotine did not develop purple membrane after induction: the species absorbing at 412 nm was much less abundant than in non-inhibited cells, but a new fraction was present with a sharp peak at 345 nm consisting mainly of lycopene.


The sarcoplasmic calcium pump - a most efficient ion translocating system.

In contrast to the sodium-potassium transporting plasma membranes, the sarcoplasmic membranes (SR) are highly specialized structures into which only two major intrinsic proteins, a calcium transporting protein and a calcium binding protein are embedded. The calcium transporting protein is a highly asymmetric molecule. It binds two calcium ions with a very high affinity at its external, and two calcium ions with low affinity at the internal section of the molecule. ATP is bound with high afffinity to an external binding site, inducing a conformational change. When the vesicular membranes are exposed to solutions containing Ca++, Mg++ and ATP, ATP is hydrolyzed and simultaneously calcium ions are translocated from the external medium into the vesicular space. When calcium ions are translocated in the opposite direction, ATP is synthesized. The calcium-ATP ratio for ATP cleavage as well as for ATP synthesis is 2. Thus, the SR membranes can transform reversibly chemical into osmotical energy. Inward and outward movements of calcium ions are relatively slow processes connected with the appearance and disappearance of different phosphorylated intermediates. One phosphorylated intermediate is formed by phosphoryltransfer from ATP when calcium ions are present in the medium. In contrast, when calcium ions are absent from the external medium, two different intermediates can be formed by the incorporation of inorganic phosphate. Only when calcium ions present in the internal space of the vesicles are released, the incorporation of inorganic phosphate gives rise to an intermediate who phosphoryl group can be transferred to ADP.


Drosophila mutant with a transducer defect.

The trp is a conditional phototransduction mutant of Drosophila. Direct electrical measurements and shot noise analysis suggest that a prolonged intense light causes in the mutant a reduction in the quantum efficiency for quantum bump production that does not arise from bleaching of the visual pigment. This effect depends on the duration of the light and only weakly on its intensity. In the normal fly, an intense blue light that shifts the visual pigment from rhodopsin to metarhodopsin, induces an excitatory process manifested by a prolonged depolarizing after potential (PDA). In the mutant, the PDA has a small amplitude and bump noise is superimposed on the response. It can thus be shown that the excitatory process underlying the PDA is also present in those trp mutants where the PDA voltage response is small or absent. It is suggested that the absence of the PDA voltage response in the mutant is probably due to a defect in an intermediate process, which links the excitatory process to the membrane conductance change.


Assimilation of liquid hydrocarbon by microorganisms. II. Growth kinetics.

The growth kinetics of a microorganism with high affinity for liquid hydrocarbon which has a low solubility in water was investigated for Candida intermedia IFO 0761 in our previous work. The microorganism contained a hydrocarbon pool in and/or on the cell. The transfer of water-soluble substrates to the cell was not the rate-limiting step in the growth of C. intermedia accompanied by clump formation with liquid hydrocarbon. The operating conditions necessary for the oxygen supply for the growth were adequate for the growth of C. intermedia on n-tetradecane. The saturation kinetics was valid for the specific growth rate of C. intermedia and specific concentration of hydrocarbon per unit cell mass; the specific growth rate was expressed by the following equation: (formula: see text).


[State of nutritive circulation in rats following long-term limitation of motor activity].

Experiments were conducted on rats. It was shown that long (80-100-day) hypokinesia led to the following significant disturbances of the nutritive circulation: reduction of the true capillary plexus, emptiness of the terminal arterioles and arteries, the direction of the terminal circulation chiefly through the arterio-venous anastomoses, avoiding the nutritive vessels. A tendency to reduction of the buffer bases in the venous blood and to the respiratory alkalosis in the arterial blood was noted. Hemorheological changes were insignificant.


[Effectiveness and adaptation to food quality of the starch-glucose conveyor following section of the bile and pancreatic ducts in rats].

Membrane hydrolysis and carbohydrate transport were determined 2 hours after bread or meat feeding of rats on the 4th, 7th, and 14th day after the ligation of the bile-pancreatic duct in various segments of the small intestine. Even with a sharp reduction of the amylolytic activity of the mucosa surface, the transport of glucose released during the membrane hydrolysis of starch was retarded much less. During observation for two weeks the intensity of starch glucose transport increased sharply in preparations of the intestine in bread-fed rats, but fell in meat-fed rats. The differences between the sucrose hydrolysis level and the free-glucose transport were insignificant in both groups of rats. The data obtained are discussed in the light of A. M. Ugolev's hypothesis on plastic organization of the digestive-transport conveyer.


[Response of the body to a combination of severe cranio-cerebral trauma and acute massive blood loss].

It was shown in acute experiments on 54 dogs that peculiarities of body reaction to the combination of a severe craniocerebral trauma with acute blood loss depended on the sequency of the extreme influences inflication. The craniocerebal trauma inflicted at the early period of hemorrhagic shock failed to influence significantly the dynamics and results of the main pathological process. Against the background of the craniocerebral trauma hemorrhagic shock appeared with a much less blood loss and its course was more severe.


[Role of cerebral nitrogen metabolism in the mechanisms of the therapeutic action of oxygen under elevated pressure in hemorrhagic shock].

The hyperbaric oxygen therapy (3 ata O2, 60 min exposure) of hemorrhagic shock in cats (arterial pressure--60/50 mm Hg) prevents a sharp increase of ammonia, urea, alanin, gamma-aminobutyric acid content in the brain, typical of oxygen deficiency, and activates glutamine formation. The processes of ammonia detoxication connected with glutamate and glutamine synthesis intensification course more intensively in the sensory-motor cortex than in the limbic brain structures.


[Possiblity of using immobilized urease for degradation of urea in blood plasma].

As revealed the velocities of urea decomposition in the citrate donor plasma with soluble urease and urease immobilized by addition to carboxymethyl ester of cellulose, 2-(3'-amino-4'-methoxyphenyl)-sulphonylethyl ester of cellulose, diethylaminoester of cellulose, stained with dichlortriazine stain, or graft copolymere of cellulose and polyglycidylmetacrylate were sufficiently close to one another. Preparations of immobilized urease can be repeatedly used for urea decomposition in the citrate donor blood. Periodical treatment of the mentioned preparations with cystein solution led to a lesser decrease of enzymatic activity of the immobilized urease after repeated use.


[Influence of psychotropic substances on the self stimulation response].

D-amphetamine and cocaine were found to facilitate septal self-stimulation. In low doses morphine, imipramine, benactyzine, meprobamat, diazepam, chlordiazepoxide, phenobarbital, LSD-25 failed to influence this index, but in high doses they depressed the self-stimulation. It is supposed that d-amphetamine and cocaine exerted a direct stimulating action of the positive reinforcement system of the septum. Inefficiency of the other above-mentioned agents was due to the absence of the nervous substrate of negative reinforcement at the septum level. A comparative study of the above results and the influence of neurotropic agents on the hypothalamic self-stimulation (literature data) indicated that the activating effect of psychotropic agents on the systems of positive reinforcement depended upon their  influence on the emotiogenic brain structures and not on the structures responsible for the formation of motivations.


[Directed attacks on the specific activity of leukemia cells].

Different means of attenuating the leukemogenic activity were studied comparatively with a continuous strain of bone marrow cells derived from mice infected with Rauscher leukemia virus. Treatment of leukemic cells with neuraminidase, like their cultivation at suboptimal temperature resulted in a complete loss of leukemogenic activity, as evidenced by 100% survival of experimental animals and the absence of splenomegaly. In parallel experiments with concanavallin A and 5-bromdeoxyuridine treatment, development of splenomegaly was retarded and lethality of the animals reduced by 70 and 20%, rnd 20%, respectively. These results permit to raise the problem of using the above-memtioned methods for the attenuation of leukemogenic activity as an approach to prepare material with the immunizing activity.


[Characteristics of hematopoiesis in athymic nude mice].

The bone marrow, peripheral blood and spleen state was studied from the age aspect in Nude mice characterized by recessive nu mutation. A number of blood system peculiarities were revealed in homozygotic (nu/nu) mice by which they differed from the heterozygotic (nu/+) animals: low peripheral blood, bone marrow and spleen lymphocyte count, erythropoiesis depression in the bone marrow and erythroid element hyperplasia in the spleen.


[Effect of poly-4-vinylpyridine on the quantity of colony-forming cells in the bone marrow and spleen of mice].

Administration of poly-4-vinylpyridine to mice in the maximal tolerance dose doubled the colony-forming units (CFU) and decreased the cell count by 22% in the bone marrow. The increase of CFU number takes place from 2 to 7 days. The cell count in the bone marrow began to decreased 2 hours after the administration of the polymer and remained below the initial level for 5 days; however it began to return to the normal from the second day, reaching the normal figures by the 7th day. In the spleen the CFU rose 9-fold, with a simultaneous development of splenomegaly on account of increased cell count.


[State of the pituitary--thyroid gland system in pregnant rabbits, fetuses and newborn rabbits].

The paper deals with the study of the functional condition of the hypophysis-thyroid system in pregnant rabbits and their fetuses during last trimester of gestation. The 131I uptake and the hormonal synthesis proved to be increased in the maternal and fetal thyroid glands during the 12rd--27th days of pregnancy; this was confirmed by the changes in the histostructure of the gland and was apparently connected with the increased thyroxin requirement in the maternal-fetal system at this period. These changes were preceded by the enhancement of the thyrotropic function of the hypophysis. No such correlation was present in the fetus. These data served as the evidence that the functional activity of the maternal hypophysis-at the functional activity of the maternal hypophysis-thyroid system rapidly changed depending on the requirement of the pregnant organism in thyroid hormone.


[Relationship between the muscle and connective tissue components of the heart in rats with experimental myocardial infarct].

As soon as the first 24 hours of experimental myocardial infarction in rats the "intact" portions of the left ventricle displayed an increase in volume and mean amount of the elements of the connective tissue stroma of the heart. On the second day of the experiment these indices reached the maximum, and on the 20th day--they were similar to those in the control animals myocardium. On the 3rd day the number of muscle nuclei per standard section area proved to increase; this can be attributed to the amitotic division of the myocyte nuclei, since the number of double nuclei increased at the same periods.


[Decrease in the tissue-specific resistance of gastric epithelium to acute cell death following inhibition of DNA synthesis].

Administration to mice of hydroxyurea caused an acute death of an insignificant number of DNA-synthesizing cells in the epithelium of the glandular stomach. When actinomycin D was injected 8 hours prior to hydroxyurea there occurred a sharp intensification of cell degeneration: about 80% of the cells which synthesized DNA during the hydroxyurea administration proved to degenerate. No intensifying action was produced when actinomycin D was injected simultaneously with hydroxyruea. It is supposed that actinomycin D intensified the protein synthesis in the cells of the stomach (the "superinduction" effect), this increasing their sensitivity to the inhibition of the DNA synthesis.


[Ultrastructural analysis of the cortical substances of the adrenal glands of rats under conditions of bilateral subdiaphragmatic vagotomy].

Electron-microscopic study of zona fasciculata and zona reticularis of the adrenal cortex showed that 7 days after bilateral subphrenic vagotomy there occurred a diatation of the perinuclear space of adrenocorticocytes and endothelial cells; there was also a dilatation of the tubules in the smooth cytoplasmic reticulum, edema of the mitochondria, reduction of their cristae, and increase in the number and size of lipid drops. In 45 days there are encountered mitochondria with the beginning myelinization, lipid drops form accumulations, and electron-transparent vacuoles are revealed in them. The functional activity of adrenocorticocytes in zona fasciculata and zona reticularis of the adrenal cortex diminished in rats after vagotomy.


The management of cardiac arrhythmias.

The first and most important aspect of the management of a cardiac arrhythmia is its precise identification, and intracardiac recordings should be made in cases of doubt. When confronted with a patient with an arrhythmia the first decision is whether it needs treating at all. If it does the next problem is to decide whether to begin with electrical or pharmacological treatment, and here the important feature is the clinical state of the patient: the more ill the patient and the larger the haemodynamic disturbance, the greater the relative indication for electrical (cardioversion or pacing) therapy. Although there has been some progress in the understanding of the pathological physiology of arrhythmias and attempts have been made to rationalize the classification of cardioactive drugs, these sciences have not so far provided information of clinical value. The drug therapy of arrhythmias is largely arbitrary, but few drugs are involved. The most important thing of all is for an individual clinician to become familiar with the most significant of these drugs and not to indulge in polypharmacy, because all antiarrhythmic drugs can lead to either depression or overexcitation of the myocardium.


Directional variations in the extensibility of human skin.

When an extending load is applied to human skin in vivo or in vitro the skin stretches with small increases in load until a limit is reached (limit strain) after which much greater increases in load are required to extend it further (terminal stiffness). At many sites there are marked directional variations, the direction of maximum limit strain being the same as that of minimum terminal stiffness and vice versa. The directional variations of extensibility at constant load on a young male body have been measured and recorded.


Effects of N-methylamino acids and convulsants on spontaneous action potentials in guinea-pig cerebellar slices.

1. N-methyl-gamma-aminobutyrate (N-methylGABA), N-methylglycine, N-methyltaurine and N-methylbeta-alanine diminished the frequency of spontaneous spike discharges in guinea-pig cerebellar slices. Usually a weak excitatory effect preceded the inhibition. 2. The inhibitory effects of N-methylGABA and N-methylbeta-alanine were competitively antagonized by both picrotoxin and strychnine. 3. The inhibitory action of N-methyltaurine was competitively suppressed by strychnine and by low concentrations of picrotoxin. 4. The inhibitory action of N-methylglycine was suppressed by strychnine but not by picrotoxin. The suppression was competitive at low concentrations of strychnine. 5. N-methylDL-glutamate brought about a strong inhibition followed by a strong excitation of the neurones. The inhibitory effects were competitively suppressed by both picrotoxin and strychnine. Neither convulsant affected the excitation. 6. Whereas L- or D-glutamate caused only excitation in the majority of cells examined, a small proportion of the cells exhibited inhibition preceding the excitation by L- or D-glutamate. Such inhibitory effects were suppressed by picrotoxin but not by strychnine. 7. Kinetic analyses of the dose-response curves for the N-methylamino acid in the presence or absence of the convulsant indicated that the number of molecules of the amino acid combining with the receptor site to produce a response was 3 for N-methylGABA, 2 for N-methylglycine, 3 for N-methyltaurine, 3 for N-methylbeta-alanine. The corresponding value was 1 for N-methylDL-glutamate (inhibition). The number of molecules of convulsant combining with the receptor site was calculated to be 2 for picrotoxin with N-methylGABA, N-methylbeta-alanine and N-methylDL-glutamate and 1 for strychnine with all N-methylamino acids examined.


Renal blood flow autoregulation and renal venous prostaglandins in the pump-perfused canine kidney (in situ).

1. Renal autoregulation of blood flow was re-examined in the pump-perfused canine kidney and concentrations of prostaglandins E and F in the renal venous plasma were measured by radioimmunoassay. 2. At low perfusion pressures, below the range of autoregulation, prostaglandin E and F concentrations rose and calculated prostaglandin E secretion rate fell. 3. Meclofenamate (10 mg/kg i.v.) reduced renal blood flow and prostaglandin E and F secretion rates, but did not abolish autoregulation. 4. Renal prostaglandins do not appear to mediate autoregulation in the kidney but may affect the level at which flow is controlled.


The effect of systemically and topically applied drugs on ultraviolet-induced erythema in the rat.

1. Exposure of the skin of rats to u.v. light (greater than 295 nm) for 30 s or longer elicited a delayed erythema response, the rate of onset increasing with period of irradiation. The erythema was still present at 24 h and was replaced by scab formation in 48 hours. 2. Both topically applied steroidal and non-steroidal anti-inflammatory drugs reduced the erythema formation when administered immediately after u.v. exposure. Propyl gallate, an antioxidant with sun screening properties in man, also possessed topical anti-erythemic activity. 3. Both steroidal and non-steroidal anti-inflammatory drugs, systemically administered 1 h before u.v. exposure, reduced and erythema. However, the steroidal compounds were less effective than the non-steroids and reduced the intensity of erythema by less than 50%. Antagonists of 5-hydroxytryptamine (5-HT) reduced the erythema but several other drugs with different pharmacological activities were ineffective. 4. Neither topical nor systemic treatments of any of the drugs examined suppressed the scab formation at 48 hours. 5. These results and those using other selective blocking agents indicate that in the mediation of the erythema reaction prostaglandins may play a major role and 5-HT perhaps a minor one but that H1 histamine receptors and alpha- and beta-adrenoceptors have no significant role.


Effects of sympathetic innervation and temperature on the properties of rat heart adrenoceptors.

1. The pharmacological characteristics of adrenoceptors at different temperatures were assessed on the basis of the effects of various alpha- and beta-adrenoceptor agonists and antagonists on electrically-driven left atria and spontaneously-beating pairs of atria from rats. 2. Phenoxybenzamine (Pbz) potentiated inotropic responses of left atria to noradrenaline (NA) at 31 degrees C, produced significantly less potentiation at 24 degrees C and inhibited responses at 17 degrees C; it had little effect on responses to CaCl2. Both Pbz and phentolamine inhibited responses to phenylephrine more effectively at 17 than at 31 degrees C. N-cyclohexylmethyl-N-ethyl-beta-chloroethylamine hydrochloride (GD-131), a haloalkylamine with negligible alpha-adrenoceptor blocking activity, caused only potentiation of responses to NA at 17 degrees C. 3. The presence of phentolamine during incubation with Pbz eliminated block of responses to NA and revealed a potentiation that was equivalent at all three temperatures tested. Phentolamine did not alter the block of responses to 5-hydroxytryptamine by Pbz. Protection of alpha-adrenoceptors by phentolamine during exposure to [3H]-Pbz significantly decreased the amount of label bound to the myocardium at 17 degrees C, but did not alter binding at 31 degrees C. 4. Inhibition of responses to NA by propranolol decreased with temperature, and the magnitude of the change increased with the concentration of propranolol. Compared to 31 degrees C, the effect of the highest concentration of propranolol. (4.0 micronM) was significantly decreased at 24 degrees C, and the effects of all except the lowest concentration (0.04 micronM) were significantly decreased at 17 degrees C. 5. The potency of isoprenaline decreased and that of phenylephrine increased at low temperatures, and their potency ratio was much lower at 17 than at 31 degrees C for both the inotropic and chronotropic responses of spontaneously-beating atria. However, the ratio was unaffected by temperature in electrically-driven left atria. A similar difference between spontaneously-beating and driven preparations is apparent in the data of other workers, but its basis is not clear. 6. Atria from rats pretreated with 6-hydroxydopamine (6-OHDA) were sensitized to the effects of NA, and there was no increase in alpha-adrenoceptor properties at low temperatures. Little alpha-adrenoceptor activity could be demonstrated in chemically denervated atria at any temperature, 6-OHDA pretreatment did not alter the binding of [3H]-Pbz at 31 degrees C, but decreased it significantly at 17 degrees C. Pretreatment with reserpine caused some sensitization, but not significantly after the characteristics of the adrenoceptors or their responses to temperature. 7. It is concluded that the adrenoceptors of atria are affected by temperature in much the same way as those of frog hearts, although the transition from beta- to alpha-adrenoceptor properties may begin at a slightly higher temperature...


The sympathomimetic activity of fenfluramine hydrochloride on rat vas deferens.

1. The peripheral, pharmacological effects of the anorexigenic agent, fenfluramine hydrochloride, have been investigated on rat isolated vas deferens. 2. Characteristic spiked contractions were observed within 2 to 3 min after exposure to fenfluramine; these contractions reached a rate of around 13 per min and were of variable height. 3. Pre-treatment of vasa with the indirectly acting sympathomimetic amine, tyramine, greatly reduced both the height and rate of contraction induced by fenfluramine. 4. The uptake inhibitor, desipramine, required a concentration in excess of 10 micronM to affect fenfluramine-induced contractions. Effects of desipramine on fenfluramine contractions were of equal magnitude whether desipramine was administered before fenfluramine or at the height of the fenfluramine-induced contractions. 5. Pre-treatment with debrisoquine (0.5 mM), reduced the contractions in response to fenfluramine over a period of time. 6. Fenfluramine, added to vasa from rats which had been injected intraperitoneally with 5 mg/kg reserpine 24 h and 48 h previously, failed to induce its characteristic contractions. 7. It is concluded that fenfluramine can be classed as an indirectly acting sympathomimetic amine on peripheral adrenergic nerve terminals.


Evidence for a visceral smooth muscle abnormality in Okamoto spontaneous hypertension.

1. In order to discover whether the changes in reactivity are related to the primary cause of hypertension in spontaneously hypertensive rats (SHR) or are just an adaptation induced by the high arterial blood pressure we tested the contractile response of a visceral smooth muscle from such rats. 2. Longitudinal strips of the fundus from 20 week old male and female SHR and  Wistar normotensive (NW) rats were used. Dose-response curves to Ba2+ in SHR strips were displaced to the right as compared to NW rats. Maximal responses were identical. Male SHR fundus strips contracted much more with Sr2+ (SHR: 42+/-3% of maximum response to Ba2+, n=10; NW: 19+/-4%, n=10, P less than 0.01) than NW strips. There was no difference in the response to both BaCl2 and SrCl2 between female SHR and NW fundus strips, and MnCl2 and LaCl3 were relaxant in all cases. 3. Dose-response curves to Ca2+ of depolarized SHR and NW fundus strips were obtained and the effect of diazoxide on Ca2+ contractions was observed. The contractile action of Ca2+ in depolarized preparations was enhanced in  both male and female SHR strips. The effect of diazoxide was more marked in SHR strips than in NW fundus strips. 4. SHR fundus smooth muscle shows the same modification of reactivity to Ba2+, Sr2+, Ca2+ and diazoxide that was previously described in arterial smooth muscle. This indicates that the cellular modification responsible for the increase of vascular tonus in SHR is not an adaptive reaction to high blood pressure. The differences between female SHR and male SHR responses are not unexpected, considering the natural evolution of hypertension in Okamoto rats which is milder in the female.


A comparison in rabbit isolated hearts of the dysrhythmogenic potential of amitriptyline, maprotiline and mianserin in relation to their ability to block noradrenaline uptake.

1. In isolated hearts of rabbits, perfusion with (-)-noradrenaline (0.0059 to 5.9 micronM) resulted in chronotropic and inotropic responses and a shortening of the interval between peak atrial and peak ventricular tensions (the A-V contraction interval). No dysrhythmias developed but at higher concentrations (590 micronM) 2 out of 7 hearts developed dysrhythmias (extrasystoles). 2. Perfusion with the antidepressants amitriptyline or maprotiline (4.8 micronM) or mianserin (28.8 micronM) reduced ventricular force, did not change heart rate and only amitriptyline reduced atrial force and lengthened the A-V contraction interval. At 4.8 micronM mianserin produced only a marginal shortening of the A-V contraction interval. 3. At these concentrations no dysrhythmias developed but at higher concentrations (amitriptyline 8 micronM, maprotiline 8 micronM, mianserin 60 micronM) all the agents produced dysrhythmias involving an interference with atrio-ventricular synchronization. 4. In the presence of mianserin (4.8 micronM) perfusion with noradrenaline (0.0059 to 5.9 micronM) shortened the A-V contraction interval and did not produce dysrhythmias. In the presence of amitriptyline or maprotiline (4.8 micronM) or mianserin (28.8 micronM) the A-V contraction interval generally lengthened and most hearts developed dysrhythmias (usually involving interference with atrio-ventricular synchronization). 5. [3H]-(-)-Noradrenaline uptake in perfused rabbit hearts and in mouse isolated atria or vasa deferentia was inhibited by the antidepressants to a similar extent, amitriptyline being marginally most potent (molar potency taken as 1.0), maprotiline being less potent (1.5) and mianserin least potent (2.0)). 6. It is concluded that of these three antidepressants, mianserin is least cardiotoxic in this preparation and that the ability of these antidepressants to predispose to noradrenaline-induced dysrhythmias is not related to blockade of noradrenaline uptake.


Treatment settings in psychiatry. The use of hospital services: long term follow-up.

Ninety-eight matched pairs of patients treated respectively on first admission in a general hospital psychiatric unit or a psychiatric hospital have been followed-up for from five to eight years. The periods of treatment received in either setting remained comparable, and only 3 per cent of patients were in-patients or day patients at five years, while 14 per cent were out-patients and 20 per cent attended their family doctor. There was no evidence in these patients of a transfer of chromicity to the psychiatric hospital, but patients with longer periods of treatment were noted to change hospital from both the general hospital unit to the psychiatric hospital, and in the reverse direction.


Suicidal intent in self-injury.

The difficulties in measuring suicidal intent in cases of self-injury are discussed, and a scale is described to measure this intent. This scale has been used in 500 cases of self-injury. It is practical and reliable. Results show that the scores derived from it are closely related to the similar Beck Scale; they are also related to age, sex, social isolation method of self-injury, previous history of self-injury or of psychiatric treatment, physical health at the time of self-injury and alcohol abuse. These results are discussed with particular reference to suicide prediction and the future validation of the scale by long-term follow-up.


Reflective processes and anxiety.

Three hypotheses are abstracted from Laing's writings on the inter-relationships between self-consciousness, anxiety, and self-evaluation. Indicators of self-consciousness are obtained from an open-ended instrument which requires the respondent to provide twenty answers to the question (asked of himself): "Who am I?" Statements indicative of self-consciousness are those on which the respondent inserts a self-reference into the statement made about himself and are distinguished from statements in which no explicit self-reference is found. The data indicate that the mark of the self-conscious person is anxiety and that anxiety is largely restricted to those statements devoid of self-consciousness. The structure of a system likely to produce this configuration is considered by drawing upon Kierkegaard's conjectures on the joint emergence of consciousness and anxiety. Equating self-consciousness with "reflection", it is suggested that while reflection brings about anxiety in a system as a whole, the act of reflection deters the emergence of anxiety at the particular points within the system upon which it is deployed. Thus the function of reflection is two-fold, creating anxiety at the same time (but not at the same place) as anxiety is reduced.


A comparison of radiography and radioisotope scanning in the detection of Paget's disease and in the assessment of response to human calcitonin.

In eight patients with symptomatic Paget's disease, radiographs and radioisotope bone scans have been compared before treatment by high dose synthetic human calcitonin (CIBA 47175-Ba) and three months and 12 months later. In six patients quantitative radioisotope scans allowed calculation of relative radioactivity in normal bone, Paget's disease and bone adjacent to osteoarthritic joints. Comparison of radiographs and scans showed 69 sites diagnosed as Paget's disease on both examinations; nine sites showed radiographic changes of Paget's disease but had negative radioisotope scans and two sites were abnormal on the scan but radiologically normal. One of these two reverted towards normal during treatment with calcitonin. Comparison of analogue scans done at three months and 12 months with initial scans showed diminished uptake in Paget's disease compared with normal bone, and quantitation showed this was due to decreased absolute uptake in Paget's disease and a slight increase in normal bone. Osteoarthritic bone showed no significant response to calcitonin.


The "negative chest radiograph" in acute pancreatitis.

Twenty-six patients with acute pancreatitis and normal chest radiographs on admission were studied. Twelve (46%) developed abnormalities in the first five days of illness. Twenty-four of the 25 patients tested developed an arterial pO2 of less than 10.6 kPa (80 mmHg) but severe hypoxia was present in a greater percentage of patients whose chest radiology became abnormal. Blood gases and chest radiographs should be checked on a regular basis in patients with acute pancreatitis.


Tumour regression as a guide to prognosis: a study with experimental animals.

An analysis has been made of the relationship between regression during therapy and the probability of local control in a group of mice treated with single doses and fractionated doses of X rays. The tumours were first generation transplants from spontaneous mammary carcinomas in C3H mice and were irradiated with single doses 3F/4 days, 9F/10 days, 9F/18 days or 15F/18 days. The size of the smallest radiation dose approached those encountered in clinical radiotherapy. A significant correlation was observed between the shrinkage during the treatment period and the local control at 150 days, for three of the four fractionated schedules. A weaker correlation was observed for shrinkage within a week after single doses, and for shrinkage during treatment with nine fractions in 18 days. It is postulated that the rate of shrinkage is an inherent characteristic of each individual tumour and does not reflect the number of cells killed. However, the shrinkage in some tumours during the course of therapy may result in more extensive reoxygenation which makes these tumours more sensitive to subsequent doses of fractionated course. The degree of shrinkage within the period of fractionated irradiation was found to be a useful prognostic guide to ultimate local control in individual tumours.


The reconstruction and dose-rate calculation of radium needle implants.

A comprehensive computer program has been developed for carrying out the various tasks associated with radium needle implants. Data input for the program are obtained by digitizing an orthogonal pair of radiographs with an analogue tracing system. Corresponding needles in the two radiographs are automatically located, and the implant may be rotated to any desired position in three dimensional space, at the same time presenting a view of the rotation on a storage oscilloscope. Automatic determination of the mean plane of a planar implant is possible, and the dose-rate distribution may be calculated in any specified plane. A Paterson--Parker exposure rate calculation may be performed with a biological dose and treatment time correction incorporated. The program is written in Fortran and runs on a small interactive computer system.


The management of ureteric reflux in children.

A series of 262 children suffering from vesico-ureteric reflux is presented. Nearly all presented with a urinary tract infection and the age at onset of symptoms was predominantly in the pre-school years. The reflux was not diagnosed in the majority until after the children started attending school. Over one-third had symptoms for more than half their lives before reflux was diagnosed. The incidence of hypertension and renal insufficiency in the series were 4% and 7-8% respectively. Nearly 60% of the children were selected by defined criteria for operation as their initial treatment. The remainder were treated conservatively in the first instance but 30% of these subsequently required operation. Reflux was eliminated in 96.3% of the children at the first operation and 5.5% developed postoperative ureteric dilatation, none of them permanently. Reflux disappeared spontaneously in two-thirds of the children treated conservatively but took more than 3 years in 46%. Infection recurred in 24% of children treated by operation and in 29% treated conservatively. The incidence with which reflux disappeared spontaneously in the children treated conservatively was related to time, not to the infection recurrence rate.


Vesico-ureteral reflux in children.

A total of 126 children with 193 refluxing renal units were treated for vesico-ureteral reflux during the period 1966-74. The grading of the cases into mild, moderate and severe reflux revealed renal scarring in 5, 22 and 28% of the three groups. Conservative treatment was used in 10% of the patients, surgery on the urethra in 25%, and reimplantation of the ureter by the Politano-Leadbetter technique in the last two-thirds of the patients. Reflux was eliminated in 50% of the renal units treated conservatively, in 60% of renal units treated with surgery on the urethra, and in 86% of units treated with reimplantation. The overall results seemed to be better in mild than in moderate or severe reflux. The dilatation of the upper urinary tract returned to or approached normal in 85%. Of the children, 80% were without symptoms postoperatively, whereas urinary-tract infection persisted in 36%. The value of the grading of reflux is emphasised, and indications for treatment are discussed.


Guided dilatation and transurethral resection in one session for treatment of post-prostatectomy obstructive complications.

53 patients presenting with post-prostatectomy obstructive complications above the colliculus seminalis were treated in a single session by means of guided dilatation of the stricture followed by transurethral resection of the stenosing fibrotic tissue. No failures or complications with the guided dilatation technique have been observed. The results obtained after resection in cases of bladder neck stenosis were very encourging. Unsatisfactory results were observed in cases of prostatic urethral stenosis because resection is not a technique sufficient for stable resolution of the obstruction.


Hormonal pattern in prostatic carcinoma following orchidectomy: 5-year follow-up.

Clinical and hormonal findings have been collected from 14 patients affected by carcinoma of the prostate observed over a 5-year period following orchidectomy. Plasma testosterone decreased significantly after surgery (62 ng +/- 21 SD/100 ml), remaining below 100 ng/100 ml even in cases showing only a partial regression or increase of the tumour. Gonadotropin FSH increased 1 year after orchidectomy to 28 mIU +/- 5/ml and remained constantly elevated up to the 5th year, whilst LH increased progressively up to the 5th year (30 mIU +/- 12/ml). Finally, plasma cortisol increased progressively up to 23.2 microng +/- 17.4/100 ml at the 5th year post-orchidectomy, especially in those cases showing partial regression or increase of the tumoral mass, indicating an adrenal hyperfunction which is probably responsible for increased production of weak androgens that can be transformed in the prostate into more active compounds. These results suggest therefore that after orchidectomy, cortisol and adrenal androgen determination may be of value in detecting the potential activation of prostatic adenocarcinoma.


The urethral syndrome: experience with the Richardson urethroplasty.

The technique of urethroplasty described by Richardson has been used to treat 40 patients with the urethral syndrome. 38 patients have had improvement in clinical symptoms and urine flow rate following this procedure. 2 patients have had a relapse of symptoms but both have been improved by further surgery.


The interrelationship between vesico-ureteric reflux, trigonal abnormalities and a bifid pelvicalyceal collecting system: a family study.

3 investigations have been undertaken to study the interrelationship between a bifid pelvicalyceal collecting system, vesico-ureteric reflux and lateral ectopia of the ureteric orifice. Firstly, in 30 families investigated to confirm the familial incidence of bifid and double ureters there were 3 families in which siblings were found with primary reflux. Secondly, reflux was found in 62 of 110 patients investigated with a duplex pelvicalyceal system. Reflux was to the ipsilateral kidney in 48 patients but occurred to the contralateral kidney, unaffected by duplicity, in 14. This high incidence of reflux is related to lateral ectopia of the ureteric orifice which may be either bilateral or unilateral. Thirdly, the incidence of a bifid pelvicalyceal system was determined in the parents and siblings of a series of 32 patients with primary reflux. The results support the hypothesis that primary reflux may be inherited by an autosomal dominant gene of variable penetrance in a manner similar to the inheritance of a duplex urinary tract. Thus there is a direct genetic relationship between primary vesico-ureteric reflux, lateral ectopia of the ureteric orifices and a duplex pelvicalyceal system.


Death during recovery from severe malnutrition and its possible relationship to sodium pump activity in the leucocyte.

The leucocyte data on four malnourished children who died suddenly when high-energy feeding was started were retrospectively analysed. The pretreatment rate constant for sodium efflux in leucocytes was higher and the intracellular sodium concentration lower in this group than in 13 malnourished children who recovered uneventfully with feeding. Two other children with unusual leucocyte electrolyte values and sodium pump activity were identified and closely monitored when high-energy treatment was begun. They rapidly developed the syndrome of extracellular fluid overload but were successfully treated with diuretics and digoxin. Though the precise relation between the findings in the leucocytes and the development of this overload syndrome is not clear, the pretreatment leucocyte values are nevertheless valuable in predicting which malnourished children are at risk of sudden death when refeeding is started.


Oral talampicillin in the treatment of gonorrhoea.

Talampicillin was effective in eradicating the causitive organism in uncomplicated gonorrhoea, with only four treatment failures (1-6%) in 245 patients seen at least once after treatment. Symptoms, such as discharge, were seen in 61 patients at some stage after treatment but 31 of these cases were clear after one or two follow-up visits. The anitbiotic was well tolerated. One patient developed a maculopapular rash, penicillin allergy was reported in two patients, and a generalised itch (possibly penicillin allergy) in one further patient. A total of 241 out of 245 (98%) cases had the urethral gonococcus successfully eradicated with talampicillin. Talampicillin given in single doses appears to be an extremely good choice of antibiotic in the treatment of uncomplicated gonorrhoea in men.


Recurrent genital candidosis and iron metabolism.

It was thought possible that recurrent genital candidosis, like chronic mucocutaneous candidiasis, might be related to abnormalities in iron metabolism. Haemoglobin, serum iron, and serum ferritin levels of patients with recurrent genital candidosis were compared with those of patients who harboured yeasts but had no history of 'thrush', and with a control group of patients with no evidence of yeasts or history of 'thrush'. The mean haemoglobin level in patients with recurrent genital candidosis was significantly lower than in the control group (P less than 0-05) but it was thought that this difference did not have much practical meaning. There was no significant difference in the serum iron and serum ferritin levels between the three groups.


Three-day clotrimazole treatment in candidal vulvovaginitis.

The accepted modern practice is to treat each sexually transmitted disease with the shortest possible course of treatment consistent with success. In candidal vulvovaginitis, six days is the minimum period that has so far been found to be successful, but we report here a further reduction to three days. Patients were given two clotrimazole pessaries nightly for three consecutive nights; the overall success rate was 89-4% one month after treatment. This compares favourably with the 93% cure rate reported with the six-day course of clotrimazole. With both the long and short courses, patients having their first attack of genital candidosis responded better than those with a history of previous infection. Short courses of clotrimazole treatment are particularly valuable in dealing with uncooperative women who stop treatment at the earliest possible moment. Clinical and laboratory diagnostic pitfalls and their possible influence upon the therapeutic outcome are also discussed.


Hepatitis B antigen and antibody in a male homosexual population.

Sera from 600 male homosexual patients were tested for hepatitis B antigen (HBs Ag) and its antibody (HBs Ab). Thirty-one men (5-2%) were positive for HBs Ag. Testing for HBs Ab by immuno-osmoelectrophoresis 33 men (5-5%) were positive. However, sera of 85 patients negative for HBs Ab by routine methods were examined for HBs Ab by radioimmune assay. Thirty (35%) sera were found to be positive. No absolute correlation between the detection of HBs Ag, or previous history of hepatitis, jaundice, or current hepatitis was found. Similarly there was little correlation between presence of HBs Ab and this history. These observations suggest that the male homosexual population represents a pool of individuals within which the hepatitis B virus is readily transmitted, mainly as a subclinical infection although clinical hepatitis does occur in some patients. It is suggested that further work is necessary to determine whether the high antibody rate in male homosexuals is related more to sexual practice than to promiscuity.


Isolation of Chlamydia trachomatis from the male urethra.

Chlamydia trachomatis was isolated from 26% of urethral swabs taken from 509 men with urethritis. The highest yield of 68% was obtained from a selected group of men with nonspecific urethritis (NSU) who had a frank urethral discharge. This is a higher than in previous reports, and is significantly higher than the isolation of C. trachomatis from men with less severe urethritis. The higher yield was similar to C. trachomatis isolation rates reported among patients with severe trachoma in hyperendemic areas. Men with a previous history of NSU had low isolation rates. Overall, 30% of 385 men with NSU had positive chlamydial culture results, 7% of 59 men with gonococcal urethritis alone were Chlamydia-positive, 15% of 59 men with gonorrhoea followed by NSU (post-gonococcal urethritis) were Chlamydia-positive, and only 3% of 61 men without urethritis harboured Chlamydia. Swabs taken from the cervical os of 28 of 108 female contacts of men with NSU had a positive result for C. trachomatis. Significantly more pairs of sexual partners had the same chlamydial culture result than had different results. The chlamydial isolation rate was higher among men admitting a casual sexual contact than in men claiming only regular partnerships. The findings provide further evidence for the sexual transmission of C. trachomatis and for its aetiological role in NSU.


Human pituitary thyrotropin and gonadotropins: a comparative study of their in vitro lipolytic activity.

Highly purified preparations of human thyrotropin, follitropin, lutropin, and choriogondotropin were evaluated for their ability to stimulate lipolysis in isolated rat adipocytes. Thyrotropin was the most active of all while follitropin showed some activity. Lutropin and choriogonadotropin were essentially inactive. The isolated subunits of thyrotropin were inactive and 60-100% activity was regenerated upon recombination. The results provide direct evidence that lipolytic activity is intrinsic to human thyrotropin. The hybrid of human lutropin alpha + thyrotropin beta was considerably less active than the natural recombinant (thyrotropin alpha + beta), possibly indicating the presence of some differences in the alpha subunits.


Phosphatidic acid synthesis in the heart. 1. Effect of age and species difference in the mitochondrial and microsomal synthesis.

Rates of syntheses of monoacyl- and diacyl-glycerol 3-P (phosphate) were determined in the mitochondrial and microsomal fractions prepared from hearts of rats and rabbits, to compare characteristics of the acylation reactions by the two subcellular fractions. The assays were carried out with the subcellular fractions prepared from (i) hearts of hyperthyroid animals, and (ii) hearts of newborn and weanling rats. In addition, the effect of an addition of bovine serum albumin in the assay system was examined. (1) Administration of thyroid hormones increased the acyltransferase activity in rabbit hearts but not that in rat hearts. (2) Mitochondrial and microsomal fractions of hearts of newborn rats acylated glycerol 3-P at a rate 1.3-4 times greater than those of adult rats. The rate of acylation by the mitochondrial fraction of weanling rats was also high, but the rate of microsomal acylation was slightly lower than that of adult rats. By contrast, in newborn rats, diacylglycerol 3-P formation by the liver microsomes was not greater than that of the adults, although its formation by the newborn liver mitochondria was greater. (3) The accumulation of monoacylglycerol 3-P during the assay was accelerated by the addition of increasing amounts of bovine serum albumin. Therefore, the monoacylglycerol 3-P formation was less than 10% of the diacylglycerol 3-P formation in the assay containing no albumin and with the rat subcellular fractions, whereas nearly four times more monoacyl- than diacyl-glycerol 3-P was synthesized in the presence of 20 mg albumin. (4) The ratio of monoacyl- to diacyl-glycerol 3-P formation by the mitochondrial fraction was greater than that of microsomal fraction at any concentration of albumin in both rats and rabbits. At an equal albumin concentration in the assay, relatively more diacyl- than monoacyl-glycerol 3-P was formed in the mitochondrial fraction of newborn rat hearts as compared with adult hearts; (5) In conclusion, our data concerning the age and species differences in acyltransferase activities support a view that the mitochondrial fraction of both rat and rabbit hearts, in addition to the microsomal enzymes, is capable of catalyzing the de novo synthesis of phosphatidic acid.


Histone 5 from carp erythrocytes.

Histone 5 (H5) has been isolated from carp (Cyprinus carpio) erythrocytes and identified by its amino acid composition. It was found in very small relative amounts, despite the use of either 50 mM sodium bisulfite or 1.0 mM phenylmethylsulfonyl fluoride to inhibit endogenous protease activity. Carp H5 differed significantly from H5's of other lower vertebrates in its amino acid composition, position of elution from Amberlite CG-50 ion-exchange columns, and its unusually high electrophoretic mobility. The extensive quantitative and qualtitative variability of H5 from different species must be reconciled with the presumed functional homology of these cells.


In vitro and in vivo studies on the metabolism of estrogens and their sulfates in guinea pigs.

Labelled estradiol-17 beta(E2) or estrone (E1), when incubated with guinea pig liver slices, is metabolized by two main pathways. Part of each substrate is converted to estrone-3-glucuronide and estradiol-3-glucuronide. A further part of each is metabolized to estradiol-3-sulfate (E23S) and estrone-3-sulfate (E13S), which are interconverted. The latter conjugate appears to be the substrate for a 16 alpha-hydroxylase forming 16 alpha-hydroxyestrone-3-sulfate (16 alpha OHE13S). This, in turn, is further sulfurylated to yield 16 alpha-hydroxyestrone-3, 16-disulfate, accompanied by estriol-3,16-disulfate. A relatively small amount of tentatively indentified '6-hydroxyestrone disulfate accompanies these other two diconjugates. The guinea pig liver system suggests itself as a useful and relatively simple model for further study of 16 alpha-hydroxylation of E13S. The use of the latter as a natural subd E23S are present in liver, kidney, blood, gallbladder bile, intestine, uterus, and placenta after injection of labelled E2 into mature male and female guinea pigs. Some evidence has been obtained for the disulfate fraction (above) in liver and bile after injection of labelled E1.


Studies on the site of addition of sialic acid and glucosamine to rat alpha 1-acid glycoprotein.

Ultrasonic extracts of rough and smooth endoplasmic reticulum fraction and Golgi fractions from rat liver were examined by immunoelectrophoresis using antiserum to alpha 1-acid glycoprotein. Rough endoplasmic reticulum fractions contained only sialic acid free alpha 1-acid glycoprotein, whereas smooth endoplasmic reticulum and Golgi fractions also contained sialic acid containing alpha 1-acid glycoprotein. Determination of the sialic acid contents of immune precipitates isolated from the extracts suggested that the Golgi complex was the main site of addition of sialic acid to alpha 1-acid glycoprotein. Immunological studies on puromycin extracts of polyribosomes showed that polypeptide chains of alpha 1-acid glycoprotein and albumin were assemble mainly on membrane-bound polyribosomes. Evidence is presented from incorporation studies with labelled leucine and glucosamine that initial glycosylation of alpha 1-acid glycoprotein occurs mainly or entirely after release of nascent polypeptide from the ribosomal site.