Eastern equine encephalomyelitis virus in experimentally infected bats. Colonial bats (Myotis supp. and Eptesicus sp.) were infected with eastern equine encephalomyelitis virus by subcutaneous inoculation or by the bite of infected mosquitoes. Bats were maintained in an environment simulating conditions encountered in hibernacula or in summer maternal colonies. Virus was detected in the blood of hibernating bats at irregular intervals over a 42-day observation period; viremia perhaps was influenced by the amount of disturbance (arousal) involved in the blood sampling process. Target organs included brown fat, spleen, lung, kidneys, pancreas, and liver. Neutralizing antibody was not detected in sera collected from these bats between days 4 and 42 post-inoculation. In nonhibernating bats, virus was recovered from mammary glands, brown fat, pancreas, lungs, kidneys, and liver, in addition to blood. Attempts to infect bats orally or to transmit virus to suckling mice by the bite of viremic bats were unsuccessful. Virus was transmitted from viremic chickens to E. fuscus by the bite of Culiseta melanura and Aedes aegypti.


Pathology of Kawasaki disease: I. Pathology and morphogenesis of the vascular changes. Histopathological investigation of the vascular changes in Kawasaki disease was carried out on thirty-seven autopsied Japanese patients. Arterial lesions could be classified into following five stages from the view point of morphogenesis of arteritis; 1) endothelial degeneration and increased vascular permeability, 2) edema and degeneration of the media, 3) necrotizing panarteritis, 4) granulation formation, and 5) scarformation. Aneurysm with thrombus was observed mainly in the coronary artery in most patients. It is considered that the initial changes begins in the endothelial cells with increased vascular permeability. Platelet aggregation in the damaged endothelial cells seems to play an important role in the further development of the arteritic changes. Vascular lesions were observed not only in the arterial system but also in the venous system, therefore Kawasaki disease is a systemic vasculitis rather than a systemic arteritis.


Pathology of Kawasaki disease: II. Distribution and incidence of the vascular lesions. Histopathology, distribution and incidence of vascular changes were studied in tissues obtained at autopsy on thirty-seven children. Elastic and musculoelastic arteries showed a high incidence of arteritic changes, however the degree of the lesions was in general mild to moderate with the exception of the iliac artery, which revealed severe changes and a necrotizing panarteritis which was often accompanied with an aneurysm formation. A high incidence of arteritic changes, of which main histological feature was necrotizing panarteritis, was seen in extravisceral middle sized arteries. The coronary artery in particular was involved in each case and most had an accompanying aneurysm, some of which had ruptured. Intravisceral small sized arteries showed a relatively low incidence of arteritic changes and the degree of inflamation was in general mild. Phlebitis was present in over half the number of patients. The degree of lesions was mild in the small veins and mild to moderate in the large veins. Vascular lesions in Kawasaki disease should be termed systemic vasculitis rather than a systemic arteritis. There was a correlation between the caliber of involved vessels and the degree of vascular lesions. According to the histopathology, distribution and incidence of angitis, Kawasaki disease does resemble infantile periarteritis nodosa with the exception of the different manner of the coronary and iliac involvement.


Circumventing the Henderson-Hasselbalch equation. Prompt blood-gas analysis consisting of pH, PCO2, and PO2 is now almost universally accessible. PCO2 directly conveys the role of the respiratory mechanism in any acid-base disturbance. The metabolic condition, however, is hidden in a relationship between pH and PCO2 that is most faithfully expressed by the cumbersome Henderson-Hasselbalch equation (HHE). Two methods of quickly determining metabolic status from pH and PCO2, without resorting to the HHE, are discussed. The first method is well known and adjusts pH to represent only metabolic state by quantitatively cancelling the influence of hyper- or hypoventilation. The second method, not previously reported, uses measured pH to estimate the ratio [HCO3-]/PCO2, which, when multiplied by PCO2, equals bicarbonate ion concentration.


[Characteristics of the changes in the intra- and extracellular K+ and Na+ concentrations and the intra- and extracellular pH in the area of cardiac ishemia in experimental myocardial infarct complicated by ventricular fibrillation]. Localized ischemia of the heart complicated by ventricular fibrillation is characterized by a tendency to a higher rate of decrease in intra- and extracellular K+ gradient and intracellular pH in the myocardium as compared to cases without fibrillation. The higher rate of K+ escape from the ischemic cells may be linked with a sharper intracellular oxidation, evidence of which is the correlative dependence between the severity of disorders of K+ balance and decrease in intracellular pH in the myocardium in ischemia.


[Principles of the sanitary and housekeeping arrangements in space flight environments]. Significant changes in the functional state of crewmembers during prolonged space missions necessitate corrections to be made in the scope and methods of sanitation and housekeeping arrangements. Due to the fact that in space flight automicroflora representatives may acquire greater importance as possible pathogenic agents, procedures providing body cleanliness should include both physico-chemical exposures and direct skin microflora treatment. On the basis of pertinent investigations the authors have developed selection criteria and requirements for detergents and disinfectants that may be used in prolonged space flights.


Quinacrine-binding nerves: presence in the mouse ano-coccygeus muscle, disappearance after muscle transsection. The anococcygeus muscle exhibits non-cholinergic, non-adrenergic, inhibitory neurotransmission. In the present paper we describe the presence of quinacrine-binding beaded nerve fibres in the mouse anococcygeus muscle. A large number of fibres were running parallel to the smooth muscle bundles. A second distribution of fibres formed an irregular plexus. No quinacrine-binding nerve cell bodies were found within the muscle. When the anococcygeus muscle was cut close to its insertion in the rectum, the proximal part lost most of its quinacrine-binding nerves 2-7 days after transsection. The results demonstrate a correlation between presumed non-cholinergic non-adrenergic neurotransmission and quinacrine-binding nervous elements.


pH gradient electrophoresis and isoelectric focusing of lipoproteins on agarose bead thin layers. A new method of isoelectric focusing (IEF) and pH gradient electrophoresis, using thin layers of agarose gel beads, was devised to investigate chylomicrons and very low density lipoproteins (VLDL). pH gradient stability and cathodal gradient drift were similar to those of polyacrylamide gel IEF, and linearity of gradients was maintained for 23 hr. Chylomicrons and VLDL were detectable without staining. Chylomicrons from human serum and from rat lymph migrated in this system. Rat lymph chylomicrons, obtained by ultracentrifugation, migrated in several discrete bands, and this heterogeneity or rat chylomicrons was confirmed by electron microscopic demonstration of chylomicrons in each band. This new technique has permitted the first measurement of isoelectric points of some lipoproteins in the ultracentrifuged fraction of human serum chylomicrons and the first separation of multiple discrete fractions of ultracentrifuged lymph chylomicrons.


[Way of drying the skin and the renewal of skin surface activity]. The author tried to evaluate the impact of the way of drying the skin upon the renewal of its surface acidity, using a linen towel, paper towel, cellucotton and a stream of dry air. 20 persons were evaluated. Prior to the study, 1% solution of "BHP" soap had been put on their skin. PH was measured at the skin surface by a pH-meter (produced by "Radelkis") using a combined glass and calomel electrode. This method of skin drying was considered the best one, after which the average time of pH value renewal on the skin surface was the shortest, which amounted to the recovery of pH to the initial value. The way of drying the skin was found to have a significant impact on the time of pH renewal. The results were indicative of advisability of the use of linen towels.


Reactions of the sulfhydryl groups of membrane-bound bovine rhodopsin. Reactions of the sulfhydryl groups of bovine rhodopsin in rod outer segment membranes have been investigated using 4,4'-dithiopyridine. This reagent is uncharged at neutral pH and rapidly equilibrates across phospholipid bilayers. Membrane-bound rhodopsin has two kinetically distinguishable sulfhydryl groups reactive to the reagent, this stoichiometry being unchanged by bleaching provided the sulfhydryl reactions themselves are carried out in the dark. The rates of the reactions, however, are substantially increased by bleaching. Irradiation of bleached membranes, either with white light or wavelengths in the neighborhood of 475 nm, results in an increase in the number of reactive sulfhydryls relative to that found for bleached membranes in the dark. A component of the light-driven reaction is dependent on the Ca2+ content of the medium.


Effects on mitochondrial K flux of pH, K concentration, and N-ethyl maleimide. Based on published evidence that cation transport in mitochondria is not significantly dependent on a membrane potential, it is suggested that the process of mitochondrial cation transport may be nonelectrogenic. These experiments focused on the possibility that K+ flux into rat liver mitochondria may be directly coupled, via an energy-linked carrier mechanism, to OH- influx or H+ efflux. The dependence of the unidirectional K+ influx on the external K+ concentration indicates involvement of a saturable mechanism. Increasing the external pH from 7.0 to 8.0 increases the apparent V max of the K+ influx without significantly altering the apparent Km for K+. The pH dependence is greater in the presence of N-ethyl maleimide, a known inhibitor of the mitochondrial Pi/OH- exchange mechanism. N-Ethyl maleimide decreases the apparent V max at pH 7.0 and increases it at pH 8.0. Evidence indicates that both N-ethyl maleimide and a high external Pi concentration may stimulate the K+ influx at alkaline external pH (8.0) by preventing net exchanges between endogenous Pi and external OH-. An apparent first-order dependence of the K+ influx on the external OH- concentration is observed in the presence of N-ethyl maleimide. These results are consistent with a possible role of external OH- as a cosubstrate of the K+ transport mechanism.


Effect of carbohydrates and control of culture pH on beta toxin production by Clostridium perfringens type C. Clostridium perfringens type C strain CN 5384 produced a higher level of beta toxin in a controlled pH medium containing 1% glucose, starch, or sucrose than in media with dextrin, fructose, or raffinose. Toxin synthesis was not related to the growth yield. The effect of glucose on beta toxin production by 11 strains was investigated with and without control of the culture pH at 7.5. Strain CN 5386 produced distinctly higher toxin when the pH of the culture was maintained at 7.5, compared with uncontrolled pH.


Effect of trichloroacetic acid treatment on certain properties of spores of Bacillus cereus T. Spores of Bacillus cereus T treated with trichloroacetic acid (6.1--61.2 mM) were compared with untreated spores, and as the concentration of the chemical increased, the following alterations in spore properties were found: (1) the extent of germination decreased irrespective of the germination medium used; (2) the spores became sensitive to sodium hydroxide (1 N) and hydrochloric acid (0.27 N), but not to lysozyme (200 micrograms/ml); (3) loss of dipicolinate increased on subsequent heating; and (4) the spores became more sensitive to heat. However, trichloroacetic acid-treated spores were still viable and there was no significant change in spore components. The mechanism of action of trichloroacetic acid is discussed.


Interferon and cytotoxic factor (cytotoxin) released in the blood of mice infected with Mycobacterium bovis BCG. I. Enhanced production of interferon and appearance of cytotoxin stimulated by capsular polysaccharide of Klebsiella pneumoniae or bacterial lipopolysaccharide. Interferon production stimulated by the active substance (neutral fraction) of the capsular polysaccharide of Klebsiella pneumoniae (neutral CPS-K) in BCG-infected mice was compared with that by bacterial lipopolysaccharide (LPS). Prior infection with BCG increased the responsiveness of mice to the lethal effect of neutral CPS-K as well as to that of LPS. Associated with this, BCG-infected mice showed a markedly enhanced ability to produce interferon after stimulation not only by LPS but also by neutral CPS-K. In addition, a cytotoxic factor (cytotoxin) was found to be released in the serum of BCG-infected mice after injection of these inducers. The kinetics of production of interferon and cytotoxin stimulated by neutral CPS-K were very similar to those stimulated by LPS. The time pattern of cytotoxin production was not in parallel with that of interferon production. Interferon reached a peak 2 hr and cytotoxin 3 hr after injection with these inducers. Interferon and cytotoxin produced by neutral CPS-K showed essentially the same stabilities to heating at 56 C and to treatment at pH 2 respectively as those produced by LPS. Interferon was inactivated by heating at 56 C more rapidly than cytotoxin. Cytotoxin was inactivated by treatment at pH 2 for 24 hr, whereas interferon activity was well preserved after this treatment. These results suggest that both activities are the result of different substances.


Effects of tannic acid and its related compounds upon Chikungunya virus. The present report describes not only the effects of tannic acid (TA; belonging to hydrolyzable tannins) and its related compounds upon the infectivity of Chikungunya virus (CHIKV) but also the mechanism involved in this phenomenon. Our data show that TA inactivates CHIKV in vitro. Since the inactivating effect turned out to be pH-dependent and was suppressed by bovine serum albumin, it is most probable that the virus-inactivating capacity of TA is attributable to its preferential binding to proteins of virus particles. Examination on the virus-inactivating capacities of some TA-related compounds and comparison of their structures indicated that the active site of TA and its analogues might be the phenolic hydroxyl groups in their molecules. It seems that the active groups interact with the proteins of virus particles, resulting in a reduction or loss of viral infectivity. Discussion is made on the specificity of the actions of tannins and the possibility of application thereof to chemicals which are useful to investigate the nature and properties of viral proteins.


[Proteolytic enzymes produced by Aspergillus ochraceus in relation to their plasma coagulating and fibrinolytic activities]. The paper describes some properties of coagulases and fibrinolytic enzymes isolated from a proteolytic complex produced by Aspergillus ochraceus HP-19 during submerged cultivation on a synthetic medium. Proteolytic enzymes with the plasmocoagulating activity hydrolyzed casein at the maximum rate at 45 degrees C and pH 8.0--10.0. The coagulases were stable at pH 5.0--7.0 and were rather resistant to low pH values. The enzymes were entirely inactivated at 55 degrees C within 20--30 min. The activity of the coagulases was inhibited with the ions of Cu, Co, Ag, Pb, Mn, Zn and Hg (1.10(-3) M) by 100, 91, 85, 50, 50, 38 and 25%, respectively. The coagulases were entirely inhibited with EDTA whereas PCMB and PMSF inhibited their activity only to a small extent. The mechanism for blood clotting with the coagulases of Aspergillus ochraceus HP-19 is presumed to consist in the activation of protrombin via its limited specific proteolysis. The fibrinolytic enzymes of Aspergillus ochraceus HP-19 had the optimal pH 8.5 for casein, were stable at pH 6.0, and entirely inactivated at 55 degrees C within 5 min. In contrast to coagulases, they were resistant to the action of heavy metal ions. The enzymes were stabilized by the ions of Ca. The activity of the fibrinolytic enzymes of Aspergillus ochraceus HP-19 was completely inhibited with PMSF. Therefore, they belong to the class of serine proteases.


[Effect of cultivation conditions on lipase biosynthesis by Serratia marcescens]. The effect of cultivation conditions (the composition of the growth medium, pH, temperature) on lipase biosynthesis was studied with Serratia marcescens 345 and Serratia marcescens 348. The composition of the medium was found to be of key importance for lipase synthesis; the highest lipolytic activity of S. marcescens 345 was detected on the medium with 5% soybean flour broth while that of S. marcescens 348 was displayed on MPB. Sodium salts of cholic acids added at different concentrations to the growth medium had different effect on the biosynthesis of exo- and endolipases by the two strains.


Squalene synthetase.

Squalene synthetase. In the first part of the review the background to the discovery of the asymmetric synthesis of squalene from two molecules of farnesyl pyrophosphate and NADPH is described, then the stereochemistry of the overall reaction is summarized. The complexity of the biosynthesis of squalene by microsomal squalene synthetase demanded the existence of some intermediate(s) between farnesyl pyrophosphate and squalene. This demand was satisfied by the discovery of presqualene pyrophosphate, an optically active C30 substituted cyclopropylcarbinyl pyrophosphate, the absolute configuration of which at all three asymmetric centers of the cyclopropane ring was deduced to be R. Possible mechanisms for the biosynthesis of presqualene pyrophosphate and its reductive transformation into squalene are presented. In the second part of the review the nature of the enzyme is discussed. The question whether presqualene pyrophosphate is an obligate intermediate in the biosynthesis of squalene is examined, with the firm conclusion that it is. It is as yet uncertain whether the two half reactions of squalene synthesis, i.e. (i) 2 x farnesyl pyrophosphate leads to presqualene pyrophosphate; (ii) presqualene pyrophosphate + NADPH (NADH) leads to squalene, are catalyzed by one or two enzymes or by a large complex with two catalytic sites. Evidence is cited for the existence on the enzyme of two distinct binding sites with different affinities for the two farnesyl pyrophosphate molecules. The types of enzyme preparations available at present are described and types of experiments carried out with these are critically examined. The implications of the properties of a low molecular weight squalene synthetase solubilized with deoxycholate from microsomal membranes is discussed and a model for the enzyme in an organized membrane structure is presented.


Development of psychiatric illness in drug abusers. Possible role of drug preference. The origin of the psychiatric illnesses observed in drug abusers is often unclear. This study examines the causal relation between drug abuse and specific psychiatric disorders. Fifty-one male veterans first seen in 1972, who were admitted at least once per year for six consecutive years for inpatient drug-abuse treatment, underwent psychiatric assessments at each admission. Eleven men mainly used stimulants, 14 depressants, and 26 opiates. Initial psychiatric examinations showed low symptom levels in all groups but no statistically significant differences among them. By the end of six years, five of the stimulant users had psychoses, and eight of the depressant users had serious depression. The narcotics users showed no change in psychopathology. Differences between the groups were significant at the 0.01 level. These changes were not due to acute toxic reactions, but our data suggest that abuse of particular drugs has a major role in the development of specific psychiatric illnesses. The possibility that different preexisting personality disorders lead to different kinds of drug abuse cannot be excluded.


Intracellular pH and the sodium requirement at fertilisation. Several lines of evidence suggest that ionic messengers are primary agents in the metabolic derepression which occurs at fertilisation. The derepression at fertilisation or parthenogenetic activation of the sea urchin egg occurs in two main phases. The first phase, which triggers the early events of fertilisation, is mediated by transitory increase of intracellular free calcium. The second, which triggers the late events of fertilisation, is mediated by a rise in the intracellular pH (refs 4-6). The transition from the early events of fertilisation of sea urchin eggs to the late events requires a minimal concentration of sodium in the external medium. External Na+ is required for the acid effux which follows fertilisation. Na+ requirement and the acid effux have been correlated in a hypothesis which proposes that internal protons are exchanged for external Na+ (refs 8, 9). By using pH-sensitive microelectrodes, we have examined the relationship between external Na+ and internal pH more closely. We demonstrate here that the increase of the intracellular pH following egg activation does require external Na+. However, the relative insensitivity of the alkalisation of the egg cytoplasm to large reductions of external Na+ is evidence against the Na-H exchange hypothesis.


Stimulation of the pituitary-adrenocortical axis by nerve growth factor. Nerve growth factor (NGF) is a protein essential for the development and maintenance of the peripheral sympathetic nervous system, causing responsive neurones to increase in size and to extend neurites. Biochemically, the selective induction of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase key enzymes in catecholamine biosynthesis is one of its most characteristic effects. Both the morphological and biochemical effects are modulated by glucocorticoids, suggesting a close relationship between specific effects of NGF and hormone action. NGF has been shown to induce an increase in adrenal cyclic AMP in intact but not in hypophysectomised rats, and so we have looked directly at the effect of systemic administration of NGF on the hypothalamo-pituitary-adrenal axis. We report here that NGF induced an enhanced secretion of adrenocorticotropin (ACTH) and a prolonged increase in plasma glucocorticoid concentration after intravenous (i.v.) injection. Such effects could have important implications for the biological activity of NGF.


Antidepressant and antipsychotic agents. Numerous studies have been reviewed concerning the relationship of the catecholamines and indoleamines to depression and its treatment with antidepressant drugs. The profusion of data on the subject suggests the amines may be implicated in the disease, however no convincing hypothesis is available which unites the two. A similar situation seems to exist concerning the antipsychotic agents, dopamine and schizophrenia. The evidence appears to be overwhelming relating dopamine in the action of these agents and yet serious inconsistencies have been reported suggesting the need for a revision of the hypothesis regarding the mode of action of antipsychotic agents.


Foot-shock stress accelerates non-striatal dopamine synthesis without activating tyrosine hydroxylase. Electric foot-shock stress (20 min) increases DOPAC content in the frontal cortex (by about 80%) and in the nucleus accumbens (by 35%) but not in the striatum. However, foot shock stress failed to modify the kinetic properties of tyrosine hydroxylase (Vmax, Km for DMPH4 cofactor) in any of the above areas. Similar results were obtained in rats in which noradrenergic terminals in the n. accumbens and in the frontal cortex had been eliminated by injection of 6-OH-dopamine into the ascending dorsal noradrenergic bundle. The results support the hypothesis that limbic and cortical DA is involved in emotional states and indicate that DA synthesis may be regulated independently from changes in the kinetics properties of tyrosine hydroxylase.


The importance of choice of agonist in studies designed to predict beta 2 : beta 1 adrenoceptor selectivity of antagonists from pA2 values on guinea-pig trachea and atria. 1. pA2 values have been obtained for propranolol, butoxamine, H35/25 and atenolol on guinea-pig isolated trachea and atria (rate) using noredrenaline (beta 1-selective), isoprenaline (non-selective) and fenoterol (beta 2-selective) as agonists. 2. pA2 values varied with the agonist used on trachea but not on atria and, therefore, trachea : atria selectivity values varied with the agonist used. 3. It is suggested that the best estimate of the selectivity of an antagonist between beta 2- and beta 1-adrenoceptors is obtained by comparing its pA2 value obtained on trachea using a beta 2-selective agonist with that obtained on atria using a beta 1-selective agonist. The reasons for this are discussed. 4. The quantitative values for beta 2 : beta 1 selectivity obtained using the above pA2 values were butoxamine 17.0 H35/25 13.5, propranolol 2.75 and atenolol 0.036, i.e. butoxamine and H35/25 were beta 2-selective, propranolol was non-selective and atenolol was beta 1-selective. 5. The results support the hypotheses that guinea-pig trachea contains a mixture of beta 1- and beta 2-adrenoceptors and that guinea-pig atria contain only beta 1-adrenoceptors.


Demonstration of alpha-adrenoceptors in the rabbit heart by [3H]-dihydroergocryptine binding. For direct identification of alpha-adrenoceptors in a membrane fraction of the rabbit heart the potent alpha-adrenoceptor antagonist [3H]-dihydroergocryptine ([3H]-DHE) was used. 1. The binding of [3H]-DHE was saturable with 80 fmol of [3H]-DHE bound/mg protein and of high affinity with an equilibrium dissociation constant (KD) of 11.5 nM. Binding of [3H]-DHE (6 nM) was rapid (t 1/2 = 2 min) and readily reversible. From the ratio of the rate constants for forward (K1 = 1.97 X 10(7) M-1 min-1) and reverse (K2 = 0.206 min-1) reactions a KD-value of 10 nM was calculated, which is in good agreement with that obtained by equilibrium studies. 2. Adrenergic agonists compete for [3H]-DHE binding in an order to potency: (-)adrenaline greater than (-)phenyleprine greater than (-)isoprenaline and adrenergic antagonists in the order: phentolamine greater than yohimbine greater than (-)propranolol. Binding is stereospecific as indicated by the greater potency of (-)adrenaline than (+/-)adrenaline in displacing [3H]-DHE from the binding sites. 3. For comparison the binding of the potent beta-adrenoceptor antagonist (-)[3H]-dihydroalprenolol ((-)[3H]-DHA) was measured in the same membrane fraction. The number and affinity of beta-adrenoceptors amounted to 115 fmol of (-)[3H]-DHA bound/mg protein at saturation and KD = 7.9 nM. Adrenergic agonists compete for (-)[3H]-DHA binding in an order of potency: (-)isoprenaline greater than (-)adrenaline greater than (-)phenylephrine; and adrenergic antagonists in the order: (-)prapranolol greater than phentolamine. 4. It is concluded that in a membrane fraction of the rabbit heart there exist binding sites for [3H]-DHE which have characteristics indistinguishable from alpha-adrenoceptors. Thus the present results are in agreement with previously reported data on the existence of cardiac alpha-adrenoceptors in the rabbit heart (Schümann et al., 1974; Endoh et al., 1976b).


Alpha adrenoceptors in rat brain: direct identification with prazosin. Tritiated prazosin was used to characterize high affinity binding sites with characteristics similar to alpha 1 adrenoceptors in rat brain membranes. These sites were compared with alpha 2 adrenoceptors labeled with tritiated clonidine. The prazosin sites had an association constant of 2 nM-1 and bound to ligand optimal around pH 7.0. The density of the sites was 300 fmoles per mg of protein; the half time of dissociation of prazosin was 7 min at 30 degrees C. The order or potencies of agonists, determined from binding-inhibition experiments with labeled prazosin, was: naphazoline greater than clonidine greater than adrenaline greater than noradrenaline greater than phenylephrine greater than alpha-methylnoradrenaline greater than dopamine. The order of potencies of antagonists was: prazosin greater than phenoxybenzamine greater than phentolamine greater than clozapine greater than yohimbine. Sodium ions and divalent cations as well as guanyl nucleotides have little or no effect on the binding of the labeled antagonist. This is in contrast to the binding of the labeled agonist clonidine (Glossmann and Presek, 1979a, 1979b). Labeled prazosin may be a useful tool to characterize alpha 1 adrenoceptors.


Adenosine effects upon transmitter release parameters in the Mg2+-paralyzed neuro-muscular junction of frog. The influence of adenosine upon the process of transmitter release was investigated in Mg2+-paralyzed frog neuromuscular junction by using conventional microelectrophysiological techniques and binomial statistical analysis. The statistical parameters used were: m (mean number of quanta released per impulse), p (the probability of quantal release) and n (store of quanta available for release). Adenosine decreased the mean quantal content (m) by reducing n. This effect appeared to be dependent upon the free intracellular Ca2+. The nucleoside competed with the transmitter releasing effects of ouabain and increased the amount of effective Ca2+ necessary for quantal release. It did not change the slope of release parameters modification during low-frequency facilitation. The data are discussed in terms of a physical model of release.


Lipid metabolism in uremic and nonuremic acidosis. In bilaterally nephrectomized rats and in normal rats treated with 0.1 N HCl elevated levels of plasma total glycerol and lowered FFA and free glycerol are observed after a period of 25 h of starvation. In both cases the removal of intravenous lipid loads is slowed down, plasma LA is decreased after starvation and after lipid loads. The bicarbonate treatment of nephrectomized rats tends to normalize the plasma lipid levels and LA as well as the removal of intravenous lipid loads. The etiological importance of acidosis in uremic hypertriglyceridemia and possible clinical significance is pointed out.


Dopaminergic antagonism of L-5-hydroxytryptophan-induced myoclonic jumping behavior. We studied the effect of dopamine agonists (levodopa, apomorphine, lergotrile, and M-7 [2(dimethylamino)5,6-dihydroxytetralin] on myoclonic jumping behavior in young male guniea pigs. All these agents had a significant antagonistic effect on the frequency of this serotonin-mediated behavior. The duration of the antagonism corresponded in all cases to the duration of stereotyped chewing behavior induced by these agents alone. The dopamine antagonist haloperidol potentiated jumping behavior. Therefore myoclonic jumping behavior is influenced by dopaminergic mechanisms, and this behavior may be the result of interaction between dopaminergic and serotonergic activity. The role of dopaminergic mechanisms in human myoclonic disorders needs further clarification.


Controlled intravenous bicarbonate and fetal-maternal acid-base balance. I. The primipara. The effects of a controlled sodium bicarbonate (SB) infusion on the acid-base balance of the primiparous mother and fetus at labor and delivery were evaluated. Two identical groups of primiparas with normal labor and delivery were studied. According to acid-base parameters observed in the mothers and fetuses of a control group, the pharmacologic dynamics, and the space of distribution of SB, 2 mEq/1 kg of total body weight were administered to the mothers of the study group, beginning at a cervical dilation of 6 cm until full dilation occurred. Highly significant changes in pH, base excess (BE), and plasma bicarbonate were observed in both the mothers and fetuses. In the latter, the significant changes appeared after a time lag of about 2 hours. No adverse effects in the mothers and fetuses were observed. The significant reduction of the relative fetal acidosis by the controlled SB infusion justifies further studies on the therapy potentials of this method in high-risk deliveries and during intrapartum fetal distress.


Effect of macular ablation on vertical optokinetic nystagmus in the squirrel monkey. After macular ablation in squirrel monkeys, a significant improvement of the slow-phase eye speed of vertical optokinetic nystagmus (VOKN) was found. This findings is in contrast to the result of our previous study in which horizontal optokinetic nystagmus (HOKN) was not improved following macular ablation. The different influence of macular ablation on HOKN and VOKN could be due to the directional characteristics of vestibular and visual interaction relative to the direction of gravity and to the difference in horizontal and vertical vestibulo-oculomotor neural pathways, including vestibular end-organs. The vertical directional dominance which was observed preoperatively was similarly observed in the postablative status; the upward slow-phase nystagmus was easier to provoke than the downward nystagmus by application of an identical stimulus.


Model experiments on nitrite and nitrate in simulated primeval conditions. In experiments on the prebiotic formation of nitric oxides, anoxic mixtures of N2 and water vapour were sparked in contact with phosphate buffer solutions at various pH values. Nitrite was found in the aqueous phase, and nitrate grew from it, presumably by reaction with H2O2. In acid solutions, these anions were reduced and destroyed by Fe2+, and the same was true of nitrite in solutions kept at a pH value similar to that of the contemporary ocean (8.2) with HEPES buffer. Nitrate was not destroyed in short-term experiments, but as in sparking nitrate is formed only vianitrite, neither anion could accumulate. In further sparking experiments with alkaline sulphide, both nitrite and nitrate were reduced entirely. It is concluded that it is unlikely that the primeval ocean contained appreciable concentrations of nitrite or nitrate either at the reducing or at the redox-neutral stage.


Oxygen as a factor in eukaryote evolution: some effects of low levels of oxygen on Saccharomyces cerevisiae. A comparative study of the effects of varying levels of oxygen on some of the metabolic functions of the primitive eukaryote, Saccharomyces cerevisiae, has shown that these cells are responsive to very low levels of oxygen: the level of palmitoyl-Co A desaturase was greatly enhanced by only 0.03% (v/v) oxygen. Similarly, an acetyl-CoA synthetase associated predominantly with anaerobic growth, was stimulated by as little as 0.1% oxygen, while an isoenzyme correlated with aerobic growth, was maximally active at much higher oxygen levels (greater than 1%). Closely following this latter pattern were three mitochondrial enzymes that attained maximal activity only under atmospheric levels of oxygen.


A case of transient renal tubular acidosis type 1,4 hybrid RTA: a study of the pathophysiologic characteristics of the acidification defect. The diagnosis of transient renal tubular acidosis was made in a 16 months old boy. Bicarbonate titration studies revealed that the acidification defect consisted of an association of proximal and distal tubular acidosis. The effect of experimentally induced potassium depletion revealed that hyperkalemia contributed to the acidification defect. After correction of the acidification disorder a defect in urinary concentration was still present.


Metachromatic leukodystrophy without arylsulfatase A deficiency. Two siblings of consanguinous parents were noted to have a neurologic syndrome marked by developmental delay, regression of psychomotor performance, marked spasticity and progressive central nervous system degeneration. Markedly delayed nerve conduction times and a sural nerve biopsy which demonstrated changes typical of metachromatic leukodystrophy (MLD) were evident. Impairment of sulfated glycolipid metabolism was documented by analysis of glycospingolipid in urinary sediment. In spite of these findings, activities of arylsulfatase A and cerebroside sulfatidase in white blood cells and cultured skin fibroblasts were near normal. However, when intact growing fibroblasts were loaded with 35SO4-sulfatide a clear defect in sulfatide cleavage, comparable to that seen in MLD patients, was observed. Thus, these patients represent a new form of sulfatide storage disease -- MLD characterized by intact enzyme activity in cell homogenates but defective sulfolipid metabolism in vivo and in intact fibroblasts.


Effect of chronic reserpine administration on K+ and amylase release from the rat parotid gland. After chronic administration of reserpine, slices of the rat parotid gland incubated in vitro were found to release significantly more K+ and significantly less amylase than similar slices from untreated control animals. The extent of K+ release in the gland slices of the drug-treated animals shows no dependency on external Na+ or Ca++ and is probably enhanced because of a decreased activation of K+ uptake by the Na+-K+ pump. This effect may also involve a defective generation of cyclic GMP, which was found to enhance K+ uptake and, thus, to inhibit K+ release in control slices. The inhibition of amylase release observed after treatment with reserpine most likely results from a deficient level or utilization of the energy or nucleotide sources required for enzyme secretion, because a similar inhibition was observed in control slices after incubation in the absence of the purines inosine and adenine. These results indicate that chronic reserpine administration impairs the stimulus-secretion coupling mechanism in the rat parotid gland and results in abnormal responses from both alpha and beta adrenergic receptors.


Ontogeny of human T cells.

Ontogeny of human T cells. Current knowledge of human T cell ontogeny is reviewed in terms of appearance of T cells in central and peripheral lymphoid organs, maturation of T cell markers, and development of immune functions. Extrapolation of growth curves derived from cell counts from fetal thymus, spleen, and bone marrow indicates the appearance of lymphocytes at 3.5 weeks gestation. E-rosette-forming cells are present in thymus at 11 weeks and in peripheral organs 15 to 16 weeks gestation. beta-2-Microglobulin is associated with all lymphoid cells by 13 weeks gestation. Lymphocyte responses to the T cell mitogen phytohemagglutinin (PHA) are first detected in thymus at 10 weeks and in spleen and blood 3 to 4 weeks later. Allogeneic responses in mixed lymphocyte reactions develop at about 7.5 weeks in fetal liver and later in thymus and peripheral organs. Lymphocytotoxicity for xenogeneic cells is a property of bone marrow cells and not thymocytes. Several aspects of development of a suppressor T cell in human neonates is discussed and related to similar findings in the mouse. These studies indicate a relatively high degree of maturation of human T cells during fetal life.


Calcium inhibits urinary acidification: effect of the ionophore A23187 on the turtle bladder. The role of intracellular calcium in urinary acidification was studied in the turtle bladder with the use of the ionophore A23187. In the presence of calcium acidification was significantly inhibited in the hemibladders treated with the ionophore as compared to control hemibladders treated with dimethylsulfoxide (the vehicle used to dissolve the ionophore). In the absence of calcium both the ionophore and dimethylsulfoxide failed to cause any change in acidification. The apparent proton motive force and active conductance of H+ were unchanged in dimethylsulfoxide treated hemibladders. In the presence of the ionophore and calcium, the proton motive force and the conductance were significantly decreased when the H+ current was low (less than 30% of control values); when the H+ current was decreased, but not less than 30%, the proton motive force was unchanged. These data provide evidence for an important role of intracellular calcium in the regulation of urinary acidification.


Responses of Purkinje cells of the cerebellar vermis to neck and macular vestibular inputs. 1. The dynamic analysis of the control exerted by neck and macular vestibular receptors on the cerebellar cortex has been investigated in precollicular decerebrate cats submitted to sinusoidal rotation along the longitudinal axis of the animal at the frequency of 0.026 Hz and at peak amplitudes up to 10 degrees for the neck input and 15 degrees for the macular input. 2. Purkinje (P) cells located in the vermal cortex of the cerebellar anterior lobe, particularly in the longitudinal parasagittal zone which projects to the ipsilateral lateral vestibular nucleus (LVN), showed a sinusoidal modulation of the firing rate in response to sinusoidal stimulation of the neck receptors or the vestibular receptors, the phase of the responses being in most units related to the extreme neck or head position. Mossy fiber (MF) and/or climbing fiber (CF) responses of the same or different P-cells to the two inputs were observed. 3. The sensitivity of the MF-response of the P-cells to the neck input, elicited by sinusoidal rotation of the neck and expressed in per cent of the average firing rate per degree of neck rotation, corresponded on the average to 2.71 +/- 1.67, S.D. This value was significantly higher than that of the MF-response of the P-cells to the macular input elicited by sinusoidal tilt along the longitudinal axis of the whole animal, which correspond to 1.71 +/- 1.01, S.D. 4. Most of the MF-responses of the P-cells to the neck input were characterized by an excitation during side-down rotation of the neck  and by an inhibition during side-up rotation, whereas most of the MF-responses of the P-cells to the macular input showed just the opposite behavior, being inhibited by side-down tilt of the animal and excited by side-up tilt. 5. Units which received a convergent input from both neck and macular receptors and showed an antagonistic pattern of response to the two inputs were tested during rotation of the head alone, in order to excite simultaneously the two kinds of receptors. Due to the higher sensitivity of the neck over the macular response, the magnitude of the combined response tended to be similar to the difference between the individual ones. Moreover, the phase of the resulting response was always modified with respect to that of the response to the neck input alone, and became in some instances related to velocity of neck rotation rather than to neck position. 6. These findings indicate that opposite responses to neck and macular inputs occur at corticocerebellar level. However, a final integration of the two inputs, leading to suppression of the conflicting responses, may occur either at medullary (LVN) or at spinal cord level.


Review: ethidium fluorescence assays. Part 1. Physicochemical studies. DNA and RNA can be assayed rapidly and very sensitively by exploiting the enhanced fluorescence of ethidium intercalated into duplex regions. By assaying at different pHs and introducing a heating/cooling cycle, a great many physicochemical aspects of DNA and RNA can be studied avoiding the use of radiolabels, and often giving information not otherwise readily obtainable. Studies are described on duplex DNA which involve measurement of extinction coefficients, cross-linking by chemicals, Cot curve analysis as well as estimation of drug-DNA binding constants. The assays can be adapted to investigate multi-stranded nucleic acid structures. The use of covalently closed circular DNA also allows rapid and extremely sensitive measurements of nicking caused by irradiation or drugs.


Stability of the unique anticodon loop conformation of E.coli tRNAfMet. Initiator tRNAs have an anticodon loop conformation distinct from that of elongation tRNAs as detected by susceptibility to S1 nuclease. We now find the anticodon loop conformation of E. coli tRNAfMet to be stable under different salt conditions as detected by using S1 nuclease as a structural probe. In contrast, a conformational change is observed in the T- and D- loop of this tRNA in the absence of added Mg2+. This change can be suppressed by spermine. Even under those conditions effecting a change in T- and D- loop conformation, the anticodon loop does not change. This suggests that the conformational shift is controlled by Mg2+ and restricted to the D- and T- loop region only without affecting the anticodon domain. The use of S1 nuclease as a conformational probe requires the use of kinetic studies to determine the initial cleavage sites. Thus, the use of a strong inhibitor which immediately stops the action of this nuclease is necessary. ATP is shown to be such an inhibitor.


Tienilic acid in mild to moderate hypertension. Tienilic acid has blood pressure lowering properties alone and in combined treatment with beta-adrenergic blocking agents; 250 mg of tienilic acid seems to correspond to 50 mg of hydrochlorothiazide. Tienilic acid effectively reduces serum urate and has no marked or rapid effect on potassium balance. During short-term treatment, no impairment of glucose tolerance was found.


Physico-chemical characterization of hamster interferon. The glycoprotein nature of Syrian hamster interferon was tested on several immobilized lectins. The specific retention of a small portion (20%) of interferon activity was observed only on concanavalin A-agarose; Component I of the interferon (not retained) has an apparent molecular weight of 23,500 whereas Component II (retained) is larger, 31,500. The apparent hydrophobicity of Syrian hamster interferon was probed by its chromatography on: (a) straight chain hydrocarbons of varied length; (b) aromatic ligands (aminobenzene, benzylamine, beta-phenylethylamine, gamma-phenyl-propylamine); ligands listed in (a) and (b) were immobilized to cyanogen bromide-activated agarose (isourea linkage); and (c) phenyl-agarose (Phenyl-Sepharose CL-4B), an aromatic ligand immobilized via a 2-hydroxypropyl arm to the agarose (ether linkage).


A modified procedure for the preparation of UDP-beta-L-(U-14C)rhamnose. This paper describes the synthesis of UDP-L-(U-14C)rhamnose from UDP-D-(U-14C)glucose and NADPH using an enzyme preparation of Nicotiana tabacum var. Xanthi. A procedure to separate UDP-l-rhamnose from the other compounds in the reaction mixture is described. Optimal separation was achieved in ethanol 95%-1 M ammonium acetate (pH 3.8) (7:3, v/v) at 30 degrees C.


Effect of pentobarbital anesthesia on serotonin metabolism in hypoxic rat brain. Two hours of pentobarbital anesthesia resulted in marked elevation of serotonin (5-HT) level and decreased accumulation of 5-hydroxyindolacetic acid (5-HIAA) after probenecid, in specific parts of rat brain. In hypoxia, either no change (cortex, hypothalamus) or an increase (striatum, pons with medulla) in 5-HT content was observed. Concomitantly, the decrease of 5-HIAA level and its accumulation after transport inhibition occured. Pentobarbital pretreatment of hypoxic rats resulted in further diminution of 5-HT degradation after probenecid, having synergistic character in pons with medulla, which points to the specific sensitivity of serotonergic system in the latter area. Hypoxia-induced decrease of 5-HIAA level was prevented by pentobarbital pretreatment. The possible participation of 5-HIAA efflux inhibition in this phenomenon is discussed.


Evidence for a tyrosine residue at the active site of phosphoglucomutase and its interaction with vanadate. The rate of transfer of [32P]phosphate from [32P]-labeled phosphoglucomutase (alpha-D-glucose-1,6-bisphosphate:alpha-D-glucose-1-phosphate phosphotransferase, EC 2.7.5.1) to glucose increases dramatically between pH 8.5 and 10.5 with a half maximal rate at pH 9.8. This suggests the participation of a residue containing an ionizable group with a pK close to 10. The inhibition of enzyme activity obtained with tyrosine-derivatizing reactions--iodination, nitration, acetylation, and diazo coupling--is strongly indicative of tyrosine participation. Thiol reagents, p-hydroxymercuribenzoate and ethyleneimine, were without effect. Vanadate and arsenate augmented the transfer reaction 200- and 2.5-fold, respectively, and lowered the pH optimum of the reaction.


Hydrophilic region of lecithin membranes studied by bromothymol blue and effects of an inhalation anesthetic, enflurane. A pH-indicator dye, bromothymol blue, was used to probe the hydrophilic surface of dimyristoyl-, dipalmitoyl-, and distearoylphosphatidylcholine bilayer vesicles. The apparent pK of the surface-adsorbed dye was larger than the bulk pK value. The contribution of the choline positive charge on the dissociation constant of the dye adsorbed on the vesicle surface was estimated by screening the charge interaction with 2 M KCl. The effective surface potentials interacting with the dye were thus estimated to be 33.2, 45.6, and 46.8 mV, respectively, for the dimyristoyl-, dipalmitoyl-, and distearoylphosphatidylcholine vesicles. From the differences between the obtained effective potentials and the calculated surface potentials of the charge-determining plane of the choline head, the distances between the prototropic part of the dye and the choline charge-determining plane were estimated to be 10.5, 8.0, and 7.8 A, respectively. These values were obtained at 25 degrees C; the dimyristoylphosphatidylcholine membrane was in the liquid-crystalline phase and the other two were in the solid gel phase. Addition of an inhalation anesthetic, enflurane, decreased the distance in the dimyristoylphosphatidylcholine vesicles and increased the distance in the dipalmitoyl- and distearoylphosphatidylcholine vesicles. The increase of precessional motion of choline head by the inhalation anesthetic is apparently responsible for the changes.


pH-dependent migration of copper(II) to the vacant zinc-binding site of zinc-free bovine erythrocyte superoxide dismutase. Bovine erythrocyte superoxide dismutase (Cu(2)Zn(2)SODase; superoxide:superoxide oxidoreductase, EC 1.15.1.1) consists of two identical subunits each containing Cu(2+) and Zn(2+) in close proximity. We describe here electron spin resonance (ESR) and visible absorption spectroscopic studies of the zinc-free derivative of this protein, Cu(2)E(2)SODase (E = empty) over the pH range 6-10. The ESR spectrum of the zinc-free protein at 77 K is markedly pH dependent. At pH < 8.0 the ESR spectrum is axial in appearance. At pH > 8.0, the lineshape becomes increasingly distorted with increasing pH until, at pH = 9.5, the spectrum is very broad and resembles that of the four-copper derivative Cu(2)Cu(2)SODase and of model imidazolate-bridged binuclear Cu(II) complexes. ESR spectra at 30 degrees C are also consistent with formation of Cu(II)-Im-Cu(II). A plot of changes in the signal amplitude of g perpendicular for Cu(2)E(2)SODase as a function of pH gives an apparent pK(a) of 8.2 for the transition. The long-wavelength absorption with lambda(max) = 700 nm characteristic of Cu(2)E(2)SODase shifts with increasing pH to 800 nm and the resulting visible spectrum is identical to that of Cu(2)Cu(2)SODase. All of the above-mentioned spectroscopic changes induced by additions of NaOH are reversed when the pH is decreased with HNO(3), although the approach to equilibrium is slow in the latter case. The results of these experiments are consistent with a reversible, pH-dependent migration of Cu(2+) from the native copper site of one subunit of the zinc-free protein to the empty zinc site of another subunit. By contrast, native protein, Cu(2)Zn(2)SODase, and the four-copper protein, Cu(2)Cu(2)SODase, show no variation in visible or ESR spectral properties in this pH range. Some previous results concerning the activity of Cu(2)E(2)SODase and its thermal stability are reexamined in light of these new findings.


Leukotriene C: a slow-reacting substance from murine mastocytoma cells. Murine mastocytoma cells treated with calcium ionophore A23187 produced a slow-reacting substance (SRS) that caused guinea pig ileum to contract. The response was reversed by the SRS antagonist FPL 55712. On the basis of isotope incorporation experiments, spectroscopy, and chemical degradations, the SRS was identified as a cysteine-containing derivative of 5-hydroxy-7,9,11,14-icosatetraenoic acid. This amino acid was attached in thioether linkage at C-6. The SRS is structurally related to previously identified epoxy and dihydroxy metabolites of arachidonic acid in leukocytes. A common feature is the presence of a conjugated triene, and the name "leukotriene" has been introduced to designate these compounds. Leukotriene A (5,6-epoxy-7,9,11,14-icosatetraenoic acid) is an intermediate in the formation of leukotriene B (5,12-dihydroxy-6,8,10,14-icosatetraenoic acid) and is proposed to be a precursor also of leukotriene C, which is the SRS identified here.


Hydroperoxides can modulate the redox state of pyridine nucleotides and the calcium balance in rat liver mitochondria. When rats are fed a selenium-deficient diet, the glutathione peroxidase activity in liver mitochondria decreases within 5 weeks to 0-6% of that of control animals fed on a diet supplemented with 0.5 ppm of selenium as sodium selenite. Analysis of the temperature dependence of energy-linked Ca(2+) uptake by means of Arrhenius plots reveals two breaks (at around 11 degrees C and 24 degrees C) in mitochondria isolated from selenium-supplemented animals, whereas in selenium-deficient rats the break at 11 degrees C is absent. Ca(2+)-loaded mitochondria of selenium-supplemented rats-i.e., with active glutathione peroxidase in the matrix-lose Ca(2+) rapidly, with a concomitant oxidation of endogenous NAD(P)H, when exposed to t-butyl hydroperoxide or H(2)O(2). In contrast, in selenium deficiency, t-butyl hydroperoxide and H(2)O(2) induce neither a release of Ca(2+) nor an oxidation of NAD(P)H. The peroxide-induced oxidation of NAD(P)H is reversible in the presence of succinate when no Ca(2+) has been taken up. When Ca(2+) has previously been accumulated, however, the oxidation of NAD(P)H is irreversible. Enzymatic analysis of mitochondrial pyridine nucleotides reveals that the peroxide-induced oxidation of NAD(P)H in Ca(2+)-loaded mitochondria leads to a loss of NAD(+) and NADP(+). It is proposed that the redox state of mitochondrial pyridine nucleotides can be or is in part controlled by glutathione peroxidase and glutathione reductase and is a factor in the balance of Ca(2+) between mitochondria and medium.


Involvement of long-chain acyl coenzyme A for lipid synthesis in repression of acetyl-coenzyme A carboxylase in Candida lipolytica. Mutant strains of Candida lipolytica defective in acyl-CoA synthetase II [acid:CoA ligase (AMP-forming), EC 6.2.1.3] have been isolated. The mutants fail to grow on fatty acid as a sole carbon source but are capable of incorporating exogenous fatty acid into cellular lipids. This observation, together with our previous finding that mutant strains defective in acyl-CoA synthetase I cannot incorporate exogenous fatty acid into cellular lipids but are able to degrade fatty acid via beta-oxidation, indicates the presence of two functionally distinct long-chain acyl-CoA pools in the cell--i.e., one for lipid synthesis and the other for beta-oxidation. Unlike the wild-type and the revertant strains as well as the mutants lacking acyl-CoA synthetase II, the mutants defective in acyl-CoA synthetase I do not exhibit the repression of acetyl-CoA carboxylase [acetyl-CoA:carbon-dioxide ligase (ADP-forming), EC 6.4.1.2] by exogenous fatty acid. Measurement of the two long-chain acyl-CoA pools with the aid of appropriate mutant strains has indicated that the long-chain acyl-CoA to be utilized for lipid synthesis, but not that to be degraded via beta-oxidation, is involved in the repression of acetyl-CoA carboxylase.


A new glnA-linked regulatory gene for glutamine synthetase in Escherichia coli. Mutations in the glnA region of the Escherichia coli chromosome due to Mu prophage insertion result in two phenotypic classes. One class is Gln- and does not synthesize glutamine synthetase[L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2] under any growth condition. The other class produces a low level of glutamine synthetase under all growth conditions and is uncoupled from the regulatory effects of mutations in the glnF and glnD genes. Complementation analysis demonstrates that these two classes of insertions are in different cistrons. From these data we suggest that a regulatory gene, glnG, tightly linked to glnA, mediates both activation and repression of glutamine synthetase synthesis. An analysis of the evidence accumulated to date makes it unlikely that glnG is the only gene in the glnA region involved in the complex system of nitrogen regulation.


Nitrogen control in Salmonella: regulation by the glnR and glnF gene products. The product of the glnR gene is required for nitrogen regulation of the synthesis of glutamine synthesis (Gln synthetase) [L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2] and two periplasmic transport proteins that are subject to nitrogen control in Salmonella. Strains with mutations to loss of function of the glnR product [e.g., a strain with a Tn10 insertion or one with an ICR-induced (frameshift) mutation in glnR] have about 3% as much Gln synthetase as a fully derepressed wild-type strain and are unable to increase synthesis of this enzyme or periplasmic transport proteins in response to nitrogen limitation. The structural gene for Gln synthetase, glnA, and those for the periplasmic transport proteins are unlinked on the chromosome; thus, glnR appears to encode a diffusible positive regulatory element. Consistent with this, the mutant glnR allele is recessive to the wild-type allele with regard to expression of glnA (synthesis of Gln synthetase). Although glnR is closely linked to glnA, strains with mutations to complete loss of function of the glnR product can be distinguished from glnA strains by their ability to produce detectable Gln synthetase and to grow in the absence of glutamine. To demonstrate unequivocally that glnR is distinct from glnA, we have purified and characterized Gln synthetase from a strain with a Tn10 insertion in glnR. Because the properties of Gln synthetase from the insertion mutant, most importantly the carboxyl-terminal sequence of amino acids, are the same as those of synthetase from wild type, the Tn10 insertion cannot be in glnA (if it were, the carboxyl terminus of Gln synthetase would have to be altered); therefore we conclude that the Tn10 insertion is in a regulatory gene, glnR, which is distinct from glnA. A model for the function of the glnR product together with the previously defined glnF product in mediating nitrogen control is discussed.


Requirement for membrane potential in injection of phage T4 DNA. The first stages of infection by phage T4 may be divided into energy-dependent and energy-independent processes. Irreversible adsorption, unplugging, and initial exposure of the DNA terminus may occur at 4 degrees C, or at 37 degrees C in bacteria whose energy-yielding metabolism has been poisoned. DNA injection into the cytoplasm needs higher temperatures and energy from the host cell. The nature of this energy requirements was deduced from the use of metabolic inhibitors. Our results show that T4 DNA injection specifically requires the presence of a protonmotive force across the cytoplasmic membrane of the host. Moreover, the chemical gradient (delta pH) does not appear to be essential, but the membrane potential (delta psi) is required.


Clonal hybrid cell lines expressing cholinergic and adrenergic properties. Different cholinergic cell lines were fused with an adrenergic neuroblastoma cell line (N115-BU-8). Its fusion with a cholinergic neuroblastoma-glioma hybrid produced a "hybrid-hybrid" line containing cholinergic and adrenergic enzyme activities. Both activities were also present in subclones of this line. The presence of catecholamines in single cells was confirmed by microspectrofluorimetry. These results are discussed with respect to the possibility of a simultaneous synthesis of noradrenaline and acetylcholine in single cells. The cholinergic and adrenergic enzyme activities are influenced by cell density, by dexamethasone, and by conditioned medium.


Interaction of phencyclidine ("angel dust") with a specific receptor in rat brain membranes. [3H]Phencyclidine binds to synaptic membranes from rat brain in a saturable, reversible, and selective fashion, with a dissociation constant Kd of 0.25 microM and a maximal binding capacity of 2.4 pmol/mg of membrane protein--i.e., 250 pmol/g of brain. The binding activity is concentrated in synaptosomal fractions, is higher in cerebral cortex and corpus striatum than in other parts of the rat brain, and is not detectable in the spinal cord. Only molecules of the phencyclidine series and ketamine are able to bind to the phencyclidine receptor. [3H]Phencyclidine bound to its receptor is not displaced by the classical neurotransmitters or neuromodulators. There is a good correlation between the apparent affinities of a series of phencyclidine analogs for the phencyclidine receptor and the pharacological activities of these analogs as measured by the rotarod assay.


Selective loss of noradrenergic phenotypic characters in neuroblasts of the rat embryo. To define the fate of embryonic neuroblasts in rat gut, which transiently express several noradrenergic traits, we investigated the high-affinity uptake of norepinephrine. At 12.5 days of gestation, these cells exhibited immunoreactivity to tyrosine hydroxylase [tyrosine 3-monoxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] and endogenous catecholamine fluorescence. However, by 13.5 days these noradrenergic neurotransmitter phenotypic characters essentially disappeared. In contrast, norepinephrine uptake, which was also apparent at 12.5 days, persisted at least through 17.5 days. These observations indicate that norepinephrine uptake develops as an additional noradrenergic characteristic in these cells and persists after the disappearance of other noradrenergic traits. Consequently, neurotransmitter phenotypic characters may be transiently displayed during normal development in vivo.


The perception of music.

The perception of music. A common but none the less remarkable human faculty is the ability to recognize and reproduce familiar pieces of music. No two performances of a given piece will ever be acoustically identical, but a listener can perceive, in both, the same rhythmic and tonal relationships, and can judge whether a particular note or phrase was played out of time or out of tune. The problem considered in this lecture is that of describing the conceptual structures by which we represent Western classical music and the processes by which these structures are created. Some new hypotheses about the perception of rhythm and tonality have been cast in the form of a computer program which will transcribe a live keyboard performance of a classical melody into the equivalent of standard musical notation.


The initial heat production in garfish olfactory nerve fibres. A study has been made of the temperature changes associated with the passage of a single impulse in the non-myelinated fibres of the garfish olfactory nerve: and the time course of these temperature changes has been compared with the time course of the electrical events during the action potential. As in other non-myelinated nerves studied the observed temperature changes result from a biphasic initial heat production consisting of a transient evolution of heat (the positive heat) followed by a rapid heat reabsorption (referred to as the negative heat). There is no evidence of any additional phases of initial heat production. At 0 degrees C the measured positive initial heat is 224 mucal/g impulse (937 muJ/g impulse); and the corresponding negative initial heat is 230 mucal/g impulse (962 muJ/g impulse). The residual initial heat is very small, being about -6 mucal/g impulse (-25 muJ/g impulse). In the range 0-10 degrees C there is no significant effect of temperature on the magnitude of either the positive or the negative phases of heat production. The experimental thermal records were analysed to determine the true time course of the temperature changes in the nerve undistorted by the recording system. The time course of the temperature changes does not fit with that of the transmembrane voltage change as represented by the monophasic compound action potential recorded externally from the same point on the nerve. A better fit is obtained if the temperature changes are compared with the square of the voltage change in accordance with the view that the heat derives almost wholly from free energy changes and entropy changes in the membrane capacity. The best fit is obtained if it is assumed that the membrane potential does not discharge to zero during the action potential but that at the peak of the action potential the charge (and hence the p.d.) across the membrane capacity retains about 24% of its resting value.


Estimates of quantal content during 'chemical potentiation' of transmitter release. The number of quantal transmitter packets (m), released from motor nerve terminals in response to a single stimulus, has been estimated from the ratio of the amplitudes of endplate currents (e.p.c.) to spontaneous miniature endplate currents (m.e.p.c.), in voltage-clamped endplates of the frog. At 6 degrees C, the average value of m at normal nerve-muscle junctions was about 300. If allowance is made for the temporal dispersion of quantal transmitter release during the e.p.c., this value is increased by about 30%. After treatment with diaminopyridine or tetraethylammonium, transmitter release in response to a nerve stimulus is greatly enhanced and values of m exceeding 10(4) are frequently found. Moreover, the duration of the e.p.c. becomes much longer than that of the m.e.p.cs. The number of packets then liberated during the e.p.c. is much larger than the number of 'active zones' of the endplate and may even exceed the total number of vesicles lined up in twin-files adjacent to the presynaptic membrane.


Antigen-induced secretion in the optic gland of Octopus vulgaris. The optic gland of the octupus responds to a variety of foreign proteins--1 to 2 days after the injection--by the mass production and release of a particulated material. In the electron microscope the antigen-induced substance shows as a non-membrane bound particle of size about 30 nm. When released into the blood it does not disintegrate instantly. The chemistry of this material, which is probably identical with the optic gland hormone, is not known. However, non-vesicular storage and release as well as the abundance of tubular mitochondria suggest that the hormone is a steroid. The optic gland hormone, which is known to control sexual maturation, feeding and death in Octopus, appears to be involved in a defence mechanism against non-octopus proteins.


Nickel uptake by Flacourtiaceae of New Caledonia. Herbarium and field specimens (over 300) of all of the Flacourtiaceae of New Caledonia were analysed for nickel in order to identify hyperaccumulators (greater than 1000 microgram/g dry mass) and to assess nickel accumulation in relation to the evolutionary status of 'nickel plants' of New Caledonia. One hyperaccumulator was identified in the genus Lasiochlamys, ten among Xylosoma, one among Casearia and seven among Homalium. Although these Homalium nickel plants had previously been recorded, fresh data for these and other Homalium are presented. The remarkable tolerance of Flacourticeae to ultrabasic rocks is shown by the fact that 75% of the species are found on such substrates. The number of hyperaccumulators was greatest in the genera Xylosoma and Homalium. The Flacourtiaceae are among the most primitive of all angiosperms and in common with other primitive hyperaccumulators, contain nickel as a complex with citric acid. The only advanced New Caledonian nickel plant (Psychotria douarrei) has most of its nickel bound with ligands other than citric acid, a feature of other advanced hyperaccumulators. It is postulated that nickel complexing with citric acid may be a primitive character. Most of the New Caledonian nickel plants belong to the order Violales of subclass Dilleniidae. It is suggested that hyperaccumulation of nickel is an evolutionary character which occurs in long-indisturbed floras such as that of New Caledonia.


Is genetically transmitted obesity due to an adipose tissue defect? 1. The aim of this investigation was to ascertain of a variety of obese rodents whether the primary cause of fat cell enlargement lay in the fat cell itself, or in its environment. Rodents studied were the mutant mice 'diabetic' (db/db), 'adipose' (dbad/dbad), and 'yellow obese' (Ay/+), New Zealand obese mice, CBA mice made obese with gold thioglucose, and obese BIO 4.24 hamsters. 2. Gonadal fat of obese or lean genotype was transplanted under the kidney capsule of an obese or lean host. Grafts were left in place for at least one month, then examined histologically to measure fat cell diameters, from which fat cell masses were calculated. 3. Immunological rejection of grafts was avoided either by using mice syngeneic except for the obesity producing mutation (db/db, dbad/dbad or Ay/+) or by transplanting into F1 hybrids (NZO X BALB/c) made by mating the strains acting as donors of obese or lean fat. Transplantation of fat between lean BIO 4.22 hamsters and obese BIO 4.24 hamsters was possible because these had common histocompatibility antigens. 4. In all the forms of murine obesity studied, 'lean' fat cells enlarged in an obese recipient to the size typical of cells in 'obese' fat whilst 'obese' fat cells shrunk in a lean recipient to, at least, the size typical of 'lean' fat. Lean hamster fat cells also enlarged in an 'obese' environment and 'obese' hamster cells shrunk in a 'lean' environment. 5. Environment therefore contributes to the determination of fat cell size in all the rodents studied, and in several rodents (db/db, dbad/dbad, Ay/+, and gold thioglucose obese mice) our results showed that environmental factors are of paramount importance in determining cell size, and factors associated with the fat cell itself make a negligible contribution.


The influence of the brain hormone on retention of blood in the mid-gut of the mosquito Aedes aegypti (L.). III. The involvement of the ovaries and ecdysone. Most female mosquitoes require a blood-meal in order to produce mature oöcytes. An egg development neurosecretory hormone (EDNH), which is produced in the medial neurosecretory cells (m.n.c.) of the brain and stored in the corpus cardiacum, is released into the haemolymph following the ingestion of blood and is essential for the promotion of ovarian development to maturity. It has been shown that a factor from the m.n.c., presumably EDNH, is necessary if the blood-meal is to be retained in the mid-gut until the oöcytes approach maturity. The present paper shows that retention is not a direct result of the action of EDNH, but is dependent on the ovaries and may well involve ecdysone. Removal of the ovaries before a blood-meal leads to early haem-defaecation, but delay can be restored by injection of ecdysterone. Sub-threshold feeders and mosquitoes decapitated immediately after the intake of blood, each of which would be expected to eliminate the blood-meal early, also show a delay in the onset of haem-defaecation when injected with ecdysterone. Further, both in ovariectomized insects and sub-threshold feeders the time of onset of haem-defaecation is associated with the dose of ecdysterone given.


Specific protection of the binding sites of D-Ala2-D-Leu5-enkephalin (delta-receptors) and dihydromorphine (mu-receptors). Phenoxybenzamine causes a long-lasting inactivation of the opiate receptors of the mu- and delta-type in homogenates of guinea-pig brain. The effect is selectively prevented when, before exposure to phenoxybenzamine, the homogenate is pre-incubated with ligands of high affinity for either of the two binding sites, i.e. dihydromorphine for the mu-receptor and Tyr-D-Ala-Gly-Phe-D-Leu for the delta-receptor. In contrast, Tyr-D-Ala-Gly-Phe-L-Leu amide, which has high affinities for both binding sites, protects both receptor sites.


Effects of MIF-I and melatonin on novelty-induced defecation and associated plasma 11-OHCS and brain catecholamines. In two experiments the effects were investigated of MSH-inhibiting factor-I (MIF-I) and of Melatonin on step-down latencies, defection, plasma 11-OHCS levels, whole brain DA and whole brain NE concentrations on Days 1, 3 and 5 of novelty exposure. Treatment with MIF-I led to a significant habituation of novelty-induced defecation over 5 days, whereas plasma 11-OHCS level was reduced only on Day 1. The concentrations of whole brain DA and whole brain NE also showed a significant increase over days of MIF-I and novelty treatment. Melatonin treatment, on the other hand, significantly inhibited novelty-induced defecation and reduced plasma 11-OHCS level on Day 5 of novelty exposure. Melatonin treatment led to a significant increase of whole brain DA in animals exposed to novelty for 5 days. Neither MIF-I nor Melatonin was found to significantly affect the step-down activity of treated animals. The overall results suggested a possible relationship between novelty-induced defecation and brain DA levels of MIF-I and Melatonin treated animals.


Effects of some anti-epileptic, neuroleptic and gabaminergic drugs on convulsions induced by D,L-allylglycine. The antagonism of various seizure and time-related components of the convulsions resulting after IV injection of D,L-allylglycine into male Wistar rats were assessed in a standard test procedure. Trimethadione and ethosuximide did not antagonize the seizure components, whereas clonazepam, phenobarbital, diphenylhydantoin, primidone, valproate sodium, aminoxyacetic acid, etomidate, acetazolamide, flunarizine, pipamperone and baclofen did. The allylglycine test may thus represent a relatively specific method of differentiating between drugs effective against partial or generalized convulsive seizures from those effective against absence seizures. The neuroleptics haloperidol and pimozide were completely inactive in contrast to their reported antagonism of bicuculine seizures. The spectra of the active substances are discussed with respect to Principal Component and Cluster Analysis. Noteworthy are the similarities between baclofen and etomidate; between aminoxyacetic acid, phenobarbital and valproate sodium; and between diphenylhydantoin and flunarizine.


The influence of cholinergic transmitter substances on the incorporation of (14C)-leucine and (3H)-fucose into the total proteins of hippocampus in vivo and in vitro. The incorporation of (14C)-leucine into the total proteins of the hippocampus is inhibited by high concentrations of cholinergic agonists, with nicotinic substances (such as 1,1-dimethyl-4-phenyl-piperazine) being more effective than muscarinic compounds (such as arecoline and pilocarpine). Under these conditions the incorporation of (3H)-fucose is not influenced.


Partial anxiolytic action of morphine sulphate following microinjection into the central nucleus of the amygdala in rats. In the social interaction test of anxiety, bilateral microinjections of morphine sulphate (10 microgram) into the central nucleus of the amygdala counteracted the reduction in social interaction normally seen when the test arena is unfamiliar to rats. However, these injections did not counteract the decrease in social interaction that is observed as illuminance of the arena is increased. Morphine injections into the medial site depressed social interaction below the levels shown by control animals. In the open field test, morphine produced a facilitation of peripheral activity when injected into the central nucleus whilst a decrease in rearing was observed following similar injections into the medial nucleus. Overall, these data indicate a partial anxiolytic action of morphine in the central amygdaloid nucleus. Results are discussed in relation to possible differences in opioid peptide innervation of these two amygdaloid nuclei.


Actions and interactions of amphetamine on self-stimulation in rats. The dose-response relationship for d-amphetamine (0.125-2 mg/kg, IP) and its l-isomer (0.125-3 mg/kg, IP) was studied in self-stimulation behavior of rats each with an electrode at posterior hypothalamus (PH, mainly monoaminergic) or area ventralis tegmentum (A10, dopaminergic). The drug effects increased with the dose reaching a peak (at 0.5 mg/kg with d-amphetamine and at 1.0 mg/kg with 1-amphetamine) and then decreased. The d-isomer was approximately twice as potent as the l-isomer in enhancing intracranial self-stimulation (ICSS) rate with electrodes at either site. Azaperone (mainly an alpha-adrenergic blocker) and haloperidol (an antidopaminergic neuroleptic) used in small doses (0.05 and 0.008 mg/kg respectively) which did not affect the baseline responding, blocked amphetamine-induced enhancement of ICSS in both groups of rats. Thus, amphetamine-induced facilitation of ICSS at both PH and A10 areas and its blockade by an alpha-adrenergic blocker as well as an antidopaminergic neuroleptic show the involvement of both noradrenergic and dopaminergic mechanisms in self-stimulation behavior.


Stability of indomethacin solubilized system. Physical and chemical stability testing was carried out for indomethacin in the form of solution, syrup and elixir solubilized by Tween 80 and Brij 99. It was found that sucrose and alcohol markedly stabilize the indomethacin solution.


The effect of a second laminated gelatin film on the release rate of sulfadiazine from gelatin-gel matrix. The influence of a second laminated gelatin film without the drug over a gelatin-gel matrix containing sulfadiazine on the release rate was investigated. It was found that increasing the thickness of the second laminated layer markedly decreased the release of the drug incorporated in the gelatin layer.


Characterization of the pH-dependent dimer-to-protomer transformation of cytochrome C oxidase at alkaline pH. The pH-induced dissociation of cytochrome c oxidase from dimer to protomer has been studied in the pH range 7 to 11. Findings are as follows: The heme A:copper ratio is 1.0 at both pH 7.4 and 10.6. The relative enzymatic activity is preserved at all pH values at which the dimer or protomer are found. The fraction of protomer, determined from sedimentation velocity profiles, increases from 0 to 1 as the pH is raised. The absorption and circular dichroism spectra in the Soret region change in ways indicating that the contributions of cytochrome a in typical cytochrome aa3 spectral patterns are progressively lost as pH increases. At pH values more alkaline than the above, denaturation occurs. The fraction of protomer, and certain parameters defined to quantitate the changes in spectral form, exhibit similar pH profiles for a given preparation; but these concerted changes occur over different pH ranges for different preparations. Nevertheless the optical parameters are linearly correlated with the fraction of protomer for each preparation. It is concluded that the spectral properties of the dimer and the protomer are intrinsic attributes of each species and are not directly affected by changes in ambient pH.


Intraventricular self-administration of acetaldehyde, but not ethanol, in naive laboratory rats. For 11 consecutive days, naive rats were maintained in operant chambers where they were given the opportunity to self-administer acetaldehyde (1,2, or 5% v/v), ethanol (2 or 10% v/v), or pH control solutions directly into the cerebral ventricles. Only the animals that had access to the 2 and 5% acetaldehyde solutions showed rates of lever pressing significantly higher than controls. It is suggested that acetaldehyde rather than ethanol itself may mediate the positive reinforcing effects of ethanol in the brain.


Discriminative stimulus properties of pentylenetetrazol and bemegride: some generalization and antagonism tests. In an operant procedure of lever pressing on a FR 10 schedule of food reinforcement, male hooded rats were trained to respond with a lever on one side of a food cup following a drug injection, and to respond with a lever on the alternate side following a 1 ml/kg saline injection. All of 14 subjects learned to discriminate reliably between the effects of 20 mg/kg pentylenetetrazol (PTZ) and saline. Seven of eight rats learned to discriminate between the effects of bemegride (5 mg/kg) and saline. None of 14 rats learned to discriminate between 5mg/kg PTZ and saline. The bemegride discriminative stimulus generalized to PTZ (20mg/kg) and was antagonized by chlordiazepoxide (10 mg/kg). Chlordiazepoxide, diazepam, flurazepam, clobazam, and meprobamate were all effective antagonist of PTZ in a dose-dependent manner. Bemegride and cocaine generalized to the PTZ discriminative stimulus in a dose-dependent manner, but d-amphetamine, methylphenidate, and nicotine did not. Since bemegride and PTZ are convulsants at higher doses, the discriminative stimulus properties of these drugs might be based on a subtle convulsive brain state. The anxiolytic properties of benzodiazepines and meprobamate suggest that the discriminative stimulus produced by these convulsants is related to an "anxiety-inducing" action.


Chronic effects of mianserin on noradrenaline metabolism in the rat brain: evidence for a pre-synaptic alpha-adrenolytic action in vivo. Chronic administration of the pre-synaptic alpha-adrenoreceptor agonist clonidine decreases the concentration of the extra-neuronal metabolite of noradrenaline normetanephrine in the amygdaloid cortex and increases it in the mid-brain. Conversely, blockade of these pre-synaptic receptors by yohimbine increases the normetanephrine concentration in the amygdaloid cortex and decreases it in the mid-brain. Mianserin had a qualitatively similar action to that of yohimbine. When given clinically to rats in combination with clonidine, mianserin antagonizes both the depression of behaviour of the rats in the 'open field' apparatus and also the effects of the alpha-agonist in reducing the concentration of normetanephrine in the amygdaloid cortex. It thus appears that the chronic effects of mianserin are due to an increase in noradrenaline release as a consequence of the inhibitation of pre-synaptic alpha-adrenoreceptors.


LSD-induced alterations of investigatory responding in rats. The effects of lysergic acid diethylamide (LSD) on investigatory responses of rats in a novel hole-board were assessed in a series of experiments. LSD (40-160 micrograms/kg) altered the temporal distribution of "nose-poke" response during a 24-min session; LSD-treated rats responded less than controls initially, yet increased their response rates late in the session. This dose-dependent effect was not related to the time course of the drug's action nor to alterations in general locomotor activity. Only partial tolerance was found after eight daily injections of 100 micrograms/kg LSD. When handling stress was minimized by placing the animals in an anteroom for 10 min before starting the test, the distribution of responding was normal although the overall frequency was still reduced. Conversely, vigorous handling potentiated the LSD effect. These results are interpreted as indicating an increased sensitivity of the LSD-treated rats to the stimuli associated with being handled and placed into the novel hole-board rather than a direct effect on investigatory tendencies. This LSD-induced potentiation of defensive responses appears to compete with the active exploration of the novel environment.


[Small pulmonary nodular opacities in inflammatory and granulomatous lung disease (author's transl)]. Small pulmonary nodular opacities are usually visible only due to the summation of several small foci. The size of a single focus and its number per area determine their radiographic appearance either that of small nodular lesions, or of a reticular pattern or of diffus opacification. The small nodular pattern is primarily observed in lung disease beginning in the interstitium rather than the alveoli. Pattern and course of small nodular lung lesions give important differential diagnostic clues.


Modification by histamine H2-receptor blockade of acid secretion stimulated by histamine, pentagastrin and methacholine in the isolated whole mouse stomach. The direct influences of the blockade of the gastric histamine H2-receptors on the secretory actions induced by histamine, pentagastrin and methacholine, have been studied on the isolated perfused whole mouse stomach. According to the results cimetidine did not modify the spontaneous basal acid secretion. The interactions of cimetidine with the secretagogues were of a competitive nature with histamine and non-competitive with pentagastrin, while no modification of methacholine stimulated acid secretion.


Effects of adrenergic blockers on platelet aggregation. The action of orciprenaline, tolazoline, propanolol and inpea on platelet aggregation induced by ADP epinephrine and norepinephrine was studied in vitro in human platelet-rich plasma. Orciprenaline did not significantly affect aggregation induced by ADP. Tolazoline inhibits the aggregation induced by epinephrine and norepinephrine more intensely than the beta-blockers. Inpea blocks the platelet aggregation induced by epinephrine and norepinephrine to a greater extent than propanolol at similar concentrations. The beta-blockers inhibit platelet aggregation non-specifically.


Comparison of Mg2+ vs Ca2+, K+ and actin-activation of myosin after trinitrophenylation. While modification of six lysyl residues causes a near maximal decrease in Ca2+, K+, and actin + Mg2+ -activated myosin ATPase activities in rabbit skeletal muscle myosin, it takes nearly twice this number of modified lysyl groups to cause a similar alteration in canine cardiac myosin where trinitrophenylation is nonspecific. It appears that there are several rapidly reacting lysyl residues in cardiac myosin;  the active site of cardiac myosin is protected by ATP after modification of a limited number of these rapidly reacting lysyl groups. In both myosins, after a charge modification of these rapidly reacting lysyl groups, 6 in rabbit skeletal muscle myosin and 10 in canine cardiac myosin, there is a decrease in Ca2+, K+, and actin + Mg2+ -stimulation of myosin but an activation of Mg2+ -stimulated myosin ATPase activity, thus making actin + Mg2+ -stimulated myosin ATPase activity more like activation with K+ or Ca2+ as compared to activation with Mg2+ alone.


The binding of lactate and chloride ions to human adult hemoglobin. The effects of sodium lactate (Lact) on the oxygen affinity and the Bohr effect of purified human adult hemoglobin solutions have been compared to the effects of sodium chloride (Cl). Changes in the affinity for oxygen have been estimated from the variations of log(O2)50 with pH and at various salt concentration from 0.005 up to 2.0 mol.l-1. (O2)50 was calculated as alpha.P0.5 where alpha is the solubility coefficient of oxygen in the solutions at various salt concentrations. Variations of log(O2)50 with pH at constant salt concentration and variations of log(O2)50 with anion concentration at constant pH have been studied according to the linked-functions theory (Wyman, 1968). Bohr curves and salt binding curves were calculated from standard iterative curve fitting procedures and various parameters relevant to the effects of salts on hemoglobin function were estimated. It is shown that Lact and Cl increase (O2)50 and the alkaline Bohr effect in a comparable way at low salt concentration. At high concentration the effect of Lact predominated over that of Cl. The amount of oxygen linked Lact was larger than that of Cl. Binding constants for both anions to deoxy and oxy Hb were estimated. Lact and Cl have comparable binding constants to deoxy hemoglobin. By contrast Lact binds to oxy hemoglobin to a lesser extent than Cl. This may account for the differences observed in the effects of Lact and Cl on the function of hemoglobin. The reason for the low affinity of oxy hemoglobin for Lact may be related to steric differences between the two anions.


Temperature and acid-base status of human blood at constant and variable total CO2 content. The influence of temperature on the acid-base status of normal human blood was studied in closed systems (constant CCO2) and open systems (variable CCO2). pH-T coefficients of true plasma and erythrocytes in closed systems were similar to coefficients for water (dpH/dT = -0.017 U/degrees C at 25 degrees C). Between 26 degrees C and 42 degrees C there was no significant variation in the relative alkalinities [OH-]/[H+], the charge state of proteins in plasma and erythrocytes or the proton Donnan ration. The equations established enabled calculation of the pH of true plasma and erythrocytes and of blood PCO2 and temperature, using only one of these four parameters. Under open-system conditions, temperature was shown to cause a rise in the apparent buffer power of whole blood non-bicarbonate systems (28.8 and 34.8 mM.l-1.u-1 at 26 degrees C and 42 degrees C respectively). These results show (1) that erythrocytes in closed systems seem very well able to maintain proton distribution regardless of temperature fluctuations and (2) when blood temperature rises, it cannot be excluded that vital organs are better protected against respiratory disturbances.


The peripheral platelet count and the isoprenaline-induced splenic platelet pooling in response to beta-adrenoceptor blockade. Previous reports have shown that a single, oral dose of 50 mg metoprolol (a selective beta-1-receptor antagonist) causes a significant increase in the peripheral platelet count by releasing platelets from the spleen. In the present study 20 healthy volunteers received 50 mg metoprolol and 40 mg propranolol orally. Both drugs induced a statistically significant increase in the platelet count lasting more than 5 h. In addition, the effect of metoprolol and propranolol on beta-adrenoceptor mediated splenic platelet trapping was studied on 7 healthy subjects who received intravenous infusions of isoprenaline before and after the ingestion of these 2 beta-blocking drugs. It was demonstrated that the isoprenaline mediated decrease in the venous platelet count was diminished by both propranolol and metoprolol but the former compound appeared to be more potent in this respect. We conclude that both selective and non-selective beta-receptor blockade causes an increase in the peripheral platelet concentration during rest as well as during beta-adrenoceptor stimulation.


Flatworm control of mosquito larvae in rice fields. We describe some flatworms (some in the genus Mesostoma) that kill mosquito larvae and may account for the variability in the population densities of Culex tarsalis and Anopheles freeborni in rice fields. When mosquito larvae brush against these worms, the larvae immediately become paralyzed and die. When C. tarsalis larvae are placed inside floating cages that exclude flatworms (50-micromter mesh), there is a fourfold increase in the their survival. Rice fields that have abundant mosquito populations lack flatworms. Most such fields have only recently been turned over to rice production, suggesting that the flatworms have difficulty dispersing to new fields but, once established, are able to overwinter and control mosquitoes for the subsequent years of rice production.


[Value of quantitative evaluation of bacterial species in bronchitic infections (author's transl)]. A quantitative evaluation of the bacterial species in the sputum collected from 93 patients was conducted over a period of 7 months. The patients were divided into five different pathological categories, while separating those patients who had not received antibiotic therapy before samples were taken from those who had been administered this type of treatment. Samples were repeated after one week in 30 cases. The most frequently encountered germs were hemophilus (10(6)) and pneumococcus (10(7)). In simple bronchopathies, Gram-negative germs were only isolated at levels of 10(5) from patients previously treated with antibiotics, which demonstrates their low pathogenic role. In most severe bronchopathies they were isolated more frequently, but at levels which rarely exceeded 10(6). Their pathogenic role has to be evaluated as a function of the progression in clinical signs. The authors underline the frequency with which pneumococcal strains are resistant to tetracyclines.


The treatment of anxiety states by drugs and other means. The place of pharmacotherapy, behaviour therapy and biofeedback techniques in the general strategy of treating anxiety states is critically discussed. The dangers and disadvantages of barbiturates are described and the value and limitations of other drugs are considered. Beta-adrenergic receptor blocking drugs have a limited but valuable role in some patients, neuroleptics have a strictly limited place in treatment, and the role of antidepressants of various kinds is considered when anxiety is part of a depressive illness. The benzodiazepines are the most important group of drugs available for the treatment of anxiety states. The differences between various benzodiazepines are presented, with particular reference to their onset of action, half-life and the relevance of active metabolites of some of these drugs. A knowledge of the pharmacokinetics of the benzodiazepine drugs is of practical importance to the clinician. Emphasis is placed on the doctor-patient relationship and psychotherpeutic management in which drugs and other treatment serve as tactical aids in the general strategy of care.


[Clinical study of benorilate. Co-ordinated multi-centre test (author's transl)]. During a co-ordinated test realized by four centres of rheumatology, 91 patients had been treated by benorilate,giving so the possibility to appreciate its efficiency and its tolerance as well in the articulary fits of degeneration as in the inflammatory rheumatisms. The observations had been collected during ambulatory treatments and consequently in the conditions of daily practice. The efficiency of benorilate has been estimated by the two big disadvantages of the rheumatic person: importance of pains and capacity to do principal movements of daily life. The almost totality of patients having been preliminarily treated by another anti-inflammatory and/or antalgesic, the judgement aimed at the amelioration given by the benorilate : 41% of patients affected by arthropathy of degeneration feeled better or much better after taking benorilate. The amelioration reaches 61% for cases of rheumatoid polyarthritis. About the four criterions which could have been numerically quoted, the amelioration that had been observed is statistically significant. No anomaly had been pointed out as for the biological tolerance. The suspension form is well accepted by the rheumatic persons.


Potentiating effect of adrenaline on adenosine diphosphate-induced reduction in rabbit circulating platelet count: inhibition by dihydroergotoxine. The number of circulating platelets was monitored in anaesthetized rabbits by a continuous flow technique, using a Technicon Autocounter. Transient reductions in circulating platelet count induced by a submaximal dose of adenosine diphosphate (ADP) were potentiated by concomitant infusion of adrenaline at doses (1-25 microgram/kg) i.v.) which did not influence platelet count when infused alone. The adrenaline effect was dose-dependent. Repeated infusions of adrenaline at 15 min intervals resulted in reproducible reductions in circulating platelet count during an observation period of at least 105 min. Dihydroergotoxine (DHET), administered either i.v. (2.5-10 microgram/kg) or intraduodenally (i.d.; 25-100 microgram/kg), inhibited adrenaline-induced potentiation dose-dependently; ADP-induced effects were not influenced. Duration of action was relatively long, and significant inhibitory activity was still apparent 50 (i.v.) and 115 (i.d.) min after drug administration. DHET doses inhibiting adrenaline-induced potentiation of platelet aggregation in the rabbit are similar to doses used in the treatment of impaired human cerebral function. It is conceivable that DHET could prevent activation of human platelets by catecholamines released into the blood stream in clinical stress situations.


The isoelectric point of thrombin and its behaviour compared to prothrombin at some solid surfaces. We have shown that the isoelectric point of thrombin is high and that thrombin is a cation at the pH of blood. On the other hand, prothrombin has a low isoelectric point, being more anionic at the pH of blood. It was also found that thrombin adsorbs readily to surfaces, especially negatively charged surfaces, like behenic acid surfaces at pH 8.2. Furthermore, thrombin adsorbed onto behenic acid was active in the sense that it coagulated fibrinogen. The significance of the electric charge of the thrombin molecule in the mechanism of atherosclerosis is discussed.