[A double blind cross-over study with mepindolol-sulfate in patients with coronary heart disease (author's transl)]. The effect of mepindolol-sulfate (2 x 2.5 mg) was tested in 20 patients with coronary heart disease in a controlled study against placebo resp. propranolol (3 x 40 mg). 1. The frequency and intensity of anginal attacks were reduced significantly under mepindolol-sulfate therapy.-- 2. There was a significant improvement of ergometric exercise tolerance under mepindolol-sulfate therapy.--3. Additionally a reduction of ST-depression in ECG was found under mepindolol-sulfate.--4. In patients with coronary heart disease the clinical effect of mepindolol-sulfate in a daily dose of 2 x 2.5 mg was equal to propranolol in a daily dose of 3 x 40 mg.


7-Bromo-5-(2'-chlorophenyl)-1,3-dihydro-2H-1,4-benzodiazepin-2-one (I), a new tranquillizing agent: metabolism in rats. 1. After intraperitoneal injection of rats with the new benzodiazepine (compound I), four metabolites (compounds II, III, IV and V) were found in the urine. 2. Compound II was identified as 7-bromo-5-(2'-chlorophenyl)-1,3-dihydro-3-hydroxy-2H-1,4-benzodiazepin-2-one by comparison of g.l.c. and mass spectral properties of the metabolite and synthetic compound. 3. Mass spectra of compound III and its acid hydrolysis products indicate that compound III contains a hydroxyl group in the C(5)-phenyl ring. 4. Compounds IV and V were identified by mass spectrometry as products of simultaneous aromatic hydroxylation and methoxylation of the diazepine I. 5. The major urinary excretion products are compounds III, IV and V. Only very small amounts of compounds I and II were detected.


Metabolism of mesocarb in the rat.

Metabolism of mesocarb in the rat. 1. Rats treated orally with [14C]mesocarb (I; 3-(1-methyl-2-phenyl[2-(14C]ethyl)-N-(phenylaminocarbonyl)sydnone imine) (50 mg/kg) excrete 35% of the radioactivity in 24 h urine and 51% in 48 h urine. 2. Only traces of unchanged drug were found in urine. Hydroxy-mesocarb (II), dihydroxy-mesocarb (III), amphetamine (VII) and the conjugates of II and III account for 86% of the urinary radioactivity. 3. Cannulated male rats excrete about 40% of the radioactivity in 30 h in bile, mainly as conjugates of II and III.


[Photodynamic effect of quinacrine on bacteria]. The acridine dye quinacrine (QA) was tested with regard to the photodynamic action on bacteria (Proteus mirabilis, Escherichia coli). The absorption maximum of the yellow dye QA ist in contrary to the photodynamically active dyes methylene blue (MB) and thiopyronine (TP) situated in the short wave region of the visible spectrum. Using for illumination a common light source--they have in general a weak emission in the short wave region--relatively high concentrations of QA are necessary for photodynamic action, and the difference between photodynamic inactivation and toxic effect is small. Using that light source XBO 500 with nearly equal emission in the range from 400 to 700 nm, a distinct photodynamic action of QA results. Comparing the photodynamic action of QA with those of MB and TP, QA has a low photodynamic effect, and the kinetics of inactivation of bacteria with QA is completely different from those obtained with the dyes MB and TP.


[Contribution of the medical-scientific association for bronchopneumonology of the GDR and of its sections to development of bronchopneumonology (author's transl)]. Report on the historical development of the medical-scientific associations in GDR, engaged in tuberculosis and pulmonary diseases. Important tasks of the Association for Bronchopneumonology and Tuberculosis of the GDR are promoting and propagating scientific knowledge and progress, arranging national and international conferences in GDR and taking part in conferences in other countries. The association takes influence on the profile of the speciality, and on its future development. The Zeitschrift für Erkrankungen der Atmungsorgane is the scientific organ of the association. Sections, working- and study groups are essential parts of the association dealing with special problems of the discipline. There exists good relations to medical associations of other countries; the association of bronchopneumonology and tuberculosis is member of the International Union against Tuberculosis.


Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC 1.14.18.1) from potato tubers (Solanum tuberosum). IV. Association and dissociation phenomena. The soluble phenol oxidase of various potato juices (adjusted from physiological pH to pH 4.5, 7.0 and 7.8) was separated by gel chromatography into multiple molecular forms. In acid or neutral and alkaline potato juices, low-mol.-wt. (less than 150,000 daltons) or high-mol.-wt. (greater than 150,000 daltons) enzyme forms predominate, respectively. Conversion of the low-mol.-wt. enzyme forms into high-mol.-wt. enzyme forms, and vice versa, was achieved by changing the pH values from acidic to neutral or alkaline pH, and vice versa. This substantiated our previous idea that the enzyme multiplicity arises from association of various subunits. In alkaline potato juice, considerable loss of monophenol oxidase activity (assayed at pH 6.0) occurred. This confirmed our previous findings that o-diphenol oxidase is more alkali-stable than monophenol oxidase.


[Sugar-substitutes in the prevention of dental caries (author's transl)]. Investigations performed with sugar-substitutes showed that from the partial substitution of carbohydrate in the food a significant cariesreduction may be expected. According to the data of the microbiological, pH, animal experiments and clinical examinations xylit proved to be the least cariogen agent and sorbit too has a reduced cariogen effect.


[The effects on caries in animal experiments when reducing sugar in cola-drinks(author's transl)]. The effect of stepwise sugar reduction in cola drinks was tested in a programmed animal experiment using Wistar rats. In a parallel study the pH-decrease at different sucrose concentrations was measured in vitro using a plaque model system which involved s. mutans (strain OMZ 176). The reduction of sugar contents by 20%, 33% und 42% results in a lower acid production by s. mutans strain connected with a decrease of the experimental produced caries. For this reason the sugar restriction in cola drinks will be seen as a contribution towards an improvement of dental health.


[Mechanism of the protective effect of antiserum against isologous aggregated mouse immunoglobulins in transplantation immunity]. The injection of antiserum against isogeneic aggregated mouse immunoglobulins prolonged the survival time of skin allografts in mice by 5-6 days. The study of the possible mechanisms of this phenomenon revealed that the injection of aggregated immunoglobulin antiserum decreased the ability of the lymphocytes of the animals receiving the antiserum to induce local "graft versus host" reaction and to proliferate in the blast transformation test in response to phytohemagglutinin treatment and in the mixed lymphocyte culture test. When added in vitro, the antiserum did not influence either the mixed lymphocyte, or the specific cytotoxic effect of immune lymphocytes, but stimulated 3H-thymidine incorporation into the suspension of lymphocytes incubated with syngeneic irradiated cells. The latter fact may be indicative of the mitogenic action of the antiserum. The analogy between the mitogenic action of the antiserum and that of nonspecific mitogens, such as concanavalin A and phytohemagglutinin which also show opposite effects in vivo and in vitro, is discussed. The conclusion has been made that the increase of the survival time of the skin allograft after the multiple injection of aggregated immunoglobulin antiserum was primarily due to its action on the recognizing and proliferative functions of T lymphocytes, and later due to an increase in IgG synthesis.


[Participation of vaccinia virus in the pathogenesis of different clinical forms of postvaccinal complications. I. Frequency of vaccinia virus detection in the vaccinted who have usual and complicated reactions to vaccination]. The virological examination of 1365 samples taken from 469 children vaccinated against smallpox revealed considerable differences in the frequency and the time of vaccinia virus detection in different clinical forms of postvaccinal pathology as compared with uncomplicated vaccinal process. During the postvaccinal period taking its normal course vaccinia virus was isolated from 7.3% of children only from the pharynx till day 8 following vaccination. In generalized and creeping vaccinia the virus was isolated from 71.4% of children, in postvaccinal encephalitis from 57.1% of children, in vaccinal angina frove-mentioned complications vaccinia virus was detected in the samples obtained from the patients till days 24, 35, 15 and 24 respectively. The etiopathogenetic role of vaccinia virus in a number of postvaccinal complications is discussed.


[Electron microscopic study of presumptive photoreceptor cells in the aboral organ of the ctenophore, Beroë cucumis]. Presumptive photoreceptor cells are located in the receptor epithelium of the aboral organ and exhibit some common morphological characters with flagellar photoreceptors of some invertebrates, as well as with photoreceptors of vertebrates. These cells are specifically distinguished by the presence of a special membrane structure in their epinuclear zone, this structure being the derivative of a cilium. Presumptive photoreceptor cells which have a short central projection and form in their basal part the synaptic contacts ("triads") with the adjacent nervous endings, may be classified as pseudosecondary receptor cells. Acetylcholinesterase was found in the synaptic regions. Cytoplasm of the presumptive photoreceptor cells is rich in granular vesicles which contain biogenic amines.


[Psychopharmacotherapy of slowly progressive hypochondriacal schizophrenia]. The efficacy of psychopharmacological treatment modalities and comatose methods was studied in the therapy of patients with slowly progressive schizophrenia, the clinical picture of which was characterized by cenestho-hypochondriacal disturbances (155 cases). The use of high doses of neuroleptics, atropine and insulin comas in these cases were ineffective. A positive compliance to therapy was attained in the treatment by tranquilizers of the benzodiazepine series, especially by a parenteral administration. The best results were demonstrated in the treatment of pseudoneurotic hypochondria. The therapeutic reluctancy increased in the cenestho-algic syndrome and in the syndrome of "rigid" hypochondria. The most effective in these cases was a combined therapy by tranquilizers (parenteral administration) and  small doses of neuroleptics.


[Effect of neurotoxins on mediator release from the motor nerve endings of the Drosophila melanogaster larva]. Spontaneous miniature junctional potentials and induced by motor nerve stimulation junctional potentials were recorded in muscle fibres of the third instar Drosophila larvae. Presynaptically active neurotoxins isolated from the snake venoms (beta-bungarotoxin 4.5 x 10(-7) M, notexin and notechis-II-51 X 10(-7) M, Crotoxin 9 x 10(-8) M, phospholipase from the venom of cobra 4 x 10(-6) M did not induce any changes in transmitter release during 60--120 min of contact with Drosophila preparation, although these concentrations exceed those used in frog or mice preparations. On the contrary, bee venom phospholipase (1.25 x 10(-7) M) turned out to be even more active than in frog or mice junctions. A similar sequence of three phases of the presynaptic blocking effect was found: initial decrease followed by a period of increase and finally -- depression of both spontaneous and evoked release. Bee venom phospholipase was ineffective in Ca2+-free solution but its irreversible binding to nerve endings took place evidently in spite of Ca lack.


Kinetics of elementary steps in the cytochrome P-450 reaction sequence. III. NADPH reduction of cytochrome P-450LM at different integrational levels. The aerobic NADPH reduction of cytochrome P-450LM has been investigated on microsomes, as well as on the solubilized enzyme system in the associated, disintegrated, and reconstituted state, respectively. P-450 exhibits biphasic reduction kinetics of about 70/30% phase distribution and rate constants differing 10-fold. The partial reactions are due to organizational asymmetries, the cytochrome being either incorporated into P-450/reductase associates (cluster) or localized outside (randomly distributed, homoassociated, weakly cluster-associated). Triton N-101 disintegrates the different associate structures, consequently followed by the disappearance of the rapid reaction phase. The enzyme system can be reconstituted; at microsomal stoichiometry the respective standard parameters are approached, depending on the composition and structural organization of the phospholipid. The reorganization without any membrane matrix is obviously thermodynamically determined.


Kinetics of elementary steps in the cytochrome P-450 reaction sequence. IV. Mechanism of the NADPH reduction reaction of cytochrome P-450LM. The aerobic NADPH reduction of the Triton N-101 disintegrated cytochrome P-450LM system has been studied. At this organization level--the components being dispersed in solution--a first-order monophasic reaction is exhibited. Neither the complex formation of cytochrome and reductase, respectively, nor preceding diffusion is rate limiting. The initial rate follows the ratio reductase/P-450 (mole/mole) thus indicating a Michaelis-Menten type enzyme mechanism. A model treatment of the reaction fits the systems behaviour as a whole. A multiparameter equilibrium state and different specified time function equations were developed for the determination of individual step rate constants and other system parameters as well.


NADPH reduction of cytochrome P-450 at different integrational levels of the enzyme system. The complex monooxygenatic enzyme exhibits different functional behaviour at different integrational levels, thus indicating distinct organizational states. The aerobic NADPH reduction of microsomes, solubilized and reconstituted systems follows a biphasic kinetics, the two phases are attributed to associated state (cluster) and random cytochrome P-450 reduction. States of different cytochrome P-450/reductase ratio (associates) could not be differentiated in rate. Detergents (Triton N-101, cholate) are capable of disintegrating the system, at last only monophasic slow reduction is observed. The hydroxylation activity follows the respective reduction behaviour. Sedimentation analysis proves the distinct structural states. Reconstitution of the system can be achieved by means of detergent dilution as well as by combining the constituents. The activity of the reconstituted system depends on the composition of the phospholipids as well as on its organizational state. The reassociation of the solubilized enzyme system at nearly microsomal components stoichiometry (Triton N-101 dilution) proves to be thermodynamically governed leading to self-organization of the system without matrix prerequisite. Individual step rate constants of the reduction reaction and other system parameters are accessible by means of a model treatment of the disintegrated system. Further application to mixed kinetics systems is in progress.


NADPH-dependent electron transport chain in microsomes and lipid peroxidation catalyzed by metal ions. Like iron ions copper ions are also able to stimulate the NADPH-dependent lipid peroxidation in rat liver microsomes. This effect is strongly dependent on the concentration of Cu2+ added. Initial concentrations of Cu2+ above 50 microM completely inhibit the formation of malonaldehyde. The activator and inhibitor functions may be interpreted by a simultaneous participation of Cu+ ions formed in the chain branching and termination reaction of the free radical lipid peroxidation process. Inhibition studies with pCMB and the His-reagent diethyl pyrocarbonate indicate an essential role of cysteine and histidine residues in the Cu+-NADPH-dependent lipid peroxidation process.


Mechanistic studies with purified components of the liver microsomal hydroxylation system: spectral intermediates in reaction of cytochrome P-450 with peroxy compounds. Recent investigations in this laboratory on the mechanism of action of liver microsomal cytochrome P-450 (P-450 LM) and its interaction with other components of the hydroxylation system are presented. Two electrophoretically homogeneous forms of the cytochrome, phenobarbital-inducible P-450 LM2 and 5,6-benzoflavone-inducible P-450 LM4, so designated according to their relative electrophoretic mobilities, were used in these studies. Phosphatidylcholine is required in the reconstituted enzyme system for rapid electron transfer from NADPH to P-450 LM, catalyzed by NADPH-cytochrome P-450 reductase, as well as for maximal hydroxylation activity with either molecular oxygen or a peroxy compound serving as oxygen donor to the substrate. The phospholipid facilitates the binding of both substrate and reductase to P-450 LM and apparently causes a structural change in the cytochrome as shown by an increase in alpha-helical content, determined by circular dichroic spectrometry. P-450LM3 and LM4 are one-electron acceptors under anaerobic conditions, in accord with previous potentiometric titrations and product yield data, but in disagreement with previous titrations with reducing agents. The cause for the discrepancy between the present and earlier results is not yet fully understood. Stopped flow spectrophotometry was employed to detect intermediates in the reaction of peroxy compounds with P-450LM2. With m-chloroperbenzoic acid the intermediate formed has absorption maxima at 375, 425, and 540 nm in the absolute spectrum and at 370, 436, and 540 nm in the difference spectrum (intermediate minus oxidized form). A study of the magnitude of the spectral change at various peracid concentrations indicated that with this oxidant the reaction shows a dependence resembling a binding curve. These and other experiments with various oxidants, including cumente hydroperoxide, suggest a reversible two-step mechanism according to the reaction: P-450 LM + oxidant equilibrium C equilibrium D, where C may be an enzyme-oxidant complex and D is a spectral intermediate of unknown structure. A scheme is proposed for the mechanism of action of P-450 LM based on these and earlier studies, including evidence from deuterium isotope experiments for the formation of a substrate carbon radical prior to oxygen transfer.


Kinetics of reduction of purified liver microsomal cytochrome P-450 in the reconstituted enzyme system studied by stopped flow spectrophotometry. Stopped flow studies were undertaken to examine the kinetics of reduction of 5,6-benzoflavone-inducible P-450 LM4 by NADPH in the presence of NADPH-cytochrome P-450 reductase and phospholipid under anaerobic CO at 25 degrees C. The reaction exhibited biphasic kinetics irrespective of NADPH concentration or of the molar ratio of reductase to P-450 LM4. The apparent first order rate constants for the fast and slow phases were determined to be 0.9 to 1.0 and 0.25 s-1, respectively. With the reductase and P-450 LM4 present in equimolar amounts, the total amount of P-450 LM4 reduced increased linearly with NADPH concentration; the titration gave a stoichiometry of 2 mol of NADPH per mol of reductase-cytochrome complex. The NADPH concentration had no appreciable effect on the magnitude of the first order rate constants for the fast and slow phases. The kinetics obtained in the presence of benzphetamine were essentially indistinguishable from those seen in the absence of this substrate, while the amount of P-450 LM4 reduced in the fast phase, but not the rate constant for this phase, decreased when phospholipid was omitted from the reaction mixture. Nearly maximal rates of NADPH oxidation by P-450 LM2 OR LM4 were obtained with a molar ratio of reductase to P-450 LM of 1.0. Benzphetamine enhanced the oxidation of NADPH by P-450 LM2 but had no effect on the activity of P-450 LM4. Rates of NADPH oxidation in the presence of P-450 LM2 and LM4 decreased by 80 and 40%, respectively, when phospholipid was omitted from the reconstituted enzyme system. These studies provide evidence for the formation of a catalytically functional 1:1 complex between the reductase and P-450 LM4, and indicate that P-450 LM2 and LM4 differ in their dependence on phospholipid.


Electrochemical investigations on the oxygen activation by cytochrome P-450. The application of cytochrome P-450 in substrate conversion is complicated both due to the limited stability and the cofactor regeneration problems. To overcome the disadvantages of NADPH consumption the transfer of the reduction equivalents from an electrode into the cytochrome P-450-system was studied: 1. NADPH was cathodically reduced at a mercury pool electrode. By immobilization of NADP on dialdehyde Sephadex the reductive recycling was possible. 2. Different forms of reduced oxygen were produced by the cathode: a) The reaction of O2- with deoxycorticosterone yields a carboxylic acid derivative. In contrast the cytochrome P-450 catalyzed NADPH-dependent reaction with the same substrate gives corticosterone, O2- represents only an intermediate in the activation of oxygen and is not the "activated oxygen" species. b) Molecular oxygen was reduced to HO2- and H2O2, respectively. The interaction of adsorbed cytochrome P-450 on the electrode surface with the reduced oxygen species in the absence of NADPH was studied. The electrochemically generated peroxide seems to be more active than added H2O2. 3. In a model of electro-enzyme-reactor several substrates were hydroxylated by microsomal cytochrome P-450 with cathodically reduced oxygen which substitutes NADPH.


[Non arrhythmogenic sudden death as complication of coronary heart disease]. In a cohort of 417 patients admitted consecutively to the Coronary Care Unit for acute myocardial ischemia (unstable angina pectoris in 121, acute myocardial infarction in 296 patients) 21 cases of non arrhythmogenic sudden death occurred within 24 hours after admission. 16 of these patients suffered from acute myocardial infarction and 5 from unstable angina pectoris. Cause of death was cardiac rupture in 12 and pump failure in 4 patients with acute myocardial infarction, whereas all patients with unstable angina pectoris died from pump failure. Patients with cardiac rupture within 24 hours after admission, had significantly higher systolic and diastolic blood pressure in comparison with the other groups and with patients dying from cardiac rupture on the third day, or later. All patients dying from pump failure with unstable angina pectoris and one of the patients dying from pump failure with acute myocardial infarction had beta blocker therapy. Beta blockers were given to 68 of the patients with unstable angina pectoris. Acute pump failure occurred in this group only. The risk of pump failure with beta receptor blocking drugs is indicated by angina decubitus, marked dyspnea during anginal attacks (even in patients free of signs of cardial insufficiency outside their attacks) and a lack of responsiveness to beta blocking therapy. In these patients rapid coronary angiography and bypass surgery seems to be the prefered method of management. Beta blockers should not be given to these patients or discontinued in cases which lack responsiveness.


pH and temperature dependence of the double inhibition of D-glyceraldehyde-3-phosphate dehydrogenase by ATP and quinaldate. ATP and quinaldate, two inhibitors of D-glyceraldehyde-3-phosphate dehydrogenase, act antagonistically if [Pi] greater than 1 mM, at pH 7.5 and 25 degrees C. However, the type of interaction of the two inhibitors changes with pH and temperature. Antagonism is strongest at pH values near neutrality and decreases at higher pH. Above pH 9.5 the two inhibitors act nearly additively. At pH 8.5 and [Pi] = 5 mM there is antagonism above 20 degrees C between the two inhibitors whereas there is synergy below 20 degrees C. A lag period in the time course of the enzyme reaction was detected when both inhibitors were present. The lag period is a function of pH. Below pH 8.5 the pH-dependence of the lag period resembles a titration curve with a pKapp greater than 8.


An adenosine triphosphate dependent deoxyribonuclease with adenosine triphosphatase, activity from Bacillus cereus. An adenosine triphosphate-stimulated deoxyribonuclease was purified to about 4200 fold from Bacillus cereus. The enzyme activity of the crude extract increased by a factor of about 5 after dialysis. One of the low molecular weight inhibitors of the crude extract was found to be inorganic phosphate. During enzyme purification two nucleases were identified. One of them was specific to denatured DNA and the other one degraded both denatured DNA and native DNA. The activity towards native DNA could be increased several times by ATP. Through all steps of purification the ATP-independent DNase always accompanied the ATP-dependent one and the ratio of their activity was found to be constant. The ATP-dependent DNase also possessed ATPase activity stimulated both by native and denatured DNA. The fact that ATPase was stimulated by DNA and went together with ATP-dependent DNase during purification suggests that these functions belong to the same enzyme complex. Maximal activity of ATPase had broader pH, Mg2+ and ATP concentration ranges than that of DNase. Cooperation of the two functions may be limited only to a narrow range of ATP concentration. Km for ATPase was 1.6x10-4 M ATP.


Isoelectric focusing of macrophage culture media and the effect of the fractions on the synthesis of DNA and collagen by fibroblasts. Macromolecules from rat peritoneal macrophage culture media were separated into 30 fractions by flat bed isoelectric focusing (IEF). The fractions were tested for their influence on thymidine and proline incorporation into cultured rat granuloma fibroblasts. Three fractions, stable after freezing and lyophilization, were of interest: one inhibiting thymidine incorporation (focusing at pH 7.3-7.6), another stimulating thymidine incorporation (focusing at pH 4.4-5.3), and the third stimulating proline incorporation into cells and medium collagen (focusing at pH 10.2-10.7). The last also exhibited a ribonuclease (RNase) activity with a pH-optimum of 7.0-7.5.


Effects of GABA-analogues on the high-affinity uptake of GABA in astrocytes in primary cultures. Employing primary cultures of astrocytes which seem to constitute a valid model of their in vivo counterparts, it has been demonstrated that this cell type is likely to be of importance in terminating the transmitter activity of GABA. It has been shown that the transport carrier in astrocytes is stereospecific for the C-4 hydrogens of the GABA molecule and that its structural requirements are distinct from those exhibited by the neuronal GABA carrier. beta-Proline was a selective inhibitor of GABA transport in astrocytes, whereas R-trans-4-methyl-4-aminocrotonic acid and S-nipecotic acid seemed to be selective inhibitors of neuronal GABA transport, as studied using very thin slices ("prisms") of brain cortex. These findings may be important for studies on the relative significance of the two transport systems in GABA-mediated neurotransmission, and thus for future pharmacological manipulations of the GABA system. This may eventually be beneficial for the treatment of neurological disorders such as epilepsy, Huntington's chorea and Parkinson's disease in which the GABA system seems to be disturbed (34,60,62).


The Schultz-Dale reaction of the guinea-pig ileum: influence by beta-adrenoceptor agonists and theophylline. One of the organs that can be used to study the Schultz-Dale reaction, is the guinea-pig ileum. The reaction is characterized by a specific pattern: a quick contraction, followed by a quick relaxation and a second, more slow contraction. A description is given of this typical reaction. Selective inhibition of the slow contraction could be obtained using beta-adrenoceptor agonists. Also theophylline inhibited the slow contraction, but this inhibition was only selective within a narrow range of concentration. These findings encourage further investigation about a possible involvement of cyclic nucleotides in the liberation of the mediators involved in the Schultz-Dale reaction of the guinea-pig ileum. Furthermore they provide more information about this preparation which could be used as a screening model for anti-allergic drugs of distinct pharmacological groups.


A review on methods of phospholipase A determination. A short review is presented on methods to determine phospholipase A activities. On purpose, only those methods are covered which can, at least potentially, be used for assaying the low activities generally associated with intracellular phospholipases A. A selection was made of procedures which apply non-radioactive or radioactive substrates or chemically modified substrate analogs which can be used for spectrophotometric assays.


On the regulation of tryptophan metabolism "via" kynurenine. The tryptophan leads to nicotinic acid pathway is inhibited in the nicotinic acid administered rats; in this case tryptophan chiefly metabolizes "via" serotonin. The serotonin pathway is inhibited in the excess phenylalanine administered rats, especially due to the inhibition of the tryptophan-5-hydroxylase reaction. Xanthurenic acid inhibits kynurenine-3-hydroxylase in vitro, which is a very important regulatory enzyme for xanthurenic acid and nicotinic acid production.


Properties of influenzavirus nucleocapsids in nonpermissive cells. The properties of fowl plague virus (Influenzavirus A) nucleocapsids isolated from the cytoplasm of infected Ehrlich ascites carcinoma cells and chick embryo cells were compared. Nucleocapsids isolated from both systems possessed similar polypeptides (P and NP) but differed in their biophysical characteristics. Nucleocapsids from ascites cells sedimented in velocity sucrose gradients slower (from 25 to 50 S) and the majority of them banded at higher density in CsCl gradients (rho 1.38 as compared to 1.34 g/ml) than nucleocapsids from chick embryo cells. In the electron microscope they appeared as thin threads 3--4 nm in diameter.


Variola virus strains of 1960--1975: the range of intraspecies variability and relationship between properties and geographic origin. A comparative study of 61 variola virus strains recovered from patients in various geographic regions of the world revealed significant intraspecies variability in some biological properties, established the range of this variability and demonstrated the existence of strains with atypical properties. There was a certain correlation between some laboratory markers of variola virus and the degree of its pathogenicity for man. The prevalence of more virulent and thermoresistant strains in the Hindustan subcontinent and the peculiarity and heterogeneity of the strains circulating in Africa were demonstrated. Alastrim virus was detected among the African isolates.


Development of tumours in mice chronically infected with herpes simplex virus. Tumours developed in chronic infection lasting for 150--180 days in 39 (60%) of 65 mice infected with strain L2 of herpes simplex virus (HSV) type 1 and in 12 (20%) of 60 mice infected with strain 333 of HSV type 2. Similar results were obtained in 150 immunosuppressed mice chronically infected with HSV types 1 and 2. Pathomorphologically, the neoplasias in the first group (strain L2) were similar to adenocarcinoma and malignant lymphoma and in the second (strain 333) to lymphoma and angio- or fibrosarcoma. The respective HSV strains were isolated by cocultivation of blood leukocytes from chronically infected animals and cultures of the tumour cells with human embryo fibroblasts (HEF). HSV and Gross murine leukaemia virus antigens were detected in tumour cells by immunofluorescence and radioimmunoassay, respectively, and HSV antigen by immunofluorescence also in cultures of tumour cells and in cells of the brains, spinal cords, livers and spleens of the animals. HSV antibody was demonstrable in the blood serum from chronically infected tumour-bearing mice in a titre of 32.


Isolation and characterization of influenza A viruses from wild ducks in northern Japan: appearance of HSW1 antigens in the Japanese duck population. Twenty-six influenza A viruses were isolated from cloacal and tracheal samples of 235 resident and 396 migratory ducks in Miyagi prefecture, Japan, in 1977--78. Of these, twelve were antigenically related to the avian-origin HSW1 virus, A/duck/Alberta/35/76 (HSW1N1), but their neuraminidase antigens were characterized as Nav2-3, Nav4 or N2. These antigenic configuration have not previously been reported. In addition, one strain in which the neuraminidase antigen was identified as Nav4, was demonstrated to be a mixture of two haemagglutinins, HSW1 and Hav7. Two distinct strains were separated from the mixture and characterized as HSW1Nav4 and Hav7Nav4. The antigenic identification of an additional 13 influenza A viruses revealed the presence of six haemagglutinin subtypes (Hav1, Hav3, Hav4, Hav6, Hav7, and Hav8) and five neuaraminidase subtypes (Nav1, Nav2-3, Nav4, Neq2, and N2) in various combinations. The results suggest that the avian influenza A viruses among feral ducks may be isolated in various combinations of haemagglutinins and neuraminidase subtypes in Japan, and that feral ducks may be the site of genetic recombination occurring as a result of dual infection with different subtypes of influenza A virus.


Selective resistance to togaviral superinfection in mice with tolerant lymphocytic choriomeningitis virus infection. Mice infected neonatally with lymphocytic choriomeningitis virus (LCMV) developed partial and complete resitance to cerebral superinfection with tick-borne encephalitis virus (TEV) in 10 and 20 days after birth, respectively. This resistance lasted at least till the age of 40 days. LCMV tolerant mice neither succumbed to TEV infection, nor circulated TEV in their blood. Moderate, gradually decreasing TEV titres were detected in the brains and TEV-induced brain interferon was lower than in control mice of the same age. TEV superinfection caused a significant depression of the blood titre of tolerated LCMV while the titres in the brains remained equal to those in tolerant but not superinfected mice. LCMV tolerant mice showed a similar resistance to another togavirus (chikungunya) but not to encephalitogenic picorna-, herpes- and rhabdoviruses.


Morphological characterisitcs of the pathological process in the central nervous system of monkeys infected with variants of measles virus strain L-16. The neurovirulence of L-16 vaccine measles virus strains and its 9 clonal variants was investigated in intracerebrally infected Macaca mulatta monkeys. Two groups of clones were distinguished: one comprised variants that caused destructive changes of neurons in the brain and diffuse proliferative gliosis, and the second variants that caused changes of a transitory character, connected with the traumatic exposure.


Effect of incubation temperature on infectivity titration of mouse brain-passaged avian infectious bronchitis virus in laboratory host systems. Mouse brain-passaged infectious bronchitis virus (IBV) could not be assayed for its infectivity at 39.5 degrees C, but could be so at 37 degrees C and 39.5 degrees C. Antigen accumulation was not detected by immunofluorescence in CEK cells infected with mouse brain-passaged IBV at 39.5 degrees C, perhaps due to the difference in the cellular environments.


Effect of 2-deoxy-D-glucose on the binding of preimmune and immune sera to herpes simplex virus-infected cells as measured by quantitative immunofluorescence. The binding of immune and preimmune rabbit sera to fixed VERO cells infected with herpes simplex virus type 1 (HSV 1) was measured by quantitative immunofluorescence. False positive binding of the preimmune serum (PS) occurred from 8 hr post infection (p.i.), while the immune serum (IS) revealed positive fluorescence starting with 4 hr p.i. The false binding of PS decreased significantly (P = 0.01) if the cells were cultured in the presence of 2-deoxy-D-glucose (DOG). Statistical analysis of data on the binding of IS to infected cells kept in the presence or absence of DOG showed that the mean values of immunofluorescence were not significantly different (P less than 0.1). Cells cultured for 8 hr p.i. in the presence of 6 mM DOG seem to be a suitable antigen for detection of the actual level of specific antibodies by indirect immunofluorescence.


Antiphytoviral activity of 2,4 dioxo hexahydro triazine. Three treatments with 2,4 dioxo hexahydro triazine (DHT) significantly reduced the concentration of potato virus X (PVX) in systemically infected tobacco plants. In hypersensitive plants DHT caused a reduction in the number of local lesions produced by PVX. In systemic and hypersensitive hosts, treatment with DHT resulted in a more or less marked reduction in the concentration of, and in the number of local lesions caused by, potato virus Y (PVY), potato virus A (PVA), tobacco mosaic virus (TMV), and cucumber mosaic virus (CMV). Three days after treatment with DHT the incorporation of 32P into TMV RNA was significantly reduced, while that into host RNA was less affected.


Quantitative study on the reproduction of virulent and vaccine Rickettsia prow azeki strains in cells of different origin. Vaccine and virulent strains of Rickettsia prowazeki differ by the degree of reproduction in McCoy, B, and chick embryo cells but replicate to similar levels in FL cells. As distinct from the virulent Breinl strain, the vaccine E strain rickettsiae permanently lost their capacity for long-term reproduction in McCoy cell cultures but retained their capacity to adsorb on to and penetrate into these cells. Consequently, the reproduction of rickettsiae is limited at later stages of intracellular infection. The E strain of R. prowazeki has been defined as a conditional lethal, host-dependent (hr) mutant.


Investigation of the ecology of Soldado virus on Puffin Island, North Wales. The prevalence of Soldado (SOL) virus and SOL virus antibodies was investigated on immature sea birds and the argasid tick Ornithodoros (Alectorobius) maritimus collected on Puffin Island, North Wales. No SOL virus was recovered from 133 bird sera, but 2 of the birds exhibited neutralizing antibodies against SOL virus. Nine of 27 tick pools (226 individuals) and 34 of 173 ticks tested individually proved to be infected with SOL virus.


The effect of immunization on polymorphonuclear leukocyte chemotaxis in influenza. Challenge of human subjects with influenza virus produced a significant fall in the chemotaxis of polymorphonuclear leukoyctes (PMN's) obtained from their peripheral blood. This was seen by 3 days with some return toward normal function at 7 days but with a persistent defect still found at 20 days after challenge.


Attempt to transfer priming by cocultivation of L cells and chick embryo fibroblasts. Interferon-treated L cells did not transfer priming to chick embryo fibroblasts (CEF) during cocultivation. Nor could transfer be observed when CEF were treated with homologous interferon in the mixed cultures. The results indicate that the lack of transfer by cocultivation is another characteristic of priming different from antiviral resistance.


Suitability of Coxiella burnetii strains for preparation of phase II artificial diagnostic antigen for microagglutination test. Sensitivity in the microagglutination (MA) test of artificial (prepared by potassium periodate treatment) phase II Coxiella burnetii antigens depended on the C. burnetii strain used and the number of its chick embryo yolk sac passages.


Vaccination of an adolescent group with A (H3N2) + A(HSW1N1) bivaccine before an A (H1N1) influenza epidemic. A group of 35 adolescents aged 18--22 years were vaccinated intradermally with 0.2 ml of a mixed A (H3N2) + A (Hsw1N1) formolized vaccine and revaccinated with A (Hsw1N1) monovaccine 10 days before an A (H1N1) influenza epidemic. The vaccination had no effect on morbidity or the clinical course of disease. Serology revealed a primary immune response to A (Hsw1N1) and a booster response to A (H3N2). Apparently, prevention of a new influenza subtype by formolized vaccine possessing only the corresponding neuraminidase type is ineffective.


Adsorption properties of U.I.C.C. Rhodesian chrysotile and crocidolite in aqueous solution -- effects of cation depletion. Adsorption properties of U.I.C.C. Rhodesian chrysotile and crocidolite were tested in aqueous solution with acid and basic dyes (Eosine Y. Picric Acid, Acridine Orange and Methylene Blue). Rates of adsorption showed a diffusion of only basic dyes in micropores of fibers. The strong adsorption of basic dyes on the surface of the fibers led us to suppose that the active sites have a negative charge. Our hypothesis was supported by the verification of an increase in adsorption capacity whick depends on depletion of Mg++ from chrysotile and of Fe+++ from crocidolite.


Hemoglobin Cowtown (beta 146 HC3 His-Leu): a mutant with high oxygen affinity and erythrocytosis. A new mutant, hemoglobin Cowtown, has been found in a white man and his father, both of whom have erythrocytosis. The father had previously been treated with 32P for polycythemia vera. The abnormal hemoglobin is not detectable on electrophoresis in alkaline buffers, but it resolves distinctively on electrophoresis in citrate agar, pH 6.0; similarly, the abnormal beta-globin chain does not separate from beta-A in urea 2-mercaptoethanol buffers of pH 8.9, but it moves anodically to beta-A at pH 6.0. Peptide chromatography and amino acid analysis of the beta chain reveal that the C-terminal histidine residue (beta 146) has been replaced by leucine. Like several other hemoglobins substituted at this residue, Hb Cowtown has a high oxygen affinity and a diminished Bohr effect.


Immunosuppressive and corticosteroid therapy of polyarteritis nodosa. The clinical manifestations, treatment and survival of 64 patients with polyarteritis nodosa seen from 1955 to 1977 were evaluated. In general, the patients had multisystem involvement. No patient with cutaneous vasculitis alone was accepted into the study. The clinical diagnosis was confirmed by biopsy in 34 patients, by autopsy in 13 and by angiography in 10. The patients were treated at the discretion of the physicians responsible for their care. Eight of the 64 patients received only supportive therapy (group 1), 34 received corticosteroids alone (group 2), and 22 received both corticosteroids and an immunosuppressive agent (group 3). Five patients in group 2 and one patient in group 3 were excluded from survival studies because of insufficient length of therapy. Patients in the three treatment groups were very similar with respect to 18 clinical and laboratory variables. Median survival times for the three groups were three months, 63 months and 149 months, respectively; 5 year survival rates were 12 per cent, 53 per cent and 80 per cent (p less than 0.05). Despite difficulty in precisely defining polyarteritis nodosa, the data suggest a better prognosis for treated patients than has previously been appreciated, with improvement in outcome when an immunosuppressive agent is added to corticosteroid therapy.


The effect of multiple psychotropic drugs on the accommodation of pre-presbyopes. Among 1,000 psychiatric patients in a Veterans Administration hospital, there were 15 in the age group 21--41 years who complained of difficulty at near and who showed reduced accommodation. All 15 were receiving benztropine mesylate (Cogentin) and most also received a phenothiazine or a butyrophenone. The reduction in accommodation ranged from 40 to 100%. Data are insufficient to correlate doses of specific drugs with the precise extent of the impairment of accommodation.


Influence of mucosal and serosal pH on antidiuretic action in frog urinary bladder. Mucosal acidification to pH 6.5 reduced by 88% the oxytocin- (2.2 x 10(-8) M) elicited increase of water permeability in frog urinary bladder. Mucosal alkalinization (pH 10.5) increased by as much as 200% the response to the same concentration of oxytocin. These effects were not observed when supramaximal concentrations of oxytocin were imployed. Similar changes were found when the serosal pH was modified. The hydrosmotic responses elicited by serosal hypertonicity or cyclic AMP plus theophylline were also affected by mucosal or serosal changes of the hydrogen in concentration, suggesting an effect at a post-cyclic AMP level. Important interactions were found between luminal pH and serosal hypertonicity when experimental conditions were employed similar to those observed in the collecting duct of mammalian nephron. Freeze-fracture studies showed that the number of intramembranous aggregates of particles induced by ADH in the luminal membrane was reduced by mucosal acidification and augmented by an increase in medium pH.


Neurobiologic correlates with defense mechanisms and psychotherapy. It is proposed that repression, regression, and other defenses are associated with various transmitter systems under the guidance of the predominantly serotonergic, aversive, and the noradrenergic reward centers. The alteration of total and base composition of glial and neuronal RNA and the inferred encoding of memory are applied to the formation of neurosis and to psychotherapy.


Detection of alcoholism relapse: comparative diagnostic value of MCV, GGTP, and AANB. Mean red blood cell volume (MCV), plasma gamma glutamyl transpeptidase (GGTP), and plasma alpha amino-n-butyric acid (AANB) were measured in 245 patients undergoing treatment for alcoholism at the V.A. Medical Center, Bronx, N.Y. and Raleigh Hills Hospitals, Denver, Colo. AANB and GGTP, but not MCV, decreased following withdrawal from alcohol and during a period of abstinence. Among patients who relapsed, AANB and GGTP, but not MCV, increased in a majority of subjects. The combined use of AANB and GGTP identified 28/33 heavy drinkers, while 4/33 patients in remission had positive tests. In conclusion, measurement of AANB and GGTP are useful in documenting the success of treatment for alcoholism and in identifying patients who relapse following treatment for alcoholism.


Alterations in neurotransmitter activity after acute and chronic ethanol treatment: studies of transmitter interactions. Acute and chronic ethanol treatment has multiple effects on the neurotransmitter systems in the nigrostriatal complex. A single dose of ethanol increases striatal dopamine release at low doses, but depresses it at high doses. In ethanol-dependent rats, dopamine release is accelerated during intoxication, but is reduced during a withdrawal syndrome. Concomitantly, high-affinity choline uptake, an index of cholinergic activity, is elevated at times when dopamine release is depressed. Changes in dopaminergic or cholinergic receptor activity do not induce or result from these effects. Neither has a role for GABA or substance P yet been implicated. The data suggest that interactions between at least two trasmitters in the caudate nucleus may occur after acute and chronic ethanol treatment.


Adverse reactions following fazadinium-thiopentone induction. Three cases of hypersensitivity to an anaesthetic induction with fazadinium and thiopentone are described. One reaction was fatal. Two patients had documented and the third presumptive previous exposure to thiopentone, and all had a history of allergy (skin rash) to penicillin. None of the patients had received fazadinium before. The possibility that fazadinium contributed to the severity of the reactions is conjectural.


Cytochemistry of oxidoreductases in spermatozoa: the technique revisited. A revised cytochemical procedure for demonstrating sperm oxidoreductases is presented in an attempt to standardize its use in comparative and experimental studies. Its detailed description includes improved incubation techniques, preparation of media, preservation of cytochemical activity on smears and evaluation of cytochemical preparations. Special attention is paid to the competing activities which may affect the specificity of the results.


Determination of the minimum alveolar concentration (MAC) of aliflurane in dogs. The minimum alveolar concentration (MAC) of aliflurane was measured in ten dogs. A value of 1.84 volumes per cent was determined, which correlates well with predictions based on lipid solubility. Induction of anesthesia and recovery were rapid, as would be anticipated with an agent of relatively low solubility in blood (blood-gas partition coefficient = 1.7). Circulatory responses over a relatively narrow range of aliflurane concentrations (0.8 to 1.4 MAC) remained stable, but the development of tachypnea, irregular ventilatory patterns, and increased muscle tone were frequently encountered during aliflurane anesthesis.


Hydrogen ion concentration versus pH.

Hydrogen ion concentration versus pH. There appears to be a tendency to convert pH values into "hydrogen ion concentrations" using the antilog of negative pH values. The present communication describes the thermodynamic basis of pH to explain that the above procedure is erroneous and that pH values should be treated as primary variables. Acidity expressed by the hydrogen ion concentrations measured by titrations (base excess or base deficit) has no bearing with the "hydrogen ion concentrations" derived by the antilog of negative pH.


The effects of H1 and H2 antihistamines on histamine inhalation challenges in asthmatic patients. This study was designed to examine the effect of an H1 antihistamine, chlorpheniramine, an H2 antihistamine, cimetidine, and the combination of chlorpheniramine and cimetidine on histamine-induced bronchoconstriction in a double-blind, randomized protocol on 11 patients with asthma. Each patient underwent a histamine inhalation challenge on 5 separate days. After a control day, histamine inhalation challenges were performed 2 h after the administration of a single oral dose of 8 mg of chlorpheniramine, 300 mg of cimetidine, the combination of chlorpheniramine and cimetidine, or placebo. Baseline pulmonary function measurements were not significantly altered by the 4 treatments. Body plethysmography data and measurements from the forced vital capacity maneuver were obtained before and after the histamine inhalation challenges. The provocation dose of histamine that produced a 20% decrease in forced expiratory volume in one second, a 35% decrease in mean forced expiratory flow during the middle half of the forced vital capacity, and a 50% decrease in specific airway conductance was significantly increased after administration of chlorpheniramine (p less than 0.002) and decreased after administration of cimetidine (p less than 0.02), where as no significant effect was noted after the combination of chlorpheniramine and cimetidine. The results suggest the presence of both H1 and H2 receptors in the airways of asthmatic patients.


Lung lysyl oxidase activity: relation to lung growth. The activity of lysyl oxidase (LOX), the extracellular enzyme responsible for initiating crosslinking of collagen and elastin, was measured during 3 types of postnatal lung growth. Lung parenchymal and pleural LOX activity was high in the first 3 wk of of life, decreasing by 50 % to stable amounts by 4 to 10 wk. In contrast, airway and aortic LOX activity remained high during the first 10 wk of life, decreasing by 50 to 75 % thereafter. After pneumonectomy, lung LOX activity doubled within 24 h, decreasing to control values thereafter. Hypoxia (12 to 13 % O2) resulted in a prompt and sustained increase in lung but not pleural, airway, or aortic LOX activity. Thus, LOX activity can be controlled precisely within specific tissues and appears to be related to early phases of connective-tissue synthesis. Further studies of the synthesis and degradation of LOX should provide important information about the control of connective-tissue formation within the lungs.


Pneumococcal infections after human bone-marrow transplantation. Seven of 26 long-term survivors (greater than 7 months post-transplant) of bone-marrow transplantation developed penicillin-sensitive pneumococcal infections more than 7 months after transplantation. One patient had two infections. Six of eight infections were associated with pneumococcal bacteremia, and Streptococcus pneumoniae type 6A was isolated in three cases. Two infections were fatal. All patients had normal nematopoietic function, and none was receiving immunosuppressive therapy. The development of pneumococcal infection was significantly associated with males and with abnormally low or high serum IGG and IgM levels but not with graft-versus-host disease. Serum opsonic activity for S. pneumoniae type 6A was decreased in six of the seven patients when compared to normal pooled serum in an in-vitro bactericidal assay. Four of the six patients with impaired opsonic activity had low serum antibody levels for S. pneumoniae type 6A capsular polysaccharide, while the other two patients had low serum CH100 complement activity. Bone-marrow transplant recipients have an increased susceptibility to pneumococcal infections and should be evaluated for prophylactic penicillin or pneumococcal vaccination.


Home parenteral nutrition in treatment of severe radiation enteritis. Ten patients with radiation enteritis unresponsive to conventional medical and surgical therapy were put on long-term parenteral nutrition at home. Six of the patients are alive at home; four patients died, two from recurrent cancer. Some of the patients have been able to resume oral intake, but none have been able to discontinue parenteral nutrition. Fistulas healed or had a marked decrease in output. Two patients in our series were given prednisone and slinical improvement of radiation enteritis with this therapy.


An interlaboratory survey of hydrogen ion and blood gas determinations. A survey of [H+], Pco2, and Po2 analyses in 360 laboratories was conducted using three commercial control materials: two aqueous gas-equilibrated buffer solutions (General Diagnostics and IL) and one whole blood material (DADE). There was little difference in precision or accuracy between instruments, classified according to manufacturer into four groups, and materials for [H+] and Pco2, or in precision for Po2. There were, however, differences in accuracy between instrument groups for Po2 analysis on the aqueous materials, and in some cases the mean values lay outside the range assigned by the material manufacturers. The 35 instruments outside clinical chemistry departments yielded results similar to those of all participants. The relation between inter- and intra-laboratory precision was similar to that for many analytes, suggesting that neither these materials nor the state of these analyses is unsatisfactory. Aqueous materials should, however, be used with caution as accuracy controls for Po2.


An improved radioenzymatic assay for histamine in human plasma, whole blood, urine, and gastric juice. A radioenzymatic method suitable for the assay of histamine in human blood, urine, plasma, and gastric juice is described. It differs from earlier methods by use of a histamine methyltransferase preparation from pig brain, of high activity tritiated S-adenosylmethionine, and of a heat precipitation step to reduce the previously noted interference from plasma constituents. The method is simpler than those requiring solvent extraction and concentration of histamine, gives recoveries in the range 80-120%, and so sliminates the need for internal standardisation. The method is sensitive and precise with coefficients of variation for blood, urine, and plasma of 5%, 6%, and 13% respectively. The mean +/- standard deviation for normal human plasma histamine is 5 +/- 4 nmol/l, for whole blood 559 +/- 193 nmol/l, and for urine 229 +/- 128 nmol/24h.


Vein of the vestibular aqueduct.

Vein of the vestibular aqueduct. The vein of the vestibular aqueduct (VVA) was investigated in a series of 40 human temporal bones. The processing included vascular injection with a colored medium, decalcification and cutting in serial, thick sections, which were put in a clear fluid and studied with a stereo-microscope. The labyrinthine roots of the VVA are the single veins of the ampulla and simple limbs of the semicircular canals  and of the posterior wall of the utricle. They drain the rich capillary bed of the simple endolymphatic walls of the canals and the utricle, as well as a small peripheral area of the cristae and the utricular macula. The VVA leaves the vestibule through an individual bone canal running parallel to the vestibular aqueduct up to the dura of the posterior side of the petrosa in the area of the endolymphatic sac. It then opens in the inferior petrosal sinus or the jugular bulb. The vein receives other branches from the bone, dura and sac. Correct information on the course of this vein appears to be lacking in contemporary textbooks and articles, although it has been correctly described since the last century.


Clinical pharmacology of mephenytoin and ethotoin. Effective prescribing of anticonvulsants requires foreknowledge of baseline pharmacokinetic data. Little such information is available about the hydantoins other than phenytoin, although one of them, mephenytoin, is widely used. Useful pharmacokinetic data should be derived from patients already exposed to anticonvulsants to reflect the induction of hepatic oxidative enzymes. Single-dose studies of mephenytoin (Mesantoin) and ethotoin (Peganone) were performed in adult inpatients on stable regimens of other anticonvulsants. Five patients received mephenytoin, 7 mg per kilogram of body weight. Serial blood sampling was performed rigorously. The time to peak concentration (Tmax) for mephenytoin was 1 hour, with a half-life (T 1/2) of 7 hours; the T 1/2 of its metabolite, 5-ethyl-5-phenylhydantion, was 96 hours. Ethotoin administration was 25 mg per kilogram in 5 patients. Ethotoin Tmax was 2 hours, with a T 1/2 of 5 hours. Saliva accurately represented the unbound fraction for all three agents. Mean salivary levels (as percentage of total levels) were 61% for mephenytoin, 73% for its metabolite, and 54% for ethotoin. The implications for therapy are that following mephenytoin administration, the metabolite 5-ethyl-5-phenylhydantoin will provide anticonvulsant effectiveness, with its long half-life producing stable blood levels on simple dose schedules. Ethotoin, in contrast, has a short half-life and would require divided daily doses to achieve a steady state. This, rather than pharmacological ineffectiveness, limits its usefulness.


Absence of otoconia in a human infant.

Absence of otoconia in a human infant. Temporal bone specimens were obtained at autopsy from a six-week-old victim of sudden infant death syndrome. The inner ears were microdissected and studied by a combination of light and scanning electron microscopy. Otoconia were completely absent from the sensory organs of the saccule and utricle on both sides. However, the gelatinous otoconial membranes, neuroepithelia, and innervation were normal. In parallel with unusually light pigmentation of the skin in this Negro infant, relatively few melanocytes were found in the membranous walls of the saccule and utricle. No abnormalities were present in other inner ear structures. This anomaly, which we believe was congenital in nature, is strikingly similar to a genetically determined inner ear defect which has been extensively studied in experimental animals.


[Solubility and the complex-forming properties of the zwitterlytes, 6-aminopenicillanic and 7-aminodesacetoxycephalosporanic acids]. Relation between solubility of the zwitterlites 6-aminopenicillanic acid (6-APA) and 7-aminodesacetoxycephalosporamic acid (7-ADCA) and the ionic strength and pH of the solution was studied. A specific effect of benzylpenicillin (BP) and 7-phenylacetamidodesacetoxycephalosporanic acid (7-PADCA) on solubility of the above zwitterlites was found and complex formation was suggested. An explanation of the complex formation mechanism and a method for estimation of the instability constants of the complexes are presented. The method is based on determination of the changes in the zwitterlite solubility in the presence of the complex forming compound. The values of the characteristic solubility of the ampholyte zwitter ions and the instability constants of the complexes "zwitterlite-complex forming compounds" were determined. A mathematical description of the relation between the zwitterlite solubility within a wide range of pH and concentrations of the complex forming compounds is presented. Coincidence of the calculated and experimental data was observed, which provided a supposition that the substances involved in complex formation were charged.


[Effect of additions of the carbon and nitrogen sources on the NADP+ and NAD+ level in the mycelium of a producing strain of P. nigricans Thom]. The levels of NADP+ and NAD+ in the mycelium of a cultured strain of P. nigricans Thom grown on the mineral medium with 2 per cent of glucose were studied as dependent on the time and method of addition of glucose. NaNO3 or their mixture to the medium, i. e. a single addition simultaneously with the spore inoculation or on the 7th day of the culture development, divided additions on the 5th--7th days. It was shown that addition of the above sources simultaneously with inoculation affected the levels of both dinucleotides by the 5th and 8th days. Analogous additions on the 7th day in 24 hours decreased the level of NAD+, while the NADP+ concentration increased after addition of a mixture of glucose and NANO3 and remained unchanged on their separate addition. When the above sources were added in divided doses the level of NADP+ increased till the 7th day and that of NAD+ till the end of the experiment (the 8th day).


Hyperproduction of tryptophan by Escherichia coli: genetic manipulation of the pathways leading to tryptophan formation. Conversion of glucose and ammonium salts into tryptophan by mutants of Escherichia coli was examined as part of a feasibility study on the manufacture of tryptophan. This involved construction, largely by transduction, or a variety of multiple-mutation strains with defined genotypes. By comparing the properties of these strains, we were able to define in biochemical terms several changes that significantly enhance process productivity, namely (i) release of the first enzyme of the common pathway of aromatic biosynthesis and the first enzyme of the tryptophan pathway (3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and the anthranilate aggregate, respectively) from inhibition by end products, (ii) blockage of the diversion of chorismate to phenylalanine and tyrosine biosynthesis, and (iii) presence of highly elevated tryptophan pathway enzyme levels, such as result from interference with both repression and attenuation, combined with gene amplification. By using strains carrying appropriate mutations to effect all of these changes, high values of specific productivity were obtained in bath culture (approximately 80 mg/g [dry weight] per h). Furthermore, a pronounced decay in the level of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase activity was implicated as a cause of declining process producitivity during stationary phase, emphasizing the value of having derepressed levels of this enzyme.


Oxidation of secondary alcohols to methyl ketones by yeasts. Cell suspensions of yeasts, Candida utilis ATCC 26387, Hansenula polymorpha ATCC 26012, Pichia sp. NRRL-Y-11328, Torulopsis sp. strain A1, and Kloeckera sp. strain A2, grown on various C-1 compounds (methanol, methylamine, methylformate), ethanol, and propylamine catalyzed the oxidation of secondary alcohols to the corresponding methyl ketones. Thus, isopropanol, 2-butanol, 2-pentanol, and 2-hexanol were converted to acetone, 2-butanone, 2-pentanone, and 2-hexanone, respectively. Cell-free extracts derived from methanol-grown yeasts catalyzed an oxidized nicotinamide adenine dinucleotide-dependent oxidation of secondary alcohols to the corresponding methyl ketones, Primary alcohols were not oxidized. The effect of various environmental factors on the production of methyl ketones from secondary alcohols by methanol-grown Pichia sp. was investigated.


Rapid seven-hour fecal coliform test.

Rapid seven-hour fecal coliform test. A rapid 7-h fecal coliform (FC) test for the detection of FC in water has been developed. This membrane filter test utilizes a lightly buffered lactose-based medium (m-7-h FC medium) combined with a sensitive pH indicator system. FC colonies appeared yellow against a light purple background after incubation at 41.5 degrees C for 7 to 7.25 h. Comparison of FC test results showed that the mean verified FC count ratio (7-h FC count/24-h FC count) for surface water samples was 1.08. The mean FC count ratio (7-h FC count/24-h FC count) for unchlorinater wastewater ranged from 1.95 to 5.05. Verification of yellow FC colonies from m-7-h FC medium averaged 97%. Data from field tests on Lake Michigan bathing beach water samples showed that unverified 7-h FC counts averaged 96% of the 24-h FC counts. The 7-h FC test was found to be suitable for the examination of surface waters and unchlorinated sewage and could serve as an emergency test for detection of sewage or fecal contamination of potable water.


Comparative adsorption of human enteroviruses, simian rotavirus, and selected bacteriophages to soils. Virus adsorption to soils is considered to be the most important factor in removing viruses after land treatment of wastewater. Most of the studies on virus adsorption to soils have utilized poliovirus as the model system. In the present study, comparative adsorption of a number of different types and strains of human enteroviruses and bacteriophages to nine different soil types was studied. Under the experimental conditions of this study, greater than 90% of all viruses adsorbed to a sandy loam soil except echovirus types 1, 12, and 29 and a simian rotavirus (SA-11), which adsorbed to a considerably lower degree. A great deal of variability was observed between adsorption of different strains of echovirus type 1, indicating that viral adsorption to soils is highly strain dependent. Of the five phages studied, f2 and phi X174 adsorbed the least. In addition to being dependent on type and strain of virus, adsorption was found to be influenced also by type of soil. Thus, soils having a saturated pH of less than 5 were generally good adsorbers. From these results, it appears that no one enterovirus or coliphage can be used as the sole model for determining the adsorptive behavior of viruses to soils and that no single soil can be used as the model for determining viral adsorptive capacity of all soil types.


pH modification of the effects of detergents on the stability of enteric viruses. The effect of detergents on the stability of enteric viruses was found to be highly dependent on pH. This was demonstrated primarily with two ionic detergents, sodium dodecyl sulfate (an anionic detergent) and dodecyltrimethylammonium chloride (a cationic detergent). Both detergents were shown to be potent virucidal agents for reovirus, but the effects of sodium dodecyl sulfate were minimal near neutrality and much more pronounced at low than at high pH values. Dodecyltrimethylammonium chloride was extremely virucidal at high pH's but had little observable effect on reovirus stability at low pH values. In contrast, both detergents protected enteroviruses against heat at neutral and alkaline pH's. However, as was found with reovirus, sodium dodecyl sulfate was extremely virucidal at pH values below 5, even when the virus samples were incubated in ice. At different pH's the effects of detergents on the stabilities of coliphages T4, f1, and Q beta were qualitatively similar to those found with reovirus. Differences in viral stability in these experiments appeared to be due to the effects of pH on the ionic states of the viral capsid proteins.


[Epidemiology of measles (author's transl)]. A detailed analysis of epidemiology of measles in pre-vaccinal era is reported for both isolated, underdeveloped and developed populations, together with the most recent knowledge on epidemiological changes tooking place in those countries which introduced large-scale vaccination with live, further attenuated measles vaccines: reduction of morbidity mortality and complication rates, morbidity age shift, etc. Results obtained in U.S.A., where vaccination was introduced in 1963, are analyzed also considering that it never spanned above 66% of children 1-4 year. Suggestions derive after the american experience for a correct strategy of the incoming vaccination in this Country.


[The vaccination against measles: effectiveness and care in use (author's transl)]. The A. deals with three clinic aspects of outstanding importance of the measles and its relevant vaccination; that is: the complications of the disease, the contraindications to live virus vaccine and the possible reactions to vaccine. It is particularly stressed the possibility of neurological complications due either to disease or to vaccination posthumous.


Study of the effects of neurotransmitters on the hypothalamus-pituitary-testis function in in vitro cell suspension system. The effects of different neurotransmitters were tested in vitro on a hypothalamic tissue, collagenase-digested isolated anterior pituitary and Leydig cell suspension system by measuring the testosterone production of the Leydig cells. Neurotransmitters were used in concentrations of 0.25, 1.0, 2.5, 5.0, and 10.0 micrograms/ml incubation medium. Dopamine in doses of 1.0, 2.5, and 5.0 micrograms/ml increased the hypothalamic tissue-induced pituitary-testis activation, while it had no direct effect on pituitary and Leydig cells. Noradrenaline in the concentration range 2.5--10.0 micrograms/ml decreased the luteinizing-hormone-releasing-hormone (LHRH) sensitivity of the pituitary cells. 5.0 and 10.0 micrograms/ml 5-hydroxytryptamine decreased the testosterone production and the hCG sensitivity of the isolated Leydig cells. Carbamylcholine and pilocarpine had no action on the in vitro system at the different levels studied.


Rapid decrease of ATP content in intact cells of Saccharomyces cerevisiae after incubation with low concentrations of sulfite. Sulfite, at concentrations above 1 mM and at a pH below 4, caused cell death in Saccharomyces cerevisiae X2180 as measured by the colony-forming capacity. A rapid decrease in the ATP content was observed prior to cellular death. The depletion of ATP was reversible and the lethal effect could be prevented if the cells were exposed to sulfite for periods of less than 1 h. Extent and rate of ATP depletion were dependent on time, pH value, temperature and sulfite concentrations.


Protein synthesizing systems from spores and vegetative cells of Bacillus cereus. A system of polyphenylalanine synthesis was optimized for a comparison of the polymerizing activities of ribosomes from spores and vegetative cells of Bacillus cereus T. Ribosomes of both types react similarly, showing a magnesium optimum of about 6mM and spermidine optima of about 5mM and 4mM for vegative and spore ribosomes, respectively. These lead to optimum mono- to multivalent cation rations of 9 and 10 respectively at 100 mM ammonium ion. A comparison of the response of these ribosomes to suboptimal concentrations of magnesium and spermidine show that they differ qualitatively from each other, suggesting that they possess different structure, macromolecular or ionic components.


Characterization and synthesis regulation of Penicillium italicum 1,6-beta-glucanase. The filamentous fungus Penicillium italicum when grown in a synthetic medium, produced and secreted 1,6-beta-glucanase into the culture medium. This enzyme has been partially purified by gel filtration. After this step the active fractions were free of 1,3-beta-glucanase, alpha-amylase and beta-glucosidase activities. Only four proteins, one associated with the enzyme, were found by polyacrylamide gel electrophoresis under non denaturing conditions. The enzyme behaves as an acidic protein (pI 4.65) with an optimum pH of 5 and an endohydrolytic mode of action. The activity was also found associated with the mycelium. Its synthesis was repressed by glucose or growth-promoting sugars. Derepression in low glucose containing medium required protein synthesis. 8-Hydroxyquinoline, an RNA synthesis inhibitor, added during the derepression period did permit some increase in the specific activity but prevented it when added at the beginning of that period.


Trypotophanase from a marine bacterium, Vibrio K-7 synthesis, purification and some chemical catalytic properties. The conditions for synthesis, purification, and properties of tryptophanase by a marine organism (Vibrio K-7) were studied. Tryptophanase was induced by tryptophan and its analogs, and partially repressed by 0.5% glucose or glycerol. NaCl (0.4 M) was required for optimal growth and tryptophanase activity in whole cells. The enzyme was purified to 92% homogeneity by heat treatment, hydroxyapatite chromatography and fractionation with ammonium sulfate. This tryptophanase has been found to have kinetic properties similar to the tryptophanase from other microorganisms. It carries out both alpha, beta-elimination reactions (using tryptophan, serine, cysteine and S-methylcysteine as substrates) and beta-replacement reactions (forming tryptophan from indole and serine, cysteine or S-methyl-cysteine). The enzyme has a sedimentation coefficient of 9.2S and requires pyridoxal 5'-phosphate as a cofactor. The optimal pH for the tryptophanase reaction is pH 8.0.


Stability of the lyophilized F(ab')2 fragments of horse tetanus antibodies isolated by affinity chromatography. F(ab')2 fragments of horse tetanus antibodies were obtained from horse hyperimmune sera after peptic digestion. The digest was passed through a column of tetanus toxoid coupled with Sepharose 4B, F(ab')2 fragments were eluted with a solution of 5 mM HCl in 150 mM NaCl and the eluates were concentrated by ultrafiltration and lyophilized. Glycine and human serum albumin were used as stabilizing agents. Polyacrylamide gel electrophoretic mobility and molecular weight of the fragments remained unchanged after lyophilization. Freeze-dried preparations stored two months at 56 degrees C showed only a slight decrease in antitetanus activity. The ORD measurements and spectrophotometric determinations of unfolding over pH range 2.1-5.0 show that the F(ab')2 fragment structure is highly stable in acid medium.


Application of fine-needle aspiration biopsy for the diagnosis of dysplastic and neoplastic liver cell changes induced by N-nitrosomorpholine in rats. Male rats were treated with the hepatocarcinogen, N-nitrosomorpholine (NNM, 10 mg ad 100 ml drinking water) for 19 weeks. Repeated fine-needle aspiration biopsies of the liver were performed percutaneously. Cytomorphologic and cytochemical criteria were used for the characterization of dysplastic and carcinoma cells. The alterations seen in the smears were correlated with histopathologic findings in the punctured liver lobes. Cells showing type I dysplasia were recognized in smears obtained from day 7 on. They corresponded to the swollen, glycogen-free cells developing in zone 3 of the Rappaport acinus during the early treatment phases. In later stages type I dysplastic cells were observed in smears. This coincided with the development of neoplastic nodules seen in histopathologic preparations. Carcinoma cells were recognized first after 15 weeks. Marked gamma-glutamyl transpeptidase (gamma-GT) activity could be demonstrated cytochemically in biopsy smears and biochemically in biopsy homogenates during the early phases of NNM-treatment. Simultaneously, a rise in gamma-GT activity was also observed in the serum.


In vitro studies on Borna virus. II. Properties of the virus. Successful cultivation and titration of Borna disease virus in cell cultures enabled detailed studies of the virus properties. Borna virus is labile towards treatment with heat, pH 3.0 and lipid solvents. It is relatively stable at low temperatures and in frozen state. It is easily inactivated by ultraviolet light as e.g. vesicular stomatitis virus. After ultrafiltration studies, the size of the infectious virus unit is between 80 and 100 nm. Its buoyant density in cesium chloride is 1.165 g per ml. The one step multiplication curve shows that Borna virus has a replication cycle of about 2 days in BSC 1 cells. In growth experiments using antimetabilites it behaves like certain RNA containing viruses. As its multiplication is not inhibited by bromo- and iododeoxyuridine and actinomycin D, no DNA step seems to be involved in virus synthesis. Regarding these properties and the intracellular antigen distribution as shown by fluorescent antibodies, it is not possible to attribute Borna virus to any of the established virus groups.


[Study of seminal fluid. Critical evaluation of various laboratory parameters]. The various humoral and morphological parameters on which examination of seminal fluid is based are considered. Two groups of causes that may modify normal fertility are pointed out: one concerns functional changes of the gonad and/or of the accessory glands, the other the ejaculatory phase and specifically the modalities for collecting the ejaculte. In this regard, stress is laid on how the first fraction of split ejaculate not only possesses biochemical characteristics different to those of the second fraction, but also presents a much higher sperm concentration and, hence, greater fertilizing capacity. It is therefore clear that separate examination of the two fractions may point to states of "latent" subfertility which global examination cannot spot.


Production, purification and characterization of rat interferon. Priming with heterologous mouse interferon, increased production of an antiviral substance induced in rat diploid fibroblasts by Newcastle disease virus. This substance was characterized as an acid stable interferon. This rat interferon exhibited marked cross-species antiviral activity when tested in mouse L929B cells, guinea pig embryo fibroblasts, human fibroblasts and bovine cells but was not active on chick embryo cells.


[Plasma and intra-erythrocyte variations in acid-base equilibrium in acute acidosis due to acetazolamide loading]. Plasma and red cell acid-base equilibrium was studied in 14 healthy subjects following the induction of acidosis by means of i.v. acetazolamide. It was found that changes in red cell values were significantly less marked than those in plasma.


Biochemical differences between cytoplasmic malate dehydrogenase allozymes of Drosophila virilis. Two allozymes (MDHf and MDHs) of cytoplasmic malate dehydrogenase of Drosophila virilis were partially purified and their biochemical properties were compared. MDHf has a pH optimum of 9.75 and MDHs one of 9.25 for malate oxidation. Optimal pH for oxaloacetate reduction is 6.75 and 8.0 for MDHf and MDHs, respectively. The Km value for oxaloacetate of MDHs is approximately twice as that of MDHf. No differences were found with respect to thermostability and Km's for malate, NAD+, or NADH. These results are discussed in terms of the physiological role of cytoplasmic malate dehydrogenase of D. virilis.


Isolation and partial characterization of three acidic proteinases in erythrocyte membranes. 1. The distribution of proteolytic activity in membranes from human erythrocytes and from rabbit reticulocytes and erythrocytes was investigated, after removal of leucocytes and platelets from the cell suspensions. 2. All membrane preparations displayed proteolytic activity in the acidic pH region only. Membranes from human and rabbit mature erythrocytes showed latent activity, which could be increased when extracted with a number of detergents. 3. Three active fractions were resolved either by gel chromatography of solubilized membrane extracts or by standard polyacrylamide-gel electrophoresis. The three proteinase activities (designated proteinases I, II and III) were purified from solubilized extracts of human erythrocyte membranes. 4. The relevant mol.wts. were around 80000, 40000 and 30000, respectively, and each of the three proteinases appeared to be composed of a single polypeptide chain. 5. Distinctive pH optima (in the range pH2.8-3.9) and different saturation profiles with globin as substrate were observed for proteinases I, II and III. 6. Dithioerythritol, Hg(2+) and Cu(2+) inhibited each of the three human enzymes, but more selective inhibitory effects were exerted by other modifiers of proteolytic enzymes and by haemin. Similar effects were observed with the three proteinases from rabbit cells. 7. The activity of the three human proteinases seems to be restricted to naturally occurring protein substrates, although with poor specificity, and none of them was active on synthetic substrates. 8. Digestion of globin by each of the three enzymes yielded similar polypeptide fragments in all cases, this indicating an endopeptidase type of activity.


Native cross-links in collagen fibrils induce resistance to human synovial collagenase. A model system consisting of highly purified lysyl oxidase and reconstituted lathyritic chick bone collagen fibrils was used to study the effect of collagen cross-linking on collagen degradation by mammalian collagenase. The results indicate that synthesis of approx. 0.1 Schiff-base cross-link per collagen molecule results in a 2--3-fold resistance to human synovial collagenase when compared with un-cross-linked controls or samples incubated in the presence of beta-aminopropionitrile to inhibit cross-linking. These results confirm previous studies utilizing artificially cross-linked collagens, or collagens isolated as insoluble material after cross-linking in vivo, and suggest that increased resistance to collagenase may be one of the earliest effects of cross-linking in vivo. The extent of intermolecular cross-linking among collagen fibrils may provide a mechanism for regulating the rate of collagen catabolism relative to synthesis in normal and pathological conditions.


Reinvestigation of the reaction of chymotrypsin with N-furylacryloyltryptophan derivatives at acidic pH. The reaction of alpha-chymotrypsin with N alpha-3-(2-furyl)acryloyl-L-tryptophan methyl ester (FA-Trp-OMe) and amide has been investigated in aqueous and dimethylsulphoxide cryosolvent solutions from pH2 to 7 and over a wide temperature range. Previous reports have suggested that an intermediate preceding the acyl-enzyme can be detected spectrophotometrically in the reaction with methyl esters of FA-Trp and FA-Tyr at low pH [Yu & Viswanatha (1969) Eur. J. Biochem. 11, 347--352), and that this intermediate is an oxazolinone [Coletti-Previero et al. (1970) FEBS Lett. 11, 213--217]. We show that the previous interpretations of the time-dependent spectral changes were incorrect, and that the only detected intermediate is the acyl-enzyme. This may be isolated by gel filtration at pH less than 2.5, 1 degree C, owing to its relative stability. The pH-dependence of the rates of acylation and deacylation from pH 8.5 to 2.0 are consistent with a single ionization of pK congruent to 7.0 in both aqueous and cryosolvent solutions.


Dodecyl sulphate/polyacrylamide-gel electrophoresis at low pH values and low temperatures. A simple method is described for dodecyl sulphate/polyacrylamide-gel electrophoresis of pH- and temperature-labile biological intermediates. The method is based on a catalyst system that works at temperatures of 2--4 degrees C and pH values of 2--4 and an appropriate buffer system containing Li+ or Tris [CH2OH--C(CH2OH)2--NH3+] instead of Na+. This system does not lead to the precipitation of 1% dodecyl sulphate.


Stimulation of phosphatidylinositol turnover in various tissues by cholinergic and adrenergic agonists, by histamine and by caerulein. Studies are reported of the biochemical and pharmacological characteristics of the stimulation of phosphatidylinositol metabolism that is produced in appropriate target tissues by stimulation of various receptors that use Ca(2+) as their second messenger. (1) Muscarinic cholinergic and alpha-adrenergic phosphatidylinositol responses were observed in rat lacrimal gland, and a response to caerulein was detected in the longitudinal smooth muscle of guinea-pig ileum. (2) The muscarinic cholinergic phosphatidylinositol response of rat lacrimal gland, like that of several other tissues, is not dependent on the availability of extracellular Ca(2+). (3) Three phosphatidylinositol responses, namely to histamine in guinea-pig ileum smooth muscle, to alpha-adrenergic stimulation in rat vas deferens and to muscarinic cholinergic stimulation in rat lacrimal gland, were all found to involve phosphatidylinositol breakdown. (4) The stereospecificity of the muscarinic receptor responsible for the phosphatidylinositol response of guinea-pig pancreas was tested by using the two stereoisomeric forms of acetyl-beta-methylcholine; the S-isomer was very much more active than the R-isomer in provoking both phosphatidylinositol breakdown and its labelling with (32)P, as it is in provoking other physiological responses such as contractility or secretion. (5) Pilocarpine, a muscarinic partial agonist, provoked a significantly smaller phosphatidylinositol breakdown in rat parotid fragments than did carbamoylcholine, a potent muscarinic agonist. (6) All of these results are consistent with, but do not prove, a previously offered hypothesis that suggests that phosphatidylinositol breakdown is a reaction essential to stimulus-response coupling at a variety of cell-surface receptors that mobilize Ca(2+) from and through the plasma membranes of target tissues.


Quantitative analysis of proton-linked transport systems. The lactose permease of Escherichia coli. Evidence is presented that lactose uptake into whole cells of Escherichia coli occurs by symport with a single proton over the range of external pH 6.5--7.7. The proton/lactose stoicheiometry has been measured directly over this pH range by comparison of the initial rates of proton and lactose uptake into anaerobic resting cell suspensions of E. coli ML308. Further, the relationship between the protonmotive force and lactose accumulation has been studied in E. coli ML308-225 over the range of external pH 5.9--8.7. At no point was the accumulation of the beta-galactoside in thermodynamic equilibrium with the protonmotive force. It is concluded that the concentration of lactose within the cell is governed by kinetic factors rather than pH-dependent changes in the proton/substrate stoicheiometry. The relevance of these findings to the model of pH-dependent proton/substrate stoicheiometries derived from studies with E. coli membrane vesicles is discussed.


Mechanism of the stimulation of serine and alanine transport into isolated rat liver cells by bicarbonate ions. 1. Bicarbonate ions stimulate the transport of serine and alanine into isolated hepatocytes. 2. The effect of bicarbonate is to increase the Vmax. of the transport process without changing the apparent Km. 3. The intracellular pH was estimated from the distribution of the weak base methylamine and the weak acid 5,5'-dimethyloxazolidine-2,4-dione (DMO) across the plasma membrane. 4. The addition of bicarbonate to a cell suspension caused the internal pH to become more acid. 5. The initial rate of serine, alanine and glycine transport was a linear function of the initial difference in pH across the membrane. 6. It is concluded that bicarbonate activates the transport of these amino acids primarily by increasing the pH difference across the plasma membrane. 7. It is suggested that the uptake of serine together with Na+ ions occurs in exchange for H+ ions, which are translocated outwards on the same carrier system. Some preliminary evidence consistent with this model is presented.


Biosynthesis of ethylene from methionine. Isolation of the putative intermediate 4-methylthio-2-oxobutanoate from culture fluids of bacteria and fungi. Methods are described for identifying the 2,4-dinitrophenylhydrazones of 4-methylthio-2-oxobutanoate by means of t.l.c., n.m.r. and mass spectroscopy. By using these methods 4-methylthio-2-oxobutanoate, a putative intermediate in the biosynthesis of ethylene from methionine, has been identified in culture fluids of Aeromonas hydrophila B12E and a coryneform bacterium D7F grown in the presence of methionine. Relative to 4-methylthio-2-oxobutanoate, the yield of 3-(methylthio)propanal (methional) from the same cultures was less than 1%. Because 4-[2H]methylthio-2-oxobutanoate was obtained from cultures grown on [Me-2H]methionine, the 4-methylthio-2-oxobutanoate must be derived from methionine. By means of t.l.c. alone, 4-methylthio-2-oxobutanoate was identified in the culture fluids of a range of bacteria, the yeast Saccharomyces cerevisiae and the fungus Penicillium digitatum. A photochemical assay developed for 4-methylthio-2-oxobutanoate shows it to be a product of the metabolism of methionine by Escherichia, Pseudomonas, Bacillus, Acinetobacter, Aeromonas, Rhizobium and Corynebacterium species.


A controlled study of pneumococcal polysaccharide vaccine in systemic lupus erythematosus. The immunogenicity and potential for disease modification of pneumococcal polysaccharide vaccine in systemic lupus erythematosus were evaluated in a controlled, double-blind study. Forty patients were randomly chosen to receive an intramuscular injection of either vaccine or placebo. Changes in mean antibody concentrations (nanograms antibody nitrogen per milliliter serum) to 12 type-specific pneumococcal capsular antigens from prevaccination to one month after vaccination were 177 to 1045 in the vaccine (P less than 0.001) and 164 to 153 in the placebo-treated patients. In the month after vaccination, neither vaccine nor placebo-treated patients had a significant change in lupus disease activity as assessed by a composite clinical, laboratory, and serologic index. We conclude that patients with systemic lupus erythematosus can be successfully immunized with pneumococcal vaccine without detectable alterations of the underlying disease.


[The influence of benzoctamine on dopamine and norepinephrine contents in substantia nigra and on the muscle stretch reflex of the cat (author's transl)]. The concentration of dopamine (DA) and norepinephrine (NE) are decreased to a maximum of 40% and 20%, respectively, in the substantia nigra of precollicular and prenigral decerebrated cats previously treated with benzoctamine (Tacitin) (0.7 mg/kg i.v.), while the tyrosine-hydroxylase and MAO activities remain unchanged. Muscle stretch reflexes recorded simultaneously showed a significant decrease of stretch tension which is evoked by a clear reduction of Ia discharges of the primary muscle spindle afferents. It would appear that benzoctamine suppresses reflexly evoked gamma-motoneurone discharges.


Pharmacological study of [2-chloro-11-(2-dimethylaminoethoxy) dibenzo[b,f]thiepine] (zotepine), a new neuroleptic drug. 2-Chloro-11-(2-dimethyl-aminoethoxy)dibenzo [b,f]thiepine (zotepine) is a new neuroleptic drug with a chemical structure different from known neuroleptics. The psychopharmacological effects of zotepine in mice, rats and dogs were studied and compared with those of commercially available neuroleptics. Haloperidol and perphenazine were the most active and thioridazine was the least active in hibiting apomorphine-induced gnawing and circling movement, methamphetamine-induced gnawing and circling movement, conditioned avoidance response, motor activity, dopamine-induced pancreatic secretion and apomorphine-induced vomiting. These drugs also had the same order of potency in inducing catalepsy and increasing dopamine turnover and prolactin release. Chlorpromazine, propericiazine and thiothixene were intermediate in potency. Zotepine equalled chlorpromazine in most activities, however, it was clearly less active than chlorpromazine in potentiation of barbiturate sleep and cardiovascular effect.


Pharmacokinetics and metabolism of 2-chloro-11-(2-dimethylaminoethoxy)-dibenzo[b,f]thiepine (zotepine) in rat, mouse, dog and man. The absorption, distribution, metabolism and excretion of 2-chloro-11-(2-dimethylaminoethoxy)dibenzo [b,f]thiepine (zotepine), a new neuroleptic agent, were investigated in rat, mouse, dog and man. Zotepine was absorbed rapidly and almost completely from the gastrointestinal tract in all species after oral dosing. The serum level of the unchanged drug in man was comparatively higher than in animals. The serum half-lives of zotepine after i.v. dosing were approximately 3.2 h in rats, 1.5 h in mice and 3.0 h in dogs. The drug and radioactive metabolites were rapidly distributed to the tissues of rats and mice, and the brain levels of the unchanged drug were about 20 to 30 times higher than the serum levels. Only small amounts of the unchanged drug were excreted in the urine in all species; faecal excretion through the bile was the main route of elimination of the drug and metabolites. Extensive biliary excretion and enterohepatic circulation of the radioactive compound were observed in rats. Zotepine was well metabolized in the body. Besides N-demethylation and oxygenation of the N and/or S atoms, hydroxylation of the aromatic ring and consecutive conjugation were important metabolic pathways of zotepine.


Toxicological and teratological studies of 2-chloro-11-(2-dimethylaminoethoxy)-dibenzo[b,f]thiepine (zotepine), a new neuroleptic drug. Toxicological and teratological studies of 2-chloro-11-(2-dimethylaminoethoxy)dibenzo[b,f]thiepine (zotepine) were performed in mice, rats, rabbits and dogs. There was no essential difference among mice, rats, rabbits and dogs in the acute toxicity of i.v. given zotepine. The rather small variation between intravenous and oral acute toxicity suggests the good absorption of zotepine from the gastrointestinal tract. In the subacute and chronic toxicity studies in rats, significant changes attributed to the drug were impairment of growth, alveolar proliferation in the mammary gland, decrease in uterine weight and increased number of diestrous rats. These changes were dose-dependent in the animals given 16 mg/kg or higher. Incidence of tumor in the treated groups in the 12-month study was almost the same as that in the control. In the subacute and chronic toxicity studies in dogs, abnormal quietness and abnormal gait occurred. Enlarged breasts and galactorrhea also occurred in females given 16 and 64 mg/kg in the 6-month study. Apart from these changes, one dog given 64 mg/kg had reversible hepatic dysfunction. In the teratological studies, zotepine had no adverse effects on pregnant animals and their fetuses in rats and rabbits, or reproductive performance of the F1 rats.


Antihypertensive effect of intravenously and orally administered bunitrolol (Kö 1366). 1. Hypotensive effect of O-[3-(tert, butylamino-2-hydroxypropoxy]-benzonitrile hydrochloride (bunitrolol, Kö 1366) was examined by intravenous administration of 5 mg to 20 cases of essential hypertension and by longterm oral administration of 15-30 mg to 27 cases of essential hypertension. 2. It was found that bunitrolol given either i.v. or p.o., produced a significant decrease of systolic, diastolic and mean blood pressure at rest. 3. Inhibitory effect of bunitrolol on exercise-induced increase of blood pressure was remarkable, while there was little inhibitory effect on the cold pressor-induced one. 4. Bunitrolol decreased blood pressure and lowered plasma renin activity in high renin hypertensive patients, suggesting that the hypotensive mechanism based, at least in part, on its capacity of inhibiting renin secretion. 5. However, the main hypotensive mechanism of bunitrolol seemed, in the case of i.v. use, related to the cardiac inhibitory action. Regarding the hypotensive mechanism of this drug in the long-term use, further investigation will be required.


Spectrum analysis of the EEG in schizophrenic patients. A method is presented for the harmonic analysis of the human EEG. Its use for the study of schizophrenic patients under phenothiazine treatment is discussed and some examples are given. Correlations are observed with clinical evolution and plasma level of the drugs.


Pharmacokinetics of fazadinium in patients with renal failure. The serum concentrations of fazadinium and its metabolites were measured in 14 surgical patients with end-stage renal failure and in 11 patients free from kidney disease undergoing abdominal surgery. A two-compartment open model was used in the pharmacokinetic analysis of the data. The elimination half-life (T 1/2 beta) was prolonged by 60% in patients with renal failure (from 85 to 140 min) and corresponded to a 30% decrease of the plasma clearance. The plasma concentration of the metabolites never exceeded 7% of the unchanged fazadinium. These results suggest that the duration of action will be less prolonged for fazadinium than for other non-depolarizing neuromuscular blocking drugs in patients with renal failure. A supplementary biliary pathway appears to be a possible explanation for the rapid elimination of fazadinium, even in patients with renal failure.


Cimetidine in the prevention of the pulmonary acid aspiration (Mendelson's) syndrome. Cimetidine 200 mg was given i.v. 1 h before induction of anaesthesia to 20 patients about to undergo emergency surgery. There was an increase in pH of gastric aspirate in all patients (P less than 0.001) and in 80% pH was greater than 2.5 units at the time of induction of anaesthesia. The pH of gastric aspirate continued to increase after induction of anaesthesia and was greater than 4 units in all patients at the time of tracheal extubation and in the early recovery period.