[Infestation of mosquitoes from the Maritime Territory by the entomopathogenic fungus, Coelomomyces iliensis, under laboratory conditions]. The infection of larvae of Culex pipiens molestus, C. vagans and C. tritaeniorhynchus (the vector of the virus of Japanese encephalitis) with the entomopathogenic fungus Coelomomyces iliensis from Kazakhstan was carried out in the Primorje Territory in 1973--1974. At the infection of larvae of C. tritaeniorhynchus with the suspension of infected larvae the mortality was recorded only from 60% of individuals, while at the infection with a content of sporangia--from 36.6% of larvae. Somewhat less mortality (27.5%) was observed among larvae of C. pipiens molestus and C. pipiens. In control the mortali larvae did not exceed 5%.


Interaction of peripheral and central respiratory drives in cats. I. Effects of sodium cyanide as a peripheral chemoreceptor stimulus at different levels of CSF pH. In cats anesthetized with chloralose-urethane, the central respiratory chemoreceptors were exposed to mock CSF of pH 7.02, 7.02, or 7.57. The right carotid body was simultaneously stimulated by intracarotid injections of 40, 80, or 160 microgram sodium cyanide in 200 microliter Ringer solution. The left carotid nerve and, in some animals, both vagosympathetic truncs were dissected. It could be demonstrated that the increase in ventilation produced by application of NaCN to the peripheral chemoreceptors is significantly larger at high than at low mock CSF pH (i.e. at low than at high central stimulus intensity). In vagotomized cats the responses of VT and V to NaCN similarly depend upon CSF pH; they are somewhat larger, though, than in intact animals. These results are discussed as compared with results reported by different authors.


[Gastric acid secretion by vagal electrostimulation (author's transl)]. In the operative verification of vagotomy, electrical stimulation of the vagus nerves with the study of pH should seem to be a reliable method. On the basis of an experiment in the cat, the authors emphasise the effectiveness of electrical stimulation of gastric acid secretion, confirmed by 16 clinical cases. Stimulation of the proximal end of the divided right nerve may be used to detect a nervous strand arising above the level of the section and to confirm the completeness of left vagotomy.


Protonated polynucleotides structures - 22.CD study of the acid-base titration of poly(dG).poly(dC). The acid-base titration (pH 8 --> pH 2.5 --> pH 8) of eleven mixing curve samples of the poly(dG) plus poly(dC) system has been performed in 0.15 M NaCl. Upon protonation, poly(dG).poly(dC) gives rise to an acid complex, in various amounts according to the origin of the sample. We have established that the hysteresis of the acid-base titration is due to the non-reversible formation of an acid complex, and the liberation of the homopolymers at the end of the acid titration and during the base titration: the homopolymer mixtures remain stable up to pH 7. A 1G:1C stoichiometry appears to be the most probable for the acid complex, a 1G:2C stoichiometry, as found in poly(C(+)).poly(I).poly(C) or poly(C(+)).poly(G).poly(C), cannot be rejected. In the course of this study, evidence has been found that the structural consequences of protonation could be similar for both double stranded poly(dG).poly(dC) and G-C rich DNA's: 1) protonation starts near pH 6, dissociation of the acid complex of poly(dG).poly(dC) and of protonated DNA take place at pH 3; 2) the CD spectrum computed for the acid polymer complex displays a positive peak at 255 nm as found in the acid spectra of DNA's; 3) double stranded poly(dG).poly(dC) embedded in triple-stranded poly(dG).poly(dG).poly(dC) should be in the A-form and appears to be prevented from the proton induced conformational change. The neutral triple stranded poly(dG).poly(dG).poly(dC) appears therefore responsible, although indirectly, for the complexity and variability of the acid titration of poly(dG).poly(dC) samples.


Protonated polynucleotides structures - 23. The acid-base hysteresis of poly(dG).poly(dC). The large hysteresis observed during the acid-base titration of poly(dG). poly (dC) was studied by CD and potentiometric scanning curves. Intermediate scanning loops as well as the equilibrium and metastable branches of the hysteresis loop have been determined. The potentiometric titrations showed, however, that the various complexes were not discrete entities, but were linked in "polycomplexes" as had been already suggested. This prevented a thermodynamic study of the system. The acid-base titration was further investigated as a function of ionic strength and temperature. The pK's showed considerably lower ionic strength dependence than observed for polyribonucleotide complexes. The thermal transitions permitted to establish the relative stabilities of the various complexes between pH 2.5 and pH 12.0.


Cross-linking of nucleosomal histones with monofunctional imidoesters. Cross-linking experiments with the MONOfunctional imidoester methyl-acetimidate, in the pH range 7.0 - 8.0, on rat liver nucleosomes generate a cross-linking pattern almost identical with the one observed for much longer BIfunctional reagents (e.g. dimethylsuberimidate). Combined cross-linking and trypsin digestion experiments suggest that all or at least the great majority of this cross-linking occurs on trypsin digestible segments (or "tails") of the histones. The formation of oligomers over such extremely short distances and especially the observation of an H3 homodimer suggests a very close proximity of half-nucleosomes.


An efficient in vitro total reconstitution of the Escherichia coli 50S ribosomal subunit. A new, relatively simple technique for the total invitro reconstitution of E. coli 50S ribosomes has been developed. It is a two-step procedure like that previously reported by Nierhaus and Dohme [Proc. Natl. Acad. Sci. 71, 4713 (1974)], but it differs in a number of important aspects. Ribosomal RNA is prepared by direct phenol extraction of 70S particles to minimize nuclease fragmentation. A mixture of 50S proteins is prepared by acetic acid extraction and immediate removal of the acetic acid by thin film dialysis. The resulting protein mixture is soluble and stable. Separate RNA and protein fractions are mixed, incubated first at 44 degrees C in 7.5 mM Mg(2+), and then at 50 degrees C in 20 mM Mg(2+). The resulting 50S particles comigrate with native 50S particles in analytical gradients. They range from 50 to 100% active in five different functional assays. This is a fairly stringent test of the effectiveness of reconstitution since 50S particles derived from highly active vacant couples were used as a control.Images


The isolation of duplex DNA fragments containing (dG.dC) clusters by chromatography on poly(rC)-Sephadex. Duplex DNA containing oligo(dG.dC)-rich clusters can be isolated by specific binding to poly(rC)-Sephadex. This binding, probably mediated by the formation of an oligo(dG.dC)rC+ triple helix, is optimal at pH 5 in 50% formamide, 2 M LiCl; the bound DNA is recovered by elution at pH 7.5. Using this method we find that the viral DNAs PM2, lambda and SV40 contain at least 1, 1 and 2 sites for binding to poly(rC)-Sephadex, respectively. These binding sites have been mapped in the case of SV40; the binding sites can in turn be used for physical mapping studies of DNAs containing (dG.dC) clusters. Inspection of the sequence of the bound fragments of SV40 DNA shows that a (dG.dC)6-7 tract is required for the binding of duplex DNA to poly(rC)-Sephadex. Although about 60% of rabbit DNA cleaved with restriction endonuclease KpnI binds to poly(rC)-Sephadex, no binding is observed for the 5.1 kb DNA fragment generated by KpnI digestion, which contains the rabbit beta-globin gene. This indicates that oligo(dG.dC) clusters are not found close to the rabbit beta-globin gene.


Histone H1--DNA interaction. On the mechanism of DNA strands crosslinking by histone H1. Crosslinking of DNA fibers by histone H1 or phosphorylated on Ser-37 histone H1, and by the individual fragments of the H1 polypeptide chain was studied by the method of turbidimetry. The dependence of the turbidity of DNA-protein complexes on the ionic strength in solution suggests that the condensation of H1.DNA complexes in vitro is apparently due to both specific histone-DNA interactions with the contribution of hydrogen and/or hydrophobic bonds and the formation of polycationic "bridges" fastening the DNA fibers. The effectiveness of the condensation is postulated to be a function of a proportion between the two mechanisms which in turn can be controlled by slight changes in ionic surroundings. The sharp dependence of shrinkage of H1.DNA complexes on ionic strength at "physiological" salt concentrations could provide a mechanism to regulate density and consequently the total activity of chromatin in the cell nuclei. The phosphorylation of histone H1 on Ser-37 by a specific histone kinase does not noticeably affect the pattern of DNA crosslinking by the H1.


In vitro transcription in E. coli crude lysates prepared on cellophane discs. An in vitro RNA-synthesizing system consisting of gently lysed E. coli cells on cellophane discs is described. The system has been optimalized with respect to total RNA synthesis. Under certain standard conditions DNA dependent RNA polymerase (EC 2.7.7.6) is responsible for the majority of the RNA synthesis. The extensive rifampicin sensitivity of the synthesis indicates that most of the transcripts are initiated in vitro. The RNA synthesizing system described here has been developed with the aim of studying phage transcription in vitro. We show here that lysates of a P4 infected P2 lysogen support initiation and propagation of transcription from the P2 prophage.


Hospital phobia: a rapid desensitization technique. The less disabling phobias do not normally present a problem in that the stimulus may be avoided. This would also apply to hospital phobia until an acute medical or surgical problem might arise, when avoidance could constitute a direct threat to life. Although phobic illness is a common problem the small number of cases of hospital phobia recorded may represent the tip of the iceberg beneath which could be many phobic patients who deny their symptoms and risk their health because of their irrational fear. A case of hopsital phobia in a pregnant patient with suspected disproportion was treated by a rapid desensitization technique using hypnosis. After five sessions of 30 min each, the patient was symptom free. This simple method of desensitization, if more widely known would considerably minimize the risk caused by concealment of the phobic problem.


[Effect of cultivation temperature and pH on the biosynthesis of extracellular phosphohydrolases by the culture Penicillium brevi-compactum]. The effect of cultivation temperature and pH on growth of the culture Penicillium brevi-compactum and biosynthesis of extracellular phosphohydrolases (acid and alkaline RNases and acid PMEase) involved in RNA degradation was studied. The maximum growth of the culture was seen at 30 degrees and pA-7.0 and its maximum physiological activity with respect to the above enzymes was noted at 24 degrees and pH = 3.0, i.e. optimal conditions for the biosynthesis of phosphohydrolases did not coincide with those for the culture growth. The cultivation temperature and pH produced an effect on the oxygen consumption and synthesis of extracellular phosphohydrolases by the culture.


Hydrolysis of poly (A) to adenine nucleotides by purified poly (A) polymerase. Highly purified poly(A) polymerase (polynucleotide adenylyltransferase, EC 2.7.7.19), which synthesizes poly(A) from ATP substrate, can also catalyze hydrolysis of poly(A). The enzyme, designated as poly(A) hydrolase, requires either Mn2+ or Mg2+ for activity. Although AMP is the predominant product of the reaction, ADP and ATP are also formed. The enzyme is a 3'-exonuclease that does not degrade poly(A) associated with poly(A) poly(U) helical structure. AMP, ADP, and ATP inhibit the hydrolytic reaction. These data suggest that (i) the levels of adenine nucleotides regulate synthesis and degradation of poly(A), (ii) poly(A) itself is a storage form of adenine nucleotides, (iii) the hydrolytic reaction is responsible for poly(A) shortening or turnover observed in vivo, and (iv) the synthetic and hydrolytic activities are functions of the same protein molecule.


[Examination of the action of effectors after storing glutamine synthetase from the fodder yeast Candida tropicalis at room temperature]. The pattern of action of effectors (AMP, IMP, CMP, anthranilic acid, tryptophane, alanine, glutamine, glycine, histidine) and glutamine synthetase (GS) extracted from the fodder yeast Candida tropicalis and stored at room temperature for 1.5-2 hrs was examined. As regards the action of effectors on GS after its exposure at room temperature, they can be subdivided into four groups: 1) the effector loses completely its inhibitory effect (glutamine, CMP, anthranilic acid in the synthetase reaction); 2) the inhibitory effect on the enzyme increases (AMP, IMP, anthranilic acid in the transferase reaction); 3) at low concentrations of the effector the peak of activation appears (tryptophane, GMP, alanine, glycine); 4) at low concentrations of the enzyme two peaks of activation appear (histidine). Similar results were obtained with the purified preparation of GS.


[Kinetics and stability of glucooxidase from Penicillium vitale]. The kinetics, thermal stability and ultrasonic resistance of glucosoxidase from Penicillium vitale have been investigated. Its oxidative constant is close to, catalytic constant is 1.7 times less and the reductive constant is 5.4 times more than the respective constants of the enzyme from Aspergillus niger. The relationship between the thermal inactivation constant (k(i)=2.1 pT 15.3. The same relationship is true of the ultrasonic inactivation constant and pH in the pH range of 6.5-8.0. In a more acidic medium the rate of ultrasonic inactivation shows an insignificant dependence on pH. The mechanism of enzyme inactivation is discussed.


Isolation of a low molecular weight Ca2+ carrier from calf heart inner mitochondrial membrane. A protein was isolated from calf heart inner mitochondrial membrane with the aid of an electron paramagnetic resonance assay based on the relative binding properties of Ca2+, Mn2+, and Mg2+ to the protein. The molecular weight of this protein has been estimated to be about 3000 by urea/sodium dodecyl sulfate gel electrophoresis and amino acid analysis. The protein is shown to have two classes of binding sites for Ca2+ by flow dialysis studies and can extract Ca2+ into an organic phase. The selectivity sequence of this protein determined from the organic solvent extraction experiments shows that it favors divalent cations over monovalent cations. Also, the relative selectivity sequence for divalent cations is Ca2+, Sr2+ greater than Mn2+ greater than Mg2+. Ruthenium red and La3+ are shown to inhibit the protein-mediated extraction of Ca2+ into the organic solvent. The calcium translocation in a Pressman cell by this protein is selectively driven by a hydrogen ion gradient. Control experiments indicate that the Ca2+ trnsport properties of the protein are not due to the contaminating phospholipids. It appears that we have isolated from the inner mitochondrial membrane a calcium carrier, which we have named "calciphorin."


31P nuclear magnetic resonance studies of bioenergetics and glycolysis in anaerobic Escherichia coli cells. 31P nuclear magnetic resonance spectra of glycolyzing, anaerobic Escherichia coli cells and their perchloric acid extracts were obtained at 145.7 MHz. Time-dependent intracellular concentrations of nucleoside di- and triphosphates, Pi, and sugar phosphates were measured during glycolysis with 2-min resolution, while intracellular and extra-cellular pH values were monitored simultaneously. Upon glucose addition, anaerobic E. coli cells rapidly produce acids and develop a transmembrane pH gradient (delta pH). Glycolysis rates were calculated from the changes in the external pH. It was found that glycolysis rates are strongly dependent on internal pH, sharply decreasing when the pH drops below approximately 7.2. The ATPase inhibitor, dicyclohexylcarbodiimide (DCCD), prevented NTP hydrolysis and inhibited delta pH formation. The uncoupler, carbonyl cyanide p-triflouromethoxyphenyl hydrazone (FCCP), drastically reduced both the delta pH and the NTP level. When the cells were previously treated with DCCD, FCCP collapsed the delta pH while the NTP levels remained high. It is concluded that ATP produced by glycolysis is hydrolyzed by the membrane ATPase to generate a delta pH and that FCCP stimulates ATP hydrolysis by ATPase and collapses the proton gradient.


Selective labeling of alpha-adrenergic receptors in caudate nucleus by [3H] dihydroergocryptine in the presence of spiperone-blocked dopamine receptors. Because it was known that [(3)H]dihydroergocryptine can label alpha-adrenergic receptors as well as dopamine receptors, this study was done to establish the conditions under which [(3)H]dihydroergocryptine would be a reliable ligand for selective labeling of alpha-adrenergic receptors. The calf caudate was chosen because it contains both dopamine and adrenergic receptors, and 5 nM spiperone (spiroperidol) was used to block the neuroleptic/dopamine receptors. Thus, in the presence of spiperone, [(3)H]dihydroergocryptine exhibited saturable binding with a K(d) of 0.73 nM and a total number of sites of 150 fmol/mg of protein. The catechol neurotransmitters competed for [(3)H]dihydroergocryptine binding in the potencies order epinephrine > (-)-norepinephrine > dopamine, indicating that [(3)H]dihydroergocryptine (in the presence of 5 nM spiperone) was revealing alpha receptors. The alpha-adrenergic antagonists also competed for binding in the appropriate order: phentolamine > phenoxybenzamine > dibenamine. Finally, chlorpromazine was more potent than haloperidol in competing for [(3)H]dihydroergocryptine, also in accord with the properties of alpha receptors. These results with [(3)H]dihydroergocryptine as an alpha-adrenergic receptor ligand correlate well with those published by others for [(3)H]WB-4101.


Multiple glycosidase deficiencies in a case of juvenile (type 3) Gaucher disease. Biochemical investigations were performed on autopsy tissues obtained from an 11-year-old girl who died with the juvenile, subacute neuropathic form of Gaucher disease. In addition to the expected deficiency of glucocerebrosidase activity, extracts of both liver and kidney from this individual displayed a profound (greater than or equal to 90%) deficiency of "soluble" beta-glucosidase, beta-xylosidase, and beta-galactosidase activities. Fibroblasts obtained from this individual also contained markedly reduced levels of beta-xylosidase activity but normal levels of beta-D-fucosidase and beta-galactosidase activity. Because the soluble beta-glucosidase, beta-xylosidase, and a portion of the beta-galactosidase activities from control human liver all cochromatographed on a gel filtration column of Sephadex G-200, it is suggested that these activities all reside in a single enzyme, analogous to the situation described in a number of nonhuman, mammalian tissues. This demonstration of multiple glycosidase deficiencies in addition to the deficiency of glucocerebrosidase in a case of subacute neuropathic Gaucher disease suggests that other biochemical aberrations, in addition to a deficiency of glucocerebrosidase, might contribute to pathology in some cases of Gaucher disease.


Reduction of membrane potential, an immediate effect of colicin K. Colicin K causes a rapid and drastic reduction of the membrane potential of Escherichia coli cells, as measured by the uptake of the lipophilic cation triphenylmethylphosphonium. The colicin causes no major changes in the pH gradient, as measured by the uptake of butyric acid. The decrease in membrane potential following addition of colicin K to the cells is a prompt response that parallels or precedes known physiological effects such as efflux of accumulated substrates. Hence the loss of membrane potential qualifies as the primary action by which the colicin uncouples membrane-associated function from respiration. Certain peculiarities of bacterial cells pretreated with EDTA in their response to uncoupling agents and lipophilic ions are described.


Neurochemical and operant behavioral studies of a strain of alcohol-preferring rats. The levels of serotonin (5-HT), 5-hydroxyindoleacetic acid, tyrosine (TYR), norepinephrine (NE), acetylcholine, GABA, glutamate (GLU), aspartate (ASP), alanine, glycine (GLY) and taurine were measured in the CNS of adult male rats selectively inbred to the F8 generation for alcohol preference (P) and non-preference (NP). With respect to the values found in the NP group, higher levels of 5-HT, GABA, GLU and GLY and lower levels of ASP were found in the diencephalon-mesencephalon (D-M) and higher levels of NE were found in the telencephalon (TEL) of the P group. The animals in the P and NP strains were further subdivided into two additional groups, one given only H2O (W) and the second given 10% ethanol (A) during a one week period, thereby producing four groups (NP-W, NP-A, P-W and P-A). With these conditions, the level of (a) TYR in the D-M was higher in the P-A and NP-A animals than in the P-W and NP-W groups respectively; (b) 5-HT in the TEL was higher in the NP-A group than NP-W group; and (c) GABA in the TEL was higher in the P-A than P-W animals. No differences were observed in the cerebellum between the two strains or between the subgroups within each strain. The present study also demonstrated that the P animals will work in an operant situation to obtain 10% ethanol, even when H2O is freely available, and will voluntarily bar-press up to 6-7 times for each ethanol reinforcement.


[Anxiety- an approach to defining terms and formulating a methodological concept for measuring anxiety states within the framework of clinical trials with anxiolytic substances (author's transl)]. A methodological concept for measuring anxiety states and changes in anxiety states in clinical studies with anxiolytic substances is presented: Effectivity should be evaluated on several levels which include emotions, objects of anxiety, psychophysiology, biochemistry, and observed behavior. This test concept is based on new knowledge from behavior psychology, learning theory, psychology of testing, psychophysiology, biochemistry, neuroendocrinology, and psychiatry. In diagrams, important mechanisms of thr origin of experimental, stress-induced and pathological anxiety states are depicted. The point is stressed that anxiety is situational and fluctuates.


Determination of mepyramine, aminophenazone, nialamide and chloroquine using the deltapj method. Mepyramine maleate, aminophenazone, nialamide and chloroquine phosphate are examples of compounds which do not fulfil the requirements of the delta A method. By the proper choice of polynomial and wavelengths, the deltapj method has been successfully applied to their determination. The mean percentage recoveries were found to be 100.2 +/- 0.8, 99.6 +/- 0.6, 99.1 +/- 1.7 and 100.4 +/- 1.3, respectively.


Autonomic perception and responses in anxiety-inducing situations. The anxiolytic and sedative properties of a thienodiazepine (clotiazepam) were investigated in 40 healthy subjects with varying degrees of arousal and with the aid of specific anxiety-inducing conditions. A cross-over design with a double blind technique was used. EEG, heart and respiratory rate, peripheral pulse volume and skin resistance were measured during the anxiety-inducing conditions with the visualization of neutral and fearful situations as well as with the threat of shock. The mood, physical sensations and behaviour tendencies of the test subjects were determined by self-rating questionnaires. The anxiolytic and sedative effects of the thienodiazepine under study were able to be differentiated as far as subjective perception and autonomic responses were concerned. The development of sedative and anxiolytic effects is dependent on the subject's state of arousal. Situations involving the anticipation of anxiety are sufficient as proof of pharmacogenic anxiolysis. The tranquilizer reduces anxiety and physical tension, but does not influence the timing of autonomic readiness responses, and thus does not affect the coping strategies.


[Treatment of pain with psychotropic drugs (author's transl)]. 30 patients suffering from therapy-resistant chronic pain were divided into three groups according to drive and mood: I. psychomotor inhibition combined with depressive mood, II. agitation and depressive mood, III. agitation combined normal mood. The psychopharmacological treatment of pain showed sufficient results, when the drugs were directed to the target symptoms "agitation", "lack of drive", and "depression". The differentiated administration of activating or sedative antidepressants and neuroleptics caused reduction or absence of pain while the routine administration of neuroleptics (i.e. chlorpromazine) was less effective.


Early diagnosis of crepitant gangrene caused by Bacteroides melaninogenicus. A rapid non-invasive test for the presence of B. melaninogenicus in the wounds of crepitant non-clostridial gangrene is described. The wounds are viewed under an ultraviolet light, and the presence of bright red fluorescene indicates the probable presence of B. melaninogenicus.


The effects of TAB and of adrenergic blocking drugs on the course of the experimental irradiation disease. The experiment is aimed at establishing the consequences of the endotoxic stress (TAB) associated with alpha- or beta-adrenergic selective blocking drugs on the course of the acute irradiation disease in rats (700R, gamma rad Co60). TAB administered before or after irradiation, alone or associated with propranolol, increases the surviving rate at 30 and 60 days from exposure as compared to the controls (LD 95/30 days). Generally, dibenzyline has unfavourable effects on mortality and survival rate at the mentioned intervals. Based on the data obtained, an adrenergic component is analysed in achieving the survival increase by TAB and beta-adrenergic blocking substances; a hypothesis about the mechanisms involved is advanced.


Some neural effects of adenosin.

Some neural effects of adenosin. The possible neural effects of adenosine were investigated by using electrophysiological techniques at the level of some central and peripheral synapses. The evoked potentials in the somatosensorial cerebral cortex are influenced according to both the type of administration and the level of the electrical stimulation. While the local application does not induce significant alterations, the intrathalamic injections and the perfusion of the IIIrd cerebral ventricle do change the distribution of activated units at the level of different cortical layers especially during the peripheral stimulation. The frequency of spontaneous miniature discharges intracellularly recorded in the neuromuscular junction (mepp) is significantly depressed by adenosine. This effect is calcium- and dose-dependent. The end plate potentials (EPP) were also depressed. The statistical binomial analysis of the phenomenon indicated that adenosine induces a decrease if the presynaptic pool of the available transmitter. The data obtained demonstrate a presynaptic inhibitory action of adenosine beside its known vascular and metaholic effects.


Use of antianxiety agents in anxious outpatients. The present paper reviews the role of antianxiety agents in the treatment of nonpsychotic anxious patients. Major emphasis is placed on findings from clinical trials conducted by this research group. Consideration is given to the relative efficacy of available antianxiety agents, to patients response in relation to patterns of symptomatology, and to the degree of symptom relief experienced by drug-treated anxious patients. Also discussed are some of the many nonspecific factors that affect improvement with drug treatment in general and with the benzodiazepines in particular. It is concluded that many anxious patients are not currently receiving adequate drug treatment, and may require longer periods of drug therapy or supplemental nondrug intervention.


Neuropharmacological profile of clobazam, a new 1',5'-benzodiazepine. The effect of the new 1,5-benzodiazepine clobazam on visual evoked potentials (VEP) and spontaneous EEG in the conscious rabbit and on spinal polysynaptic reflexes in the decerebrated cat was studied in comparison to the 1,4-benzodiazepine diazepam. Clobazam was half as potent as diazepam in depressing the amplitude of visual evoked potentials in the nonanaesthetized rabbit, whereas the depressing effect on spinal polysynaptic reflexes in the decerebrated cat was only 1/7 --1/30 of the diazepam effect. The action of clobazam and diazepam on VEP also showed differences in time course, i.e., the peak effect of clobazam lasted from 1 to 6 h after application, whereas the effect of diazepam appeared after 10 min and declined already after 1 h. Both compounds had similar effects on computer-analyzed spontaneous EEG in the rabbit (power spectrum analysis), with an increase of power in the beta-band (13-39Hz) and a decrease in the alpha (8-13 Hz) and theta (4-8Hz) bands.


Studies on cholesterol esterification in human tissues. In vitro cholesteryl synthesis from oleic acid [1-14C] was studied with enzyme preparations from human thoracic aorta and liver. Results from studies on the properties of the esterifying system provide good evidence that the mechanism involves fatty acyl-CoA-cholesterol acyl transferase. In studies on human thoracic aorta with varying degrees of atherosclerotic disease, and pairs of normal and diseased aorta from the same subject, there was no obvious relationship between aortic cholesteryl esterifying activity and severity of atheroma. Normal aorta from two young males, presumably free of atherosclerosis, had relatively very low esterifying activity. In the six liver samples tested, there was negligible esterifying activity, in contrast to the high activity seen in the case of rat liver. For the thin layer chromatographic isolation of the labeled cholesteryl oleate a solvent system of isooctane:diethyl ether (100:6) was found to give a better separation of the ester than the petroleum ether: diethylether:acetic acid system generally used.


Inhibition of acute anaphylaxis in the chicken by anti-allergic drugs. The anti-anaphylactic effects of four potential anti-allergic agents (DSCG: disodium cromoglycate, PRD-92-EA, M&B 22,948 and DECC: diethylcarbamazine citrate) were studied on cardiovascular responses of acute systemic anaphylaxis and chemical mediators of immediate hypersensitivity in chickens. Compounds M&B 22,948, PRD-92-EA, DSCG and DECC all inhibited cardiovascular manifestations of systemic anaphylaxis in domestic fowl. High doses of Compounds PRD-92 EA, M&B 22,948 and DECC also exhibited a nonspecific receptor blocking activity towards exogenously administered chemical mediators of anaphylaxis.


Comparison of the mechanisms controlling intracellular pH and sodium in snail neurones. Ion-sensitive microelectrodes were used to record intracellular pH, Na+ and Cl- in snail neurones. NaCl or HCl was injected iontophoretically to compare the Na pump with the pHi regulating system. The Na pump was inhibited by ouabain, carbonyl cyanide m-chlorophenyl hydrazone and increasing the membrane potential, whereas the pHi regulating system was relatively unaffected. Activation of the Na pump had no effect on pHi whereas activation of the pHi recovery process increased internal Na+. Activation of the pHi recovery process by CO2 application increased internal Na+ and also decreased internal Cl-. The results show that there is no direct connexion between the Na pump and the pHi recovery process, and that the pHi recovery process is electroneutral, and appears not to require metabolic energy. The results also confirm that the pHi recovery process involves the influx of Na+ ions and the efflux of Cl- ions.


Intracellular acid-base state at a variable temperature in air-breathing vertebrates and its representation. When temperature changes are superimposed on changes in the control variables of acid-base state (PCO2, strong ion difference), the understanding of acid-base changes becomes difficult. A solution has recently been proposed for blood (Malan, 1977); it was based on the assumption that closed system conditions correspond to a minimal change in the overall acid-base state when temperature varies. The feasibility of extending these concepts to muscle intracellular acid-base vs temperature relationships is evaluated on the basis of a model study; the errors made by replacing closed conditions (which require knowledge of chemical composition) by more convenient approximations are estimated. A representation of both extracellular and intracellular acid-base data on a temperature-corrected bicarbonate-pH diagram is derived. It allows the interpretation of variable-temperature intracellular acid-base changes in terms of changes in control variables, 'respiratory (PCO2) or 'metabolic' (strong ion difference).


Regulation of blood pH in teleost fish.

Regulation of blood pH in teleost fish. The nature of pH changes with temperature is similar in both air-breathing and water-breathing poikilotherms having a pH-temperature slope of -0.015 to 0.020 U/degrees C. The means by which this is accomplished are quite different in the two groups. The air-breather behaves in vivo as a Rosenthal, or constant CO2 content system, with PaCO2 increasing at higher temperatures. The increase in PaCO2 is brought about by a reduction in the ration VE/MCO2. The water-breather, on the other hand, does not change VG/MO2, so as not to sacrifice oxygen delivery, and consequently does not control PaCO2 by ventilation and perhaps does not control it at all. Instead, the ratio of CT/SCO2 is controlled by altering blood bicarbonate concentration. Since alteraction of Na+ and Cl- uptake rates affects blood pH, and alterations of blood pH affect uptake of Na+ and Cl- at the gills, the carrier-mediated Na+/H+(NH4+) and Cl-/HCO3-(OH-) mechanisms now known to occur in gills appear to be the primary pH-regulating mechanism in water-breathers. Our knowledge of the role of gut and kidney in pH regulation is incomplete, but on the basis of preliminary information the kidney does not appear to respond to acid challenge.


Weak acids, weak bases and intracellular pH. Against the background of classical observations made 50 years ago, a brief review is offered of some of the work performed in the author's laboratory on the behavior of weak acids and bases towards living animal cells. The significance of membrane permeability of the charged partner of these electrolytes is pointed out, and the existence of an active process of H+ extrusion (or its equivalent) in response to acid loading is demonstrated. The effect of intracellular inhomogeneity on weak acid and base transmembrane distribution is examined. The significance of these variables for weak acid- or base-derived intracellular pH is discussed.


Evaluation of the DMO method for measuring intracellular pH. Intracellular pH in single muscle fibres from Balanus nubilus was determined, both by the weak acid distribution method (using 5,5-dimethyl-2,4-oxazolidine dione) and by means of a glass pH microelectrode, following prolonged equilibrations in normal, acid or alkaline Ringer solutions. In the DMO method [3H]inulin was combined with [14C]DMO so that an extracellular space could be calculated for each 'DMO space' on each fibre. Less than ideal uptake curves were obtained both for [14C]DMO and [3H]inulin, yet the calculated pHi compared favourably with the microelectrode pHi, especially when the DMO-pHi was calculated from [14C]DMO and [3H]inulin values obtained by extrapolating the 'plateaus' of the uptake curves to time zero. The Waddell and Butler equation was examined to determine why acceptable pHi values should be forthcoming from less than ideal uptake data.


Intracellular pH transients induced by Co2 or NH3. Experiments are reviewed in which intracellular pH (pHi), during exposure of a cell to CO2- or NH3-containing solutions, not only undergoes an acidification of alkalinization, respectively, but tends to return toward its original value and, upon removal of the test solution, rebounds to a value more alkaline or acid, respectively, than the initial one. A simple physiochemical model is discussed which interprets these observations both qualitatively and quantitatively. In the case of NH3-induced transients, only passive movements seem to take place; in the CO2-induced transients one must postulate an active 'proton pump'. Experimentally verifiable predictions can be made from these models. It is suggested that many physiological effects follow exposure to CO2 or NH3 may be subject to similar transients and rebounds.


Intracellular buffering.

Intracellular buffering. In the quantification of buffering the distinct roles of bicarbonate and of other buffers must be taken into account. The determination of the total non-bicarbonate buffer value, betaa, in intact tissues is complicated by active pH regulation and by heterogeneity of cytoplasm with respect to betaa, while heterogeneity with respect to pH in vivo could cause errors in estimates made with homogenates. Available estimates of betaa are discussed, as are the individual contributions of proteins, dipeptides and phosphates. A high betaa is appropriate in cells which sometimes have high rates of glycolysis, or which buffer extracellular fluid, but non-protein buffer concentrations can be well below the limits imposed by osmolarity, perhaps because buffering can upset ionic gradients.


Active control of intracellular pH.

Active control of intracellular pH. When the interior of a squid giant axon is loaded with acid, the cell responds by extruding that acid in an energy-requiring process. Our results indicate that this acid extrusion is accomplished by exchanging external HCO3- for internal Cl- at the expense of ATP.


Measurement of intracellular pH and aspects of its control in higher plant cells cultivated in liquid medium. The 5,5-dimethyloxazolidine-2,4-dione (DMO) method for the determination of intracellular pH has been adapted to Acer pseudoplatanus cells cultivated in liquid medium. DMO is metabolized by these cells and this parameter has to be taken into account in the measurement of the intracellular concentration of free DMO. As the cells contain a large acidic vacuolar compartment, the intracellular pH estimated from the DMO technique is an overall pHi', the dependence of which on various cytoplasmic and vacuolar characteristics has been discussed. Important modifications of these overall pHi values under the influence of CO2 or NH4+ have been measured. These pHi modifications strongly affect the regulation of cell division through the control of the intracellular concentration of hormones of the auxin type.


Independent and dependent variables of acid-base control. Basic physical principles and concepts plus computer-implemented numberical techniques now make possible a thorough quantitative analysis of acid-base systems. Some important conclusions from that analysis are presented: 1. Acid-base balance for physiological solutions hould be defined as the value of [OH-]/[H+]. 2. pH is a dangerously misleading indirect representation of [H+]. 3. Strong electrolytes affect [H+] and other deendent acid-base variables primarily through their resultant, the strong ion difference. 4 Hydrogen ion concentration in biological solutions is determined by the strong ion difference, the carbon dioxide partial pressure, and the total weak acid present. Changes in hydrogen ion concentration can be broght about only by changing one or more of these three independent variables. The same statements apply to all the other dependent variables, notably bicarbonate ion concentration. None of the dependent variables determines any other dependent variable, although their quantitative behaviors are necessarily correlected. 5. Solutions separated by membranes can interact in acid-base terms only by processes which alter the values of their independent variables. Interaction of intra- and extracellular acid-base balance can only occur by the cell membrane altering these independent variables in the extracellular fluid and in the cytosol.


[Pathological and compensatory processes in the cerebral circulation during blood transfusion shock]. In experiments on dogs the intravenous injection of heterogenous blood resulted in a decrease of total arterial pressure, weakening of the brain blood flow, fall of Po2 and pH in the brain cortex. A simultaneous constriction if inner carotid arteries is depending on direct action on the vascular wall of heterogenous proteins and on a release in it of physiologically active substances, such as serotonin. Fine pial arteries were dilated by the compensatory mechanism that was not associated with a decrease of intravascular and with direct action of hypoxia or acid metabolites on vascular walls. It was proposed that the trigger mechanism of this vasodilatation is hypoxic changes of metabolism in the nervous tissue.


[Gastric secretion and gastrin values following portocaval and splenocaval shunts in the rat]. The basal and gastrin-stimulated gastric acid output was measured in rats with a distal splenocaval shunt. In addition, serum gastrin levels were determined in rats with splenocaval and portacaval shunts. The rise in gastric acid output with increasing gastrin dosage was the same in rats with splenocaval shunt as in the controls. Serum gastrin levels were not influenced by portocaval or splenocaval shunting.


[Preventive practice in dentistry and its current meaning]. Improvements in the field of sealants and of adhesive restorations based thereon represent the most important new development in preventive dentistry. Caries prevention by means of better nutritional habits is greatly enhanced by newly developed and marketed sugar-free and tooth sparing products. Statistical mortality studies have again demonstrated that prevention by means of fluorides constitutes no danger to public health. Considerable advances have been made in the field of fluoridation of domestic salt. Newer Scandinavian studies revealed that intensive care programs with children including dental hygiene sessions every 3-5 weeks resulted in moderate decreases of caries incidence. By contrast, intensive care programs with adults were extremely successful. By means of 4-6 recall sessions with dental hygienists it was possible to completely stop periodontal destruction for 4, even for 7 years. Apart from initial restorative and conservative dental work, the success of periodontal treatment may be guaranteed only by prevention of recurring gingivitis. Extensive dental statistics from Switzerland reveal a marked success of applied preventive dental medicine. The work of dental practitioners is by now greatly influenced by the new scientific insight gained from research in the prevention of dental disease.


Lymphocyte ecto-5'-nucleotidase deficiency in agammaglobulinemia. Fresh peripheral blood lymphocytes from eight patients with congenital agammaglobulinemia demonstrate reduced ecto-5'-nucleotidase activity when compared to the mean activity of normal subjects and patients with other forms of immunoglobulin deficiency. A specific defect of ecto-5'-nucleotidase is further suggested by normal values for lymphocyte ecto-adenosinetriphosphatase and ecto-nonspecific phosphatase. The data provide evidence for an enzyme deficiency in this X-linked, B lymphocyte deficiency syndrome.


The abnormalities of beta-galactosidase in GM1-gangliosidoses. The activity of GM1 beta-galactosidase in the brain and liver of patients with GM1-gangliosidosis was assayed using GM1-ganglioside tritiated in the terminal galactose. In the cases of GM1-gangliosidosis Types 1 and 2A the activity was less than 0.5% of the control. In the liver of GM1-gangliosidosis Type 2B the activity was observed to be much higher than that of Types 1 and 2A. On Sephadex G-150 gel filtration, three active fractions (I, II and III) for 4-methylumbelliferyl beta-galactopyranoside (4MU) and two active fractions (I and II) for GM1-ganglioside were obtained in the control liver. There was no active fraction for GM1-ganglioside in spite of the preserved fraction I for 4MU in the liver of GM1-gangliosidosis Type 1 or Type 2A. In any of the three cases fraction II for both 4MU and G71-ganglioside was not detected.


Effects of phenobarbital in cerebral ischemia. Part I: cerebral energy metabolism during pronounced incomplete ischemia. Changes in cerebral cortex concentrations of high-energy phosphates, glycolytic metabolites, citric acid cycle intermediates, associated amino acids, and ammonia, were studied after 5, 15 and 30 min of incomplete ischemia in rats anesthetized with 70% N2O or 150 mg.kg-1 of phenobartibal. Previous results have shown that with this type of ischemia (bilateral carotid artery occlusion combined with reduction in blood pressure to 50 mm Hg) cortical blood flow is reduced to below 10% of nitrous oxide values, whether animals are anesthetized with 70% N2O or 150 mg.kg-1 of phenobarbital. In animals under 70% N2O, changes in tissue concentrations of phosphocreatine, ATP, ADP and AMP were similar to those previously obtained in complete ischemia. However, some glucose remained in the tissue, and the lactate concentrations gradually rose to reach excessive values. Changes occuring in glycolytic and citric acid cycle intermediates were similar to those seen in complete ischemia but, after 30 min, there was some reduction in the pool size of amino acids. In those animals given phenobarbital and which lost all EEG activity during ischemia, changes in cerebral metabolites were virtually identical to those observed in nitrous oxide-anesthetized animals. However, some animals exposed to 5 or 15 min of ischemia had some remaining EEG activity. In these, cerebral energy state was significantly less deranged, and levels of glycogen, glucose and pyruvate were higher.


Inhibition of 3'-methyl-4-dimethylaminoazobenezene-induced hepatocarcinogenesis by portacaval shunt. Adult male Sprague Dalwey rats on which end-to-side portacaval shunt (PCS) operation was performed did not hyperplastic nodules and hepatoms when they were fed 3'-methyl-4-dimethylaminoazobenzene in semisynthetic basal diet for periods of up to 169 days. In contrast, all the intact rats fed the same diet for only 75 days, developed hyperplastic nodules in the liver. Transferred to normal pellet for another 25 days, hepatomas developed in 100% of these animals. The amount of protein-bond 3'-Me-DAB was found to be much smaller in operated rats than in intact animals. The glutathione (GSH) level in PCS-operated rats was lower than in intact controsl. A single large dose of 3'-Me-DAB led to the increase of only about 30% in the concentration of GSH during the period of 24-48 h, compared to the increase of 50-100% in non-operated rats. No clear tendency to a gradual increase in the activity of gamma-glutamyl transpeptidase was noted in PCS-operated rats during the period of 5 1/2 months of 3'-Me-DAB feeding. The increase in GT-ase activity never exceeded 30% above the level of GT-ase in the livers of PCS-operated rats fed basal diet without the carcinogen. This striking inhibibiton of GT-ase increase induced by 3'Me-DAB in PCS-operated rats contrasted with an increase of GI-ase activity by 5,000% found in livers of non-operated rats with hyperplastic nodules after 75 days of 3'-Me-DAB feeding and the increase by up to 10,000% in developed hepatomas. These effects and the inhibition of 3'Me-DAB-induced hepatocarcinogenesis, manifested by lack preneoplastic morphologic changes and the absecnce of hepatomas in rats after PCS, can best be explained by functional deficiency of the liver to metabolize the procarcinogen 3'-Me-DAB into an activated carcinogen.


[Neurohumoral correlations and the effect of hydrocortisone on respiratory function in experimental pancreatitis]. From the observations on 18 patients and the experiments on 77 dogs with experimental pancreatitis neuro-humoral interrelations and hydrocortisone effect upon the respiratory function have been studied. Both in the clinic and in the experiment the authors have found the intermediate metabolism disturbed: adrenaline-like substances blood level rises, while the cholinergic complex blood level drops dramatically. The conclusion on a vital importance of a consecutive neuro-humoral correction with the use of cholinomimetics and corticosteroids in pancreatitis is drawn.


[Comparative assessment of the methods for surgical treatment of cryptorchism]. The experience with the surgical treatment of 305 cases of cryptorchism is presented. 108 Keetly and Torek's operations and 22 Mixter's operations have proved that in surgical technic the main emphasis should be transferred to the mobilization of the spermatic cord elements, but the orchoifixation should be simple and atraumatic. The Fowler's method of the descent of the testicles (107) in superficial inguinal ectopy and various modifications of the Petriwalsky's method of the descent of the testicles (43) in high inguinal retention and abdominal cryptorchism have made a favorable impression.


[The localisation of endocrine cells in the distal esophagus (author's transl)]. At the esophago-cardiac junction squamous epithelium is intermingled with gastric epithelium. Within the lamina propria of the distal 3 cm of the esophagus we found cardiac glands with many argyrophil cells in 60% of cases. Electron microscopy revealed seven well knonwn types and two further forms which we could not identify: EC, ECL, D, D1, AL, I eg. M, S. It is supposed that these cells are modified neurotransmitters with a local activity. Their function is local regulation of the lower esophageal sphincter (LES).


Maintenance of 2,3-DPG and ATP levels in blood stored for 30 days at a constant pH obtained by variation of CO2. A novel procedure for storing blood at a controlled pH consists in collecting blood in a pH 8.20, Tris-CPD solution and storing it in a special recipient including a gas permeable membrane under a CO2 atmosphere. The recipient is placed in an atmosphere of variable CO2 content, so that the initial alkalinity of the preservative is balanced by dissolved CO2, the proportion of which is diminished when lactate production increases with storage. 2,3-DPG and ATP were studied at three different pH levels of approximately 7.25, 7.45, and 7.65 at 4 degrees C. The best pH for the simultaneous maintenance of 2,3-DPG and ATP was 7.65. Under these conditions, 2,3-DPG is maintained at its initial level and ATP at 55% of its initial level at the 30th day. Lactate production is linear and hemolysis moderate.


Production of valine by a Bacillus sp.

Production of valine by a Bacillus sp. A bacterium isolated from Burdwan (India) soil was found to accumulate L-valine in the growth medium and identified to be a strain of Bacillus subtilis. The strain is able to grow and accumulate valine in a purely synthetic medium, but supplementation of the synthetic medium with either Casamino acids or yeast extract or with both, significantly improves the yield. The entire fermentation period can be divided into a growth phase and a production phase, which can be prolonged by adjustment of pH to the neutral range. Among the different carbon and nitrogen sources tested glucose at 8.5% and L-glutamic acid at 0.8%, respectively, were found most suitable. Cane sugar molasses tested as a substitute for glucose significantly stimulated growth but valine production was less. Different vitamins tested stimulated growth and valine yield and an inoculum level of 10% (v/v) of the medium was found to be optimal. The yield of valine under optimal conditions was found to be 4.53 g per litre of the medium. Valine has been isolated in crystalline form from the fermented broth by ion exchange resin chromatography and found to be a pure sample of the L-isomer.


[Circulating immune complexes in acute viral hepatitis, chronic active hepatitis, and periarteritis nodosa (author's transl)]. Circulating immune complexes were determined in patients with acute viral hepatitis, chronic active hepatitis and periarteriitis nodosa with the Raji cell technique. Circulating immune complexes were found in 11/18 HBsAg positive and HBsAg negative cases of acute viral hepatitis. In HBsAg positive chronic active hepatitis immune complexes were detectable in 42/43 cases but only in 1/27 HBsAg negative cases. Ten healthy HBsAg carriers demonstrated no detectable immune complexes. Using FITC conjugated antisera against HBs, HBc, and e antigen, immune complexes could not be found in any case of acute viral hepatitis or chronic active hepatitis. Elution of immune complexes from Raji demonstrated IgG, C'3 and a lack of HBsAg, HBc or e antigen. Immune complexes were present in 4/8 cases with periarteriitis nodosa. Viral components were detectable in one case.


[Psychophysiologic characteristics of the individual psychotropic activity of benzodiazepine derivatives under experimental conditions]. In 156 experiments performed on 41 cats with adequate experimental models of pseudoneurotic conditions, states of anxiety due to an electrostimulation of the hypothalamus through implanted electrodes and an emotional stress due to a conflict situation the authors compared the individual traits of the psychotropic activity of lorazepam, diazepam, chlordiazepoxide, oxazepam, nirazepam. These experiments permitted to display significant differences of psychophysiological structures in the tranquilizing effect of 5 benzodiazepine derivatives.


A direct lead technique for histochemical demonstration of leucocyte alkaline phosphatase activity in blood smears. A histochemical technique for demonstrating leucocyte alkaline phosphatase activity (LAP), based on direct precipitation of lead phosphate at pH 9.5, is described. The effect of fixative, temperature and stability of the medium on the activity of the enzyme and stability of the colour reaction was thoroughly studied. Peripheral blood smears obtained from both normal humans and pathological cases were studied and the results were compared with these obtained by the azo-dye method.


Effect of beta-sympatholytic agents on vascular responses to noradrenaline and potassium chloride. Vasoconstrictory responses to noradrenaline (NA) or high potassium chloride (130 mM) usually show a biphasic behaviour: an initial peak is followed by a lower steady state level. The influence of three beta-sympatholytic agents (propranolol, pindolol, practolol) and a "Ca-antagonist" (verapamil) on this particular behaviour of KCl-induced vasoconstrictions was compared with responses to NA. Studies were performed on the intact vascular bed of an isolated intestinal preparation of the rat. 1. Increases in the concentration of propranolol and pindolol from 10(-10) to 10(-6) M attenuated the vascular responses to high KCl. Practolol, however, distinctly enhanced the KCl-responses. NA (1.3 microgram/ml)-elicited vasoconstriction was found to be influenced in a similar manner by the beta-receptor antagonists used, though lower concentrations of pindolol had an enhancing, higher concentrations a reducing effect on the constrictory responses to NA. 2. The extent to which the biphasic response adjusted to the steady state level was concentration-dependently increased KCl-responses and significantly decreased to a monophasic response to NA, in the presence of the beta-sympatholytic agents. 3. By using a quotient of deltaPs (steady-state phase) to deltaPi (initial vasoconstriction) and by comparing the effect of the three beta-sympatholytic agents with that of verapamil on this quotient showed a concentration-dependent decrease in presence of the beta-sympatholytic as well as of the Ca-antagonistic agents. These myotropic actions, i.e. negative influences on the mechanism of vascular smooth muscle activation ranged in the following sequence: Verapamil greater than propranolol greater than pindolol greater than practolol. 4. The mechanism underlying the biphasic responses to high potassium chloride are concluded to be basically different from the NA-responses because they are inversely affected by beta-sympatholytic agents. The steady-state response to KCl was found to be very sensitive to non-specific pharmacological actions of beta-sympatholytic substances. The myotropic action of the beta-receptor antagonists is compared with that of the Ca-antagonist verapamil and the role of calcium in the mechanism of activation of vascular smooth muscle is discussed.


Effects of alpha-adrenergic stimulation and blockade of plasma parathyroid hormone concentrations in cows. Experiments were designed to investigate responses of immunoreactive parathyroid hormone (PTH) during alpha-adrenergic stimulation and blockade in cows. Alpha-adrenergic agonists (methoxamine, phenylephrine and noradrenaline, the beta-adrenergic action of which was blocked by propranolol) did not change PTH and free fatty acid levels, whereas they characteristically increased the blood pressure and decreased the heart rate. In contrast, alpha-adrenergic blockade by phentolamine progressively increased PTH levels. The elevated PTH concentrations, associated with increased plasma noradrenaline and free fatty acid levels, rising heart rate and decreasing blood pressure, indicated that all these changes can be related to a beta-adrenergic stimulatory mechanism. Beta-adrenergic stimulation was presumably responsible for the initial elevation of PTH concentrations, whereas, during the later phase of the phentolamine infusions, a concomitant hypocalcaemia probably also produced a stimulatory effect.


Testicular blood flow and testosterone concentrations in the spermatic venous blood in rats with experimental cryptorchidism. Testicular blood flow and testosterone concentrations in the spermatic venous plasma were measured on unilaterally cryptorchid rats. Blood flow to the cryptorchid testis was 31.4 +/- 11.7 (SD) ml/100 g X min which was significantly higher thant that of the scrotal testis (17.7 +/- 4.4 ml/100 g X min). Stereological analysis showed a relative increase of blood vessel containing interstitial tissue in the cryptorchid testis, which was probably the main factor responsible for the relative increase of blood flow to the cryptorchid testis. The increase of interstitial tissue was greater than the increase of blood vessels and thus, the interstitium in the cryptorchid testis contained a number of vessels which was smaller than that of the interstitium in the scrotally located testis. The concentration of testosterone in the spermatic vein of the abdominal testis was 18.0 +/- 5.5 (SD) ng/ml and the corresponding value for the scrotal testis was 41.2 +/- 7.0 ng/ml. Calculations based on functional and morphological data indicate that the function of the Leydig cells in the abdominal testis was impaired. It was concluded that the outflow of testosterone from the cryptorchid testis was highly reduced.


Erythrocyte acetylcholinesterase activity changes in the early stages of anemic and hypobaric hypoxia or after CoCl2 treatment in rabbits. The authors studied the changes of erythrocyte acetylcholinesterase (AChE) activity in rabbits respectively subjected to bleeding, hypobaric hypoxia and CoCl2 treatment; in such conditions a rise of the enzyme activity was observed. Similar increases take place also as a consequence of intravenous infusions carried out by using lactic or hydrochloric acid. The authors suggest that these increases in the erythrocyte AChE activity are connected with the decrease of the pCO2 that occurs in the various experimental conditions achieved.


The adrenergic mechanism of the haemodynamic effects of massive doses of hydrocortisone in controlled haemorrhagic shock in the dog. Haemorrhagic shock was induced in anaesthetized dogs by bleeding them into a blood reservoir system. By adjusting the blood level of the reservoir at a certain distance over the heart level the mean arterial blood pressure was kept at 6.7 kPa (50 mmHg). As has been found earlier, massive doses of hydrocortisone (80-560 mg.kg-1 body weight) caused a dose-dependent decrease in the total peripheral resistance. The degree of vasodilation distinctly increased during concomitant alpha-receptor blockade induced by phenoxybenzamine. The beta-receptor blocking drug propranolol efficiently inhibited the vasodilation caused by hydrocortisone and phenoxybenzamine. The findings fit with the hypothesis that massive doses of hydrocortisone induce an increased stimulation of the adrenergic beta2-receptors of the vascular smooth muscles.


Neuromuscular blockade of fazadinium bromide (AH 8165) in renal failure patients. Fazadinium bromide (AH 8165) is a new non-depolarizing neuromuscular blocking agent with rapid onset of action and intermediate duration of effect. The characteristics of the neuromuscular blockade induced were compared in normal and renal failure patients by monitoring the mechanical responses of thumb adductor muscle to ulnar nerve stimulation. The data suggest that renal failure, even in anephric patients, does not influence the depth of neuromuscular relaxation when judged from the identical dose-effect curves in both groups of patients. Moreover, the duration of the neuromuscular block was not significantly increased in end-stage kidney failure patients, whether the initial or late phases of the spontaneous recovery of neuromuscular transmission were considered, and this occurred over the entire therapeutic range. However, additional incremental injections of fazadinium bromide should be used cautiously, as evidence appeared of prolonged duration of action. This agent, fazadinium bromide, may therefore present useful advantages to other non-depolarizing neuromuscular blocking agents as to clinical safety and effects in patients with renal insufficiency.


Cerebral metabolic rate of oxygen (CMRO2) in the acute phase of brain injury. In 22 comatose patients with acute brain injury, the cerebral metabolic rate of oxygen (CMRO2) was calculated as the product of the hemispheric cerebral blood flow (CBF) and the arteriovenous oxygen content difference. All patients were subjected to moderate sedation without barbiturates, normothermia and respiratory treatment. The CBF was calculated by the 133xenon washout method as the average of 16 regions. The results indicate high jugular venous oxygen tension and, in some studies, very low oxygen consumption. A critical, low CMRO2 was not found, and values of about 0.4 ml/100 g/min were compatible with restitution of intellectual function. The CMRO2 was unrelated to the clinical outcome and to the time after the trauma. In bilateral studies, the lowest values were measured in the most severely injured hemisphere.


Cerebral autoregulation in unconscious patients with brain injury. In 18 unconscious patients with traumatic brain injury, the cerebral autoregulation was tested during the first 2-3 weeks after the acute trauma. Regional cerebral blood flow (rCBF) was measured by the intra-arterial 133xenon washout method before and after an increase of about 20% in the mean arterial blood pressure (MABP) by angiotensin. The difference between MABP and intraventricular pressure (IVP) was used as cerebral perfusion pressure (PP). Simultaneously, ventricular fluid pH, lactate and pyruvate were measured. Regional loss of autoregulation indicated by a 20% flow increase was observed in 29 out of 35 studies (83%), while hemispheric loss of autoregulation was observed in only one study. The results of the autoregulation tests were unrelated to the clinical outcome, the presence of brain-stem lesion, and the ventricular fluid pH, lactate and lactate/pyruvate ratio. In repeated studies, a gradual normalization of the autoregulation was observed about 5 days after the acute trauma.


Haemodynamic effects of different corticosteroids in controlled haemorrhagic shock in the dog. The haemodynamic effects of massive doses of hydrocortisone (80-320 mg.kg-1), methylprednisolone (4-32 mg.kg-1), betamethasone (1.6-12.8 mg.kg-1) and aldosterone (0.1-0.8 mg.kg-1) and the interaction with phenoxybenzamine and propranolol have been studied during controlled haemorrhagic shock in the anaesthetized dog. Hydrocortisone was the only steroid which showed any significant vasodilating ability when given alone. The alpha-receptor blocking agent phenoxybenzamine distinctly decreased the total peripheral resistance. The effect of the phenoxybenzamine was increased in combination with hydrocortisone or methylprednisolone, especially if the steroid was given as the first drug. The vasodilation found was efficiently abolished by the beta-receptor blocking agent propranolol. The ability of hydrocortisone or methylprednisolone to potentiate phenoxybenzamine was not shared by betamethasone or aldosterone. Thus, the haemodynamic effect of the steroid does not seem to be correlated to either a glucocorticoid nor a mineralocorticoid effect. It is suggested that the steroid effect studied is related to the ability of hydrocortisone or methylprednisolone to block the extra neuronal amine uptake which decreases the rate of elimination of the sympathetic transmitter from the vicinity of the adrenergic receptor of the vascular smooth muscle.


Effects of isoprenaline and dibutyryl-C-AMP on the electrical and mechanical activity of the rabbit myometrium. Mechanical and electrical responses of spontaneously contracting muscle strips from oestrogen dominated rabbit uterus were investigated by the sucrose gap method. The effects of isoprenaline and dibutyryl-c-AMP in the presence and absence of propranolol were tested. It has been shown previously that the beta-adrenoceptor agonist isoprenaline inhibits the spontaneously contracting uterus even in the presence of the beta-blocker propranolol. The rise in c-amp which is caused by isoprenaline, is, however, blocked by propranolol. The aim of this investigation was to study further the role of c-AMP in the isoprenaline-induced relaxation of the uterus. Isoprenaline (10(-6) M) consistently abolished spikes and mechanical contractions, usually accompanied by hyperpolarization. Propranolol (3 X 10(-6) M) markedly reduced the incidence of hyperpolarization, but did not affect the other actions of isoprenaline. Hyperpolarization does not seem to be a prerequisite for inhibition of uterine contraction. Dibutyryl-c-AMP mimicked the actions of isoprenaline. No positive evidence to support a hypothesis of a c-AMP-independent mechanism was found. Therefore a possible explanation of the results could be that isoprenaline increases c-AMP in a small intracellular pool, mediating electrical and mechanical responses but not influenced by propranolol.


The release of serotonin from rat duodenal enterochromaffin cells by adrenoceptor agonists studied in vitro. The serotonin (5-HT) content of enterochromaffin cells (EC) was studied by a cytofluorimetric method in biopsies from rat duodenal mucosa after in vitro incubation with different adrenoceptor agonists and antagonists and acetylcholine (ACh). Noradrenaline (NA), Adrenaline (A) and Isoprenaline (IP) caused a decrease of 5-HT in EC down to 40--60% and for NA and A this effect was concentration-dependent. The effect was antagonized by d,1-propranolol but not by d-propranolol, metoprolol, phentolamine or phenoxybenzamine indicating that the 5-HT release from EC is probably mediated via a true beta-adrenoceptor mechanism possibly of the beta2 type. ACh also decreased the 5-HT content of EC but was much less potent than the adrenergic substances. Dopamine (DA) had no effect.


Comparative evaluation of hypnotic efficacy of flunitrazepam in psychiatric in-patients. Flunitrazepam in 2-mg and 4-mg doses was compared with flurazepam 30 mg and nitrazepam 10 mg for hypnotic efficacy in a double-blind, multiple cross-over trial involving 41 psychiatric in-patients. In the vast majority of comparisons involving a number of sleep parameters differences did not reach significance level. Flunitrazepam 4 mg tended to produce the shortest latency time and the longest duration of sleep but also the most side-effects. Nitrazepam 10 mg appeared to have a slight advantage over the other drugs in terms of patient preference. For severe sleep disturbance flunitrazepam 2 mg tended to be the most satisfactory drug in terms of efficacy and paucity of side-effects.


Neuroendocrine disorders in depressions and their significance for the monoamine hypothesis of depression. This article discusses the results of recent neuroendocrinological research in depressions. The abnormalities found in a given category of vital depressive patients--cortisol hypersecretion, decreased growth hormone response to insulin hypoglycaemia and decreased luteinizing hormone secretion in menopause--are believed to be due to deficient noradrenalin-(NA)-ergic activity in the hypothalmus. Thus explained, they support the so-called MA (monoamine) hypothesis, which postulates that a functional NA deficiency in the brain plays a role in the pathogenesis of certain types of vital depression. Disorders in certain central MA-ergic systems are predictive of disorders in the hormone secretion of the anterior pituitary. Inversely, disorders in the hormone secretion of the anterior pituitary can be indicative of disorders in the MA-ergic transmission in the hypothalamus. Consequently we can expect a convergence of transmitter research and neuroendocrinological research--two lines of research which have so far been largely separated in studies of human individuals.


Primary culture of adult rat liver cells. III. Hormonal effects on cytological and biochemical properties of primary cultured cells. The effect of various hormones on cytological and biochemical properties of cultured hepatic cells were investigated in order to obtain long-term survival of the hepatocytes with adult liver functions in primary culture. Insulin supplementation of the culture medium enhanced the attachemnt efficiency of cells in primary culture without affecting either maintenance of morphological characters of epithelial cells or retention of liver-specific functions in cultured cells. A combination of dexamethasone and insulin was apparently effective in stimulating the formation of a monolayer of polygonal cells with granular cytoplasm and in maintenance of liver-specific functions for relatively longer periods. Supplementation with either dexamethasone or hydrocortisone alone enhanced tyrosine aminotransferase activities in cultured cells for at least 4 days postinoculation. These steroid hormones also allowed growth of small epithelial cells with clear cytoplasm and maintenance of increased albumin production for 8 days after inoculation. The roles of these hormones in the primary culture of isolated hepatic cells are discussed in the present paper.


[Actual knowledge on beta blocking agents in psychiatry (author's transl)]. A new group of substances blocking central beta receptors, ie beta blocking agents among which oxprenol and pindolol, have been used in psychiatry, mainly in anxious or depressive-anxious patients. The anxiolytic effect, particularly on somatic anxiety, is interesting, and is present in double blind studies with benzodiazepines, where it is superior to these classical agents. The author has observed this therapeutic activity in series of one hundred patients and also a dynamising effect in some of these patients. Without having observed by himself and possibly because of the low doses experimented, the author states that these drugs have an antipsychotic activity.


Resonance Raman studies on the ligand-iron interactions in hemoproteins and metallo-porphyrins. The resonance Raman spectra (RRS) were measured for various hemoproteins, as well as for free hemes, and analysed in terms of the interactions between the porphyrin ring and the heme iron and between the axial ligand and the heme iron. To establish vibrational assignments, the RRS of metallo-octae-thylporphyrin (M(OEP)), its 15N substituted and meso-deuterated derivatives were measured. It was found that the Raman line corresponding to the oxidation state marker of hemoproteins involves appreciable displacement of the pyrrolic nitrogen atoms toward metal ion but not that of methine bridges and that the frequencies of methine-bridge CC stretching vibrations are altered by the conjugation interaction between the porphyrin ring and the metal ion present. The prominent Raman lines of Ni(OEP) were assigned on the basis of the normal coordinate analysis. The Raman spectra of heme a bis-imidazole complex (Fe(A)(Im)2) in the presence of cyanide changed when pH was reduced to neutral. It implied formation of cyanhydrin at the peripheral formyl group of heme a. The Fe(A)(Im)2 formed an addition compound with NaHSO3 in a neutral solution but the added NaHSO3 was eliminated when the heme iron was reduced. The Raman spectra of hemoproteins were classified into four groups on the basis of the relative intensities and frequencies of the four key bands. It demonstrated the existence of two kinds of ferrous low-spin states. The difference between the two species was attributed to the difference in the nature of chemical bond between the heme iron and its sixth ligand (L). The Fe-L bonds of ferrous low-spin molecules in Groups A and C are mainly associated with the dz2(Fe)-lone pair and dz(Fe)-II(L) interactions, respectively. This interpretation of the Raman spectra appears consistent with the experimental facts that the ferrous low-spin molecules in Group C photodissociate upon illumination of light at the Soret band and also that the internal stretching frequencies of the sixth ligand are shifted to lower frequency upon coordination to the heme iron. The frequency of the oxidation state marker of reduced cytochrome P-450 (P-450cam) was unusually low in comparison with those of other hemoproteins. It was ascribed to delocalization of electrons from the thiolate anion to the porphyrin II*(ring) orbital through the pi-type molecular orbital (Eq.3). The RRS of reduced P-450cam.metyrapone adduct was quite close to that of reduced cytochrome b5. In conclusion, the Raman frequencies of hemoproteins may depend primarily upon the amount of electrons delocalized to the II* orbital of the porphyrin ring. The coordination of lone-pair electrons of the axial ligand to the iron dz2 orbital yields bond energy but scarcely affects the Raman frequencies. If the pi orbital of the axial ligand interacts with the iron dpi orbital, it perturbs the delocalization of the dpi(Fe) electrons to the II*(ring) orbital of porphyrin ring and thus affects the Raman frequencies.


[Flunitrazepam and memory (author's transl)]. This work is based on the observation of memory troubles before sleep in patients with sleep problems treated with flunitrazepam. Eight subjects are tested in two sessions, flunitrazepam and placebo, in double blind, with memory tests given the evening after medication and controlled the next morning. One additional test is controlling vigilance. The results show that memory is significantly disturbed by flunitrazepam, both for direct and delayed memory, the next day. On the contrary, recognition is not altered, neither vigilance.


Nuclear magnetic resonance studies of high-spin ferric hemoproteins. 220 MHz proton Fourier transform (FT) NMR with quadrature phase detection (QPD) technique is applied to observe largely hyperfine-shifted signals of various hemoproteins and hemoenzymes in ferric high-spin state. The binding of F-, OCN-, SCN-, and CH3OH to the ferric heme iron in high-spin state in various hemoproteins has been studied by the use of FT/QPD technique at 220 MHz. The binding of formate ion to metmyoglobin (metMb) has also been studied. The spectrum of the formate complex was compared with that of hemoglobin M Milwaukee where carboxylate groups are bound to the hemes of the beta subunits. The acid-base transition of ferric myoglobin (Mb) was confirmed by monitoring the pH-dependent shift of the heme side methyl signals with the reflection point at pH 9.1. This finding is analyzed on the basis of rapid exchange between alkaline (low spin) and acidic (high spin) forms accompanied by the dissociation and association of one proton in the ferric Mb. The structure of the heme environment of ferric horseradish peroxidase (HRP) was studied. The pH-dependent features of NMR spectra of the ferric enzyme and its complexes with cyanide and azide were discussed in terms of heme environmental structures, comparing with the case of metMb. The results were interpreted as follows: There exists an ionizable amino group near the heme responsible for the ligand binding reactions of the enzyme, which modulates the entry of external azide to the heme iron through protolytic equilibrium of this group. The pK value of this group was determined to be 5.9 by monitoring the pH-dependent shift of the heme peripheral methyl signals of the native enzyme, indicating that the group is probably a histidyl residue. Acid-alkaline transition of metMb was confirmed to associate with the proton dissociation of an iron-bound water molecule, whereas in HRP, pH-dependent spin state change characterized by pK 11 is attributed not to the simple protolytic reaction of the iron-bound water but to the direct coordination of an amino acid residue of the polypeptide chain to the ferric heme iron. Histidyl imidazole is a possible candidate for the new sixth iron ligand in alkaline peroxidase above pH 11. Interaction of HRP with electron donor(indolepropionic acid, IPA) was also studied. The hyperfine-shifted proton signals of the heme peripheral groups of the enzyme showed a small but significant shift with stepwise additions of IPA, indicating that the donor binds at a specific site of HRP. There results are interpreted in terms of the interaction between the enzyme and the donor at the heme edge site.


Analysis of acid-base properties of peroxidase and myoglobin. Two heme-linked groups of horseradish peroxidases, characterized by pKa = 5.8 (isoenzyme A) and 7.3 (isoenzyme C), are believed to be distal amino acid residues and, judging by their acid-base properties in various derivatives, influence the functions of these isoenzymes. In contrast, in myoglobin ionization of the distal histidine does not influence its reactivity. Accordingly, we conclude that the interaction of the distal base with a 6th ligand is weak in myoglobin but very strong in peroxidases.


Conformational change and cooperative ligand binding in hemoglobin. Although many studies have been carried out on the cooperativityly of ligand binding to hemoglobin, its molecular mechanism is not yet completely confirmed. Various models have been proposed on one hand, but on the other hand the direct judgement of the validity of each model is confronted with experimental difficulties. With this situation in mind, we re-examine carefully various kinds of experimental data, with the intention of finding any hints for future studies on the molecular mechanism of the cooperativity of ligand binding not only to hemoglobin but also to other allosteric proteins composed of subunits. As a result of these re-examinations most of the experimental data can be consistently explained from the viewpoint that the strain due to ligation is transmitted to the interfaces of the subunits and the degree of the strain stored in the interfaces between the subunits controls the cooperativity of of ligand binding. The subjects selected to the present study are molecular structural differences between the deoxy- and oxy-hemoglobin, temperature dependence of oxygen equilibrium constants, and recombination curves of carbon monoxide in flash photolysis experiments. In the first section, the difference of the interfaces of the subunits between deoxy- and oxy-hemoglobin is investigated on the basis of the atomic coordinates determined by Perutz et al. and we examine energetically which of the segment's pairs contributes mainly to the difference in the molecular structure between the two forms. On the basis of this investigation, it is shown in the next section that the oxygen equilibrium constants of human adult hemoglobin under normal physiological conditions are well explained by the following sequence of the conformational changes. The local changes in the interfaces of the subunits are reflected directly on the small differences among the first three Adair constants and the succeeding rearrangement to the four subunits causes the outstandingly larger value of the fourth Adair constant. In more alkaline pH, this rearrangement begins to occur at the earlier stages of ligation. In the succeeding section, it is pointed out that the property of this rearrangement can be studied in more detail through the recombination curves followed by flash photolysis under special conditions.


The effect of inhibitors of cellular RNA synthesis on stimulation of mouse encephalomyocarditis virus reproduction by poliovirus in HeLa and MIO cells. Inoculation of HeLa and MIO cells with a guanidine-dependent mutant of poliovirus (gd-polio) in the absence of guanidine resulted in stimulation of mouse encephalomyocarditis (EMC) virus reproduction. No stimulation was found to occur in the presence of actinomycin D, on pre-treatment of the cells with cordycepin, or on their UV irradiation or enucleation. In contrast to HeLa cells, the enhancement of EMC virus reproduction by gd-polio in MIO cells required no current synthesis of cellular RNAs: it occurred in the presence of actinomycin D, upon UV irradiation of the cells or their enucleation. The possible mechanisms of the stimulating effect of poliovirus in HeLa and MIO cells are discussed.


Marked difference in electrophoretic migration rates between two influenza A viruses. Comparative moving boundary electrophoresis revealed that influenza virus A/PR/8/34 (H0N1) has a 2.5 times higher electrophoretic migration rate at pH 7 than influenza virus A/Singpore/1/57 (H2N2). This difference was the same whether the compared viruses were purified first by either ammonium sulphate precipitation or adsorption onto and elution from red blood cells and then by density gradient centrifugation. The same electrophoretic methods was used for testing the homogeneity of influenza virus preparations purified by either method.


Multiplication of herpes simplex virus types 1 and 2 in macrophages of NMRI and C57/BL mice. The multiplication of 8 strains of herpes simplex virus of type 1 and 2 in normal and stimulated macrophages of NMRI and C57/bl mice was tested. Only 3 strains multiplied in normal NMRI macrophages, whereas all strains multiplied in stimulated cells. No multiplication except of one strain was observed in C57/bl macrophages. The multiplication rates of the virus strains correlated well with the determination of the acid-soluble and acid-insoluble thymidine (dTR) radioactivity and autoradiographic studies of infected (normal and stimulated) NMRI and C57/bl macrophages. The influence of the age of mice used for obtaining the macrophages was also studied. In stimulated macrophages more viral DNA was synthesised than in non-stimulated cells.


Haemagglutination inhibition antibodies in nasal secretions of persons after natural parainfluenza virus infection. In 7 adults and 7 children with upper respiratory illness of parainfluenza aetiology, the virus was isolated in the acute phase from nasal washings or nasal smears in spite of high titres of haemagglutination inhibition (HI) antibodies in serum. By contrast, secretory HI antibodies were not demonstrated at the onset of illness in any of the patients, but their formation started early and the antibodies reached maximal levels about 10 days after onset of illness. The individual patients showed considerable differences in the dynamics of secretory antibody formation and especially in their persistence. In some of the patients, secretory antibodies were demonstrated as late as 12 months after the illness.


Cell- and antibody-mediated responses to measles and mumps viruses in experimental animals. Monkeys and guinea pigs were immunized with various doses of measles and mumps viruses. A cell-mediated response measured by leukocyte migration inhibition occured during the first two weeks after immunization. In case of measles virus, marked migration inhibition occurred already on the 7th day in monkeys and guinea pigs immunized with high virus doses. The response declined as early as 14 days after immunization in contrast to the group of guinea pigs inoculated with a lower virus dose. In these animals the response sustained for up to 50 days. Immunization of animals with various doses of mumps virus was similar and positive responses were obtained in the absence of detectable antibodies. Even 2.5 TCID50 of mumps virus induced the response although more irregularly than 250 or 2500 TCID50 virus.


Morphological characteristics of the infection of animals with tick-borne encephalitis virus persisting for a long time in cell cultures. The pathogenicity of the Soph-K strain of tick-borne encephalitis (TBE) virus produced by the persistently infected HEp-2-Soph cell culture was investigated in monkeys, white mice and hamsters. The persistent infection of this culture with TBE virus has been maintained for 17 years. The Soph-K strain was shown to retain its pathogenic properties for mice of different ages and hamsters. Morphological examinations of the brains and spinal cords of these animals infected with the Soph-K strain revealed acute and subacute forms of encephalitis. Monkeys inoculated with the Soph-K strain intracerebrally showed an asymptomatic infection with the pathomorphological picture of subacute disseminated meningoencephalitis with a progredient course for 3 months of observation. This chronic TBE infection in monkeys offers an interesting model for investigations on progressive degenerative disease of the nervous system.


Sepcific immunofluorescent IgG, IgM, and IgA antibodies in lymphocytic choriomeningitis. In postnatally acquired lymphocytic choriomeningitis (LCM), specific immunofluorescent (IF) IgM antibody was found nearly in all the examined sera from 23 patients for 6-8 months after onset of the disease, IgA antibody in two-thirds of the patients and only during the first 5 weeks of the disease. In the first weeks, the titres of IF IgG and IgM antibodies reached values from 256-512 in some patients; IgA antibody titres were considerably lower. No specific IF IgM and IgA antibody was found in sera from 5 infants with prenatal LCM virus infection (3 infants with congenital hydrocephalus and chorioretinitis, 1 with cerebral palsy and chorioretinitis, and 1 with asymptomatic infection). IF IgM antibody in a titre of 4 was detected only in 2 specimens of the cerebrospinal fluid from one of the infants with hydrocephalus. It is suggested that in prenatal LCM virus infection the production of specific IgM antibody is more or less inhibited.


Accurate determination of volumes injected intracerebrally into suckling mice. Inoculum with 125I added as a marker was used to determine the volume actually administered to suckling mice on intracerebral (i.c.) inoculation. In 1-2 days old animals, this volume corresponded to approximately 75% of a 20 microliter dose.


Effect of interferon and poly I:C on replication of influenza virus in organ cultures of human origin. Interferon and poly I : C inhibited the growth of influenza virus in organ cultures of foetal nasal mucosa, trachea, oesophagus and adult nasal mucosa. No antiviral effect was observed in decidua organ cultures. The combined use of interferon and poly I : C resulted in the enhancement of the antiviral effect.


Quantitative immunoelectrophoresis of human interferon. A new approach to characterization of interferon preparations. The electroimmunoassay (quantitative "rocket" immunoelectrophoresis) method was adopted for analysis and characterization of interferon preparations. Ammonium sulphate-precipitated anti-interferon globulin also containing unknown antibodies against antigens that contaminate interferon preparations was used in the tests. By comparing the results of the electroimmunoassay of fractions obtained by polyacrylamide gel electrophoresis of human leukocyte interferon with the antiviral activity of the fractions, an excellent dissociation of molecules with interferon activity from the bulk of contaminating antigens was achieved. The method is extremely sensitive and requires very small volumes for assay.


Transmission of Kyasanur Forest disease virus by Rhipicephalus haemaphysaloides ticks. Larvae of Rhipicephalus haemaphysaloides were infected with Kaysanur Forest disease (KFD) virus by feeding on viraemic rodents and reared into next generation larvae. Fed larvae, nymphs, unfed adults, fed adult males, and females after oviposition were found infected, while the larvae were found free from infection. Nymphs and adults transmitted the infection by bite to rodents and rabbits respectively. The virus was also passed through a second rodent-tick cycle. Adult ticks showed a titre of 3.1 to 4.5 dex mouse LD50/0.03 ml, and the virus was also detected 245 days after infection.


Failure of 5-bromodeoxyuridine to influence the activation of latent herpex simplex virus infection in explants of the rabbit regional sensory ganglia. The occurence of virus-specific antigens in explants of the Gasserian ganglia of rabbits with healed keratitis induced by the Kupka strain of human herpesvirus type 1 was tested by indirect immunofluorescence. In explants, the neurons and satellite cells revealed specific fluorescence from the 3rd day of culturing. The pattern of virus activation in the ganglion tissue was similar irrespective of whether the explants were cultured in the presence or absence of 5-bromo-2'-deoxyuridine.


Khasan virus, a new ungrouped bunyavirus isolated from Haemaphysalis longicornis ticks in the Primorie region. Strain LEIV-776P, possessing no haemagglutinating activity was isolated in 1971 in the Primorie region (U.S.S.R.) from Haemaphysalis longicornis Neumann 1091 tick. Complement fixation (CF) tests revealed no antigenic relationship with 24 antigenic groups of arboviruses or 21 ungrouped viruses isolated from ticks. The virus isolate contains RNA and is relatively sensitive to ether and sodium deoxycholate; it is pathogenic for suckling mice and two-week-old white mice by the intracerebral route, and replicates in primary cultures of chick and duck fibroblasts and green monkey kidneys and in a continuous line of pig embryo kidney cells without any cytopathic effect. According to electron microscopy, its size is 90-110 nm. Its morphological properties permit to classify it as a member of the family Bunyaviridae.


Correlation between foetal pH, cord blood glucose level and Apgar score in a foetal intensive care unit--preliminary report. The foetal pH (the term 'foetal pH' is used for the pH of foetal capillary blood obtained by making a small incision in the foetal scalp or buttocks) and the umbilical cord blood glucose levels in pregnant women with foetal distress were estimated in 19 babies as a prospective study in Ile-Ife, Nigeria. The Apgar Scores on delivery were also estimated. There was a significant correlation coefficient between foetal pH and Apgar Score r = 0.38 and the coefficient of correlation between umbilical artery blood glucose and foetal pH was 0.08. Estimation of the pH of blood taken from the foetal scalp is now well established as a diagnostic measure in the early diagnosis of foetal distress due to hypoxia. The results obtained from this small series of nineteen patients highlights the importance of the estimation of foetal pH, blood glucose and the Apgar Score. This is the first account in Ile-Ife, Nigeria.


Adjunctive drug use by methadone patients. Clinical impressions, reinforced by reports in both professional journals and the lay press, suggested that use and abuse of minor tranquilizers and sedative-hypnotics was extensive among patients in our drug treatment clinics and required some change in our policies. We surveyed a randomly selected national sample of methadone program physicians and found they shared our assumptions concerning the use of these drugs. A pilot program of urine screening for the presence of adjunctive drugs was completed, and we found the incidence of use to be below expected levels, except for opiates and alcohol. These results suggest we should reconsider plans to alter clinic policies concerning the regulation of these substances. A rigorous examination of serum and/or urine in a national research program to determine both the incidence of adjunctive drug use by clients in drug treatment programs and correlations of such data with national surveys would be an expensive undertaking, but would provide a more rational basis for policy making at both local and national levels.


Esophageal epithelial response to gastroesophageal reflux. A quantitative study. Exposure of the distal esophageal mucosa to acid gastric juice was quantitated by 24-hr pH monitoring in 100 individuals and was correlated with morphologic data derived from esophageal biopsies. The degree of acid exposure to the distal esophagus correlated directly with increases in both relative and absolute length of the subepithelial papillae and to relative basal zone hyperplasia. Both papillary length and basal zone hyperplasia decreased after antireflux surgery had reduced acid exposure to normal. Reflux in the recumbent position resulted in prolonged exposure of the mucosa to acid because of poor acid clearing from the esophagus. This caused longer papillae than did upright reflux, where there were more frequent reflux episodes, but with rapid acid clearance. The presence of a hiatal hernia was associated with longer papillae, lower DES pressure, increased reflux frequency, and prolonged recumbent acid clearance. Twenty-four hour pH monitoring correlated better with papillary length than did symptoms or other clinical measures of gastroesophageal reflux.


NADPH-oxidation activities in subcellular fractions isolated from resting or phagocytozing human polymorphonuclears. Using a fluorometric assay for the determination of oxidized pyridine nucleotides (NAD[P]+), total and cyanide-resistant NADPH-oxidative activities have been measured in subcellular fractions isolated from resting and phagocytosing human polymorphonuclears. Enzymatic activies responsible for the oxidation of the NADPH have been recovered in the heavy particles (15,000g/15 min), the low-density particles (100,000g/30 min), and the cytosolic fraction. Stimulation of the cells with opsonized zymosan had a different effect on the NADPH-oxidative activities of these subcellular fractions, which suggests the involvement of various types of enzymatic systems in the oxidation of NADPH. The cytosolic fraction interacted strongly with the enzymatic activities occurring in the sedimentable fractions and is therefore thought to play a central role in the regulation of the activation of the oxidative metabolism associated with phagocytosis.


Presence of arylsulfatase A (ARS A) in multiple sulfatase deficiency disorder fibroblasts. Multiple deficiency disorder fibroblasts cultured in MEM-CO2 showed deficiencies of arylsulfatase A(ARS A) comparable to the deficiency in metachromatic leukodystrophy fibroblasts. However, the MSDD fibroblasts cultured in MEM-HEPES contained near normal levels of ARS A. Moreover, the enzyme from the latter fibroblasts was indistinguishable from ARS A of control fibroblasts on DEAE-cellulose chromatography, ratio of activity with several substrates, thermal inactivation, sensitivity to inhibitors, and precipitation by antiserum to human ARS A. These data support the conclusion that the ARS A genome is intact in MSDD fibroblasts and, by extension, in MSDD patients. Other sulfatases were present at levels ranging from mildly deficient to near normal but never as low as seen in the corresponding specific sulfatase deficient disorders.


Renal response of fetal lamb to complete occlusion of umbilical cord. The renal response of the fetal lamb to repeated complete occlusion of the umbilical cord was studied in nine chronically instrumented animals. Five episodes of occlusion of the umbilical cord, each lasting for two minutes, produced a twofold rise in fetal urine osmolality and sodium, chloride, and potassium concentrations. Output of urine and glomerular filtration rate remained essentially unchanged while free water clearance decreased from a control of +0.10 to -0.02 ml. per kilogram per minute at the end of the fifth episode. Electrolyte concentrations in urine remained elevated for at least two hours following the occlusions. In addition to changes in urine composition, there was a 50- to 200-fold increase in the fetal plasma concentration of vasopressin. These studies indicate that complete interruption of the umbilical circulation, even though of short duration, produces disturbances in fetal renal function that can lead to loss of electrolytes in the urine. They provide an explanation for the low sodium levels reported in asphyxiated newborn infants in renal failure.


Effects of pH, Ca, ADH, and theophylline on kinetics of Na entry in frog skin. The short circuit current as a function of Na concentration in both solutions was found to obey Michaelis-Menten kinetics under a variety of experimental conditions. Values of maximal transport rate (Im) and half-maximal Na concentrations (Kt) were determined from these experiments. Three type of results were obtained: 1) Im and Kt both decreased by approximately the same fraction when the pH of both solutions was reduced by increasing PCO2, 2) Im decreased and Kt increased when the external pH was decreased, and 3) Im increased with ADH and theophylline, decreased with external Ca, and Kt remained unchanged. Various criteria were utilized to determine that these were properties of the entry barrier for Na into the "transport pool." The results are explained in terms of a model that separates three different types of actions on the entry barrier: 1) competition of Na with other ions in the external solution for entry, 2) modulation of the number of sites available for Na translocation by changing the cytoplasmic pH, and 3) alterations in the rate of Na translocation caused by changes in the Na permeability or the electrochemical gradient across the entry barrier.