Studies of acidosis in the ischaemic heart by phosphorus nuclear magnetic resonance. 1. Phosphorus-nuclear-magnetic-resonance measurements were made on perfused rat hearts at 37 degrees C. 2. With the improved sensitivity obtained by using a wide-bore 4.3 T superconducting magnet, spectra could be recorded in 1 min. 3. The concentrations of ATP, phosphocreatine and Pi and, from the position of the Pi resonance, the intracellular pH (pHi) were measured under a variety of conditions. 4. In a normal perfused heart pHi = 7.05 +/- 0.02 (mean +/- S.E.M. for seven hearts). 5. During global ischaemia pHi drops to 6.2 +/- 0.06 (mean +/- S.E.M.) in 13 min in a pseudoexponential decay with a rate constant of 0.25 min-1. 6. The relation between glycogen content and acidosis in ischaemia is studied in glycogen-depleted hearts. 7. Perfusion of hearts with a buffer containing 100 mM-Hepes before ischaemia gives a significant protective effect on the ischaemic myocardium. Intracellular pH and ATP and phosphocreatine concentrations decline more slowly under these conditions and metabolic recovery is observed on reperfusion after 30min of ischaemia at 37 degrees C. 8. The relation between acidosis and the export of protons is discussed and the significance of glycogenolysis in ischaemic acid production is evaluated.


The effect of reduction of perfusion rate on lactate and oxygen uptake, glucose output and energy supply in the isolated perfused liver of starved rats. 1. Lactate and O2 uptake and glucose output were studied in isolated livers from starved rats at perfusate flow rates varying from 100 to 7% of "normal" (11.25-0.75 ml/min per 100 g body wt.). 2. With moderate diminution of flow rate, lactate and oxygen uptake fell more slowly than would be expected if uptake purely depended on substrate supply. 3. Use of a mathematical model suggests that the intrinsic capacity of the liver for lactate uptake is unaffected until the flow rate falls below 25% of "normal". 4. Some lactate uptake was always observed even at 7% of the "normal" flow rate. 5. At flow rates below 33% of the "normal", lactate was increasingly metabolized by pathways other than gluconeogenesis, which became a progressively less important consumer of available O2. 6. ATP content decreased with diminution of flow rate, but substantially less markedly than did lactate uptake and glucose output. 7. Intracellular pH fell from a mean value of 7.25 at "normal" flow rate to 7.03 at 7% of the "normal" flow rate.


Regulation of p-nitroanisole O-demethylation in perfused rat liver. Adenine nucleotide inhibition of NADP+-dependent dehydrogenases and NADPH-cytochrome c reductase. Perfusion of rat livers with 10 mM-fructose or pretreatment of the rat with 6-aminonicotinamide (70 mg/kg) 6 h before perfusion decreased intracellular ATP concentrations and increased the rate of p-nitroanisole O-demethylation. This increase was accompanied by a decrease in the free [NADP+]/[NADPH] ratio calculated from concentrations of substrates assumed to be in near-equilibrium with isocitrate dehydrogenase. After pretreatment with 6-aminonicotinamide the [NADP+]/[NADPH] ratio also declined. Reduction of NADP+ during mixed-function oxidation may be explained by inhibition of of one or more NADPH-generating enzymes. Glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, isocitrate dehydrogenase and "malic" enzyme, partially purified from livers of phenobarbital-treated rats, were inhibited by ATP and ADP. Inhibitor constants of ATP for the four dehydrogenases varied considerably, ranging from 9 micrometer for "malic" enzyme to 1.85 mM for glucose 6-phosphate dehydrogenase. NADPH-cytochrome c reductase was also inhibited by ATP (Ki 2.8 mM) and by ADP (Ki 0.9 mM), but not by AMP. Concentrations of ATP and ADP that inhibited glucose 6-phosphate dehydrogenase and the reductase were comparable with concentrations in the intact liver. Thus agents that lower intracellular ATP may accelerate rates of mixed-function oxidation by a concerted mechanism involving deinhibition of NADPH-cytochrome c reductase and one or more NADPH-generating enzymes.


Effects of glucose on the cytosolic ration of reduced/oxidized nicotinamide-adenine dinucleotide phosphate in rat islets of Langerhans. The maximal extractable activity of "malic" enzyme (EC 1.1.1.40) in rat islets of Langerhans was similar to that reported for liver. Thus "malic" enzyme may catalyse a near-equilibrium reaction in the cytosol of islets of Langerhans. Measurements of islet content of malate and pyruvate, the metabolite substrate and product of "malic" enzyme, were therefore used to calculate the cytosolic ration of [NADPH]/[NADP+]. This ratio was higher in islets incubated with 20 mM-glucose than in islets incubated with 2 mM-glucose.


[A review of pharmacological studies on nicergoline]. 10-Methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) shows a strong alpha-blocking activity both in vitro and in vivo. Various studies (dog, cat, rabbit, rat, mouse, guinea-pig) show that nicergoline affects only slightly blood pressure and heart rate and increases the blood flow in brain and hind limb without affecting the splanchnic and aortic flow in normal animals. Nicergoline does not interfere with CNS functions unless applied in high doses. It stimulates the muscle oxidative metabolism and function and lacks any emetic and hallucinogenic activity. Its acute and chronic toxicity in different animal species is very low.


[Influence of nicergoline on the cerebral blood flow and alpha-sympatholytical properties (author's transl)]. The cerebral vasodilatory effect of 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) was examined by recording the cardiac output of the vertebral artery in the dog. As from a dosage of 25 micrograms/kg nicergoline increases permanently the cardiac output to a normal or nearly normal level which had been decreased by phenylephrine reducing the increased local vascular resistance. The alpha-sympatholytical properties of nicergoline were more precisely proved by reducing the reactive increase in blood pressure after epinephrine and norepinephrine (3 micrograms/kg) by administration of nicergoline in 3 doses (25, 50 and 100 micrograms/kg). The same method was used with phentalamine in dosages of 125, 250 and 500 micrograms/kg. The statistical comparison of the results obtained with both substances shows that the ratio of equiactive doses is about 3:1 in favour of nicergoline.


Non-destructive measurement of metabolites and tissue pH in the kidney by 31P nuclear magnetic resonance. Phosphorus nuclear magnetic resonance (31P NMR) can be used as a non-destructive method for the simultaneous observation of the major phosphate-containing metabolites (ATP, ADP, nucleotide monophosphate, Pi, sugar phosphate) and intracellular pH in isolated rat kidney. The time course of changes in these metabolites and in cellular pH in the ischaemic kidney are examined at two temperatures and in the presence of different flushing media. ATP is rapidly depleted while the pH change is slower and shows biphasic behaviour. Pi production and total nucleotide (ATP and ADP) depletion also occur on the same time-scale as the tissue acidification. The relation of these observations to tissue viability is discussed and the possibility of extending the measurements to human organs is considered.


Changes in transglutaminase activity in an experimental model of pulmonary fibrosis induced by paraquat. An experimental model of pulmonary fibrosis has been developed by dosing rats with one-fifth the LD50 dose of the herbicide paraquat on 5 consecutive days. Approximately 50% of the rats died within 4 days of the completion of dosing, showing macroscopic changes and wet weight increases in the lung consistent with severe oedema. Those animals which died between Days 4 and 10 had markedly increased levels of hydroxyproline in the lung, maximum at Day 6, and increased prolyl hydroxylase activity, maximum at Day 4. These changes, together with an increase in thymidine incorporation into DNA, and increased lung DNA content, were consistent with the development of fibrosis. Measurement of transglutaminase activity in the lung showed marked increases at Days 4 and 10 after completion of dosing. This activity paralleled closely the changes in prolyl hydroxylase activity and became increasingly associated with particulate protein present in the "nuclear pellet" fraction. The presence of zymogen plasma transglutaminase trapped in lung homogenates could not be demonstrated but the contribution by the active plasma transglutaminase (Factor XIIIa) to increases shown at Day 4 cannot be ruled out.


The twenty aminoacyl-tRNA synthetases from Escherichia coli. General separation procedure, and comparison of the influence of pH and divalent cations on their catalytic activities. A general separation procedure of the twenty E. coli aminoacyl-tRNA synthetases including either a 105 000 g centrifugation or a poly-ethyleneglycol-dextran two-phases partition fractionation, and chromatographies on DEAE-cellulose, phosphocellulose and hydroxyapatite is described. The specific activities of the synthetases have been determined after each chromatographic step and compared to their respective activities in the 105 000 g supernatant. Some aminoacyl-tRNA synthetases were obtained at 80 per cent purity. The presence of phenylmethylsulfonyl fluoride does not significantly modify either the elution patterns of the synthetases during the various chromatographic steps or their specific activities. Thus, contrarily to enzymes from various eukaryotic organisms no significant inactivation of the E. coli aminoacyl-tRNA synthetases occurs via proteolytic processes during the purification procedure. The effects of various factors: pH, magnesium, and other bivalent cations including spermidine, were tested on the aminoacylation and the [32P] PPi-ATP isotope-exchange reactions, and the optimal aminoacylation and isotope-exchange conditions determined for 18 of the 20 E. coli aminoacyl-tRNA synthetases.


Thermal stability of soluble mitochondrial H+-ATPase. ATPase melting has been studied by circular dichroism and differential scanning microcalorimetry. Decomposition of the alpha-helix of H+-ATPase (in which about 80% of the peptide groups of the enzyme are involved) following thermal treatment is shown to proceed gradually, beginning with room temperature. Effect of nucleotides upon melting is detected in the range of 20 degrees--40 degrees C. Above 40 degrees C, the pattern of thermal decomposition of the three-dimensional structure of H+-ATPase is independent of the nature of nucleotides present. Highly stable alpha-helical sites have been found in the enzyme molecule. Possible mechanism of formation of such sites is discussed, and the results obtained are compared with data on thermal stability of ATPase from thermophilic bacteria. Structural changes in the molecule following thermal treatment are compared with ATPase activity changes under similar experimental conditions.


Kinetics of ion transport in lipid membranes induced by lysine-valinomycin and derivatives. Lysine-valinomycine and two N epsilon-acyl derivatives are compared with respect to their potency to transport Rb+ ions across thin lipid membranes. Lysine-valinomycin acts as a neutral ion carrier only above a pH of about 7 of the aqueous solutions, while at lower pH the molecules seem to be positively charged due to a protonation of the epsilon-NH2 group of the lysine residue. A kinetic analysis based on voltage jump relaxation experiments and on the nonlinearity of the current-voltage characteristics showed that the conductance increment delta per carrier molecule for uncharged lysine-valinomycin is similar to that of natural valinomycin. The attachment of a rather bulky side group such as the dansyl or para-nitrobenzyloxycarbonyl group reduced delta by approximately one order of magnitude. Some of the relaxation data of the valinomycin analogues were influenced by an unspecific relaxation of the pure lipid membrane. This structural relaxation represents a limitation to the possibility of analyzing specific transport systems in thin lipid membranes by the voltage jump or charge pulse techniques. It is shown that the time dependence of this structural relaxation--which was first published by Sargent (1975)--is at variance with a three capacitor equivalent circuit of the membrane, which was suggested by Coster and Smith (1974) on the basis of a.c. measurements. A modified equivalent circuit has been found to represent a satisfactory analogue for the current relaxation in the presence of valinomycin. It turned out, however, that such an equivalent circuit provides little insight into the molecular mechanism of transport.


[Characterization of neutral proteinases from human spleen]. Several proteinases hydrolyzing histone and caseine in neutral media were obtained by Sephadex G-100 fractionation of water and salt (1 M KCl) extracts of human spleen. The level of the activity of proteinases in the extracts was very low as a result of the presence of an inhibitor. Neutral proteinases were found in two protein fractions. The "high-molecular-weight-" proteinases were inhibited by DFP and therefore they were attributed to a group of serine proteinases. The "low-molecular-weight" fraction contained neutral SH-dependent proteinase(s) and DFP-inhibited enzymes. In this fraction, the kininogenase activity and the hydrolysis of Boc-1-ananine p-nitrophenyl ester, N-benzoyl-L-tryosine ethyl ester and N-benzoyl-DL-arginine p-nitroanilide were observed.


[Influence of neurotransmitters on the antinociceptive effect of midbrain stimulation]. In experiments on rats with implanted electrode-cannules there were studied the effects of L-tryptophane (25 mg/kg intraperitoneally) and microinjections of serotonin (20 micrograms), dopamine (10 micrograms) and proserine (5 micrograms) into the area of periaqueductal central gray on the antinociceptive effect caused by stimulation of the same "points" of the midbrain. L-tryptophane, serotonine and proserine (in the presence of methylatropine) potentiated the effect of subthreshold antinociceptive stimulation which could be tested from the modifications of thresholds of the development of some complex pain reaction components under electrical stimulation of the rat tail. Dopamine did not have such an effect. The potentiating effect of serotonine is not eliminated by naloxone.


[Increased resistance of (CBAxC57BL/6)Fl hybrids to the graft-versus-host reaction during spleen regeneration]. Partial splenectomy was performed in CBAXXC57BL/6/F1 hybrids 2, 12, 15 and 20 days before the graft-versus-host reaction induction (GVHR) and 2, 10 days after administration of parenteral immunocompentent cells. The recipients with the intact spleen or complete splenectomy and sham operated recipients served as control. Hybrids with the regenerating spleen were more resistant to GVHR within 12 days of regeneration, while splenectomy increased hybrid resistance to GVHR only when performed 2 days before or 2 days following the GVHR induction.


Somnambulistic-like episodes secondary to combined lithium-neuroleptic treatment. Ten of 114 psychiatric patients undergoing combined lithium-neuroleptic treatment exhibited somnambulistic-like episodes. These episodes are differentiated from nocturnal wanderings and epileptic attacks during sleep; they occurred within two to three hours after sleep onset and were characterized by the patients appearing confused and walking about in a quiet, detached and clumsy manner. Generally, there was amnesia for the event. Since sleepwalking occurs out of slow wave sleep, the increase in slow wave sleep induced by lithium and certain neuroleptics may represent a neurophysiological mechanism responsible for these patients' somnambulistic behaviour. The occurrence of grand mal seizures in two patients was probably unrelated to the somnambulistic-like episodes. However, persistence of the latter appears to be associated with drug-induced EEG irregularity.


N-acylation of tyramines: purification and characterization of an arylamine N-acetyltransferase from rat brain and liver. The N-acylation of tyramine isomers and other biogenic amines has been studied. The liver exhibits the highest activity towards tyramines, while the brain exhibits a low but significant activity. In the brain, tyramine N-acylation activity was heterogenously distributed. The arylamine N-acetyltransferase has been partially purified from both rat liver and brain, the two enzymes being quite similar with respect to their chromatographic properties, optimal pH requirement (pH 7.8), and their kinetic parameters. The product N-acetyltyramine is not oxidized by liver amidohydrolase or monoamine oxidase.


The kinetic parameters of trehalase in whole and disrupted mitochondrial preparations from two insects with asynchronous muscle. The kinetic parameters of trehalase in honey bee and flesh fly mitochondria were compared. The studies were carried out with whole mitochondria and with mitochondria disrupted in various ways and to different degrees. Honey bee mitochondrial trehalase was significantly activated by Lubrol WX treatment (30.0-fold), by high pH treatment (20.8-fold), and by a treatment consisting of 10 passes through a French press (37.9-fold) but not by the other treatments tried (salt, proteases, Waring blender, and sonication), despite the fact that these treatments also disrupted the mitochondria significantly. The activation effect was on the Vmax. The Km value did not change. Simple breakage of either the outer or inner (or both) membranes was not sufficient to activate trehalase from honey bees, which showed that the activation was not an indirect result of a change in the case with which trehalose can pass through the membranes. Honey bee trehalase is the first trehalase from insects with asynchronous muscle which has been shown to be activatable by physical and chemical methods. Flesh fly mitochondrial trehalase behaved quite differently from the honey bee enzyme in that it could not be activated by any of the techniques tried, even when there were significant amounts of disruption.


Stability of microtubule protein over the pH range: 6.9--9.5. The pH stability range of a microtubule protein preparation has been investigated between 6.9 and 9.5. Microtubule protein was exposed to various pH values in this range and then returned to pH 6.9. The appearance of microtubules as verified by electron microscopy and sedimentation analysis under polymerizing conditions was taken as an indication of a conformationally stable protein. Between pH 6.9 and pH 8.0 the loss in the ability to form microtubules was found to be reversible, at pH 8.2 it was partially reversible, above pH 8.2 it was irreversible. Tubulin and the microtubule-associated protein fraction were separately exposed to high pH. It was observed that tubulin exposed to high pH can still form microtubules in the presence of untreated microtubule-associated protein. On the other hand, microtubule-associated protein exposed to high pH could not initiate microtubule assembly with untreated tubulin. It was concluded from these observations that the loss in the ability of a microtubule protein preparation to assemble at high pH is due to a change in the microtubule-associated protein fraction and that tubulin is conformationally stable even after exposure to pH 9.5.


The action of tributyltin chloride on the uptake of proline and glutamine by intact cells of Escherichia coli. Tributyltin chloride inhibits growth and uptake of glutamine and proline into intact cells of Escherichia coli. It causes efflux of the accumulated amino acids. A pH gradient generated in intact cells and everted membrane vesicles is dissipated by this compound. These effects do not require lipoic acid but are dependent on the presence of chloride, bromide, or iodide ions. We conclude that tributyltin chloride can catalyse a transmembrane OH- -anion exchange exchange reaction and that this is its mode of inhibition of the uptake of these amino acids. The response of proline and glutamine uptake to the inhibitor is similar and is consistent with the transport of both amino acids requiring an electrochemical gradient of protons.


The action of tributyltin chloride on energy-dependent transhydrogenation of NADP+ by NADH in membranes of Escherichia coli. Respiration- and ATP-dependent transhydrogenation of NADP+ by NADH in everted membrane vesicles from Escherichia coli is inhibited by nigericin but is relatively insensitive to valinomycin. The sensitivity to nigericin is enhanced 30-fold in the presence of valinomycin. It is concluded that both the transmembrane pH difference and the membrane potential constitute the driving force for energy-dependent transhydrogenation. Respiration- and ATP-dependent transhydrogenation are inhibited by tributyltin chloride. Although effects on the energization system have not been excluded, the inhibitor appears to react with a sulfhydryl group on the transhydrogenase enzyme. This inhibition is not dependent on the presence of a permeant anion and can be reversed by mono- and particularly di-thiol compounds. The transhydrogenase is also inhibited by 5,5'-dithiobis(2-nitrobenzoic acid), N-ethylmaleimide, p-chloromercuriphenyl sulfonic acid, and Zn2+, but these reagents are less effective than tributyltin chloride. Energy-independent transhydrogenation is inhibited at high concentrations (20 mM) of cysteine. The reason for this is unknown.


Oxidation of reduced nicotinamide adenine dinucleotide phosphate by plant mitochondria. Mitochondria isolated from various plant tissues (leaves, etiolated shoots and hypocotyls, and stem tubers) oxidize exogenous NADPH with respiratory control values and ADP:O ratios similar to those obtained with exogenous NADH as substrate. In all the mitochondria investigated, the electron-transfer inhibitors rotenone and amytal each had the same effect on the oxidation of NADPH as they had on the oxidation of NADH. The oxidation of exogenous NADPH by white potato tuber mitochondria was much more sensitive to inhibition by citrate or ethylene glycol bis-(beta-aminoethyl ether)-N,N-tetraacetic acid than was the oxidation of NADH. Mitochondria isolated from aged beetroot slices showed an increased capacity for the oxidation of exogenous NADH (compared with mitochondria from fresh tissue) but no such increase in the capacity to oxidize exogenous NADPH. These results suggest that exogenous NADPH and NADH are oxidized via different flavoproteins in plant mitochondria.


Characterization of a stable intermediate in the unfolding of diazoacetylglycine ethyl ester--pepsin by urea. The irreversible unfolding of covalently inhibited swine pepsin by urea was studied by spectrophotometric and viscosity measurements. At pH 4.5 and 25 degrees C in 8 M urea, a stable intermediate form of the protein was detected. It differed from the native protein by a slight loss of secondary structure and an increased intrinsic viscosity ([pi] = 7.5 mL g-1), indicating the intermediate to have an increased molecular volume or to be more asymmetric in shape. The protein was transformed into a random coil form by increases of temperature and pH. Comparison with other results suggested that at pH 6 pepsin is less stable than its inactive precursor, pepsinogen, by about 3 Kcal mol-1 (1 cal = 4.187 J).


Factors affecting germination, mycoparasitism, and survival of Sporidesmium sclerotivorum. Macroconidia of Sporidesmium sclerotivorum, a mycoparasite of Sclerotinia spp., germinated after 3 days in soil adjacent to sclerotia of S. minor and on membrane filters placed on soil containing sclerotia. Germination increased with time up to 18 days and with concentration of sclerotia. Conidia as distant as 9 mm from single sclerotia germinated. Germination of conidia was maximum on a sclerotial agar medium in the range of pH 5 to pH 7. Cultivation of S. sclerotivorum parasitically on living sclerotia proceeded optimally in moist, fine quartz sand amended with 1 to 2% (w/w) sclerotia and 0.07% (w/w) CaCO3, at 25 degrees C. Infection of sclerotia in sand reached 100% by 5 weeks. Conidia production paralled infection resulting in logarithmic increase in numbers; a maximum of 3 x 10(5) to 4 x 10(5) conidia/g was reached in 6 to 12 weeks. Viability of air-dried sand-sclerotial cultures of S. sclerotivorum was reduced after 1 and 6 days, but viability was undiminished in air-dried soil. Sporidesmium sclerotivorum survived in moist and air-dried soils stored at room temperature for 15 months.


Low molecular weight substitutes for beef extract as eluents for poliovirus adsorbed to membrane filters. Basic solutions of beef extract and casein were able to elute poliovirus adsorbed to four membrane filters with different chemical compositions. Hydrolyzed protein and individual amino acids were able to elute virus adsorbed to certain filters but were unable to elute virus adsorbed to other filters efficiently. A solution of 4 M urea buffered at pH 9 with 0.05 M lysine was able to elute greater than 60% of the virus adsorbed to each of the filters tested. Certain solutions of amino acids were capable of eluting virus adsorbed to one filter but permitted adsorption of virus to another filter with a different chemical composition. Acidic amino acids could interfere with elution of virus from membrane filters. Aromatic compounds with an amino group attached to the ring were good eluents for virus adsorbed to epoxy-fiberglass membrane filters. In contrast, aromatic compounds with other substituents were generally poor eluents.


A liquid medium for the production of Kabatiella zeae conidia. A liquid medium, designated as Kabatiella zeae medium (KZM), containing 10.0 g of carboxymethylcellulose, 5.0 g of maltose, 1.5 g of peptone, 1.0 g of monobasic potassium phosphate, 1 L of distilled water, is described. Peptone was found to be the component most influential in stimulating sporulation. The optimum temperature for conidial production was 25 degrees C. In shake culture about 10(7) conidia/mL were produced in 5 days at room temperature (about 21 degrees C) when KZM was seeded with 4-day-old colony plugs. The optimum for conidial production was pH 5 with only a slight gradual reduction to pH 9. However, fungal development was abnormal at pH values of 4 and 5. Of the 29 isolates tested, only two did not sporulate in KZM or in any other medium. Because of the high yield of conidia in a short time, the ease of preparation and use, and its low cost, KZM is especially useful where large amounts of inoculum are needed.


A medium for commercial production of the halophilic Micrococcus nuclease. A simple synthetic medium (glutamate-sucrose medium) was devised for production, during growth in shaken flasks, of extracellular halophilic nuclease (nuclease H) by a moderate halophile, Micrococcus varians subsp. halophilus. A simple medium consisting of 0.7% ammonium sulfate, 1.0% glucose, minerals, three vitamins, and 2 M NaCl gave good growth and excellent production of nuclease H in a jar fermentor when the pH was adjusted to 7.5 to 8.0 during cultivation.


Effect of baroreceptor deafferentation on central catecholamines in the rat. 1. Sinoaortic deafferentation in the rat leads to increased blood pressure and heart rate. 2. Early increases in tyrosine hydroxylase activity both in brain stem and hypothalamus suggest that increased noradrenaline synthesis may contribute to the development of neurogenic hypertension. 3. After 4 weeks, phenylethanolamine-N-methyltransferase activity was reduced in the hypothalamus. 4. Noradrenaline- and adrenaline, but not dopamine-containing neurones may participate in regulation of sympathetic efferent activity.


Treatment of hypertension: state of the art in 1979. 1. The results of the Veterans Administration Co-operative Study have been extended by the subsequent clinical trials, which included patients of both sexes and with less vascular disease. The later studies confirm the effectiveness of treatment in preventing most complications except myocardial infarction and sudden death. Furthermore, the lower diastolic blood pressure in which treatment has been shown to have a significant beneficial effect has been lowered from 105 mmHg as indicated by the Veterans Study to 100 mmHg as shown by the much larger Australian trial. The possibility of reducing the incidence of sudden death and fatal myocardial infarction has been suggested by other recent control trials using beta-adrenoreceptor-blocking drugs, an approach that needs further exploration. 2. A number of interesting and useful new drugs have appeared which include tienilic acid, minoxidil, saralasin and captopril, and in addition recent controlled trials have re-emphasized the effectiveness of the old drug, reserpine, when combined with a diuretic. The art of treatment of hypertension therefore appears to be in a healthy state and we should expect more advances in the future.


Treatment of mild hypertension in elderly males. 1. Males, born between 1900 and 1925, with mild hypertension have been treated for periods varying from 300 to 2000 days. 2. The life and death status of all patients (except two) was known on 1st November 1978. 3. A group of patients with mild hypertension receiving treatment based on a thiazide diuretic had a greater mortality than the other drug-treated group. 4. The increased mortality was caused by an increased number of myocardial infarcts. 5. Elderly male patients with mild hypertension probably have preexisting vascular disease and therapy should not automatically be started. If therapy is started, beta-adrenoreceptor-blocking drugs may be a preferred therapy.


SK&F 92657, a novel antihypertensive acting by precapillary vasodilatation and beta-adrenoreceptor blockade. 1. The properties of a new antihypertensive agent, SK&F 92657, DL-3-[2-(3-t-butylamino-2-hydroxypropoxy)phenyl]-6-hydrazinopyridazine, have been studied. 2. The compound caused a sustained fall in blood pressure in several species as a result of precapillary vasodilatation, particularly in the renal and coronary vasculatures. 3. The beta-adrenoreceptor-blocking actions of SK&F 92657 prevent reflex cadiac stimulation.


Plasmin can activate plasma prorenin but is not required for the alkaline phase of acid activation. 1. Plasma prorenin is an inactive form of renin that is converted into active renin at alkaline pH in previously acidified plasma; this conversion of prorenin into renin is mediated by Hageman factor-dependent activation of prekallikrein, which, in turn, leads to prorenin activation. 2. Since plasma kallikrein can activate plasminogen, the present studies were designed to evaluate whether alkaline-phase activation of prorenin by plasma kallikrein is mediated via plasminogen activation. 3. We demonstrated that plaminogen is present in acid-treated plasma in sufficient quantity to convert prorenin into renin after activation by streptokinase. 4. However, alkaline-phase activation was completely normal in plasminogen-free plasma. 5. Therefore alkaline-phase activation of plasma prorenin is mediated by plasma kallikrein but is not dependent on kallikrein activation of plasminogen.


Chromatin protein methylation in proliferating liver and hepatoma cells. Chromatin protein methylation in proliferating liver cells and hepatomas was examined in vivo and in vitro. Methylation in vivo was estimated using 2-14 C-L-methionine and 3H-methyl-L-methionine. 3H/14C ratios were calculated for crude histone and nonhistone chromatin protein fractions and compared to those calculated for serum albumin. From this, determination of a methylation index was calculated using serum albumin as a standard. Results of this study indicate that both histones and nonhistone chromatin proteins are methylated to a greater extent in proliferating liver cells and in hepatomas than in control preparations. These observations are generally supported by our in vitro methylation studies. The results are discussed in terms of chromatin protein methylation and its possible relationship to DNA replication.


Environmental effects of increased coal utilization: ecological effects of gaseous emissions from coal combustion. This report is limited to an evaluation of the ecological and environmental effects of gaseous emissions and aerosols of various types which result from coal combustion. It deals with NOx, SOx, fine particulate, photochemical oxidant and acid precipitation as these pollutants affect natural and managed resources and ecosystems. Also, synergistic effects involving two or more pollutants are evaluated as well as ecosystem level effects of gaseous pollutants. There is a brief summary of the effects on materials and atmospheric visibility of increased coal combustion. The economic implications of ecological effects are identified to the extent they can be determined within acceptable limits. Aquatic and terrestrial effects are distinguished where the pollutants in question are clearly problems in both media. At present, acid precipitation is most abundant in the north central and northeastern states. Total SOx and NOx emissions are projected to remain high in these regions while increasing relatively more in the western than in the eastern regions of the country. A variety of ecological processes are affected and altered by air pollution. Such processes include community succession and retrogression, nutrient biogeochemical cycling, photosynthetic activity, primary and secondary productivity, species diversity and community stability. Estimates of the non health-related cost of air pollutants range from several hundred million dollars to $1.7 billion dollars per year. In general, these estimates include only those relatively easily measured considerations such as the known losses to cultivate crops from acute air pollution episodes or the cost of frequent repainting required as a result of air pollution. No substantial nationwide estimates of losses to forest productivity, natural ecosystem productivity which is tapped by domestic grazing animals and wildlife, and other significant dollar losses are available.


The development of inducibility for glutamine synthetase in embryonic neural retina: inhibition by BrdU. The hydrocortisone-mediated induction of glutamine synthetase (GS) in the neural retina of the chick embryo is a characteristic and unique feature of differentiation of this tissue. The induction involves genomic activity elicited by the inducer resulting in synthesis and accumulation of the enzyme. We describe correlations between the growth of embryonic retina tissue in vivo and in vitro and the development of its inducibility for GS, and demonstrate that this development proceeds through two phases: competence-acquisition phase (before the 7th day of development), and maturation phase. BrdU applied for 24 h to retinas of 5-day embryos irreversibly suppresses the development of induction-competence. However, BrdU does not affect the progressive maturation of inducibility when applied to retinas that already are fully induction-competent (8 days and older). The short treatment with BrdU of 5-day retinas also causes defective histogenesis resulting in drastic malformation of the tissue. The nature of the processes involved in competence-acquisition and in the maturation of inducibility for GS are examined. Possible mechanisms by which BrdU prevents the development of induction-competence for GS in the early embryonic retina and elicits defective histogenesis are discussed.


Losses in milking qualities of dairy cattle caused by mosquitoes and horseflies and reduction of such losses due to use of diethyltoluamide repellent. An experiment was carried out in South Bohemia, in which 10% water emulsion of diethyltoluamide was used as repellent against bloodsucking Diptera to protect pastured dairy cattle. The repellent was applied by means of a spraying frame and was effective for 30-36 hours. Seven mosquito species and 18 horsefly species were observed to be attacking the cattle. The average number being 120-300 mosquitoes and 50 horseflies per hour, the milking qualities in the cattle decreased by 6.2%, the milk fat content by 11.8%. After converting to milk with average fat content of 4%, the difference in milking qualities between treated and untreated animals was 9.7%.


[Effect of acebutolol, a cardioselective beta-adrenoceptor blocking agent, on the blood pressure in rats (author's transl)]. The effects of acebutolol, a cardioselective beta-adrenoceptor blocking agent, on the systolic blood pressure and heart rate were investigated in conscious Kyoto Wistar normotensive rats (WKY), spontaneously hypertensive rats (SHR) and DOCA-NaCl hypertensive rats (DOCA rats) and the results compared with those of propranolol and practolol. In WKY and DOCA rats, the intraperitoneal administration of acebutolol, propranolol and practolol (0.5 approximately 20 mg/kg) produced a hypotensive action, however, these effects were observed only with restricted doses and there was no evidence of a dose-dependency. The heart rate was decreased by acebutolol and propranolol, but was increased by practolol which possesses an intrinsic sympathomimetic activity. In SHR, propranolol produced a dual action, a slight rise followed by a slight fall, the change not being significant, while practolol induced a slight hypertension. On the other hand, acebutolol in high doses induced a dose-dependent hypotensive action. The heart rate was markedly and dose-dependently decreased by these three agents. Thus, while propranolol and practolol produced hypotensive effects in WKY and DOCA rats, acebutolol produced hypotensive effects in WKY, SHR and DOCA rats. These results suggest that acebutolol is a beta-adrenoceptor blocking agent which possesses hypotensive activity in hypertensive rats.


[Forced diuresis].

[Forced diuresis]. Forced diuresis (FD) is a frequently used method for eliminating toxins. Its therapeutic effect has yet to be evaluated by a controlled clinical trial. In the absence of such a trial its usefulness can be judged only indirectly from urinary excretion rates. In former times the usual clinical laboratory procedures could not differentiate between the unchanged toxin and its metabolites. Using more selective methods, the discussions of the effect of FD in eliminating various drugs were renewed. The problem for the indication of FD is not only a missing knowledge about the kind and amount of the ingested drug, but also when knowing it, the missing evaluation of the effect of FD on its excretion. In such cases the pharmacokinetic behaviour of the drug can be helpful. If paying enough attention to the contraindications, to the principles of electrolyte and water balance, the complication rate is low. Many infusion regimes are proposed for this treatment, but only a standardized procedure can increase the safety and efficiency of this method. More complicated and more expensive methods should be used, when there is an intoxication with a substance of high mortality or when there is no effect of FD on eliminating the toxin.


[Postoperative feeding of patients after maxillofacial surgery with the tube feeding preparation Spontavix]. 25 patients--19 to 73 years old--who underwent maxillofacial operations, received Spontavix for 10 to 12 days via a nasogastric tube. In 20% of the patients nausea, vomiting, diarrhea and/or abdominal pain occurred and disappeared after finishing nutrition with Spontavix. Mean frequency of defecation was 0.5/patient/24 hours. Body weight, serum electrolytes, blood gases, pH and base excess in the arterial blood, urea-nitrogen, hemoglobin and albumin content of the blood did not change significantly. Lipids in the serum increased insignificantly without leaving normal limits. During nutrition with Spontavix serum transaminases (SGOT, SGPT) showed a statistically significant increase which is believed to be caused by general anesthesia.


Further studies on microbiological ring-expansion of penicillin N. The rate of microbiological ring-expansion of penicillin N to deacetoxycephalosporin C using protoplast lysates of the antibiotic-negative mutant Cephalosporium acremonium M-0198 has been increased some 70-fold over that of our earlier system. We confirmed the stimulatory effects of FeSO4 and ascorbate described by Hook et al. (Biochem. Biophys. Res. Commun. 87: 258, 1979); the optimum concentrations found were 0.04 mM FeSO4 and 0.67 mM ascorbate. Adenosine triphosphate concentration was lowered to 0.83 mM; phosphoenolpyruvate and pyruvate kinase were eliminated. The optimum pH and temperature for the reaction were 7.2 and 25 degrees C, respectively. Alpha-ketoglutarate and MnCl2 showed no marked effect on the reactions, MgSO4 and KCl were mildly stimulatory, and CuSO4 and ZnSO4 were very inhibitory. Penicillin N was optimal at a concentration of 0.07 mM. Specific ring-expansion activity reached its peak 13 hours after growth ceased and then disappeared rapidly.


Somatostatin along the auditory pathway. Several structures associated with the auditory pathway have been examined by radioimmunoassay for their content of somatostatin. Of these, the medial geniculate body had the highest content, followed by the cochlear nucleus, inferior colliculus, auditory cortex and cochlea. Cochlear perilymph had no detectable somatostatin-like immunoreactivity.


Somatic cell hybrids between totipotent mouse teratocarcinoma and rat hepatoma cells. We have produced somatic cell hybrids between totipotent mouse teratocarcinoma cells and rat hepatoma cells. These hybrids were tested for the expression of liver specific functions expressed in the hepatoma cell parent and for their ability to differentiate when injected into nude mice. The results of this study indicate that hybrid cell clones do not resemble either of the parental cells, since they do not produce albumin and tyrosine aminotransferase that are expressed in the rat hepatoma parent, and are incapable of forming either teratocarcinomas or hepatomas when injected in experimental animals.


Effect of cell density on energy-dependent calcium uptake by Balb/c 3T3 membranes is independent of protein synthesis and attachment to substratum. Membranes isolated from subconfluent cultures of Balb/c 3T3 cells have low energy-dependent calcium uptake activity. Replating confluent cells at low density results in a prompt fall of energy-dependent calcium uptake by membrane fractions. The level to which uptake activity falls is a function of the density at which the cells are plated (Moore and Pastan, '77b). To determine if regulation of energy-dependent uptake of calcium by membrane fractions is dependent upon attachment to a substrate and to further characterize conditions that regulate the process, we examined calcium uptake activity of membranes isolated from cells in suspension. With cells in suspension energy-dependent calcium uptake activity of isolated membranes falls promptly if cells are diluted to a low density (less than 10(5) cells/ml) and is a function of cell density. When cells in suspension at low cell densities are concentrated to high cell densities (greater than 2 x 10(6) cells/ml), calcium uptake activity of the isolated membrane fraction is increased as a function of cell density. These changes of membrane calcium uptake activity occur promptly and do not require protein synthesis.


Highly sensitive assay for tyrosine hydroxylase activity by high-performance liquid chromatography. A highly sensitive assay for tyrosine hydroxylase (TH) activity by high-performance liquid chromatography (HPLC) with amperometric detection was devised based on the rapid isolation of enzymatically formed DOPA by a double-column procedure, the columns fitted together sequentially (the top column of Amberlite CG-50 and the bottom column of aluminium oxide). DOPA was adsorbed on the second aluminium oxide column, then eluted with 0.5 M hydrochloric acid, and assayed by HPLC with amperometric detection. D-Tyrosine was used for the control. alpha-Methyldopa was added to the incubation mixture as an internal standard after incubation. This assay was more sensitive than radioassays and 5 pmol of DOPA formed enzymatically could be measured in the presence of saturating concentrations of tyrosine and 6-methyltetrahydropterin. The TH activity in 2 mg of human putamen could be easily measured, and this method was found to be particularly suitable for the assay of TH activity in a small number of nuclei from animal and human brain.


Simplified determination of lorazepam and oxazepam in biological fluids by gas chromatography-mass spectrometry. Lorazepam and oxazepam in plasma and urine were measured by gas chromatography-mass spectrometry. Oxazepam was used as an internal standard in the assay of lorazepam and vice versa. After removal of interfering substances with n-hexane, the drugs were extracted with benzene and converted to N1,O3-bistrimethylsilyl derivatives. Glucuronide forms of the drugs were extracted after hydrolysis with beta-glucuronidase. A common fragment ion at m/e 429 was used to monitor the two drugs. The sensitivity was 2 ng/ml for both drugs, which was sufficient to determine plasma and urine concentrations after therapeutic doses to humans.


Determination of the anti-allergenic agent, 2-methoxy-11-oxo-11H-pyrido[2,1-b]quinazoline-8-carboxylic acid, in biological fluids by high-performance liquid chromatography. A rapid, sensitive, and specific high performance liquid chromatographic (HPLC) assay was developed for the determination of 2-methoxy-11-oxo-11H-pyrido-[2,1-b]quinazoline-8-carboxylic acid (I) from biological fluids. The overall recovery from blood and plasma is 69 +/- 10% (S.D.) and 84 +/- 6% (S.D.), respectively, and the sensitivity limit of quantitation is 100 ng/ml by UV absorption and 5 ng/ml by fluorescence detection using a 1 ml specimen. The assay was used in the determination of blood levels of compound in the Rhesus monkey following intravenous administration of a 10 mg/kg dose, and of blood and urine levels of compound I in a dog following intravenous and oral administration of a 1 mg/kg dose.


Chromatography of xanthines on ion-exchange resins. Short columns of a 4% crosslinked cation-exchange resin gave good chromatography of xanthines, including caffeine, theophylline and hypoxanthine, and related polar aromatic compounds. Elution volumes and sequences can be modified by changing pH, solvent composition and resin counter-ion. A macroporous cation-exchange resin showed exaggerated counter-ion effects. A method is described for determining caffeine and theophylline in blood serum, using the 4% crosslinked resin with aqueous sodium phosphate eluent of pH 7.5; the temperature was 65 degrees. Detection limits are 10 ng and less.


Elucidation of the inhibitory factors of yogurt against Salmonella typhimurium. The inhibitory nature of yogurt against contaminating microorganisms has been studied extensively. Nevertheless, the factors responsible for the death of Salmonella typhimurium in yogurt have not been elucidated. An understanding of these factors is important for the determination of yogurt's safety to consumer health. Yogurt fermented for 18 h at 42 C had a stable environment with the following conditions: pH 3.85, oxidation-reduction potential -80 mV, lactic acid concentration 158 mM, and acetic acid concentration 3.7 mM. Under these conditions, lactic acid was responsible for virtually all of yogurt's bactericidal activity against S. typhimurium at 37 C. Die-off rates were observed when these conditions were reproduced artificially in milk (artificial milk yogurt) and when lactic acid was added back to 18-h yogurt from which acids were removed by passage of the whey through a Dowex 1 (Cl-) anion exchange column (cationic yogurt). Factors that augmented lactic acid inhibition of S. typhimurium were low pH and low oxidation-reduction potential. The die-off rate of S. typhimurium was more rapid in yogurt whey (yogurt minus the casein fraction) than in whole yogurt, indicating that the casein fraction was partially able to protect Salmonella.


Determinants of the longevity of third-stage infective larvae of Ancylostoma tubaeforme. The effects of some extrinsic factors on the lipid (energy) reserves and longevity of third-stage larvae of the cat hookworm Ancylostoma tubaeforme, were investigated under controlled laboratory conditions. In nonstressful microenvironmental conditions, larval longevity was directly related to the rate of utilisation of the lipid reserves. The effects of the various environmental stresses on longevity could also be explained largely on the basis of their deleterious effects on the lipid metabolism of the larvae.


Detection of anticapsular antibodies to Bacteroides asaccharolyticus in serum from rabbits and humans by use of an enzyme-linked immunosorbent assay. A sensitive serologic test, the enzyme-linked immunosorbent assay (ELISA), was used to detect serum IgG antibodies directed specifically to a capsular antigen of Bacteroides asaccharolyticus (previously known as Bacteroides melaninogenicus subspecies asaccharolyticus). Anticapsular IgG was measured in 30 specimens of rabbit serum after the animals were immunized with whole B. asaccharolyticus, the two subspecies of B. melaninogenicus, and several other bacterial species. Species-specific anticapsular IgG was demonstrated (P less than 0.001). Levels of anticapsular IgG greater than control levels were likewise detected in serum from two humans, including one patient who had periodontitis and from whom B. asaccharolyticus was isolated, and a laboratory worker who had extensive exposure over a two-year period to B. asaccharolyticus. The ELISA was found to be a relatively simple, sensitive tool for measurement of serum IgG. Its application to detection of immunoglobulins of other classes, including secretory IgA, is anticipated, provided adequate standardization methods are used.


Serotypes of Streptococcus pneumoniae causing disease. Serotypes of Streptococcus pneumoniae from 525 infected children were examined over a four-year period. Type distribution was similar among 84 cases of bacteremia and 30 cases of meningitis, with types 6, 14, and 18 accounting for half of the illnesses. In contrast, half of 396 episodes of otitis media were caused by three other types, 19, 23, and 3. Four of eight fatalities were due to type 6. Carrier strains isolated from children had a distribution of types similar to that of the otitis media collection. Adult patients had fewer of the types that caused disease in children. Most childhood infections (80%) occurred in children less than or equal to 24 months of age; no relationship between age and infecting serotype was noted. There were no seasonal trends in type distribution.


Characterization of the autolytic enzymes of Clostridium perfringens. Clostridium perfringens and isolated walls of this organism autolysed rapidly when incubated in buffer at pH 7.0 with the release of free-reducing groups but no N-terminal amino acids. The predominant autolytic enzyme was an endo-beta-N-acetylglucosaminidase, and an endo-beta-N-acetylmuramidase was also present. The autolytic enzymes could be solubilized by extraction of the organisms with 5 M-LiCl and would then subsequently bind to and rapidly lyse walls of Micrococcus luteus and, more slowly, formamide-extracted walls of C. perfringens and walls of Bacillus subtilis. Lysis of C. perfringens walls by these extracted enzymes could not be demonstrated.


A biochemical explanation for lipid accumulation in Candida 107 and other oleaginous micro-organisms. The biochemical explanation for lipid accumulation was investigated principally in Candida 107 and, for comparison, in the non-oleaginous yeast Candida utilis. There were no significant differences between these two yeasts in their control of glucose uptake; in both yeasts, the rates of glucose uptake were independent of the growth rate and were higher in carbon-limited chemostat cultures than in nitrogen-limited cultures. There was no lipid turnover in either yeast, as judged from [14C]acetate uptake and subsequent loss of 14C from the lipid of steady-state chemostat cultures. Acetyl-CoA carboxylase from both yeasts was similar in most characteristics except that from Candida 107 was activated by citrate (40% activation at 1 mM). The enzyme from Candida 107 was relatively unstable and, when isolated from nitrogen-limited (lipid-accumulating) cultures, was accompanied by a low molecular weight inhibitor. The reason for lipid accumulation is attributed to the decrease in the intracellular concentration of AMP as cultures become depleted of nitrogen. As the NAD+-dependent isocitrate dehydrogenase of Candida 107, but not C. utilis, requires AMP for activity, the metabolism of citrate through the tricarboxylic acid cycle in the mitochondria becomes arrested. In Candida 107, but not in C. utilis, there is an active ATP:citrate lyase which converts the accumulating citrate, when it passes into the cytosol, into acetyl-CoA and oxaloacetate. The former product is then available for fatty acid biosynthesis which is stimulated by the high ATP concentration within the cells, by the activation of acetyl-CoA carboxylase by citrate and by the provision of NADPH generated as oxaloacetate is converted via malate to pyruvate. Similar characteristics were evident in oleaginous strains of Rhodotorula glutinis and Mucor circinelloides but not in non-oleaginous representatives of these species.


Cell wall-associated 1,4-beta-D-xylanase in Cryptococcus albidus var. aerius: in situ characterization of the activity. 1,4-beta-D-Xylanase (1,4-beta-D-xylan xylanohydrolase; EC 3.2.1.8) has been detected in both cell-free extracts and culture fluids of the yeast Cryptococcus albidus var. aerius grown on glucose as the only carbon source. Mild acid treatment of whole cells proved that the enzyme was extracellularly located. The activity remained almost completely linked to the wall after cell breakage, only being liberated in the presence of salt at high concentration. After release, the enzyme became very unstable and so has been characterized in situ in 'permeabilized' cells. The maximum production took place at the beginning of the exponential growth phase. The optimum pH and temperature for activity were 5.0 and 40 degrees C, respectively. The enzyme degraded xylan and xylo-oligosides by an endo-splitting mechanism giving xylobiose, xylotriose and xylose as the main end-products. Activation energy and kinetic constants for xylan degradation were determined. Several metal ions such as Ag+ and Hg2+ inhibited the enzyme. The possible function of this endo-xylanase in Cr. albidus var. aerius is discussed.


Some characteristics of early appearing isomaltase in intestinal mucosa of suckling rat. Characteristics of early appearing free isomaltase in the soluble fraction were investigated in rat intestinal mucosa. Soluble isomaltase and membrane-bound sucrase-isomaltase complex were prepared from 15-day-old rat intestine. Immunochemical properties, optimal pH and heat sensitivity of soluble isomaltase were compared with those of membrane-bound isomaltase. Optimal pH of free isomaltase in the soluble fraction was lower than that of membrane-bound isomaltase. Soluble and membrane-bound isomaltase showed different sensitivities for temperature. Furthermore, membrane-bound isomaltase in 15-day-old suckling rat intestine gave a single line with antiserum. However, soluble isomaltase gave no precipitin line. From these results, it could be concluded that soluble isomaltase is not derived from the isomaltase moiety of membrane-bound sucrase-isomaltase complex as a result of mechanical fragility and rather it would be lysosomal in origin.


Effect of fluoride on ion exchange, remineralization and acid resistance of surface enamel. In a system of constant ion activities the rates of F- exchange in enamel, under conditions of exchange alone and remineralization, depended on the concentration of F- in solutions. Acid resistance of surface minerals resulted from exchange of F- for OH- in the enamel at pH 7.0 and 4.5. The level of 0.5 mM NaF, compared to 0.05 and 6.0 mM, caused maximum rates of isotopic exchange of 45Ca and maximum acid resistance of enamel. Similarly low levels of F- may be feasible for use in caries prevention in the absence and presence of remineralization.


Sequential hepatic changes during sterigmatocystin-induced carcinogenesis in the rat. The effect of protein deficiency on the hepatotoxicity and carcinogenicity of sterigmatocystin was studied in rats. Sterigmatocystin, 500 ppm/day, fed in a regular diet induced marked toxic effects and there was a high incidence of hepatocellular carcinoma and hyperplastic nodules. The same dosage fed in a protein-deficient diet produced toxic signs followed by a high incidence of death within 27 weeks. Surviving animals showed dysplastic liver cell changes but no tumors were noted. The incidence of hepatocellular carcinoma was 87% in rats receiving 15 ppm of the substance per day for 200 days in a protein-deficient diet. Sequential histologic and histochemical studies revealed that hyperplastic and preneoplastic liver lesions appeared at 28--32 weeks after inseption of the sterigmatocystin-supplemented diet. No cirrhotic changes and only transient fibrosis were noted. The morphogenesis of the sterigmatocystin-induced lesions was compared with that of aflatoxin B1-induced lesions.


Amines and the rat exocrine pancreas: (1). Effects of receptor blockers on turnover of L-dopa. The acinar cells of the exocrine pancreas have the capacity of efficiently take up and metabolize L-dopa. In the present study, the metabolism of L-dopa by the exocrine pancreas of the rat and effects of receptor blockers on the metabolism were studied by fluorescent histochemical and chemical methods. After i.v. administration of L-dopa (50 mg/kg), a large amount of dopamine (DA) was detected in the exocrine pancreas, and in the pancreatic juice large amounts of DA and its metabolites. DA-blockers (haloperidol, sulpiride, and pimozide) and alpha-blockers (phenoxybenzamine, and phentolamine) produced a significant increase in the accumulation of DA after administration of L-dopa. On the other hand, beta-blockers (propranolol, and oxprenolol) were without effects. The excretion of DA into the pancreatic juice appeared to be associated with the secretion of zymogen granules, thus DA serves as an indicator of pancreatic secretory activity, especially of enzyme secretion. Because DA- and alpha-blockers produced an increase in the accumulation of DA, dopaminergic and/or alpha-adrenergic mechanisms probably exist in the exocrine pancreas of the rat and these mechanisms modify the enzyme secretion.


Amines and the rat exocrine pancreas: (2) Effects of receptor blockers on turnover of L-5HTP. The metabolism of L-5HTP by the rat exocrine pancreas, and effects of blockers on the metabolism were studied by fluorescent histochemical and chemical methods. Histochemically, 5-hydroxytryptamine (5-HT) blockers (methyserigide and cyprohepatdine) and dopamine (DA) blockers (haloperidol and sulpiride) produced no apparent changes in fluorescence pictures after injection of L-5HTP. alpha-blockers (phenoxybenzamine and phentolamine) and monoamine oxidase (MAO) inhibitor (iproniazide) produced an increased accumulation of 5-HT fluorescence in the apical regions of acinar cells where the zymogen granules are stored. Chemically, the 5-HT blockers decreased the 5-HT content after injection of L-5HPT. Sulpiride had no effect. Haloperidol decreased the 5-HT content. MAO inhibitor resulted in a vast accumulation of L-dopa: e.g. (1) L-5HTP was more slowly eliminated, and (2) 5-HT blockers produced a decreased content of 5-HT after injection of L-5HTP, in contrast to the finding that DA-blockers produced an incresed content of DA after injection of L-dopa. The mechanism responsible for the differences is discussed in relation to the possible pharmacological effects of L-5HTP and L-dopa on the secretion from the exocrine pancreas of rats.


Antiarrhythmic activity of dextro- and levo-isomers of 5-methyl-8-(2-hydroxy-3-t-butylamino-propoxy) coumarin hydrocholoride (bucumolol), a beta-adrenergic blocking agent, on aconitine-induced atrial and ouabain-induced ventricular arrhythmias in dogs. The beta-blocking, antiarrhythmic, and local anesthetic effects of the racemic mixture and optical isomers of bucumolol, 5-methyl-8-(2-hydroxy-3-t-butylamino-propoxy) coumarin hydrochloride, were studied in dogs, guinea-pigs and frogs. In blocking the positive chrontropic response to isoproterenol, the levo-isomer of bucumolol was about 40 times more potent in dogs, and 270 times in guinea-pigs than its dextro-isomer and twice as effective in both species as the racemic mixture. In frog sciatic nerves bucumolol was 1/10-1/15 as potent in local anesthetic action as propranolol on a weight basis. Dextro- and levo-isomers and racemic bucumolol neither elevated electrical threshold for propagated impulses nor prolonged the effective refractory period of the dog right atrium. The levo-isomer and racemic bucumolol were capable of suppressing aconitine-induced atrial arrhythmia, while the dextro-isomer was less effective. Both isomers and racemic bucumolol were capable of reversing ventricular arrhythmia caused by ouabain, but the effective dose of the levo-isomer was significantly less than that of the dextro-isomer. The results suggest that both specific beta-blocking activity and non-specific membrane action of bucumolol suppressed experimental arrhythmias in dogs produced by aconitine and ouabain.


Mitochondrial ATPase of Zajdela hepatoma. VI. Effect of extramitochondrial ATP and pH on uncoupler-sensitivity of mitochondrial ATPase activity. Coupled Zajdela hepatoma mitochondria were reported to exhibit uncoupler-insensitive ATPase activity. The results of this study show that under specific conditions the ATPase activity of Zajdela hepatoma mitochondria can be stimulated by uncouplers. These conditions include (a) the addition of ATP to the mitochondria before the uncoupler in the ATPase activity assay or (b) elevation of pH (above 8.5) of the ATPase activity assay medium.


Serum enzymes in ovarian carcinoma.

Serum enzymes in ovarian carcinoma. The following enzymatic activities were measured in serum of patients with benign and malignant ovarian tumors before treatment: alkaline and acid phosphatases, aspartyl (AspAT) and alanyl (AlAT) aminotransferases, leucyl (LAP) and alanyl (AAP) aminopeptidases, lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase, cathepsin, alkaline ribonuclease (RNase) and beta-glucuronidase. It was shown that at least three determinations (phosphatases and LAP) are practically useless in a discrimination between the examined groups. RNase in combination with AspAT (AlAT) or RNase with AAP and LDH were found to give the best results as marker enzymes.


Changes in body temperature after administration of amino acids, peptides, dopamine, neuroleptics and related agents. Drugs may alter body temperature by acting on any component of the thermoregulatory system. These components include heat production, heat conservation and heat loss effectors and their efferent pathways, thermosensors and their afferent pathways and neurons within the central nervous system that coordinate thermoregulatory effector activities. A thermostat is often thought to be involved although thermoregulation can be explained by models that do not incorporate a thermostat. An action on a particular component can be assessed by determining the effect of a drug on body temperature over a range of environmental temperatures and by observation and measurement of associated changes in effector activities. A scheme for such assessment is presented along with examples of its use. The study of drug-induced changes in body temperature has expanded greatly within the past decade. The primary purpose of this review is to provide a readily available source of information on interactions between certain drugs and the thermoregulatory system. Extensive tables are presented of body temperature changes after administration of amino acids, peptides, dopamine and related agents, phenothiazine neuroleptics and also phenothiazines that lack neuroleptic activity, butyrophenones, diphenylbutylpiperidines such as pimozide and miscellaneous neuroleptics. The information tabulated includes the species used, route of administration and dose of drugs, the environmental temperature at which the experiments were performed, the number of tests, the direction and magnitude of body temperature change and remarks on the presence of special conditions, such as age or lesions, or on the influence of other drugs, such as antagonists, on the response to the primary drug. Most of the cited literature was published since 1965.


Developmental protein malnutrition: influences on the central nervous system of the rat. Our group has been carrying out interdisciplinary studies on the effects of prenatal and postnatal protein malnutrition on the developing rat brain. Anatomical, physiological, biochemical and behavioral approaches using the same animal model have revealed that protein malnutrition affects the brain at various levels, i.e., (1) anatomical, as revealed by Golgi findings of deranged dendritic trees on analysis of cortical and subcortical areas; (2) physiological, as revealed by delayed sleep pattern maturation, disturbances in seizure thresholds, slowing of sensory cortico-cortical and thalamocortical evoked potentials, and changed power in hippocampal theta activity; (3) biochemical, as revealed by marked increases in biogenic amines dating from birth, as well as modifications in tryptophan metabolism; and (4) behavioral, as revealed by various changes in responses to different kinds of aversive stimulation. Reversal studies have revealed that many changes are permanent and not amenable to nutritional rehabilitation even at birth, which is before the brain growth spurt in the rat. Our paradigm closely mimicks the human condition of low level, chronic protein undernutrition and thus reveals the underlying disturbances due to malnutrition. The dietary reversal studies are attempts at pin-pointing critical brain growth periods, beyond which recovery of functions is not possible.


Parasites in Bulinus senegalensis (Mollusca: Planorbidae) and their detection. Isoelectric focusing studies on enzyme variation between populations of the snail Bulinus senegalensis revealed that parasitic infections in the snails contributed additional bands of enzyme activity, particularly in the glucose phosphate isomerase (GPI) and malate dehydrogenase (MDH) systems. The patterns due to the parasite enzymes were, in most cases, clearly distinct from those of the host and different from each other. Parasites encountered included Schistosoma haematobium, S. bovis, Paramphistomum microbothrium, another amphistome probably belonging to the group which infect amphibians, Echinostoma revolutum, another echinostome (probably Echinoparyphium sp.), strigeids, xiphidiocercariae (these were resolved into 3 distinct types by the enzyme data) and ciliate protozoa. The 7 host populations which were examined showed marked differences in both the prevalence and variety of their parasitic infections and these variations were tentatively related to environmental differences in their respective habitats and to the nature of human contact patterns. Seasonal changes in the parasite fauna were also noted and some of the implications of the parasite load on the host population are briefly mentioned.


Non-specific induction of increased resistance in mice to Trypanosoma congolense and Trypanosoma brucei by immunostimulants. Administration of the immunostimulants Corynebacterium parvum, Bacillus Calmette-Guérin (BCG) or Bordetella pertussis prior to, or at the same time as, challenge with Trypanosoma congolense significantly increased survival times in mice, both of trypano-susceptible (A/J) and trypano-resistant (C57Bl) strains. The increased survival time was associated with significant alterations in parasitaemia, which included lengthening of the pre-patent period, a delay in the time taken to reach the first peak of parasitaemia and a reduction in the level of parasitaemia. Similar results were obtained when these strains of mice were challenged with Trypanosoma brucei following pre-treatment with C. parvum. Thus, by the use of immunostimulants it was possible to reduce the susceptibility of mice to trypanosomiasis and the hope is that this can also be achieved with domestic livestock.


Acid phosphatase activity demonstrated by intact Angiostrongylus cantonesis with special reference to its function. Intact Angiostrongylus cantonensis is able to hydrolyse glucose-phosphate esters, mononucleotides and rho-nitrophenyl phosphate as well as beta-glycerophosphate in vitro. Reciprocal inhibition studies suggest that the hydrolysis of such substrates is due to a non-specific phosphomonoesterase. Molybdate ions, which exert no effect on either the uptake of glucose or the production of lactate, inhibit the hydrolysis of glucose-1-phosphate in the external medium and simultaneously lower the production of lactate by the intact worms in vitro.


Spontaneous generation of adriamycin semiquinone radicals at physiologic pH. Adriamycin semiquinone radicals are spontaneously generated by adriamycin solutions at physiologic pH. Rate of radical formation and equilibrium-state radical yield increase with increasing pH from 7.4 to 8.85. The radicals are oxygen sensitive, but the mechanism of radical formation is oxygen independent and associated with proton removal from the dihydroquinone of adriamycin. The less cardiotoxic and non-mutagenic (Ames test) anthracycline 5-iminodaunorubicin does not form semiquinone radicals spontaneously at physiologic pH.


Effects of propoxyphene, ethoheptazine, and azabicyclane on schedule-controlled responding: attenuation by pentobarbital but not naloxone. The effects of propoxyphene, ethoheptazine, and azabicyclane, alone and in combination with 1 mg/kg naloxone, were studied in pigeons responding under a multiple fixed ratio, fixed interval schedule of food presentation. Low doses of pentobarbital (3, 5.6, and 10 mg/kg) attenuated the rate-decreasing effects produced by 20 mg/kg propoxyphene, ethoheptazine, and azabicyclane. Naloxone antagonized the rate-decreasing effects produced by 10 mg/kg azabicyclane but did not antagonize the rate-decreasing effects of propoxyphene or ethoheptazine.


Atypical antidopaminergic properties of CI-686: a potential antipsychotic agent. The effects of the antipsychotic/antidepressant drug CI-686 on apomorphine- and amphetamine-induced stereotypies, dopamine metabolism, neuroleptic binding, and serum prolactin levels were determined. CI-686 displayed profiles of activity in each of these systems that differs markedly from those of other antipsychotics. CI-686's unique preclinical profile suggests a mechanism of action other than dopamine antagonism which could have implications regarding current thinking on the pathophysiology of schizophrenia.


Relationship of butaperazine blood levels to plasma prolactin in chronic schizophrenic patients. The relationship of plasma prolactin to plasma or red blood cell butaperazine levels was investigated in chronic schizophrenic patients treated with clinical doses of butaperazine, both after a single acute dose of the drug and during regular, twice daily butaperazine administration. Although there was a significant curvilinear relationship between peak plasma butaperazine levels after an acute single oral dose of butaperazine and the maximum prolactin response that we measured, steady-state levels of plasma or red cell butaperazine and plasma prolactin were not related. We conclude that plasma prolactin cannot be used as a substitute for or even as a rough indicator of butaperazine blood levels in chronic schizophrenic patients being treated with clinical doses of butaperazine.


Sulpiride in tardive dyskinesia.

Sulpiride in tardive dyskinesia. Tardive dyskinesia can be suppressed by drugs that block dopaminergic receptors, but often at the cost of a concomitant increase in parkinsonism. Sulpiride (400 -- 2100 mg/day), a selective type-2 dopamine receptor antagonist, was evaluated in a blind, placebo-controlled trial in 11 patients with tardive dyskinesia. It significantly (P less than 0.01) reduced tardive dyskinesia without significantly affecting parkinsonism, although three patients had a increase in preexisting parkinsonian hypokinesia and tremor. During the placebo phase, the tardive dyskinesia and parkinsonian scores returned to the pretreatment values. There was no relationship between either tardive dyskinesia or parkinsonism and eye blinking rates. These results are consistent with the hypothesis that more than one population of dopamine receptors are involved in controlling extrapyramidal function. Sulpiride is an important tool for elucidating both the practical and heuristic aspects of subtypes of dopamine receptors and is a lead in the search for compounds that selectively affect dopaminergic mechanisms.


Characterization of the two major species of slow reacting substance from rat basophilic leukemia cells as glutathionyl thioethers of eicosatetraenoic acids oxygenated at the 5 position. Evidence that peroxy groups are present and important for spasmogenic activity. The most prominent slow reacting substance from rat basophilic leukemia cells (type I) was characterized by radiochemical, chemical and physical methods and shown to contain a C20 unsaturated fatty acid oxygenated at the 5 position and a sulfur containing side chain in thioether linkage at the 6 position. Its spasmogenic action on guinea pig ileal muscle was largely inactivated under reducing conditions which suggested that a peroxy group was present and important for contractile activity. This was supported by ferrous thiocyanate analysis. The peroxy group is almost certainly at the 5 position, probably in the form of a peroxy ester or hydroperoxide. Based on amino acid hydrolysis (0.85 moles of glycine and 0.30 moles of glutamic acid per mole SRS), the sulfur containing side chain is apparently a mixture of glutathione and cysteinyl-glycine, but by chromatography the side chain is predominantly glutathione and the low yield of glutamic acid may be due to complexing of its alpha COOH group in a peroxy ester linkage. The fatty acid moiety has 3 conjugated double bonds, probably at the 7,8, 9,10 and 11,12 positions. Type II SRS, the second major species, differs in that the sulfur containing side chain is linked at the 12 or 13 position and is almost certainly glutathione and in the failure of alkaline borohydride to produce inactivation. These observations strongly implicate the lipoxygenase pathway in slow reacting substance biosynthesis.


[Pulmonary function in dogs submitted to bilateral cervical vagotomy (author's transl)]. A study of the effects of cervical vagotomy on pulmonary function in two groups of dogs was done: the first group had been vagotomized and submitted to spontaneous breathing and the second had been vagotomized and submitted to mechanical ventilation. pH, partial pressure of carbonic acid in arterial blood (PaCO2), partial pressure of oxygen in arterial blood (PaO2), plasmatic bicarbonate (HCO-3), alveolar arterial oxygen difference (P(A-a)O2) while the patients breathed room air and 100% O2 and pulmonary shunting didn't change after vagotomy. There was no difference between these two groups. Hemodynamic parameters like oxygen content difference (C(a-v)O2) between arterial and venous blood, heart hate (FC), mean blood pressure (Pam) and capillary hydrostatic pressure (Pcp) didn't change after vagotomy. None of these parameters were different between the two groups.


Effect of exogenous serotonin on intragastric pH and its influence on serum gastrin levels in rats. The effect of various doses of serotonin on the serum gastrin levels and intragastric pH in rats, was studied. After serotonin administration of 10 mg/kg i.p., a significant increase in serum gastrin levels was noted, as well as a strong increase in the intragastric pH. It was also observed that a lower dose, 5 mg/kg i.p., significantly increased serum gastrin levels, while intragastric pH was not affected, remaining at baseline values throughout the study. These results suggest that the increase observed in serum gastrin levels after administration of exogenous serotonin is not mediated by increase in intragastric pH.


Metabolic and physiological effects of adrenoceptor agonists and antagonists in the horse. In the horse the effect of the adrenergenic agonists adrenaline, phenylephrine and salbutamol on haematocrit, plasma free fatty acid, glycerol and lactate levels were investigated. Effects on heart rate, sweating and muscle tremor were also studied. The effects of administration of the adrenoceptor antagonists propranolol, metoprolol, H35/25 and acepromazine on adrenaline-induced changes were examined. The results obtained with these agonists and antagonists suggest that the lipolysis and hyperglycaemia are mediated via beta-adrenoceptors. It appears that both beta1 and beta2 subtypes are involved. Muscle glycogenolysis, muscle tremor and sweating were mediated via beta2-adrenoceptors. Although salbutamol caused an elevation in haematocrit the other results support the alpha-mediation of adrenaline induced increases in haematocrit.


pH homeostasis during cardiopulmonary resuscitation in critically ill patients. We investigated the effect of repeated administration of sodium bicarbonate on acid-base balance and serum chemistry in a group of patients who developed cardiac arrest. A mixed acidosis persisted throughout the duration of resuscitation in the majority of patients in spite of the large ventilatory volume and multiple doses of bicarbonate they received. However, the repeated administration of bicarbonate prevented a severe fall in serum pH. Our study demonstrated the beneficial role of bicarbonate in the treatment of metabolic acidosis associated with cardiac arrest of prolonged duration. Analysis of our data strongly indicated that the primary factors which determine the serum pH during cardiopulmonary resuscitation are the duration of circulatory arrest, adequacy of ventilation and circulation, pH immediately before arrest, and quantity of bicarbonate administered and its volume of distribution in the various fluid and tissue compartments.


Effect of chlorhexidine on acidogenicity of dental plaque in vivo. In order to evaluate the effect of chlorhexidine on the acidogenicity of dental plaque, pH changes in plaque were measured in situ after sucrose applications. The results showed that a 0.2% chlorhexidine mouthrinse inhibited acid production for a period of 24 h, whereas a 0.05% chlorhexidine rinse showed an inhibitory effect for 4 h. It is suggested that the mechanism involved may be related to the retention of chlorhexidine in the mouth and in plaque providing a bacteriostatic milieu which may be of importance in the observed long-term effect.


Evaluation of phagocytic and bactericidal activities of neutrophil granulocytes. Determination of viable extracellular bacteria by their incorporation of 14C-leucine and 3H-thymidine. A method for evaluation of human neutrophil granulocyte function based on the combined determination of total and extracellular bacteria is described. The total number of surviving bacteria is assessed by the determination of colony forming units (CFU) after hypotonic lysis of granulocytes. Extracellular viable bacteria can be determined by the incorporation of 14C-leucine or 3H-thymidine into bacterial macromolecules since there is a linear relationship between macromolecular synthesis and bacterial number and insignificant amounts of both 14C-leucine and 3H-thymidine is taken up by intracellular viable organisms. This method might be suitable for the differentiation of defects in phagocytosis and intracellular killing of various bacteria.


Does penicillin kill bacteria?.

Does penicillin kill bacteria?. The thesis is presented that the bactericidal action of penicillin and of other inhibitors of cell wall peptidoglycan synthesis, such as vancomycin and cycloserine, is secondary or tertiary to their ability inhibit specific reactions in the assembly of an osmotically protective cell wall. Examples are given of the inhibition of these reactions, which results in inhibition of cell growth (bacteriostatic action) in the absence of either cellular lysis or rapid loss of viability. Thus, in some instances, inhibitory concentrations of these drugs are, in effect, sublethal; this is true, for example, for Streptococcus mutans, a species of bacteria that is part of the normal flora of the oropharynx and that can cause subacute bacterial endocarditis. On the other hand, the damaging effects of the subminimal inhibitory concentrations of penicillin G on Streptococcus faecalis, a species with an active autolytic enzyme system, can be uncovered and converted to a lytic (and lethal) response by partial inhibition of fatty acid synthesis with low concentrations of cerulenin. Some theoretical and practical implications of the occurrence and inhibition of these secondary lethal consequences are discussed.


Toxic interaction between narcotic analgesics and inhibitors of catechol-O-methyltransferase. A lethal synergism between morphine and tropolone, an inhibitor of catechol-O-methyltransferase, was previously noted in adult male Holtzman rats. The present research demonstrates that this phenomenon generalizes across factors of sex, age, strain (Sprague--Dawley, Wistar) and species (Swiss albino mice). Acute toxicity was also significantly increased (1.5--1.9 times) in the case of codeine, methadone, meperidine and levorphanol, but to a lesser extent than for morphine (4.0 times) in the S-D strain. Another COMT inhibitor, 3,5-dihydroxy-4-methoxybenzoic acid, interacted with morphine in S-D rats to an equal degree as did tropolone. Post-treatment with 1 mg/kg of naloxone in rats or naltrexone in mice reduced the high lethality associated with morphine plus tropolone. There was a pronounced lowering of whole brain norepinephrine (NE) level after morphine plus tropolone in Wistar rats with doses of each component that alone caused no change in NE. Brain dopamine (DA) was elevated by tropolone and by its combination with morphine. Each drug alone caused slight lowering of brain serotonin. Enhancement by tropolone of the toxicity of (+)-amphetamine in mice and rats was of similar magnitude as for morphine. The possible role of brain NE and/or DA in the sensitivity to acute toxic effects of opioids in rodents is suggested by these data, as well as a parallel in this regard with amphetamine-type stimulants.


Determination of polyethylene glycols 4000 and 6000 in plasma protein preparations. Polyethylene glycols 4,000 and 6,000 were determined in different plasma protein preparations by the formation of complexes with barium and iodide ions after precipitation of the proteins with perchloric acid. The method, which is fast and reliable, can be used for the determinations of PEG down to 0.005% (w/w) of the total protein content.


[Catalytic properties of soluble 3':5'-AMP-dependent protein kinase of rabbit myocardium]. The results of kinetic studies are presented for two forms of soluble 3':5'-AMP-dependent protein kinase, obtained by DEAE-cellulose and hydroxyl apatite chromography. The pH optimum for both forms of the enzyme is shown to be 6.8-7.0; their activity is the highest at ionic strength of 0.023. Both forms of the enzyme at the above values of pH and ionic strength are thermoinactivated by the exponential law, their inactivation rate constants rise with temperature. Calcium ions in a concentration of 1.10(6)-2.10(-3) M have no effect on their activity.


[Oxidative phoshporylation, proton exchange and volume changes in hepatocyte mitochondria under the effect of exogenous succinate]. Long use of exogenous succinate causes changes in hepatocytes bioenergetic functions in guinea pigs. The character of cation is of essential significance in this situation: sodium succinate produces a moderate stimulating effect, potassium succinate evokes a distinct energizing influence.


[Steroid-transforming enzymes from microorganisms. IX. Affinity chromatographic preparation and studies of the apoenzyme from a 4-en-3-oxosteroid: (acceptor)-1-en-oxidoreductase from Nocardia opaca]. From the flavoenzyme, 4-en-3-oxosteroid: (acceptor)-1-en-oxidoreductase of Nocardia opaca, prosthetic group and apoenzyme were separated quantitatively by means of affinity chromatography in the presence of 2 M (NH4)2 at pH 3.0. Subsequently the apoenzyme was eluted from affinity matrix by 0.01 M phosphate buffer, pH 8.0, whereas under these conditions the intact enzyme could not be eluted. The whole enzyme activity applied could be restored by incubation of the eluted apoenzyme with FAD. The binding strength of the apoenzyme to the immobilized steroid ligand is highly decreased in comparison to the native enzyme and can be interpreted by the action of rest hydrophobicity. That indicates the essential character of FAD for both ligand binding and transformation.


[Histone-nonhistone protein complex reconstruction]. A complex of chromatin proteins composed of five histon fractions and nine nonhiston fractions and stable in 2M NaCl solution was destructed by treating with urea, changing pH and decreasing the NaCl concentration, No reconstruction of the complex was observed after the said factors has been removed and the initial solvent composition restored. Nevertheless the reconstruction was detected after dissociation of the complex by desalting in the presence of DNA. The reconstruction was successful when stimulated both dimers and tetramers, the chromatin protein complex under study cannot be reconstructed by "self-organization", the only way being through the stage of nucleoproteid formation, and is similar in this respect to some other complex protein structures of the cell.


Alcohol dehydrogenase (ADH) in yeasts. II. NAD+-and NADP+-dependent alcohol dehydrogenases in Saccharomycopsis lipolytica. In Sm. lipolytica one NAD+-dependent and three NADP+-dependent alcohol dehydrogenases are detectable by polyacrylamide gelelectrophoresis. The NAD+-dependent ADH (ADH I), with a molecular weight of 240,000 daltons, reacts more intensively with long-chain alcohols (octanol) than with short-chain alcohols (methanol, ethanol). The ADH I is not or only minimally subject to glucose repression. Besides the ADH I band no additional inducible NAD+-dependent ADH band is gel-electrophoretically detectable during growth of yeast cells in medium containing ethanol or paraffin. The ADH I band is very probably formed by two ADH enzymes with the same electrophoretic mobility. The NADP+-dependent alcohol dehydrogenases (ADH II--IV) react with methanol, ethanol and octanol with different intensity. In polyacrylamide gradients two bands of NADP+-dependent ADH are detectable: one with a molecular weight of 70,000 daltons and the other with 120,000 daltons. The occurrence of the three NADP+-dependent alcohol dehydrogenases is regulated by the carbon source of the medium. Sm. lipolytica shows a high tolerance against allylalcohol. Resistant mutants can be isolated only at concentrations of 1 M allylalcohol in the medium. All isolates of allylalcohol-resistant mutants show identical growth in medium containing ethanol as the wild type strain.


[Examinations on the pathomechanism of severe cardio-pulmonary hazards under i.v. tocolytic treatment with beta-adrenergic drugs (author's transl)]. Lungs edemas in five pregnant women under i.v. tocolytic treatment with betaadrenergic drugs between the 28.--33. week of gestation are reported. Additionally, in all cases Betamethasone was given to prevent fetal lungs dysmaturity. Within less than 72 hours the occurrence of lungs edema was observed when both drugs were given simultaneously. These findings are in agreement with those of other authors, who observed similar symptoms in as well clinically healthy pregnant women under the same therapy. As the underlying pathomechanism the coincidence of the following factors are discussed: 1. An increased pressure in the pulmonary circulatory system as found in current, not yet published heart catheter investigations. 2. A reduced colloidosmotic pressure caused by Beta-methasone.


[Forceps deliveries in epidural or general anesthesia (author's transl)]. It was the aim the present study to evaluate maternal morbidity and fetal outcome of 832 pelvic outlet forceps deliveries during the years 1974 through 1977 with special reference to the method of anesthesia (Epidural anesthesia (EA) with Bupivacain 0,5% or general anesthesia (GA) with Methohexital-Na and Succinylcholin). Maternal vaginal and/or cervical injuries were seen significantly (p less than 0,001) more often in the forceps group than in spontaneous, vacuum, or breech deliveries. Vaginal injuries occurred less often (p less than 0,01) in the EA- than in the GA-group. Forceps deliveries were not associated with increased maternal morbidity in fetuses weighing 3750gr or more. The cardiotocography scores (Hammacher et al. 1974) of the 30 minutes preceeding delivery (final scores) were better with EA than with GA (p less than 0,01). Arterial cord pH and 1 minute Apgar values were similar in both groups. The same was true for fetuses with a birthweight of 3750gr or more. The time for cervical dilatation from 7 to 10 cm and duration of the second stage of labor did not influence maternal morbidity or fetal outcome, regardless of the method of anesthesia.


[Phenylalanine-4-hydroxylase and tyrosine aminotransferase activities in rat liver tissue under different vitamin E content in the organism]. Enzymic activity of tyrosine aminotransferase is unchanged in subcellular fractions of the liver tissue of control, E-hypovitaminotic rats and of the same animals subjected to a single vitamin E administration. Enzymic activity of phenyl alanine-4-hydroxylase in the supernatant fraction determined in the incubation medium without biopterin in animals with E-hypovitaminosis is 39% lower than in the control rats. The coefficient of phenyl alanine-4-hydroxylase activation with biopterin in vitro in E-hypovitaminotic animals is thrice as high as that in the control rats. A single vitamin E administration produces a 1.8-fold decrease in the activation coefficient. The data obtained give reason to suggest the possible influence to vitamin E on the biopterin level in the liver.


Isolation and some properties of an acid protease from Fasciola hepatica. A new protease, detected in an extract of Fasciola hepatica, was isolated and partly purified. The pH optimum for the cleavage of denaturated haemoglobin by the enzyme is pH 3.0. This proteolytic activity is inhibited by diazoacetylnorleucine methyl ester, pepstatin, the pepsin inhibitor from Ascaris suum, and phenylalanine. The cathepsin D inhibitor from potatoes, EDTA, mercaptoethanol and the inorganic salts tested have no inhibitory effect. The cleavage of the B-chain of oxidized insulin by enzyme was studied and compared with the digestion of the same substrate by chicken and pig pepsin. The protease from Fasciola hepatica belongs to the carboxyl group of proteases and probably plays an important role in helminth nutrition.


Production of L-asparaginase by Streptomyces karnatakensis and Streptomyces venezuelae. Production of L-asparaginase by two soil isolates, identified as S. karnatakensis and S. venezuelae, was investigated under different environmental and nutritional conditions. The presence of carbon sources, other than starch, in the growth medium or amino acids, other than L-asparagine-inhibited the enzyme biosynthesis. L-aspartic inhibited growth and enzyme production, due to a feedback mechanism, and/or lowering the pH value. Both organisms were stimulated to produce more enzyme with increasing concentrations of starch and L-asparagine, however, the optimum starch and L-asparagine concentration depended on the tolerance of the organism to low and high pH, respectively. Aeration stimulated growth, but not enzyme production, and both organisms produced more enzyme in static cultures than in shaken cultures.


The characterization of cellulase from Paecilomyces fusisporus Saksena. The fractionation of cellulase from Paecilomyces fusisporus Saksena on DEAE-Sephadex (A-5O) resulted in separation of beta-glucosidase, cellulase (on cellulose powder), and CM cellulase activity. Cellulase activity was associated with some CM cellulase activity, whereas the latter was independent of the former.


Cellulolytic activity of some Pythium species. The cellulolytic activity of Pythium ultimum and P. oligandrum was investigated in vitro during the mycelium growth. It was found that under a constant temperature of 24 degrees C the cellulolytic activity was changed according to the pH of the medium. The optimum of cellulolytic activity in P. ultimum was recorded at neutral pH, whereas that in P. oligandrum in acid medium. The enzymatic production depended on the age of culture as well.


Steps of thiosulfate oxidation by Thiobacillus thioparus and Th. coproliticus. Chromatographic analysis was used for studying steps of thiosulfate oxidation by Thiobacillus thioparus and Th. coproliticus strains, isolated from Egyptian soil. Th. thioparus strains oxidized S2O3 to SO4 with the formation of polythionates, tetrathionate in particular, during the course of oxidation. However, strains differed in other polythionates formed. Th. coproliticus, in turn, oxidized S2O3 to SO4 without polythionate formation. However, H2S was the only intermediate compound detected during oxidation.


[Hydrotherapy pools, microbiological and chemical results (author's transl)]. The results of microbiological and chemical examinations of water samples collected from hydrotherapy pools for non incontinent patients proved that the officially agreed parameters for the surveillance of public swimming pools (Osterr. Bäderhygiene-Gesetz and Verordnung) have to be extended to meet the requirements of epidemiology. Similarly the requirements for treatment and operating conditions of those waters should be more rigorous. Minimal requirements for the treatment of water in hydrotherapy pools are recommended. The results of the study indicated also that the microbicide action of chlorine (chlorine gas, hypochlorite solution) was superior to those of DIHALO (chlorine-bromine-hydantoine). Suggestions are made for proper handling of waters which would lose their therapeutical potencies by any kind of treatment.


NADPH production in the oxidative pentose phosphate pathway as source of reducing equivalents in glycolysis of human red cells in vitro. Studies have been carried out on human erythrocytes in vitro to clarify the deficit of pyruvate formation under conditions when 2,3 DPG is degraded. The results lead to the conclusion that there exist a cross connection between the glycolytic and the oxidative pentose phosphate pathway which is mediated by the NADP/NADPH couple. NADPH serves as additional reducing equivalent in the reaction of the LDH. In the absence of glucose the pool of the metabolites of the pentose phosphate pathway is able to supply glucose-6-phosphate for the production of NADPH by recombination. The reaction of NADPH at the LDH is probably of significance under in vivo conditions.


Influence of the temperature on the MDH activity of cytosol and submitochondrial fractions of rat liver in different buffers. The temperature optimum and the temperature stability of malate dehydrogenase (MDH) in cytosol and submitochondrial fractions (matrix and inner membranes) in the buffers: Tris-HCl, potassium-phosphate and HEPEC-NaOH at pH 7.5, are determined. The temperature optimum of MDH (EC 1.1.1.37) in cytosol does not depend on the type of the buffer used, unlike the optimum in mitochondrial preparations. The mitochondrial enzyme is less resistant to temperature compared to the cytosol preparation. The effect of all three buffers used on the native and on the temperature-changed enzyme in the preparations in preserved the same. However, the presence of the buffer during the temperature action on the MDH-activity in the preparations changes the temperature stability of the enzyme. Changes are assumed in the conformation of the mitochondrial enzyme under the effect of phosphate ions, leading to activation of the native enzyme and to a rise in its temperature stability.


Quantitative changes in regional cerebral blood flow of rats induced by alpha- and beta-adrenergic stimulants. Cerebral blood flow was measured with the 14C-ethanol technique in 8 regions (frontal, parieto-temporal and occipital cortex, caudate nucleus, thalamus, cerebellum, mesencephalon, and pons) of rats. The highest flow values (83-89.5 ml/100 g/min) were found in cortical areas, whereas pons had the lowest flow (48 ml/100 g/min). Intravenous infusion of noradrenaline or adrenaline markedly reduced rCBF (by 22-48% of control levels) in all regions except thalamus, mesencephalon, and pons. The noradrenaline-induced reduction was blocked, and the effect of adrenaline reversed, after pretreatment with the alpha-receptor antagonist, phentolamine. Isoprenaline infusion markedly augmented rCBF in thalamus, mesencephalon, pons, and also in the caudate nucleus. The response was reduced by the beta-receptor antagonist, propranolol. The experiments show the presence and heterogenous distribution in the cerebrovascular bed of slpha- and beta-adrenoceptors that can be activated by sympathomimetics given systematically. If noradrenaline was allowed to pass the blood-brain barrier after osmotic opening with urea, an increased regional flow was obtained, probably due to a mechanism where the vasodilator effect secondary to activation of cerebral metabolism predominated over the direct vasoconstrictor effect of the amine.


The effect of propranolol on the serotonin concentration in the portal plasma after vagal nerve stimulation in the cat. Efferent cervical vagal nerve stimulation in the cat caused a marked increase of the portal plasma 5-HT concentration. This increase was more than two-fold within 15 min of stimulation. After cessation of stimulation portal plasma 5-HT returned to basal levels within 10 min. Treatment with the beta-adrenoceptor antagonist propranolol, in various doses (0.1-2 mg/kg b.wt.), did not abolish but significantly reduced the response to vagal stimulation, particularly during the final part of the stimulation period. The results confirm the existence of a beta-adrenoceptor-mediated release of 5-HT, but also suggest that other mechanisms for 5-HT release may be involved in the response on vagal nerve stimulation.