Effect of L-arginine on Na+,K+-ATPase activity in rat aorta endothelium

The effect of L-arginine on the Na⁺,K⁺-ATPase activity in rat aorta endothelium was studied at its physiological concentrations in the range of 10^–6-10^–3 M. The enzyme activity was 35.5% increased by low concentrations of L-arginine (10^–5 M) and its activity was 32.3-37.1% decreased at the L-arginine concentrations of 10^–4-10^–3 M. A similar inhibition (by 34.5-42.8%) was also found in the presence of a NO-donor nitroglycerol (10^–4-10^–3 M). An optical isomer of L-arginine, D-arginine, at the concentrations of 10^–5 M also increased the enzyme activity by 37.1%, but its inhibiting effect was much less pronounced and was 15.7% at the D-arginine concentration of 10^–3 M. An inhibitor of NO-synthase, L-NAME (N^G-nitroarginine, methyl ester), failed to inhibit Na⁺,K⁺-ATPase. However, the presence of L-NAME abolished the inhibition of Na⁺,K⁺-ATPase by high concentrations of L-arginine. Thus, the effect of L-arginine on the endothelial Na⁺-pump depended on its concentration, and it is suggested that the enzyme inhibition by high concentrations of L-arginine should be associated with activation of the endogenous synthesis of NO.     

Arsenite as an Inducer of Lipid Peroxidation in Saccharomyces cerevisiae Cells

The ability of sodium arsenite at concentrations of 10^–2, 10^–4, and 10^–6 M to induce lipid peroxidation in Saccharomyces cerevisiae cells was studied. Arsenite at the concentrations 10^–2 and 10^–4 M enhanced lipid peroxidation and inhibited the growth of yeast cells. Enhanced lipid peroxidation likely induced oxidative damage to various cellular structures, which led to suppression of the metabolic activity of cells. Arsenite at the concentration 10^–6 M did not activate lipid peroxidation in cells. All of the tested arsenite concentrations inhibited the activity of -ketoglutarate dehydrogenase and pyruvate dehydrogenase in cells. The inference is made that the toxicity of arsenite may be related to its stimulating effect on intracellular lipid peroxidation.     

Production of microsclerotia of the fungal entomopathogen Metarhizium anisopliae and their potential for use as a biocontrol agent for soil-inhabiting insects

Microsclerotia (MS), overwintering structures produced by many plant pathogenic fungi, have not been described for Metarhizium anisopliae. Three strains of M. anisopliae – F52, TM109, and MA1200 – formed MS in shake flask cultures using media with varying carbon concentrations and carbon-to-nitrogen (C:N) ratios. Under the conditions of this study, all strains produced MS, compact hyphal aggregates that become pigmented with culture age, in addition to more typical blastospores and mycelia. While all strains formed desiccation tolerant MS, highest concentrations (2.7–2.9 × 10⁸ L⁻¹ liquid medium) were produced in rich media with C:N ratios of 30:1 and 50:1 by strain F52. All three strains of M. anisopliae produced similar biomass concentrations when media and growth time were compared. Strain MA1200 produced higher concentrations of blastospores than the other two strains of M. anisopliae with highest blastospore concentrations (1.6 and 4.2 × 10⁸ blastospores ml⁻¹ on days 4 and 8, respectively) in media with the highest carbon and nitrogen concentrations. Microsclerotial preparations of M. anisopliae containing diatomaceous earth survived air-drying (to <5 % moisture) with no significant loss in viability. Rehydration and incubation of air-dried MS granules on water agar plates resulted in hyphal germination and sporogenic germination to produce high concentrations of conidia. Bioassays using soil-incorporated, air-dried MS preparations resulted in significant infection and mortality in larvae of the sugar beet root maggot, Tetanops myopaeformis. This is the first report of the production of sclerotial bodies by M. anisopliae and provides a novel approach for the control of soil-dwelling insects with this entomopathogenic fungus.     

Porphyrin synthesis in primary nervous tissue cultures from 10−3 M delta-aminolaevulinic acid in the presence of melatonin and neuropeptides

Of the primary neuronal tissue cultures (glia cell, neuronal cells, mixed and retina cultures), the neuronal cells of (cells + medium) display the highest total porphyrin production from 10^–3 M delta-aminolaevulinic acid (ALA). In the presence of 10^–3–10^–6 M melatonin, the quantity of total prophyrins produced by the neuronal cultures decreases in inverse proportion to the concentration. Oxytocin, lysine-vasopressin, CCK-8 sulphate ester and des-Tyr-gamma-endorphin in concentrations of 10^–5 and 10^–6 M block the porphyrin synthesis of the glia cells and display different effects on that of the neuronal cells. They enhance the total porphyrin synthesis of the cell cultures, with the exception of 10^–5 M des-Tyr-gamma-endorphin, which exerts an inhibitory effect on the glia cells.     

Regulation of Adenylyl Cyclase Signaling System in Cell Cultures of Infusoria Dileptus anser and Tetrahymena pyriformis by Peptides of Insulin Superfamily

Hormone-sensitive adenylyl cyclase signaling system (ACS) provides transduction of a wide spectrum of hormonal signals in cells of the higher eucaryotes. At the same time, ACS in the lower eucaryotes at present is practically not studied. We studied regulatory effects on ACS of the infusoria Dileptus anser and Tetrahymena pyriformis of peptide hormones of the higher eukaryotes—insulin, IGF-1, and relaxin, whose action on ACS of the higher eucaryotes was the subject of our earlier studies. The action of these hormones at concentrations of 10^–10–10^–8 M on the AC activity in infusoria had clearly stimulating character, the dose–effect curves being of a bell-shaped form with a maximum of the stimulating effect of the hormones at concentrations of 10^–9–10^–8 M. the shape of the curves and the value of the stimulating effect of the peptide hormones depended substantially on the level of the AC basal activity in homogenates of infusorian cell cultures. All the hormones (10^–8 M) stimulated GTP-binding activity of G-proteins. It was shown by the example of relaxin that its stimulating effect on GTP-binding in infusorian cells was dose-dependent and increased in the range of hormone concentrations from 10^–10 to 10^–8 M to reach its maximum at concentrations of 10^–8–10^–7 M. In the presence of suramin, an inhibitor of heterotrimeric G-proteins, the stimulating effects of the hormones on the GTP-binding and the AC activity decreased essentially or were absent completely. This indicates that the heterotrimeric G-proteins are ones of components of the signaling cascade that mediates regulatory effects of the hormones of the insulin group on the AC activity in infusorian cell cultures. Based on the obtained data, it is suggested that the basic molecular mechanisms of regulation of ACS by insulin and the related peptides that are similar to those found in the higher vertebrates already begin to be formed as early as at the level of the lower eucaryotes.     

Impact of carbon and nitrogen nutrition on the quality, yield and composition of blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus

The impact of growing cultures of Paecilomyces fumosoroseus in liquid media containing four combinations of glucose and casamino acids (8 g l^–1 or 80 g l^–1 glucose, 1.32 g l^–1 or 13.2 g l^–1 casamino acids) was evaluated, based on blastospore production, germination rate, viability after freeze-drying and short-term storage stability. When blastospores were produced using a high casamino acid concentration, blastospore yields and germination rates were significantly higher (13.2–18.5×10⁷ blastospores ml^–1, 50–60% germination after 4 h), compared to cultures grown in media containing lower casamino acid concentrations (0.4–2.3×10⁷ blastospores ml^–1, 10–20% germination after 4 h). Chemical analyses of blastospore composition showed that accelerated blastospore germination may be related to increased proteinaceous reserves rather than to glycogen or lipid accumulation. Tolerance to freeze-drying by blastospores suspended in spent medium was enhanced by a high initial casamino acid concentration in the culture medium (75% survival) and by the residual glucose concentrations in the spent medium. Under the conditions of this study, the storage stability of blastospores of P. fumosoroseus was unaffected by the nutritional condition in which they were produced.     

Possible Participation of Serotonin in the Peripheral Link of the Defensive Reflex in the Mollusc Lymnaea stagnalis

The contractions of the dorsal longitudinal muscle of the mollusc Lymnaea stagnalis L., which are evoked by electric stimulation of n. cervicalis inferior were studied. It has been shown that an increase of magnesium ion concentration in saline to 10–15 mM decreases reversibly amplitude of the evoked contractions. Application of serotonin produced a dual effect: at concentrations of 2 × 10^–5–10^–6 M, it enhanced muscle contractions, whereas at concentrations above 10-5 M, on the contrary, decreased them. The inhibitory effect of the serotonin antagonist mianserin on the evoked contraction amplitude increased with elevation of its concentrations in the studied range (from 10^–5 to 10^–3 M). The enhancing effect of serotonin on muscle contractions was blocked either by previous mianserin application or its application on the background of the already acting serotonin. A participation of serotoninergic mechanisms in the control of the contractile function of the studied muscle is suggested.     

Calcium increases the yield of somatic embryos in carrot embryogenic suspension cultures

An upward shift in the concentration of calcium present in the medium during somatic embryogenesis increased the number of embryos produced approximately two-fold. This was observed when embryogenic suspension cells grown in 2,4-D medium with the normal calcium concentration of 10^–3 M were transferred to hormone-free medium containing 10^–2 M calcium and when embryogenic suspension cells grown in 2,4-D medium containing 10^–4 M calcium were transferred to hormone-free medium with 10^–3 M calcium. At calcium concentrations between 6·10^–3 and 10^–2 M globular stage somatic embryos were found in cultures supplemented with 2·10^–6 M of 2,4-D indicating that elevated calcium counteracts the inhibitory effect of 2,4-D on somatic embryogenesis. No qualitative changes were found in the pattern of extracellular polypeptides as a result of growth and embryogenesis in media with different calcium concentrations.     

Effects of L-alanine and L-alanine peptides on the chemotaxis of tetrahymena: Evolutionary conclusions

L-alanine and its peptides (L-Ala-2–6) do not attract or repulse Tetrahymena in a 10^–8M concentration. In 10^–10M concentration there is a consistent repellent effect. Twenty four hours after L-alanine or L-alanine-peptides' pretreatment (imprinting) the progeny generation of the cells react differently to the same materials. L-Alanine, L-alanine penta- and hexapeptide in both concentrations are chemoattractant, while L-alanine tetrapeptide is repellent. L-Alanine dipeptide is inert in 10^–10M and repellent at 10^–8M concentrations, while L-alanine tripeptide is strongly repellent at 10^–10M and attractant at 10^–8M concentrations. This means, that the first encounter (imprinting) with an exogeneous amino acid or peptide is decisive to the later reaction of the protozoan cell. The chain length is important in the imprinting, however the reaction is not consistent. The experiments call the attention to the significance of imprinting in the receptor and hormone evolution.     

The effect of phosphate concentration on growth and agar content of Gelidium robustum (Gelidiaceae,Rhodophyta) in culture

Phosphate concentration of the growth medium was found to affect the growth rate and agar yield of a clone of Gelidium robustum grown in the laboratory. To study differences in growth we used phosphate concentrations from 0 to 200 µM. To determine the effect of phosphate on agar yield and its properties we used concentrations from 0 to 20 µM. Growth rates generally increased with increasing phosphate concentration, with the highest growth rate (21% d^–1) obtained at 150 µM. Agar yield as percentage of fresh weight was highest (10%) in the algae grown with low phosphate concentrations, but agar yield as percentage of dry weight was highest(43%) at 20 µM of phosphate. Gel strength increased with phosphate concentration with a maximum of 160 g m^–2 for 0.75% gels for the cultures at 20 µM. Melting and gelling temperatures of the gels were also affected by phosphate concentration of the growth medium. Starch yield was highest in algae grown in low phosphate concentrations.     

Nickel toxicity inPseudomonas tabaci: Single cell and bulk sample analysis of bacteria cultured at high cation levels

Summary The growth ofPseudomonas tabaci in nutrient medium is partially inhibited in the presence of 10^–3 M added nickel (threshold toxic concentration), with complete inhibition at 10^–2 M nickel—but no effect at 10^–4 and 10^–5 M. Toxic levels of nickel affect both cell division and cell viability.Spectrophotometric determination of intracellular levels of nickel at different external concentrations showed that the highest internal values occurred with cells cultured in 10^–4M (non-toxic) nickel medium rather than in 10^–3 (toxic) medium—suggesting that nickel toxicity does not primarily relate to internal concentration.X-ray microanalysis, carried out on whole bacterial cells, showed that toxic levels of nickel in the external medium resulted in a range of ionic changes in the cell, including a decrease in the level of K (K efflux) and an increase in the levels of Mn, Fe, Ni, and Cu (transition metal cation influx). Other changes induced by nickel toxicity included an increase in the level of soluble S (with a decrease in insoluble S), an increased cell dry mass, and a conspicuous plasmolysis—which was observed both in whole cells and in ultrathin sections.The results obtained support a primary toxic effect of nickel at the cell surface—possibly directly affecting the transport activity of the plasmalemma. The resulting changes, particularly involving the influx of a range of cations, may lead to secondary toxic activities affecting the whole metabolism, leading to plasmolysis and inhibition of division.     

Effect of vanadate on brain protein phosphorylation

The effect of vanadate on the phosphorylation of synaptosomal membrane proteins prepared from rat cerebral cortex was studied. Vanadate concentrations of 10^–6, 10^–5, and 10^–4 M increased the endogenous phosphorylation activity by 25%, 37%, and 75%, respectively. Increasing the ATP concentration in the assay medium from 50 to 500 M did not influence the above effect. A commercial preparation of the purified protein kinase was stimulated 40% by 10^–3 M vanadate. Calcium-calmodulin dependent activity was stimulated only 20% by 10^–5 M vanadate. The effect was not enhanced by further increasing vanadate concentration. Addition of calcium ions (above 50 M) suppressed the vanadate effect, while an inhibition was observed at high Ca²⁺ concentration (2.5 mM). Below 50 M calcium ions stimulated phosphorylation activity in the absence of vanadate and did not affect the stimulatory action of vanadate. Cyclic AMP-dependent endogenous phosphorylation was also stimulated by vanadate. Activation by cAMP could not be observed at vanadate concentrations above 10^–6 M. Possible mechanisms of the vanadate effect are discussed.     

Influence of salinity and temperature on the germination of Kochia scoparia

Kochia scoparia is one of the most common annual halophytes foundin the Great Basin. Seeds were collected from a population growing in asalt playa at Faust, Utah and were germinated at 5 temperature regimes(12 h night/12 h day, 5–15 ^°C, 10–20 ^°C, 15–25 ^°C,20–30 ^°C and 25–35 ^°C) and 6 salinities (0, 200, 400,600, 800 and 1000 mM NaCl) to determine optimal conditions forgermination and recovery of germination from saline conditions after beingtransferred to distilled water. Maximum germination occurred in distilledwater, and an increase in NaCl concentration progressively inhibited seedgermination. Few seeds germinated at 1000 mM NaCl. A temperatureregime of 25 ^°C night and 35 ^°C day yielded maximumgermination. Cooler temperature 5–15 ^°C significantly inhibited seedgermination. Rate of germination decreased with increase in salinity.Germination rate was highest at 25–35 ^°C and lowest at5–15 ^°C. Seeds were transferred from salt solutions to distilled waterafter 20 days and those from high salinities recovered quickly at warmertemperature regimes. Final recovery germination percentages in high salttreatments were high, indicating that exposure to high concentration ofNaCl did not inhibit germination permanently.     

Effect of Insulin and Insulin-Like Growth Factor 1 (IGF-1) on Activity of Glycogen Synthase in Skeletal Muscles of the Lamprey Lampetra fluviatilis

Activity of glycogen synthase (GS) in the muscle tissue of the lamprey Lampetra fluviatilis is studied in dynamics of the pre-spawning period as well as under effects of insulin and IGF-1. It is shown that GS exists in the muscles in two forms, the active (I-form) and inactive (D-form), the I-form prevailing during all studied time periods. With approaching the spawning, the GS activity fell 1.5–2 times due to a decrease of the I-form activity. From October to January, both insulin and IGF-1 stimulated GS at concentrations of 10^–10–10^–8 M and 10^–9–10^–8 M, respectively. The maximally effective concentrations (10^–9 M insulin, 10^–8 M IGF-1) produced a 2.5–3-fold rise of the I-form activity of GS at the period from October to December. In January the stimulating effect of these peptides decreased. In March the GS was insensitive both to insulin and to IGF-1. The obtained data indicate participation of insulin and IGF-1 in regulation of glycogen synthesis in lamprey skeletal muscles, the ability of IGF-1 to stimulate the enzyme activity being shown in the lower vertebrates for the first time. It is concluded that IGF-1 takes part in regulation of the carbohydrate metabolism already at early stages of evolution of vertebrates.     

Tryptamine as an endogenous modulator of neuronal sensitivity to serotonin

Research was performed on sensory ganglia isolated from adult rats using intracellular techniques for recording membrane potential and by measuring resistance at the membrane of individual units. It was found that tryptamine at high concentrations manifests serotoninlike activity, but, at concentrations not affecting potential and resistance at the neuronal membrane, either reinforces (at a concentration of 10^–7 M) or attenuates (at 10^–5 M) serotonin (5-HT) effects mediated by type 1A (but not type 2) 5-HT receptors. 5-HT-modulated effects were produced by tryptamine-induced changes in 5-HT sensitivity at the neuronal membrane and remained unchanged by maximum level of this transmitter. Harmane acts similarly to tryptamine, although harmane derivatives (C-412 and C-506 respectively) produce either potentiation or inhibition of 5-HT_1A over the entire concentration range used (of 10^–7 M-10^–5 M). The allosteric nature of 5-HT-modulation by tryptamine and harmane is discussed.M. Gor'kii Medical Institute, Donetsk, Ukrainian SSR. Translated from Neirofiziologiya, Vol. 21, No. 3, pp. 352–357, May–June, 1989.     

Localization of sulfate reduction in planted and unplanted rice field soil

Rates of in situ sulfate reduction (SRR) in planted and unplanted rice fieldsoil were measured by the ³⁵SO^2– ₄-radiotracermethod using soil microcosms. The concentration of ³⁵SO^2– ₄ decreased exponentially with time.However, time course experiments indicated that incubation times of10–30 min were appropriate for measurements of SRRusing a single time point in routine assays. Unplanted microcosmsshowed high SRR of 177 nmol cm^-3 d^-1 inthe uppermost centimeter where average sulfate concentrations were<33 µM. Fine scaled measurements (1 mmresolution) localized highest SRR (<100 nmol cm^-3d^-1) at the oxic/anoxic interface at 2–5 mmdepth. In planted rice field soil, SRR of <310 nmolcm^-3 d^-1 were observed at 0–2cm depth. Sulfate reduction rates were determined at a millimeter-scalewith distance to a two dimensional root compartment. The SRR was highestat 0–1.5 mm distance to the root layer with rates up to500 nmol cm^-3 d^-1, indicating a highstimulation potential of the rice roots. SRR seemed to be mainlydependent on the in situ sulfate porewater concentrations. At thesoil surface of unplanted microcosms sulfate concentration decreasedfrom <150 µM to <10 µM within the first 8 mm of depth. In planted microcosmssulfate concentration varied from 87–99 µMsulfate at the 0–3 mm distance to the root layer to48–62 µM sulfate at a root distance>4 mm from the roots.The depth distribution of inorganic sulfur compounds was determinedfor planted and unplanted rice field soil. Sulfate, acid volatilesulfide (AVS) and chromium reducible sulfide (CRS) were up to 20 foldhigher in planted than in unplanted microcosms. CRS was the majorinsoluble sulfur fraction with concentrations >1.7µmol cm^-3. Organic sulfur accounted for25–46% of the total sulfurpresent (269 µg/g dw) in an unplanted microcosm.The biogeochemical role of sulfate reduction forshort-term accumulation of inorganic sulfur compounds(FeS, FeS_{_2} and S°) in rice soil wasdetermined in a time course experiment with incubationperiods of 5, 10, 20, 30 and 60 min. The relativedistribution of CRS and AVS formation showedlittle depth dependence, whereas the formation of³⁵S° seemed to be the highest in themore oxidized upper soil layers and near the root surface.AV³⁵S was the first major product of sulfatereduction after 20–30 min, whereas CR³⁵Swas formed, as AV³⁵S and ³⁵S°decreased, at longer incubation periods of >30 min.     

Effect of urethane (ethyl carbamate) on the mitosis of phytohaemagglutinin stimulated human lymphocytesin vitro

Summary The action of urethane at concentrations between 10^–1 and 10^–8 M on the rate of mitosis of phytohaemagglutinin-stimulated human lymphocytes in culture has been investigated. At 10^–1M urethane is cytotoxic, at all other concentrations tested it has some inhibitory activity except at 10^–5M and 10^–6M where it stimulates mitosis. These results are discussed in relation to previously reported data and compared with the action of hydroxyurea in the same system.I wish to thank the Smith Kline and French Foundation for financial support and Mrs. B. Haynes and Miss C. Watson for technical assistance.     

Multiplicity of chemical mechanisms of regulation of muscle contractions in <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> L.

Effects of acetylcholine, octopamine, and their antagonists, as well as of glutamic acid were studied on contractions of dorsal longitudinal muscle of the mollusc Lymnaea stagnalis L., evoked by electrical stimulation of n. cervicalis inferior. Acetylcholine and octopamine increased amplitude of contractions evoked by applications at concentrations about 10^–8 M and decreased at concentrations higher than 10^–7 M. Glutamic acid produced only inhibitory effect on the contraction amplitude, which appeared at concentrations beginning from 10^–9 M and higher. The acetylcholine antagonists atropine and d-tubocurarine also decreased amplitude of evoked contractions. Their blocking effect rose with increase of their concentrations in the range from 10^–9 M to 10^–5 M. Specificity of the effect of the studied substances was established in experiments with a combined application of the transmitters and their antagonists. The obtained results indicate multiplicity of chemical mechanisms of regulation of contractions of the dorsal longitudinal muscle in L. stagnalis.Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 41, No. 1, 2005, pp. 44–50.Original Russian Text Copyright © 2005 by Kononenko, Zhukov.     

Emulsifying behaviour and rheological properties of the extracellular polysaccharide produced by Pseudomonas oleovorans grown on glycerol byproduct

The functional properties of a novel extracellular polysaccharide (EPS) produced by Pseudomonas oleovorans grown on glycerol byproduct, generated by the biodiesel industry, were investigated. The EPS is a high molecular weight (4.6 × 10⁶) heteropolysaccharide, composed by neutral sugars (galactose, 68%; mannose, 17%; glucose, 13%; rhamnose, 2%; fucose, 4%) and acyl groups (3.04%). This biopolymer has pseudoplastic fluid behaviour in aqueous media. The apparent viscosity was stable for the pH range 2.9–7.1 and NaCl concentrations up to 1.0 M. Though the apparent viscosity decreased at high temperatures, at alkaline conditions and at NaCl concentrations of 2.0 M, pseudoplastic fluid behaviour was retained. The EPS was capable of stabilizing water emulsions with several hydrophobic compounds, including hydrocarbons, vegetable and mineral oils. It retained its emulsifying activity during exposure to wide temperature (30–50 °C) and pH (2–12) variations, as well as to the presence of NaCl at concentrations as high as 2.0 M.     

A comparative study of the effect of various detergents on the structure and function of sarcoplasmic reticulum vesicles

Summary The effects produced by the detergents Triton X-100, sodium dodecylsulphate and sodium cholate on sarcoplasmic reticulum vesicles have been comparatively studied. In all cases, maximal effects are found 5 min after detergent addition. Triton X-100 and SDS are approximately ten times more effective than cholate in protein and phospholipid solubilization. Both Triton X-100 and SDS maintain Ca⁺⁺ accumulation in SR vesicles at detergent concentrations below 10^–3 M; higher concentrations cause a strong inhibition. On the other hand, cholate produces a gradual inhibition of Ca⁺⁺ accumulation in the concentration range between 10^–4 M and 2.5 × 10^–2 M. Triton X-100 and SDS produce a gradual solubilization of the specific Ca⁺⁺-ATPase activity up to a 10^–3 M detergent concentration, above which a strong inactivation occurs, while the enzyme solubilization increases with the presence of cholate in the whole concentration range under study. The different behaviour of sodium cholate, when compared to SDS or Triton X-100, is discussed in relation to the surfactant molecular structures. The possibility of membrane lysis and reassembly in the presence of some detergents is also considered.Abbreviations SR sarcoplasmic reticulum - SDS sodium dodecylsulphate - DTT dithiothreitol - EGTA ethyleneglycoltetraacetate - PEP phosphoenolpyruvate