H2S-Mediated Changes in Erythrocyte Volume: Role of Gardos Channels, Na+,K+,2Cl- Cotransport and Anion Exchanger.

The effect of H2S on changes in erythrocyte volume was studied by spectrophotometrical and potentiometric methods. It was found that H2S donor NaHS (2.5, 10, and 100 µM) induced an increase in erythrocyte volume in heterosmotic media. Activation of Gardos channels with A23187 or ascorbate-phenazine methosulfate system causes erythrocyte shrinkage and hyperpolarization of their membrane, while addition of NaHS restored erythrocyte volume. The decrease in erythrocyte volume upon blockade of Na+,K+,2Cl- cotransporter (bumetanide) or anion exchanger (SITS) was abolished by H2S donor NaHS, which attested to an important role of these transporters and chlorine conductivity of the membrane in the maintenance of the homeostasis of blood cells.

Cytokine Profiling of Subclinical Tick-Borne Infections in Humans.

Over many years, tick-borne infections remain one of the most serious threats to human health worldwide. The immune response to these infections in a human after confirmed bite by an infected carrier at the early stages of infection in the absence of clinical symptoms can be the first indicator of the presence of the infectious agent in the body. During viral infection, the concentration of IL-1α, IL-8, IL-10, IL-17A, and IFNγ increases; superoxide dismutase also increases, in contrast to bacterial infections. A slight decrease in the concentration is observed only for receptor antagonist IL-1Ra. During the infection caused by bacterial pathogens, very similar profiles of the innate human immune response are observed: activation of IL-1α, IL-8, and IFNα and suppression of superoxide dismutase, IL-1Ra, and IL-17A production. It has been demonstrated, that the immune response is triggered immediately after infection, and changes in the concentration of the main cytokines in the blood plasma can be detected as early as on days 2-5 after tick bite. These results can be useful in developing new methods of emergency diagnosis and prevention of tick-borne infections.

Role of Carbon Monoxide in the Mechanisms of Action of Extracellular ATP on Contractile Activity of Vascular Smooth Muscle Cells.

We studied the role of carbon monoxide (CO) in the effect of P2X and P2Y receptor agonist ATP on the tone of rat aorta segments with intact endothelium. ATP (1-1000 µM) and P2X receptor agonist α,ß-MeATP (100 µM) relaxed segments precontracted with phenylephrine (10 µM), while UTP (100-1000 µM) increased the amplitude of phenylephrine-induced contraction. The relaxing effect of ATP was enhanced by CORM II (100 µM), NO synthase inhibitor L-NAME, and guanylate cyclase inhibitor ODQ and attenuated by ZnPP IX (100 µM). The constrictive effect of UTP was weakened by CORM II (100 µM), but was not changed by ZnPP IX (100 µM). ZnPP IX (100 µM) weakened the relaxation response to α,ß-MeATP. Thus, ATP involves the CO-dependent signaling cascade through P2X receptors.

Molecular Mechanisms of Action of Gas Transmitters NO, CO and H2S in Smooth Muscle Cells and Effect of NO-generating Compounds (Nitrates and Nitrites) on Average Life Expectancy.

Gaseous signaling molecules (gas transmitters) take an especial position among the numerous signaling molecules involved in the regulation of both intracellular processes that occur in different types of cells and cell-cell interactions. At present time, gas transmitters include three molecules whose enzymatic systems of synthesis and degradation, physiological action and intracellular effectors, the change of which under the action of gas transmitters may result in physiological and/or pathophysiological effects are well- determined. These molecules include nitrogen oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H2S). They are involved in the regulation of functions of various organs and systems of the human body, including the circulatory system. Interaction of NO, CO and H2S with various enzymatic and structural components of endothelial and, especially, smooth muscle cells has a significant impact on vascular tone and blood pressure. Furthermore, the crossing of NO-, CO- and H2S-mediated signaling pathways at common effectors and interaction with each other can determine the end, resulting functional response of the cell. The knowledge of the molecular targets of gas transmitters' action, the structure of the binding centers for gas transmitters and their interaction with each other may be essential in the development of methods of regulation of these signaling systems by targeted, directed action. This review summarizes the molecular mechanisms of the NO, CO and H2S interaction with the main targets, which carry out their regulatory effect on vascular smooth muscle cells. Also we describe here different ways of cross-regulation of NO-, CO- and H2S-dependent signaling pathways. We analyzed NO-synthase and nitrite reductase systems of nitric oxide cycle and discuss the nitrate-nitrite background of the existence of modern man, which can substantially modify the signaling system, the metabolism of virtually all cell ultrastructure of neurons, neuron-neuron and neuron-glial interactions and exerts its influence on socially significant diseases that can affect the quality and the average life expectancy.

Regulation of Contractile Responses of Vascular Smooth Muscle Cells under Conditions of Hypoxia-Reoxygenation.

We analyzed the effects of hypoxia and reoxygenation on changes in contractile activity in rat aortic smooth muscles. Both hypoxia and reoxygenation induced relaxation of smooth muscle cells precontracted with high-potassium Krebs solution (30 mM KCl) or α1-adrenoceptor agonist phenylephrine. Vasodilation resulted from enhancement of potassium permeability of smooth muscle cell membranes caused by activation of voltage-gated potassium channels (triggered by both precontracting agents) or by opening of ATP-sensitive potassium channels (phenylephrine). In isolated smooth muscle cells, both hypoxia and inhibition of Na+,K+-ATPase with ouabain led to depletion of intracellular store of macroergic substances, reduced potassium concentration, and elevated the content of sodium ions.

[Child and Adolescent Interepydemic Immunity to Influenza Viruses].

OBJECTIVE: Our aim was to evaluate the child and adolescent population immunity to influenza A viruses (IAV) and influenza B virus (IBV). METHODS: The concentration and specificity of antiviral antibodies was evaluated by hemagglutination inhibition assay (HAI) that was performed using commercial HAI diagnostic kits. RESULTS: The serum samples of 254 clinically healthy children and adolescents were examined in this study. 245 participants had the antibodies to IAV, 199--to IAV and IBV and only 4 children aged between 1 and 4 years and a 12-year-old boy had no immunity to IAVor IBV. The number of children with specific immunity increased in elder groups by 43% (from 81 to 116) for N0N1, and H3N2 subtypes and by 110% (from 38 to 80) for H1N1 subtype of IAV. In children younger than 4 years the titer of specific antibodies against the H1N1pdm09 was 1:210 and against the H3N2--1:270, whereas in adolescents of 10-14 years these figures were by 1.6 and 2.4 times lower (1:130 and 1:120) respectively. Antibodies to the subtypes H2N2 and H5N1 were not detected. CONCLUSION: The results indicate that 98.4% of child and adolescent population in interepidemic influenza season are immune to the various IAV (H3N2, H1N1, H0N1) as well as to the IBV. More than half of children and adolescents (57.4%) are immune to H1N1pdm09 subtype of IAV. The strength of immune response to the recent pathogens (H3N2 u H1N1pdm09) is higher in infants than in teenagers.

[Gas Signalling in Mammalian Cells].

At the end of the last century after the discovery of signaling functions of nitric oxide (NO, II), a new class of biologically active substances was admitted. It includes so-called gas transmitters acting as intercellular and intracellular regulators of different physiological functions. Currently, this class includes such gases as NO, carbon monoxide (CO) and hydrogen sulfide (H2S). It was found that these gases regulate not only functions of the. gastrointestinal tract and the cardiovascular system, where it has been determined initially, but also affect the function of the central and peripheral nervous.systems. Apparently, they constitute a single complex of gas transmitters, which easily penetrates through the membrane and regulates numerous enzymatic and non enzymatic cells reactions. This review presents the mechanisms of gas transmitters' influence on the electrical and contractile properties of smooth muscle cells (SMC) as a possible new ways to interact with the "classical" intracellular signaling cascades (Ca2+, cyclic nucleotides) and effectors systems. On account of their interactions the role of cyclic nucleotides and calcium ions in the implementation of the signal gas molecules functions is analyzed. We summarize the literature data and the results of our own research on the role of SMC membrane ion-transporting systems in myogenic effects of NO, CO and H2S and describe possible reasons of gas transmitters multidirectional influence on the excitation-contraction coupling in SMC.

[Genetic and phenotypic differentiation of Calluna vulgaris (L.) Hull in Pritobolie and Europe].

Geographic variation and differentiation of the chloroplast DNA haplotypes and morpho-anatomical leaf parameters were assessed in a number of eastern European groups of Calluna vulgaris (L.)Hull populations and in the Pritobolien group of populations of this species in Western Siberia, which have long been isolated from the European populations. Sharply pronounced genetic and phenotypic distances and their gradients between the Pritobolien and European population groups were revealed. These distances were many times higher than those between the relatively homogeneous eastern European groups. The data obtained generally supported the hypothesis on the phenogenetic divergence of the Pritobolien marginal populations of C. vulgaris from the European, probably at the subspecies level.

[Postglacial migration and phenogeography of populations of the Scots pine (Pinus sylvestris L.) in the northeast of the Russian Plain].

The history, distribution routes, and phenogeographic structure of the Scots pine (Pinus sylvestris L.) in the northeast of the Russian Plain were studied on the basis of paleogeographic data and results of our own phenotypic and allozyme-genetic studies. It is assumed that, after the maximum Dnieper glaciation, P. sylvestris populations could successfully distribute to the northwest and north from the refugia of the South and Middle Urals as a result of seed dispersal by Belaya, Ufa, Chusovaya rivers (in Holocene, by Severnaya Dvina, Mezen', and Pechora rivers). On the basis of the hypothesis of "migration complexes" and the theory of hydrochory for coniferous species, a scheme of formation of a population structure of the Scots pine in the northeast of the Russian Plain is proposed.

[Genetic diagnostic technique for opisthorchiasis].

OBJECTIVE: To develop a test system for polymerase chain reaction (PCR) diagnosis of Opisthorchis felineus in human feces. Based on the Internet databases, the authors have developed oligonucleotide primers, as well as TaqMan tests (fluorescence label probes) specific to O. felineus genome to perform PCR with the real-time detection of a result. Conditions for PCR, including temperatures, the number of components of amplification, etc., were selected. Test systems on fecal samples from 254 subjects randomly selected and on those from 100 patients with verified opisthorchiasis, and on those from 100 healthy individuals were studies. The significance of the test systems was ascertained to be about 98%. Their sensitivity in determining O. felineus is about 10(3) copies of target DNA per ml. The efficiency of various techniques (coproovoscopy, enzyme immunoassay, PCR) was compared to detect Opisthorchis invasion.

[Nitric oxide metabolites and their significance in pathogenesis of bronchial asthma].

To study the possible potential of nitric oxide (NO) in the processes of atopic inflammation, the authors evaluated the intensity of nitrosyling and oxidative stresses in the bronchoalvelar lavage fluid and expired air condensate of patients with bronchial asthma (BA). Chronic inflammation was shown to result in an increase in the processes of nitration and oxidation in worsening BA and to reliably correlate with airway NO-producing functions, by explaining the pathological effects of NO due to the formation of 3-nitrotyrosine and malonic dialdehyde with the existing imbalance in NO metabolism, by intensifying nitrosylating stress. In the authors' opinion, nitrosothiols that are required as a NO donor may be rapidly destroyed or virtually do not form so the peroxynitrite-nitrosothiol ratio may predetermine the final effects of NO.

[Markers of gluten intolerance in children with autism spectrum disorders and Down'syndrome]. / Markery neperenosimosti gliutena u detei s rasstroistvami autisticheskogo spektra i sindromom Dauna.

AIM: To study serological and genetic markers of gluten intolerance in children and teenagers with autism spectrum disorders (ASD) and Down's syndrome (DS). MATERIAL AND METHODS: Thirty-three children with ASD (group 1) and 8 with DS (group 2), aged from 2.5 to 15 years, were examined. There were 27 boys and 6 girls in group1, 5 boys and 3 girls in group 2. Most of the children were on a regular diet and only 4 children with ASD kept gluten-free diet (GFD). Using ELI method antibodies to gliadin IgG (AntiGliadin IgG), antibodies to deamidated peptides of gliadin IgA (AntiDGP IgA), immunoglobulin A (IgA) were identified. Haplotypes HLA-DQ2 and DQ8 were determined using PCR. RESULTS: AntiGliadin IgG were identified in 12.1% (4) patients of group 1, with the exception of patients on GFD in 13.8%, and in 50% patients of group 2. One child with ASD had selective IgA deficiency. Haplotypes predisposing to celiac disease had 41.9% of patients of group 1 and 37.5% of patients of group 2. In ASD, the distribution of genotypes was as follows: DQ2 (64.3%), DQ8 (28.6%), DQ2/DQ8 (7.1%,). In DS, all patients had haplotype DQ2. AntiDGP IgA were not identified in both groups. CONCLUSION: The predominant form of gluten intolerance in children with ASD and DS is sensitivity to gluten, which can be identified in 40-50% of patients. Celiac disease, an autoimmune form of gluten intolerance, can be diagnosed in single cases, although predisposition to it is identified in 41.9% - 37.5% patients with ASD and DS, respectively. Before the start of GFD, laboratory tests should be made to identify forms of gluten intolerance and the use of GFD.

[THE INFLUENCE OF HYDROGEN SULFIDE ON COLLAGEN-INDUCED AGGREGATION OF HUMAN PLATELETS].

Study the impact of hydrogen sulfide on collagen-induced platelet aggregation from healthy donors and patients with type 2 diabetes. In healthy individuals, in contrast to patients with type 2 diabetes, NaHS significantly inhibited platelet aggregation. Activators of cAMP signaling (forskolin and phosphodiesterase inhibitor) significantly reduced platelet aggregation in both groups of examinees. NO-synthase inhibitors increased platelet aggregation in healthy volunteers, but not in patients with type 2 diabetes. The presence of H2S donor did not alter the extent of platelet aggregation at high concentrations of cAMP or decreased production of nitric oxide. It is assumed that the antiplatelet effect of H2S is not associated with the effect on the signal system, mediated cAMP or nitric oxide. Change H2S-dependent regulation of platelet aggregation in patients with type 2 diabetes is caused by disorders have been reported with this disease: the increase of intracellular calcium ion concentration, oxidative damage to proteins, hyperhomocysteinemia, glycosylation of key proteins involved in this process.

[Immune system status under effect of low levels of ionizing radiation:studies within the 10 kilometer zone of accident at Chernobyl nuclear plant]. / Sostoianie immunnoi sistemy pri vozdeistvii malykh urovnei ioniziruiushchei radiatsii: issledovaniia v 10-kilometrovoi zone avarii na ChAES.

A study was made of the influence of high radiation contamination within the ten-kilometer zone of Chernobyl disaster on the structure and function of the immune system of (DBA x C57B1)F1 and DBA mice. The cumulative radiation doses with respect to gamma-radiation, were 0.024, 0.168 and 0.336 Gy. T-lymphocyte proliferation was shown the be activated with all radiation doses mentioned above but with doses of 0.024 and 0.168 Gy, helper T lymphocytes, and with 0.336 Gy, suppressor N-lymphocytes were primarily activated. So, in the former case, some effector functions of the immunity were activated, and in the latter, inhibited. It is concluded that certain hyperstimulation of the lymphoid cell formation (including the increase in T-suppressors), that is induced by long-term exposure to low-level radiation, triggers the mechanisms of autonomous regulation of the system that suppress the radiation-induced lymphocyte proliferation and inhibit the effector functions of the immunity.

[Neutrophil phagocytic activity and the humoral factors of general and local immunity under intensive physical loading]. / Fagotsitarnaia aktivnost' neitrofilov i gumoral'nye faktory obshchego i mestnogo immuniteta pri intensivnykh fizicheskikh nagruzkakh.

The comparative examination of 91 sportsmen under physical strain of different intensity and 30 healthy adults not active in sport has been made. A decrease in the functional activity of neutrophils in response to increasing physical strain has been revealed, which coincides with a drop in the levels of normal serum and secretory antibodies, immunoglobulins and lysozyme.

[The phenomenon of the formation of universal rosette-forming cells under superextreme loads]. / Fenomenon obrazovaniia universal'nykh rozetkoobrazuiushchikh kletok pri superékstremal'nykh nagruzkakh.

A combined immunological study of cell-mediated and humoral immunity factors in sportsmen at the period of important competitions was made. The effect of the maximum bearable load was found to lead to the formation of a great number of universal rosette-forming lymphocytes and neutrophils. In the control group their number did not exceed 1-4% of the total population of these cells. Simultaneously, the suppression of the phagocytic activity of neutrophils and some of the humoral factors of systemic and local immunity with the parallel development of compensating processes were recorded.

[Oxidation and reduction of glutathione in rat organs during the administration of ethonium]. / Okislenie i vosstanovlenie glutationa v organakh krys pri vvedenii étoniia.

An intramuscular administration of ethonium in a dose of 5 and 10 mg/kg 4-6 h later decreases the level of the reduced glutathione and increases that of oxidized glutathione and the glutathione peroxidase activity in the liver and kidneys of albino rats. The duration of the preparation effect depends on the dose and organ, changes in the glutathione reductase activity in the liver, kidneys, lungs, spleen and heart are not found. Participation of ethonium in formation of peroxides in the organism tissues is shown possible.

[Morphological changes in the mucosa of the stomach and the small intestine of rats under the action of mebendazole]. / Morfologicheskie izmeneniia slizistoi obolochki zheludka i tonkoi kishki krys pod deistviem mebendazola.

The effect of mebendazole on the gastric and intestinal mucosa was studied in 52 intact white rats. It was shown that the drug caused a significant damage to the mucosa, especially of goblet cells of villi and crypts and interfere with the mucoid secretion. The alterations after mebendazole administration were more severe than those caused by naphthamon. On the 13-15th day after the treatment the complete restoration of the damage was not seen.

[Connection between changes in the nasal and mouth mucosa with immune status under the effect of environmental factors]. / Sviaz' izmenenii slizistykh obolochek nosa i rta s imunnym statusom pri vozdeistvii faktorov okruzhaiushchei sredy.

The paper summarizes the results of determination of the cytological status of nasal and oral mucosae in 789 preschool and school children from different cities and towns of the Russian Federation: Moscow, Klimovsk, Yaroslavl. A relationship between the mucosae, immunity, and morbidity, and environmental pollution is shown in the town of Klimovsk as an example.

[The interaction in the system of the environment--mucosal membrane function--immune status--morbidity]. / Vzaimosviaz' v sisteme okruzhaiushchaia sreda--sostoianie slizistykh obolochek--immunnyi status--zabolevaemost'.

The paper shows how ambient air pollution and disturbances of immunity and oral and nasal mucosal status and morbidity are related in various age groups of children.