[Effect of atorvastatin on pro-inflammatory status (in vivo and in vitro) in patients with essential hypertension and metabolic syndrome].

AIM: To study effect of atorvastatin on spontaneous production of cytokines and reactive oxygen species by mononuclear leukocytes of blood of hypertensive patients with metabolic syndrome in vivo and in vitro. MATERIAL AND METHODS: We conducted an 8-week open prospective study on 36 patients with essential stage II hypertension associated with metabolic syndrome. Along with examination made in specialized cardiological clinic we assessed spontaneous production of cytokines and reactive oxygen species by blood mononuclear leukocytes during therapy with atorvastatin (in vivo). Dynamics of these parameters under the influence of atorvastatin on suspension of mononuclear leukocytes was also assessed in vitro. RESULTS: Therapy with atorvastatin (20 to 40 mg/day) facilitated reduction of serum concentration of acute phase proteins (C-reactive protein and neopterin) and decrease of spontaneous production by blood mononuclear leukocytes of proinflammatory cytokines (interleukin [IL]-1ß, IL-6 and tumor necrosis factor [TNF]-α) and reactive oxygen species. Dynamics of cytokine concentrations in supernatants of mononuclear leukocytes obtained after incubation of the cells with atorvastatin in vitro confirmed the assumption of direct inhibitory effect of this drug on spontaneous production of some proinflammatory cytokines (IL-6 and monocyte chemotactic protein-1). Absence of significant lowering of concentrations of other proinflammatory cytokines (IL-1ß and TNF-α) and expression of reactive oxygen species in vitro evidenced for complex indirect effect of therapy with atorvastatin on their production.

[Regulation of the genes expression of CD4+ T-lymphocytes differentiation transcription factors by galectin-3 in vitro].

Now a number of CD4+ T-lymphocytes, known as Th1, Th2, Treg and Th17, is currently identified and well- studied. The methods basing on the targeted regulation of differentiation process of the Th-lymphocytes that carry out the immune response polarization attract an attention of scientists dealing with a correction of immune-mediated. In the present study, endogenous beta-galactoside-binding protein of the lectin family, galectin-3, was investigated as a regulator of T-cell homeostasis. A galectin-3 is known to be actively produced by tumor cells in malignant transformation and able to influence the processes of signal transduction, cell-cell cooperation and the implementation of programmed death. As cell differentiation processes are directly connected with the regulation of gene expression, we investigated the effect of recombinant galectin-3 on expression of mRNA of transcription.factors, which guide the differentiation of CD4+ lymphocytes. The study was performed on peripheral blood mononuclear cells of healthy individuals. The gene expression levels were evaluated by a real-time PCR. In the experiments in vitro, it has been first found the recombinant galectin-3 (0.5 mg/mL) up-regulating the expression of transcription factors Gata-3 and Rorc mRNAs and down-regulating the mRNA expression of transcription factors T-bet and FoxP3. Up to a concentration of 1 mg/mL recombinant galectin-3 stimulates Th-cells by dose-dependent manner, whereas at higher concentrations stimulating effect weakens, and inhibiting action starts prevailing. Thus, one can suppose that galectin-3 through regulation of lymphocytes differentiation promote development of allergic, autoimmune and neoplastic diseases that allows us to consider the galectin-3 as a.potential target for therapy of these diseases.

[Pleiotropic effects of atorvastatin and dynamics of quality of life characteristics in patients with hypertensive disease and metabolic syndrome].

AIM: To study pleiotropic effects of atorvastatin during 8-week therapy of metabolic syndrome and estimate their relationship with dynamics of quality of life characteristics (QLC). MATERIAL AND METHODS: This 8-week study included 36 patients with stage II hypertensive disease associated with metabolic syndrome (MS). Comprehensive clinical, laboratory and instrumental examination was supplemented by QLC assessment using the MOS SF-36 questionnaire. RESULTS: 8-week therapy of stage II hypertensive disease associated with metabolic syndrome using individually selected doses of atorvastatin (20 to 40 mg/d) significantly reduced atherogenic cholesterol fraction and serum leptin levels; it had positive effect on carbohydrate and purine metabolism and safely maintained positive dynamics of subjective assessment of most points of the MOS SF-36 questionnaire.

[Intracellular targets of proapoptotic influence of gaseous transmitters].

Main molecular targets of nitric oxide, hydrogen sulfide and carbon monoxide proapoptotic action in Jurkat cells were determined in this study. Decrease of mitochondrial transmembrane potential was shown during all three gases action. Reason of this event is the Bcl-2 family members disbalance. Proapoptotic proteins release after mitochondrion membranes permeabilisation could be abolished by protein xIAP inhibition of caspase -9 and -3 activity during NO and CO application.

[Intracellular gaseous messengers, nitric oxide, carbon monoxide and hydrogen sulfide participate in apoptosis regulation].

In this paper, participation of gases, nitric oxide, carbon monoxide and hydrogen sulfide, in cell apoptosis regulation has been analyzed according to the literature data and our own findings. Different mechanisms of nitric oxide influence on apoptotic reaction including modulation of transcription factors activity and increase in mitochondrion membrane permeabilisation are described. Brief description of the generation and signal transduction pathways of carbon monoxide is presented. Pro- and antiapoptotic mechanisms of hydrogen sulfide influence on cell fate are analyzed.

[Quality of life in patients with coronary heart disease associated with metabolic syndrome: results of factor analysis].

AIM: To identify the factors most substantially influencing the quality of life (QL) in patients with coronary heart disease (CHD) associated with metabolic syndrome (MS), by using the principal component method. SUBJECTS AND METHODS: One hundred and two male patients (mean age 48.6 +/- 1.02 years) with CHD associated with MS, who had experienced large-focal myocardial infarction not earlier than 6 months before, were examined. Estimation of QL (EORTC QLQ CORE 30) and emotional-personal sphere (brief multifactorial personality questionnaire test and Spielberg's trait anxiety inventory) was made along with the complete clinical, laboratory, and instrumental examination accepted in specialized clinical practice of cardiology. RESULTS: A factor analysis of the variables obtained after a thorough examination of the patients could identify 4 common QL determinants, such as postinfarction cardiac remodeling, neurotization, obesity, and the degree of heart and coronary failure. CONCLUSION: MS in patients with CHD appreciably determines the total score of physical, emotional, and social well-being. In the cluster of MS components, obesity is a major factor that influences QL in patients who have sustained myocardial infarction.

[Apoptosis-modulating effects of heat shock proteins: the influence of Hsp27 chaperone on TBA Bcl-2 family proteins in Jurkat cell line].

The in vitro phosphorylated and non-phosphorylated Hsp27 forms concentrations and Bcl-2 proteins affected by Hsp27 inhibition were studied in Jurkat-line tumor cells and healthy donor mononuclear lymphocytes by Western blotting technique. The Hsp27 inhibition causes the increase of intracellular Bax protein concentration and the decrease of Bcl-2 level leading to an increase of apoptotic changes in Jurkat line cells.

[Heat shock protein 90--modulator of TNFalpha-induced apoptosis of Jurkat tumor cells].

rTNFalpha-induced programmed death of Jurkat tumor cells cultured with 17-AAG, a selective inhibitor of heat shock protein (Hsp90), was studied by fluorescent microscopy with the use of FITC-labeled annexin V and propidium iodide. Caspase-3 and -8 activities were determined by spectrophotometry using a caspase- 3 and -8 colorimetric assay kit. It was shown that inhibition of Hsp90 leads to activation of Jurkat cell apoptosis while Hsp90 itself suppresses this process. 17-AAG enhances rTNFa-induced apoptosis of tumor cells.

[Population association features of interleukine gene polymorphism in khakases with gastroduodenal diseases].

AIM: To investigate association of polymorphisms of IL-1 genes and antagonist of IL-2 receptor (IL1Ra). MATERIAL AND METHODS: Patients with chronic gastritis and ulcer were examined using the method of restriction analysis. RESULTS: It was found that CCILbeta and R4/R4IL1Ra are most prevalent allel variants in khakas population. CONCLUSION: It is expedient to define population risk and protective genotypes of development of ulcer associated with Helicobacter pylori in khakases.

[The functional properties and oxidative modification of plasma and neutrophil proteins in community-acquired pneumonia].

The functional properties of neutrophils (the activity of myeloperoxidase and the production of hydroxyl radical) were studied in community-acquired pneumonia (CAP) predominantly with the alveolar and interstitial types of lung parenchymal infiltration. Protein oxidative modification was estimated from the content of protein carbonyl derivatives in neutrophilic leukocytes and plasma and from the plasma concentration of bityrosine and oxidized tryptophan in patients with CAP. The production of hydroxyl radical and the activity of myeloperoxidase in the neutrophils of patients with CAP were increased and did not depend on the type of lung tissue infiltration. The development of oxidative stress in CAP was accompanied by the substantiation activation of protein oxidative modification processes in the neutrophilic leukocytes and plasma.

[Molecular pato-physiology of chronic virus infections].

Article is based on published data and results of own original researches on development of chronic socially inportant virus infections. Special attention is paid to priority research direction--molecular-genetic markers of chronis virus infections predisposition, development of personalized therapy molecural basis and forecast of infect and man organism interrelation results.

[The role of nitric oxide synthesis induction and inhibition in regulation of blood neutrophil cell death during oxidative disbalance].

Modeling oxidative stress in vitro with 5mM H2O2 has demonstrated a protective role of nitric oxide on realization of constitutional blood neutrophil cell death. The NO-synthase inductor L-arginine and the inhibitor of nitric oxide synthesis, L-NAME, influenced on the amount of annexin-positive cells, the content of Bax protein, reactive oxygen species, cyclic nucleotides, and calcium homeostasis in neutrophils under conditions realizing programmed death during oxidative stress in vitro and under acute inflammation. During oxidative stress L-arginine normalized the increased intracellular Ca2+ level and the cAMP/cGMP ratio due to increase of cGMP level, stabilized metabolism and prolonged neutrophil life. During acute inflammation NO induction was insufficient for limitation of Ca2+ release into cytosol and for onset of the apoptotic effect; blockade of NO synthesis deteriorated this situation by activating neutrophil apoptosis due to the sharp increase in Ca2+ content and reduction of cyclic nucleotides in cytosol. The protective effect of NO on neutrophil cell death during oxidative dysbalance is not associated with regulation of apoptotic protein Bax.

[The role of p53 and NF-kappaB transcription factors in redox-dependent dysregulation of mononuclear leukocyte apoptosis].

The aim of this work was to study programmed death of blood mononuclear leukocytes taken from healthy donors and patients with acute inflammatory diseases (acute appendicitis, community-acquired pneumonia). Cellular p53 and NF-kappaB transcription factors were detected by western blotting. Active form of NF-kappaB was shown to appear in mononuclear leukocytes undergoing oxidative stress in experiment and during acute inflammation, p53 was found only under oxidative stress conditions in vitro. Despite enhanced expression of target gene mRNA of these transcription factors in oxidative stress (proapoptotic protein Bax and antiapoptotic protein Bcl-XL), the resulting vector of p53 and NF-kappaB activation is stimulation of cell's apoptotic reaction.

[The functional state of components of the hemostatic system in patients with acute intraoperative blood loss].

Thrombotic and hemorrhagic events are one of the most common and menacing complications in the postoperative period. This may be attributable to the fact that therapy is by no means always performed by keeping in mind the pattern of dysfunction of the components of the hemostatic system. The purpose of the study was to define the pattern of hemostatic disorders occurring in the intra- and postoperative periods in the presence of significant hemorrhage, the feasibilities of their monitoring and correction. Thirty-seven patients with traumatic surgical interventions for cancer were examined in 4 steps: before surgery, after final intraoperative bleeding arrest, and on days 1 and 3 postoperatively. Intraoperatively, analysis of changes in the functional state of components of the hemostatic system identified 2 types of hemocoagulative responses to surgical trauma and blood loss. These included hypo- and hypercoagulative types. This let the authors make timely and goal-oriented correction of revealed hemocoagulative disorders and achieve positive results on postoperative day 3.

[Mitogenactivated protein kinases JNK and p38 as redox-dependent molecular targets correction of programmed cell death disturbances in oxidative stress condition].

The article summarizes information from recent literature and results of the author's own investigations concerning role of mitogenactivated protein kinases JNK and p38 in disturbances of programmed cell death regulation in oxidative stress condition.

[The role of redox-dependent signal systems in the regulation of apoptosis under oxidative stress condition].

Programmed death of peripheral blood mononuclear cells from donors with acute inflammatory diseases (an acute appendicitis, a community-acquired pneumonia) was investigated under condition of oxidative stress in vitro and under effect of selective inhibitors of MAP-kinases JNK and p38. Levels of active and inactive forms of MAP-kinases, and factors of transcription were determined by immunoblotting (western blot analysis). The increase in the activity of apoptosis under condition of oxidative stress in vivo and during the acute inflammatory diseases is associated with the increase in the level of reactive oxygen species (ROS) in the cells. The action of inhibitors of MAP-kinases JNK (SP600125) and p38 (ML3403) in vitro under condition of oxidative stress prevents increase in the quantity of annexin-positive mononuclear leucocytes that testifies to involving JNK and p38 MAP-kinases in apoptosis deregulation oxidative mechanisms. The appearance of NF-kappaB in the mononuclear leucocytes under condition of oxidative stress during the acute inflammatory diseases and at the experiment was shown; p53 was registered only under condition of oxidative stress in vitro. The effect of p53 and NF-kappaB results in the increase in the quantity of apoptosis annexin-positive mononuclear leucocytes that testify to inoperativeness of antiapoptotic regulation NF-kappaB.

[Dysregulation of cooperative interactions of immunocytes and eosinophils in the mechanism of development of eosinophilia in Opisthorhis felineus invasion].

The authors studied the levels of mononuclear cell production of eosinophil-specific cytokines (IL-3, IL-5), the serum levels of eotaxin by enzyme immunoassay; the expression of the eosinophilic cell receptor apparatus by flow cytofluorometry in patients with acute and chronic Opisthorchis invasion. Eosinophilia-associated Opisthorchis invasion was found to be accompanied by a pronounced change in the serum production of the key cytokine regulators of eosinophilic homeostasis (elevated IL-3 and IL-5 levels) and eotaxin by peripheral blood mononuclear leukocytes. There was an increase in the number of receptor structures to eosinophil-specific cytokines (IL-5R-, IL-3R-, and CCR3-positive cells) in patients with opisthorchiasis. In vitro incubation of the eosinophils, obtained from patients with opisthorchiasis, with the recombinant forms of cytokines (IL-5, IL-3, and eotaxin) demonstrated the decreased expression of IL-5 and IL-3 receptors with the normal presentation of CCR3. With the developed acute helminthiasis, the revealed changes were more pronounced than those observed in chronic Opisthorchis invasion.

[Redox-dependent regulation of apoptosis as a function of intracellular reactive oxygen species in oxidative stress].

Programmed cell death of mononuclear cells in conditions of oxidative stress in vitro and selective inhibitors of MAP-kinases JNK, p38 were investigated. Levels of active and inactive forms of MAP-kinases, factors of transcription P53, NF-kB and proteins-regulators of apoptosis Bcl-X(L), Bad, Bcl-2 were determined by immunoblotting (Western blotting). The increasing of number of annexin-plus mononuclears/lymphocytes in the culture associated with enhance of the level of intracellular reactive oxygen species was shown. The treatment of selective inhibitors JNK SP600125 and p38 ML3403 in vitro prevents peroxide-induced appearance of P53 and NF-kB in blood mononuclear cells, associated with increasing of their apoptotic activity. The disturbance of the balance of pro- and antiapoptotic proteins of Bcl-2, family (the increase of the Bax level without changes of Bcl-X(L) and Bcl-2) leads to the growth of apoptosis process of mononuclear leucocytes activity in oxidative stress conditions.

[Cytokine-mediated apoptosis of granulocytes eosinophilicus in eosinophilia].

The influence of recombinant forms of cytokincs (IL-5, IL-3 and eotaxin) on programmed destruction of eosinophils obtained from patients with diseases associated with high eosinophilia of blood (malignant diseases of system of blood, opisthorchosis) was studied in vitro. It was shown that all examined categories of patients irrespective of their nosology demonstrated a low level of spontaneous apoptosis of eosinophils. Cultivation of cosinophils isolated from peripheral blood of the patients with invasion O. felineus in vitro with r-IL-5, r-IL-3 and r-eotaxin decreased the number of eosinophilic cells undergoing apoptosis in comparison with spontaneous one. At the same time, incubation of eosinophils obtained from the patients with malignant diseases of system of blood, associated with cosinophilic syndrome, with r-IL-5, r-IL-3 and r-eotaxin allowed to ascertain the absence of sensitivity of eosinophilic cells to the antiapoptotic effect of these cytocines.

[Mechanisms underlying changes in functional properties of red cells in acute action of carbon monoxide].

It has been experimentally established that a single impact of carbon mono-oxide in concentration 4000 mg/m3 during 75 minutes (CL50) in rats is accompanied with disturbance of functional activities of blood red cells in the form of a decrease of deformability and an increase of aggregative capacity of erythrocytes which are most evident to the end of the first day after carbon mono-oxide administration. These changes are combined with structural modification and changes in metabolism of erythrocytic membrane. The signs of structural modification of erythrocytic membrane in rats exposed to acute effect of carbon mono-oxide in a mean lethal concentration stay long remaining during elimination of carboxyhemoglobinemia (up to 14-21 days).