INFLUENCE OF SEASON PATTERN ON PREVALENCE AND SEVERITY OF ACUTE MYOCARDIAL INFARCTION, FUNCTION OF HEMOSTASIS IN PATIENTS WITH ISCHEMIC HEART DESEASE OF CLIMATE AREA OF MIDDLE VOLGA REGION

The aim of this study is to evaluate season pattern, severity and outcome of acute myocardial infarction in dependence of yearly patterns of function of hemostasis in middle Volga Region. Season pattern of prevalence myocardial infarction with winter peak was discovered, be accompanied of increase platelet aggregation, deterioration of blood viscosity, fibrinolysis depression. Most serious clinical course of summer myocardial infarction was observed with prosperity of hemostasis

High formation of superoxide anion and nitric oxide, and matrix metalloproteinases activity in vascular wall of rectal carcinoma vessels

Aim: To study the relationship between the level of generation of reactive oxygen species (ROS) and nitric oxide (NO) and activity of matrix metalloproteinases (MMPs) MMP-2 and MMP-9 in the vessels isolated from rectal tumors and Arteria rectalis superior. Methods: EPR at the room temperature and 77 °K, Spin Traps technology and zymography in polyacrylamide gels were applied. Results: In the vessels isolated from rectal tumors and Arteria rectalis superior high levels of ROS, NO· and formation of complexes of NO· with FeS-proteins at the sites of electron-transporting chain of mitochondria have been detected. High activities of MMP-2 and MMP-9 in vascular wall were also observed. The direct positive correlation between the rate of NO· generation and formation of complexes of NO·· with FeS-proteins as well as between ROS and NO formation and MMPs activities have been revealed. Conclusion: Altered oxidative equilibrium in mitochondria of cells in vascular wall promotes formation of cell hypoxia and its autocatalytic potentiation accompanied with activation of MMPs.Цель: изучить взаимоотношение между уровнями образования активных форм кислорода (АФК) и оксида азота (ОА) и активности матриксных металлопротеиназ (MMPs) MMP-2 и MMP-9 в сосудах, изолированных из опухолевой ткани прямой кишки, и в Arteria rectalis superior. Методы: ЭПР при комнатной температуре и при 77 °K, технология спиновых ловушек, зимофрафия в полиакриламидном геле. Результаты: в сосудах, изолированных из опухолевой ткани прямой кишки, и в Arteria rectalis superior обнаружены высокие уровни образования АФК, · и комплексов · с FeS-белками в участках цепи митохондрий. Определена также высокая активность MMP-2 и MMP-9 в стенке сосудов, находящихся в опухолевом узле. Обнаружена прямая положительная корреляция между величиной образования · и образованием комплексов · с белками, а также между образованием АФК и NO и активность MMPs. Выводы: измененное окислительное равновесие в митохондриях клеток стенки сосудов, находящихся в опухолевой ткани, усиливает формирование в них клеточной гипоксии, ее аутокаталитическое потенцирование, что сопровождается активацией в этих клетках MMP

Modulation of ROS/NO production by murine peritoneal macrophages in response to bacterial CPG DNA stimulation

Aim: To investigate the features of metabolic activation induced by bacterial CpG DNA (bCpG DNA) in peritoneal macrophages (PMjs). Methods: Electron paramagnetic resonance spin-trapping technique using respective spin traps was applied to study the generation rate of reactive oxygen species and NO production by PMjs of BALB/c mice. Results: For the first time the capability of bCpG DNA isolated from Bacillus subtilis GP1-807-03 culture medium to elevate activity of NADP·H oxidase and inducible NO synthase in PMjs of normal and tumor-bearing mice have been demonstrated. The main differences in superoxide anion generation rate and production of NO by PMjs of normal mice and mice with transplanted solid Ehrlich carcinoma were showed. The effects of bCpG DNA stimulation in vitro on ROS and NO production by PMjs depended on concentration and time exposure with bCpG DNA. Furthermore, response of PMjs from tumor-bearing mice on bCpG DNA stimulation was delayed as compared to PMjs of normal mice. Conclusion: The present findings suggest that bCpG DNA have modulatory effect on ROS/NO production by PMjs from normal and tumor-bearing animals.Цель: изучить особенности метаболической активации перитонеальных макрофагов (ПМФ) в ответ на воздействие, вызванное CpG ДНК бактериального происхождения. Методы: для исследования скорости генерирования радикальных форм кислорода (РФК) и продукции оксида азота (NO) ПМФ мышей линии BALB/c был применен метод электронного парамагнитного резонанса с использованием спиновых ловушек. Результаты: впервые в эксперименте продемонстрирована способность бактериальной CpG ДНК, выделенной из культуральной жидкости Bacillus subtilis GP1-807-03, повышать активность НАДФ·Н-оксидазы и индуцибельной NO-синтазы ПМФ у мышей с солидной карциномой Эрлиха и интактных мышей. Отмечены различия в скорости генерирования супероксидных радикал-анионов и уровне продуцирования NO ПМФ у животных с карциномой Эрлиха по сравнению с контрольными животными, что выражалось в замедлении ответа на воздействие бактериальной CpG ДНК. Выявлена концентрационная и временная зависимость влияния бактериальной CpG ДНК in vitro на продукцию РФК и NO ПМФ. Выводы: полученные данные свидетельствуют о том, что бактериальная CpG ДНК обладает модулирующим влиянием на продукцию РФК и NO ПМФ in vitro и in vivo

Pristine C60 fullereneS inhibit the rate of tumor growth and metastasis

Aim: To estimate the impact of C60 fullerene aqueous solution (C60FAS) on the rate of transplanted malignant tumor growth and metastasis. Methods: Lewis lung carcinoma was transplanted into С57Bl/6J male mice. Conventional methods for the evaluation of antitumor and antimetastatic effects have been used. Results: The C60FAS at low single therapeutic dose of 5 mg/kg inhibited the growth of transplanted malignant tumor (antitumor effect) and metastasis (antimetastatic effect): the maximum therapeutic effect was found to be of 76.5% for the tumor growth inhibition; the increase of animal life span by 22% was found; the metastasis inhibition index was estimated as 48%. Conclusion: It was found that water-soluble pristine С60 fullerenes efficiently inhibit the transplanted malignant tumor growth and metastasis

Effects of radical oxygen species and no: formation of intracellular hypoxia and activation of matrix metalloproteinases in tumor tissues

Aim: To establish the association between the radical oxygen species (ROS) and NO levels in the tumor cells mitochondria, between cell hypoxia development and activation of matrix metalloproteinases–2 and -9. Materials and Methods: Electron paramagnetic resonance (EPR) at room temperature and at the temperature of liquid nitrogen (77° K), spin traps technology, enzymography in polyacrylamide gel were applied. Results: Redox-centers in the respiration cascade of mitochondria have been revealed, multiple oxidative damage of which in breast and liver cancer tissues of experimental animals as well as in tumor tissue from patients with gastric cancer promote the development of cell hypoxia. Involvement of ROS and NO in activation of latent forms of matrix metalloproteinases in gastric tumor tissues has been shown. Conclusion: We hypothesize that superoxide radical-anions participate in development of cell hypoxia in tumors and surrounding normal tissues inducing activation of latent forms of matrix metalloproteinases.Цель: установить взаимосвязь между уровнями образования радикальных форм кислорода и оксида азота в митохондриях клеток опухолевых тканей, развитием клеточной гипоксии и активностью матриксных металлопротеиназ – 2 и –9. Методы: электронный парамагнитный резонанс при комнатной температуре и температуре жидкого азота (77 °К), технология Spin Traps, зимография в полиакриламидном геле. Результаты: выявлены редокс-центры в дыхательной цепи митохондрий, множественные окислительные повреждения, которые при канцерогенезе в молочных железах, печени экспериментальных животных и в опухолевой ткани больных раком желудка способствуют развитию клеточной гипоксии; показано участие супероксидных радикал-анионов и оксида азота в активации латентных форм матриксных металлопротеиназ в ткани рака желудка. Выводы: высказано предположение, что в формировании клеточной гипоксии в опухоли и прилегающих нормальных тканях принимают участие супероксидные радикал-анионы, которые активируют при этом латентные формы матриксных металлопротеиназ

Disordered redox metabolism of brain cells in rats exposed to low doses of ionizing radiation or uhf electromagnetic radiation

Aim: To investigate the changes of redox-state of mammalian brain cells as the critical factor of initiation and formation of radiation damage of biological structures in setting of continuous exposure to low doses of ionizing radiation or fractionated ultra high frequency electromagnetic radiation (UHF EMR) at non-thermal levels. Materials and Methods: The influence of low-intensity ionizing radiation was studied on outbred female rats kept for 1.5 years in the Chernobyl accident zone. The effects of total EMR in the UHF band of non-thermal spectrum were investigated on Wistar rats. The rate of formation of superoxide radicals and the rate of NO synthesis in mitochondria were determined by the EPR. Results: After exposure to ionizing or UHF radiation, the levels of ubisemiquinone in brain tissue of rats decreased by 3 and 1.8 times, respectively. The content of NO-FeS-protein complexes in both groups increased significantly (р < 0.05). In the conditions of ionizing or EMR the rates of superoxide radical generation in electron-transport chain of brain cell mitochondria increased by 1.5- and 2-fold, respectively (р < 0.05). In brain tissue of rats kept in the Chernobyl zone, significant increase of NO content was registered; similar effect was observed in rats treated with UHFR (р < 0.05). Conclusions: The detected changes in the electron transport chain of mitochondria of brain cells upon low-intensity irradiation or UHF EMR cause the metabolic reprogramming of cell mitochondria that increases the rate of superoxide radical generation and nitric oxide, which may initiate the development of neurodegenerative diseases and cancer. This article is part of a Special Issue entitled “The Chornobyl Nuclear Accident: Thirty Years After”

Redox-regulation of gelatinases during growth of cisplatin-sensitive and resistant Guerin carcinoma

Study was aimed to analyze the dynamics of changes and study interrelations between content of ferritin, transferrin, active gelatina­ses (MMP-2 and -9) in blood serum and tumor tissue, free iron, rate of superoxide radicals generation in tumor, activity of NADPH-oxidase and iNOS in neutrophils rats with sensitive and resistant strains of Guerin carcinoma (GC). Materials and Methods: In order to obtain resistant tumor, 12 courses of cisplatin chemotherapy have been carried out on rats bearing GC. Levels of transferrin and free iron were determined by analysis of EPR spectra from computerized radiospectrometer EPR ­RE-1307 at temperature of liquid nitrogen. Rate of superoxide radicals and nitric oxide generation in tumor and neutrophils of blood was determined by EPR using spin traps at room temperature. Content of ferritin in tumor homogenate and blood serum of rats with GC was determined by ELISA method using corresponding kits. Concentration of active forms of MMP-2 and -9 in obtained samples was determined using method of zymography. Results: Unregulated generation of superoxide radicals and NO by mitochondria of tumor cells and NADPH-oxidase and iNOS neutrophils via oxidation of iron-containing proteins causes the accumulation of “free iron” complexes in blood and tumor tissue of rats able to evoke oxide-induced damages of macromolecules. It has been shown that for resistant strain of carcinoma, as compared with sensitive one, significantly higher concentrations of active forms of MMP-2 and -9 in blood serum of rats are typical. Dynamics of gelatinases activity changes in tumor tissue corresponds in general with dynamics of changes in serum. In tumor tissue of rats the indices of gelatinases activity positively correlate with rate of superoxide radicals generation, content of “free iron” complexes, ferritin and activity of transferrin. Cytostatic agent increased levels of reactive oxygen species (ROS) and self-amplify rate of superoxide radicals generation. In turn, activation of MMPs via superoxide-depending regulation allows tumor cells to facilitate migration, invasion and finally — formation of metastatic centers. Mentioned above tumor “oxide phenotype” determines high level of its aggressiveness and forms corresponding level of drug resistance. Conclusions: Thus, high levels of superoxide radicals oxidize transport proteins and form free iron pool. Iron ions, via Haber — Weiss mechanism, initiate generation of the hydroxyl radicals, which also enhance oxidation processes. Key Words: drug resistance, redox-regulation, iron-containing proteins, matrix metalloproteinases

Superoxide- and no-dependent mechanisms of antitumor and antimetastatic effect of L-arginine hydrochloride and coenzyme Q₁₀

Aim: To study the redox-dependent mechanism of antiradical, antitumor and antimetastatic action of L-arginine hydrochloride (L-Arg) and coenzyme Q₁₀ (CoQ₁₀) in vivo. Materials and Methods: The study was performed on С57Вl mice with transplanted Lewis lung carcinoma treated by intraperitoneal injections of L-Arg at low or high doses (60 and 360 mg/kg body weight), CoQ₁₀ (0.2 and 1.2 mg/kg body weight) or their combinations. Electron paramagnetic resonance was applied for analysis of mitochondrial electron transport chain, СoQ₁₀ levels, free iron (FI), the level of NO, and the rate of superoxide radical generation. The activity of matrix metalloproteinase (MMP)-2 and -9 in tumor tissue was determined by zymography method in polyacrylamide gel. Results: Administration of L-Arg at high doses caused an inhibition of tumor growth by 48 ± 8.0%, increase of superoxide radical generation rate and NO levels to a value of 1.23 ± 0.14 аnd 2.26 ± 0.31 nm/g tissue · min, and decreased activity of MMP-2 and -9 (3.55 ± 0.8 and 4.8 ± 1.0 r.u., respectively). Treatment with L-Arg at low doses stimulated tumor growth and increased the levels of MMP-2 and -9 activities (8.44 ± 2.7 and 9.8 ± 3.1 r.u., respectively). Administration of СoQ₁₀ at high doses significantly decreased superoxide radical generation rate to the values of 0.44 ± 0.09 nm/g tissue · min, levels of free iron and NO, and caused tumor growth inhibition by 54 ± 11.3%. The combined use of L-Arg and СoQ₁₀ at high doses caused tumor growth inhibition by 51 ± 7.4% compared to Lewis lung carcinoma-bearing untreated animals (р < 0.05). Conclusions: Administration of L-Arg and СoQ₁₀ caused the dose-dependent effect on the rate of generation of superoxide radicals, level of ubisemyquinone, complexes NOFeS-proteins, levels of FI and NO. L-Arg at low doses positively modulated MMP-9 activity that promoted tumor progression. Upon combined use of L-Arg and СoQ₁₀, superoxide radicals and NO form the redox state that causes decrease of MMP-2, -9 activities with consequent inhibition of tumor invasion and metastasis

Significance of ferritin expression in formation of malignant phenotype of human breast cancer cells

Aim: The aim of our study is to investigate the disorders of ferritin functioning in breast cancer (BC) cells of different molecular subtype. Materials and Methods: The cell lines used in the analysis include T47D, MCF-7, MDA-MB-231, MDA-MB-468, MCF-10A, and 184A1. Ferritin heavy chains (FTH) expression was studied by immunohistochemical method. “Free iron” content and superoxide dismutase (SOD) activity were determined by means of EPR spectroscopy. Reactive oxygen species (ROS) level and peculiarities of microRNA expression in studied cell lines were evaluated using flow cytometry and PCR analysis, respectively. Results: It has been demonstrated that FTH expression directly correlates with proliferative activity of cells of both luminal (r = 0.51) and basal subtypes (r = 0.25), inversely correlates with expression of steroid hormones in cells of basal subtype (ER: r = −0.46; PR: r = −0.44) and does not depend on tumorigenic activity of both subtypes of studied cells (r = 0.12 and r = 0.9). Obtained data are the evidence that cells of luminal subtype B (MCF-7 cell line) and basal subtype (MDA-MB-231 and MDA-MB-468 cell lines) with high proliferative activity contain the highest level of free iron (2.9 ± 0.19·1016and 3.0 ± 0.22·1016) that can be consequence of intensive use of this element by cells, which actively divide and grow. Along with it, in cell of lines of basal subtype MDA-MB-231 and MDA-MB-468, high level of FTH (254 ± 2.3 and 270 ± 1.9) is being detected in consequence of increase of level of free iron, ROS (11.3 ± 1.05 and 7.27 ± 0.26) and SOD (9.4 ± 0.24 and 8.5 ± 0.18) as well as decrease of expression of microRNA 200b. In contrast, cells of luminal subtype B of MCF-7 line were distinguished by high expression of microRNA 200b and low ferritin level (125 ± 2.7). Conclusion: Obtained data demonstrate that tumor cells, which are referred to different molecular subtypes, are characterized by changes in system of support of balance of intercellular iron and certain associations of studied factors. Key Words: breast cancer, cell lines, luminal subtype, basal subtype, ferritin, “free iron”, ROS, SOD, miRNA 200b

Stress-overcoming behaviour of women with minor cardiac abnormalities

The purpose of this research was to determine stress-overcoming behaviour of women with minor cardiac abnormalities considering abnormality intensity. With the help of Toronto Alexithymia Scale, E. Heim`s system and S. Subbotin`s test 122 young women with different manifestation rates of the heart connective tissue dysplasia (minimal – basic group and moderate - comparison group) were examined (average age –21,46 ± 2,12 years). Significant dominance of adaptive coping strategies in all spheres was revealed when studying of stress-overcoming behaviour of healthy women with minor cardiac abnormalities of minimal intensity. Women of the comparison group used maladaptive coping-strategies in emotional and behavioural spheres more often comparing with clinically healthy women of the basic group. Significantly lower level of the stress resistance and higher alexithymia level were determined in the observed group with minor cardiac abnormalities of moderate intensity which has chosen maladaptive emotional and behavioural coping-strategies. The results are appropriate to consider for personalized arrangements for prevention of cardiovascular complications in patients with heart connective tissue dysplasia syndrome