National Repository of Grey Literature 48 records found  1 - 10nextend  jump to record: Search took 0.00 seconds. 
Proteomic analysis of myocardial integral membrane proteins
Oliva, Tomáš ; Petrák, Jiří (advisor) ; Pompach, Petr (referee)
Cardiovascular diseases (CVD) are the leading cause of morbidity and mortality in Europe. Over 4 million people die from CVDs annually and another 11 million people develops CVDs every year. These numbers show that there is a need for better diagnostic, prognostic and predictive biomarkers and, more importantly, a need for new and more efficient drugs. Integral membrane proteins (IMPs) are ideal candidates for new drug targets. However, a study of IMPs represents a major challenge in current proteomics. This challenge is associated with the low abundance of IMPs, their low solubility in aqueous solvents and the absence of trypsin cleavage sites in their transmembrane segments. To overcome these issues, methods that selectively target either N-glycosylated extra-membrane segments (CSC, SPEG, N-glyco-FASP) or transmembrane segments (hpTC) were developed. In this thesis we employed a combination of two N-glyco-capture methods (SPEG and N-glyco-FASP) performed on two different samples (membrane-enriched fraction and total tissue lysate) with analysis of membrane-embedded IMP segments by hpTC and with standard non-targeted "detergent+trypsin" approach to analyze rat myocardial membrane proteome. We also performed an evaluation of employed methods for preparation of membrane fraction by western blot...
The influence of creatine kinase system during the development of a cardioprotective phenotype in a rat adapted to a mild cold
Dzobová, Tereza ; Žurmanová, Jitka (advisor) ; Alánová, Petra (referee)
Cardiovascular diseases are still one of the most common causes of death and impaired quality of life worldwide. This is the reason why more and more researches start to focus on the possible prevention and treatment. One of the possible interventions that could help in this field is a relatively new model of a mild cold adaptation, first introduced in our laboratory, where the cardioprotective effects have already been proven as they reduced the magnitude of an ischemia-reperfusion damage without any negative side effects. One of the studied areas of the myocardium is the creatine kinase system, which represents a dynamic intracellular system of numerous isoenzymes stored specifically in the places of energy production and consumption. Its primary function lies in a cardiac energy metabolism and in an overall energy homeostasis in muscles, brain and other organs with high and rapidly changing demands for an energy supply. However, the molecular basis of these metabolic processes and their course induced by a cold adaptation are not yet fully known. Therefore, the aim of this work was to determine the changes in expression of three CK isoforms (CKB, CKM, mtCKs) after exposure to a mild cold (8±1 řC) during the period of an acute cold (1, 3, 10 days) and during the period of a chronic cold (5 weeks)...
Influence of the epicardium on cardiac regeneration after cryoinjury during embryonic development - analysis of morphological and physiological changes
Neffeová, Kristýna ; Kolesová, Hana (advisor) ; Hovořáková, Mária (referee)
Millions of people die from myocardial infarction every year. In the heart, damaged cardiomyocytes do not regenerate and fibrotic scar forms. Surface layer of the heart - epicardium, improve heart regeneration processes by producing signalling factors. Epicardial cells undergo epithelial-mesenchymal transition and give rise to various cells involved in heart regeneration (coronary vessel formation and possibly new cardiomyocytes). To study the effect of epicardium on cardiac regeneration, I introduced a novel method of myocardial cryoinjury in avian embryos. In embryonic stages, where the heart is already covered by a layer of epicardium. Probe cooled in liquid nitrogen, injured the left ventricle of the heart and simulate myocardial infarction. Cryoinjury is an appropriate alternative to induce myocardial infarction as the coronary ligation is not feasible in early embryos. After cryoinjury, were observed changes in the epicardium. Epicardium is activated (expression of WT1) and detached from myocardium and form protrusions. In subepicardium we observed a large number of migrating cells. These cells undergo epithelial-mesenchymal transition were detected by cytokeratin and vimentin antibodies. We observed α-SMA positive cells and undifferentiated cardiomyocytes (N2.261 antibody) in the...
Morphological and functional changes in microvasculature and endomysium in human atrial myocardium with atrial fibrillation
Smorodinova, Natalia ; Kučera, Tomáš (advisor) ; Čížková, Dana (referee) ; Čížková, Kateřina (referee)
This doctorial thesis was focused on morphological and functional changes in microvasculature and endomysium in human atrial myocardium with atrial fibrillation. Atrial fibrillation (AF) is one of the most common arrhythmias in the clinical practice and it is associated with an increase in mortality risk that is strongly related with old age. Its pathogenesis is still not sufficiently explored. One of the generally recognized factors contributing to the initiation and maintenance of atrial fibrillation is structural remodeling of the myocardium. Structural remodeling is reflected by changes that affect both atrial cardiomyocytes as well as endomysium. We analyzed atrial biopsies obtained from patients undergoing bypass or mitral valve surgery. The patients had a regular sinus rhythm or were suffering from AF. Immunohistochemistry was used to visualize collagen I, collagen III, elastin, desmin, smooth muscle actin and VEGF in the atrial samples.To detect capillaries UEA-lectin was used. For detection of different types of immune cells the following markers were detected immunohistochemically: CD45 as a pan- leukocyte marker, CD3 for T-lymphocytes, CD68 for monocyte/macrophages, mast cell tryptase for mast cells and DC-SIGN for immature dendritic cells. Our results document that in patients...
Mapping of the cell population expressing Sonic hedgehog during the embryonic development of heart
Břežná, Veronika ; Hovořáková, Mária (advisor) ; Šaňková, Barbora (referee)
Sonic hedgehog is one of three members of the Hedgehog family, whose signaling pathway plays a crucial role in controlling the development of vertebrates. Sonic hedgehog has an important role in organizing the developmental processes of majority of organ systems. Interestingly, its direct expression or a contribution of cell lineages expressing this signaling molecule in the myocardium has never been described. The aim of this work was to try to detect the cells that expressed Sonic hedgehog directly in the heart tissue with a focus on the myocardium. We monitored these cells from embryonic day 10.5 to 16.5 and then postnatally. We also evaluated current Sonic hedgehog expression in cardiac tissue from embryonic day 12.5 to embryonic day 15.5. We used the CreLoxP system, X-gal staining, fluorescence and confocal microscopy to detect cell lines expressing Sonic hedgehog. We also assessed the presence of cells that expressed Sonic hedgehog in the developping heart in the past using immunohistochemistry. According to our results, the presence of a descendant cell lineage expressing Sonic hedgehog in the past was demonstrated prenatally and postnatally in the mouse myocardium. The performed analysis shows that these cells can be detected in cardiac tissue from embryonic day 10.5 till the postnatal...
Myocardial phospholipases A2 and oxidative stress
Závišková, Kristýna ; Míčová, Petra (advisor) ; Chytilová, Anna (referee)
Heart and vascular diseases, in which oxidative stress plays an important role, are the most frequent cause of death in developed countries. Therefore today's researches focus on clarifying signaling pathway and their components that operates in these processes. One of the possible key factors can be phospholipases A2. These enzymes hydrolyze unsaturated fatty acids from C2 position on phospholipids, which easily subject to the oxidative modification due to their double bonds. There are five groups: the secreted, the cytosolic, the calcium independent, the platelet-activating factor acetylhydrolases and the lysosomal phospholipases A2. They differ in the requirement for calcium ions for its activity, molecular weight, position of disulfide bonds and amino acids sequences in the active site. Their main importance is to keep the stability and integrity of cell membranes and produce lipid signaling molecules (free fatty acids, lysophospholipids, eicosanoids). The phospholipases A2 play a dual role during oxidative stress in myocardium. First, they have a positive effect by protecting membranes against oxidative stress due to their higher affinity for removal of peroxidized fatty acids from membrane phospholipids. On the other hand, the phospholipases A2 can also cause destructive effects during...
Cardiac ischemic tolerance of hypertensive rats
Jelínek, Jan ; Neckář, Jan (advisor) ; Sotáková, Dita (referee)
The aim of this thesis is to summarize current knowledge about the influence of the ischemic- reperfusion injury at the myocard of hypertensive subjects. First part of this thesis is focused on the description of ischemia, reperfusion and changes in the myocardial metabolism during these processes. These changes in the myocardial metabolism are for example necrosis or apoptosis of the myocardial cells. The second part describes the currently known cardioprotective phenomena. This part also compares their effects. The signalization of preconditioning, the second window of preconditioning and the postconditioning are described here in more details. Third part is focused on the description of the risk factors connected to the ICHS and hypertension. It describes also classes of hypertension, clinical and experimental methods of hypertension treatment, description of the laboratory breeds of hypertensive rats. In the last part of this thesis I describe the influence of hypertension on the I-R injury in current laboratory studies. In the most studies spontaneously hypertensive rats (SHR) were used. As a normotensive controls Wistar-Kyoto rats were mostly used. For some other experiments transgenic genetic rats (TGR) were used. Powered by TCPDF (www.tcpdf.org)
Role of protein kinase C isoforms in cardioprotective mechanism of chronic hypoxia
Hlaváčková, Markéta ; Novák, František (advisor) ; Kopecký, Jan (referee) ; Novotný, Jiří (referee)
Cardiovascular diseases, particularly acute myocardial infarction, are one of the leading causes of death in developed countries. It is well known that adaptation to chronic intermittent hypobaric hypoxia (IHH) confers long-lasting cardiac protection against acute ischemia/reperfusion injury. Protein kinase C (PKC) appears to play a role in its cardioprotective mechanism since the administration of general PKC inhibitor completely abolished the improvement of ischemic tolerance in IHH hearts. However, the involvement of individual PKC isoforms remains unclear. Therefore, the primary aim of this study was to investigate the potential involvement of PKCδ and PKCε, the most prevalent PKC isoforms in rat heart, in the mechanism of IHH-induced cardioprotection. We showed that IHH up- regulated PKCδ protein in left ventricle, enhanced its phosphorylation on Ser643 and increased its co-localization with markers of mitochondrial and sarcolemmal membranes. PKCδ subcellular redistribution induced by IHH as well as the infarct size-limiting effect of IHH was reversed by acute treatment with PKCδ inhibitor rottlerin. These data support the view that PKCδ plays a significant role in IHH-induced cardioprotection. On the other hand, adaptation to IHH decreased the PKCε total protein level without affecting its...
Gene expression of enzymes involved in the regulation of apoptosis in rat moycardium - effect of chronic and acute hypoxia
Blahová, Tereza ; Žurmanová, Jitka (advisor) ; Kalous, Martin (referee)
Adaptation to chronic hypoxia provides myocardial protection against ischemia - reperfusion injury (IR). Cardioprotective effect of adaptation depends on the degree and duration of hypoxic exposure and daily regime of adaptation. Certain protective regimes of adaptations to hypoxia have been reported to activate proapoptotic signaling pathways and bioactive sphingolipids were recently shown to play important role in the regulation of apoptosis in the heart. We aimed to determine the mRNA level of selected genes related to apoptotic pathways and to sphingolipid metabolism in two models of hypoxic adaptation, continous normobaric hypoxia (CNH 10% O2) with different exposures (4h, 48h, 120h, 21days) and intermitent hypobaric hypoxia (IHH 7000 m, 8h/day). Both ventricles, LV and RV, were analysed after adaptation to CNH and only LV was analysed after IHH adaptation. Our results show that both types of adaptation increased mRNA of proapoptotic genes, CNH mainly in RV and IHH in LV. Furthermore, increased expressions of proapoptotic genes were accompanied by the increase of expression of enzymes producing predominantly protective kinds of sphingolipids. The exact role of apoptosis and sphingolipid signaling molecules in endogenous myocardial protection requires further research. Key words: Apoptosis,...
Effect of chronic hypoxia on antioxidative capacity of rat myocardium.
Závišková, Kristýna ; Nováková, Olga (advisor) ; Žurmanová, Jitka (referee)
Adaptation to chronic hypoxia activates endogenous signaling cascades, which lead to cardiac protection against acute ischemia/reperfusion (I/R) injury. The molecular mechanism of this phenomenon has not been fully clarified yet. However, it was proved that reactive oxygen species (ROS) take part in cardioprotective signaling pathway inducted by chronic hypoxia. The high level of ROS must be precisely regulated by antioxidative system of a cell. The aim of diploma thesis was to examine the effect of intermittent hypobaric hypoxia (IHH, 7 000 m) on relative amount of antioxidative enzymes (peroxiredoxin 6 - PRX6, thioredoxin 1 and 2 - TRX1 and TRX2, thioredoxin reductase 1 - TRXR1) and also enzymes of iron metabolism (heme oxygenase 1 and 2 - HO1 and HO2, aconitase 1 and 2 - ACO1 and ACO2), which participate in regulation of cell redox state. Moreover, we studied the effect of adaptation to IHH and an antioxidant tempol on relative amount of calcium-independent phospholipase A2 (iPLA2). iPLA2 can remove peroxidized fatty acids from membrane phospholipids. On the other hand, iPLA2 can damage cell in I/R conditions. All enzymes were studied in homogenates from normoxic and IHH adapted rat left ventricular myocardium by Western blot. Adaptation to IHH caused a decrease of PRX6 and on the opposite an increase of...

National Repository of Grey Literature : 48 records found   1 - 10nextend  jump to record:
Interested in being notified about new results for this query?
Subscribe to the RSS feed.