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Mitochondrial production of reactive oxygen species and its role in physiological regulations
Holzerová, Eliška ; Mráček, Tomáš (advisor) ; Kalous, Martin (referee)
The production of mitochondrial reactive oxygen species and the resulting oxidative stress is an important phenomenon driving long-lasting research and intense discussions. Knowledge of exact mechanisms of reactive oxygen species production and pathways leading to their formation could help us to directly affect their production, a task with potential terapeutic implications. The molecular nature of the production of reactive oxygen species by some enzymes has already been well documented, but others still remain controversial and current theories are obviously far from the truth. Much more interesting is the question of physiological importace of this production. The reactive oxygen species were considered harmful factors clearly distorting the integrity of the organism for a long time. However, recent research suggest that their existence can also be beneficial and effective. Evidently they can serve as a signaling molecules in several metabolic and regulatory pathways occurring in the organism. This bachelor thesis offers insight into the current state of knowledge. It focuses on the most detailed description of the reactive oxygen species production by mitochondrial respiratory chain enzymes. Furthermore, it deals with some signaling cascades, where involvement of mitochondrially generated...
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Association of genetic polymorphism of oxidative stress with diabetes mellitus type 1 and 2
Kloboučková, Lucie ; Kotrbová - Kozak, Anna Katarzyna (advisor) ; Daňková, Pavlína (referee)
Diabetes mellitus is a chronic autoimmune disease in which the immune system attacks the insulin-secreting ß-cells in the pancreas. It leads to an absolute deficiency of insulin. Chronic hyperglycemia induces increased production of reactive oxygen species, which leads to a decrease of natural antioxidant level in blood, and it contributes to genesis of diabetes complications (e.g. vascular or pulmonic). Moreover, the oxidative stress results in onset of pancreas inflammations and the damage of its ß-cells. Aims: Our aim was to assess whether or not certain genotypes or their combinations occur with higher frequency among groups of patients of type 1 diabetes (T1D) and type 2 diabetes and in a control group of healthy individuals. Methods: The study included groups of 40 T1D patients, 40 T2D patients and 45 healthy individuals. The polymorphisms of genes involved in the oxidative stress response were analyzedby using RFLP, PCR with TaqMan probes and allele specific PCR. The target genes involved superoxide dismutase SOD1 and SOD3 genes; glutathione-S-transferase GSTM1, GSTT1, GSTP1 genes; glutathioneperoxidase gene GPX1 and catalase gene CAT. The levels of plasma malondialdehyde were measured by using liquid chromatography. Results: Statistically significant differences were found in the...
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Physiological and hormonal mechanisms influencing ejaculate quality in birds
Mojžišová, Kateřina ; Tomášek, Oldřich (advisor) ; Svobodová, Jana (referee)
Sexual selection is an important mechanism of evolution. In addition to precopulatory sexual selection, arising when males compete for female partners, there is also postcopulatory sexual selection (sperm competition), when females copulate with more males. Even though most avian species are socially monogamous, most of them are also partially promiscuous. In such species, sperm competition is an important factor increasing variability in reproductive success between males. Male's success in sperm competition is primarily determined by sperm concentration, total sperm count and morphology and motility of spermatozoa. My bachelor thesis summarizes knowledge about the hormonal a physiological mechanisms that influence semen quality. This is especially the effect of sex and other hormones, physiological and oxidative stress, environmental conditions, antioxidant mechanism and composition of sperm cell membrane. Besides evaluating their influence on ejaculate quality on intraspecific level, I also describe their differences between avian species with respect to the differences of sperm competition.
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Thorium as an environment stressor for plant growth.
Hrdinová, Aneta ; Lhotáková, Zuzana (advisor) ; Podlipná, Radka (referee)
Thorium is an element belonging to heavy metals, which is characterized by its radioactivity similarly to uranium and radon. Thorium is not commonly used in industry, but becouse of its radioactive properties it has a great potentional for future use in nuclear energetics. Thus, increasing release of Th into the soil, water and atmosphere can be expected in near future and through plant biomass it could become a part of food chains and webs and, thus, to represent a considerable health risks to humans. Studies devoted to research on thorium and its effects on plants has not been published much yet. The majority of these studies focus mainly only on plant abilities to accumulate thorium and/or monitoring distribution of thorium in plant body. But till now, a study is still missing, which would monitor the effect of thorium on the physiological characteristics of plants. Plants of Nicotiana glutinosa (L.) medium Thorium accumulatin were hydroponically cultivated Hoagland nutrient media differedin the presence of thorium, tartaric acid, putrescine and phosphates. In first part of the present study I monitored accumulation of thorium by tobacco plants under the influence of the above-mentioned modifications of media. In the second part, I studied the Th effect on the photosynthetic apparatus (contents...
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Leaf structural changes induced by tropospheric ozone.
Češpírová, Zdeňka ; Lhotáková, Zuzana (advisor) ; Hála, Michal (referee)
Tropospheric ozone is an important pollutant formed by the photochemical reactions of ultraviolet radiation and nitrogenous. Recently, its concentration in the air is increasing due to the promoting climate change and antropogenic activities, mainly because of combustion of the fossil fuels. The ozone is highly reactive molecule degrading to the reactive oxygen species having negative effects on the leaf physiological functions and structure. Ozone penetrates the leaf through the stomata, thereafter it reacts with the cells and their singular compartments. The products formed due to ozone degradation are able to cause the disintegration of plasma membranes and other oxidative damage, for example accumulation of plastoglobuli inside the chloroplasts, changes in the shape of chloroplasts, mitochondria or peroxisomes. At the macroscopic level the visible lesions, chloroses and necroses are observed. In addition, the faster senescence of leaves is observed under the effect of ozone. The main aim of present thesis is to review knowledge of the ozone-induced changes in the leaf structure beginning on the cell structure level and ending at the whole plant body level and to explain physiological mechanisms leading to these damages. Key words: ozone, oxidative stress, leaf structure, mesophyll, reactive...
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Cell death as a result of iron-induced cellular damage
Běhounek, Matěj ; Balušíková, Kamila (advisor) ; Truksa, Jaroslav (referee)
Iron is an essential trace element for almost all living organisms. Iron overload in cells and tissues, however, leads to their disruption. Most oftenly damaged are parenchymatic organs such as the liver, pancreas and heart. The aim of this thesis was to create cellular in vitro models for the investigation of effects of excess iron on hepatocytes and pancreatic beta cells and on these models to investigate cellular processes which lead to cellular damage during iron overload. We focused on examining the presence of oxidative and endoplasmic reticulum stress and the activation of apoptotic cell death. For our experiments, we used HEP-G2 cell line which represents human hepatocytes and NES2Y cell line which represents human pancreatic beta cells. To study the mechanisms of cellular damage during iron overload, we used two approaches by which we observed both acute and long-term effects of high levels of iron on damage of the tested cell lines. When studying the acute effect of excess iron on the cells, we applied high doses of iron (using 15 mM ferric citrate in medium) that led to the activation of cell death in hours. Long-term effects of iron overload were tested on cells regularly cultivated in the presence of 50 μM and 100 μM ferric citrate over a period of several months. Iron concentrations...
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Factors affecting uptake and distribution of uranium in plants.
Krejčová, Daniela ; Lhotáková, Zuzana (advisor) ; Tylová, Edita (referee)
Uranium is a radionuclide, which naturally occurs in Earth's soil in rather an insignificant amount. It is not very dangerous in such small concentration; however, this concentration is rising due to anthropogenic activity, therefore an estimation of its increase is at hand. It is necessary to research possibilities of not only effective, but also ecological extermination of this contamination. Phytoremediation could be an appropriate solution, but this method is still in its beginning stages when it comes to uranium contaminations. That's why we need to study influences of uranium on plants and find out to what extent are they suitable for phytoremediation of areas contaminated by uranium. Phytoremediation methods are based on the abilities of plants to receive and accumulate certain pollutants. The factors that influence the amount of uranium received by a plant can be divided into four groups: plant species, uranium's oxidation levels, pH of of the medium or the substrate and ligand, which is attached to the uranium (Mitchell, 2013). Transportation of uranium to the aboveground part of the plant is also a factor here. Plants need to deal with the accepted uranium; aside from the radiological risks resulting from the characteristics of radioisotope, uranium is also a heavy metal. Chemical...
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Oxidative stress in bacteria - with an emphasis on the model organism of Escherichia coli
Moravcová, Andrea ; Lichá, Irena (advisor) ; Čáp, Michal (referee)
Most bacterial species encounter aerobic conditions during their life, which can be toxic. This leads to a state of oxidative stress. Toxicity of aerobic environment results from the chemical properties of molecular oxygen and its reactive species (ROS). Bacteria had to adapt to these conditions in the past to ensure preservation and prosperity. This thesis is preferably focused on oxidative stress adaptations in the most elaborated bacterial model - Escherichia coli. Regulation of adaptations at the regulation of transcription, translation and metabolism level are described with emphasis on molecular mechanisms. The main adaptation mechanism against oxidative stress is the deactivation of ROS, as well as the repair of damaged cell structures (macromolecules). These enzyme activities are regulated by several transcriptional regulators. The transcriptional regulators OxyR and SoxRS have been well studied in E. coli. Even though these regulators are conserved across the bacterial spectrum, they may not have the same function in all organisms. For this reason, also other, more or less studied bacterial species - Bacillus subtilis, Streptomyces coelicolor, Pseudomonas putida, Pseudomonas aeruginosa - were included in this thesis. The various strategies of how these bacteria use not only OxyR and SoxRS...
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The effect of acute cold and permanent light to left ventricular of the rat heart
Vítková, Ivana ; Žurmanová, Jitka (advisor) ; Bendová, Zdeňka (referee)
Acute cold exposure increases the risk of sudden cardiac events, similarly exposure to constant light negatively affects the cardiovascular system. However, the individual effects of these factors and the effect of their combination on cardiomyocytes are not yet known. The thesis deals with the influence of a 3 day cold exposure and constant light on the expression of β-adrenergic receptors and associated G-proteins in association with apoptotic signals in the left ventricle of the Wistar rat heart. In this work apoptotic proteins BAX, BCL2, caspase 8 and important components of β-adrenergic signalization - β1, β2, G-proteins, Gas, Gi1/2 and Gi3 were determined. The relative expression of the proteins was analyzed by Western blotting. The results confirm the detrimental effect of cold and light exposure. However, the synergistic effect of these two stressors shows surprising results.
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Mild therapeutical hypothermia and oxidative stress after cardiac arrest
Krüger, Andreas ; Ošťádal, Petr (advisor) ; Vízek, Martin (referee) ; Rokyta, Richard (referee)
Successfull cardiopulmonary resuscitation is an essential life-saving tool; nevertheless, general ischemia during cardiac arrest may trigger different pathways that could turn even into a fatal damage; this condition is called post-cardiac arrest syndrome. It has been repeatedly shown that oxidative stress (OS) plays one of the key roles in the development of ischemia-reperfusion injury. However, current evidence on the possible participation of OS in the pathogenesis of post-cardiac arrest syndrome is insufficient. We tested following hypotheses: (i) ischaemia-reperfusion injury after cardiac arrest is accompanied by OS and (ii) mild therapeutical hypothermia decreases OS cardiac arrest. In the experimental part of our work we studied the effects of hypothermia and normothermia on hemodynamic parameters, markers of organ damage and on the OS burden in porcine model of cardiac arrest. Furthermore, we compared the effects of hypothermia with ischaemic postconditioning and nitric oxide administration in the porcine model of extracorporeal cardiopulmonary resuscitation. We found protective effects of hypothermia on all major endpoints including OS in comparison with normothermia; moreover, hypothermia improved also selected variables compared to ischemic postconditioning and nitric oxide. In the...
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