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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 damage to cellular components after oxidative stress induction by specific herbicides
Kramná, Barbara ; Wilhelmová, Naďa (advisor) ; Ryšlavá, Helena (referee)
Oxidative stress is caused by overproduction and overaccumulation of ROS (reactive oxygen species). This state is responsible for cellular damage during unfavorable environmental conditions such as drought, low temperatures, salinity. In order to directly study oxidative stress at tobacco plants (Nicotiana tabacum cv. Xanthi) I used specific herbicides, MV (methyl viologen) and 3-AT (3- aminotriazole). There were several markers used for monitoring oxidative damage to cellular components: DNA damage detected by a comet assay, lipid peroxidation, carbonylated proteins and modification of activities of antioxidant enzymes CAT (catalase) and APX (ascorbate peroxidase). Fluorescent microscopy documented changes in a redox state of tobacco cells and a specific signal for peroxisomes was observed after treatment with higher concentrations of MV and 3-AT. Application of both herbicides caused significant DNA damage, while they worked in a different concentrations, MV in µM and 3-AT in mM. Another convincing oxidative stress marker for MV was protein carbonylation. The inhibition of antioxidant enzymes CAT and APX was less significant when compared to the effects of 3-AT. Decreasing membrane stability proved to be an universal oxidative stress marker for both herbicides. On the other hand, lipid...
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Oxidative stress and condition-dependence of ornamental signals of quality in socially monogamous songbird
Valášek, Stanislav ; Tomášek, Oldřich (advisor) ; Sedláček, Ondřej (referee)
6 ABSTRACT Conditional ornaments plays irreplaceable role in sexual selection in non-small part of by sexual reproduction reproducing animals. Fastidiousness of generating and later also carrying of these ornaments which show condition of their wearer, burdens also metabolism in non-small scale. This thesis tests hypothesis of mutual addiction between conditional ornaments, as the indicators of qualities of individuals and metabolism, as the most significant source of free radicals which are responsible for oxidative stress. The real weight of influence of ornament fastidiousness on organism, resp. on redox state, is tested in this thesis. Manipulations which were performed with individual males of model species should point how much the selected factors correlate each other. The barn swallow (Hirundo rustica) is the model species. Analyses of dates collected during trapping which were realized in breeding seasons in 2012 and 2013 does not show any important trends between observing variables. This fact is confirmed by minimal differences and inconsistent variability of levels of measured antioxidants - oxidoreductases, superoxide dismutase and glutathione peroxidase. The marginal effect of manipulations with one of the conditional ornaments which are presented in model species supports the hypothesis of...
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Production and elimination of superoxide radical in relation to the compatibility of snails and trematodes
Cibulková, Lucie ; Skála, Vladimír (advisor) ; Nývltová, Eva (referee)
Almost all trematodes use snails as the intermediate host in their life cycles. To survive within the host, they have to efficiently avoid defense reactions of its immune system. The most important effector cells, haemocytes, produce reactive oxygen species with the first molecule known as superoxide radical. Various snail species produce different levels of these radicals in relation to the compatibility with the invasive trematode species. The parasite decreases the levels of toxic radicals by using antioxidant enzymes including superoxide dismutase which catalyzes transformation of superoxide radical into hydrogen peroxide. This dismutation reaction is the first step during the oxidative burst and likely influences survival of trematodes within the host. Based on the current knowledge the production and elimination of superoxide radical in relation to the compatibility between snails and trematodes have been described thoroughly for a few models such as for example Biomphalaria glabrata-Schistosoma mansoni. However, this interaction appears to play a key role and, therefore, it deserves more attention in another models as well. Key words: trematodes, snails, compatibility, haemocytes, oxidative burst, antioxidant enzymes, superoxide dismutase, superoxide radical
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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...
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Antioxidant enzymes response to abiotic stress. Impact of decreased cytokinin level.
Lubovská, Zuzana ; Wilhelmová, Naďa (advisor) ; Prášil, Ilja (referee) ; Kočová, Marie (referee)
Charles University in Prague, Faculty of Science Department of Experimetnal Plant Biology Ph.D. study program: Plant physiology and anatomy Abstract Antioxidant enzymes response to abiotic stress. Impact of decreased cytokinin level. Zuzana Lubovská Supervisor: RNDr. Naďa Wilhelmová, CSc. Supervisor-consultant: RNDr. Helena Štorchová, CSc. Praha, 2015 The response of the antioxidant enzyme system to drought, heat and a combination of these stresses was followed in tobacco plants overexpressing cytokinin oxidase/dehydrogenase1 (CKX1) under the root-specific WRKY6 promoter (W6:CKX1 plants) and under the constitutive 35S promoter (35S:CKX1 plants) and in the corresponding wild type (WT). CKX1 over-producing lines developed a bigger root system, which contributed to their stress tolerance. The high stress tolerance of 35S:CKX1 plants was also associated with a dwarf shoot phenotype and changed leaf morphology. The genes for chloroplastic antioxidant enzymes, stroma ascorbate peroxidase (sAPX), thylakoid ascorbate peroxidase (tAPX) and chloroplastic superoxide dismutase (FeSOD), which are responsible for scavenging of reactive oxygen species produced via electron transfer during photosynthesis, were all strongly transcribed in control conditions. All the tested stresses down-regulated expression of these genes...
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The role of mitochondria in adaptation to chronic hypoxia in the spontaneously hypertensive and conplastic rats.
Weissová, Romana ; Kalous, Martin (advisor) ; Rauchová, Hana (referee)
Adaptation to chronic hypoxia provides cardioprotective effects. Molecular mechanism of this phenomenon is not yet completely understood, but it is known that cardiac mitochondria play an essential role in induction of protective effects. The purpose of this diploma thesis is to study effects of continuous normobaric hypoxia (CNH; 10 % O2, 21 days) on spontaneously hypertensive rats (SHR) and conplastic strain that is derived from SHR. These animals have nuclear genome of SHR strain and mitochondrial genome of Brown Norway (BN) strain. Cardiac homogenate was used to measure enzymatic activity of malate dehydrogenase (MDH), citrate synthase (CS), NADH-cytochrome c oxidoreductase, succinate-cytochrome c oxidoreductase and cytochrome oxidase (COX). Using Western blot procedure the protein amount of antioxidant enzymes was measured - manganese superoxide dismutase and copper-zinc superoxide dismutase (MnSOD, Cu/ZnSOD), catalase and chosen subunits of oxidative phosphorylation complexes (Ndufa9, Sdha, Uqcrc2, COX-4, MTCO1, Atp5a1). Under normoxic conditions the conplastic strain has lower amount of complex IV subunit MTCO1 in comparison with SHR. This subunit is encoded by mitochondrial DNA and it is one of the seven protein-coding genes in conplastic strain that differ from SHR. Adaptation to hypoxia causes an...
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Influence of supplementation with n-3 polyunsaturated fatty acids on inflammatory markers in patients on long-term parenteral nutrition
Svěchová, Hana ; Novák, František (advisor) ; Meisnerová, Eva (referee)
SMOFLipid® is a commonly used fat emulsion for parenteral nutrition. We investigated how enrichment of SMOFLipid® with n-3 polyunsaturated fatty acids (PUFA) in a form of second fat emulsion, Omegaven® , changes fatty acid composition of total plasma phospholipids and erythrocyte phospholipids, cytokine concentrations in serum and in supernatant from in vitro whole blood culture stimulated with lipopolasaccharide (LPS) and we evaluated also changes in oxido- reductive balance. Eight patients on long-term home parenteral nutrition recieved both emulsions, SMOFLipid® (6 weeks) and SMOFLipid® +Omegaven® (4 weeks), one by one. We observed no significant differences in common laboratory and clinical parameters between these two types of diet. Enrichment of SMOFLipid® with Omegaven® led to an increase in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) in total plasma phospholipids and there was also an increse in proportion of EPA in erythrocyte phospholipids, while proportion of DHA remained unchanged. These changes were in both phospholipids of plasma and erythrocyte compensated for a decrease in proportion of linoleic and arachidonic acid (n-6 PUFA). There were elevated IL-6 and TNF-α serum concentrations in patients after both diets. There was a decrease in IL-6 production by 36% with SMOFLipid®...
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Mitochondrial subpopulations in rat myocardium - effect of chronic hypoxia
Kovalčíková, Jana ; Nováková, Olga (advisor) ; Žurmanová, Jitka (referee)
Adaptation to chronic hypoxia induces endogenous cardioprotection and increases the heart resistance to ischemia/reperfusion injury. The heart mitochondria, which produce reactive oxygen species (ROS) in addition to ATP, play an important role in these processes. During ischemia/reperfusion, ROS are produced in excessive amounts and damage the cells. However, in lower concentrations, ROS are involved in the signalling pathway of cardioprotection induced by adaptation to chronic hypoxia. In the heart, two mitochondrial subpopulations have been observed, subsarcolemmal mitochondria (SSM) and intermyofibrillar mitochondria (IMFM), which differ in cell localization as well as in morphological and biochemical properties. The aim of this work was to introduce the method of SSM and IMFM isolation in our laboratory and to analyse their antioxidative capacity after adaptation to chronic hypoxia. Adult male Wistar rats were kept either under normoxic conditions or exposed to intermittent high-altitude hypoxia (IHA; 7000 m, 5 days a week/8 hours a day, totally 25 exposures). Mitochondrial subpopulations were isolated from heart left ventricle and their functionality was verified by measuring oxygen consumption and enzyme activities. The IMFM had higher oxygen consumption in comparison with SSM and activities...
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Antioxidative enzymes in heart
Kletečková, Lenka ; Hloušková, Patricie (advisor) ; Rychlíková, Jana (referee)
Cardiovascular diseases are one of the most frequently causes of morbidity and mortality in developed countries. An increase of oxidative stress is implicated by progression of heart diseases. Antioxidative enzymes protect the organism against them. The superoxide dismutases, the peroxiredoxins, the glutathione peroxidases and the catalase are advised to be used as the main antioxidative enzymes. The review is focused on the role of these antioxidative enzymes in the cardiovascular system. Antioxidative enzymes prevent the lipoperoxidation of biological membranes, the proteins inactivation, the DNA mutation and the apoptosis. It is well-known, that antioxidative enzymes positively influence a homeostasis of the vascular endothelium, prevent a burst of the cardiovascular disease and they are important for physiological functions, the morphology and the contractility of heart. The effect of antioxidative enzymes on the cardiovascular system is very complicated process and it still has not clarified all aspects of their action. A reseach of these enzymes is essential for understading the mechanisms of pathophysiological changes, which proceed into the cardiovascular system during the atherosclerosis, the myocardial infarction, the ischemic- reperfusion injury of the tissue and the calcification and...
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