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Evaluation of antibiotic-induced mitochondrial superoxide production in adherent cells
Ingrová, Kateřina ; Chmelíková, Larisa (referee) ; Zumberg, Inna (advisor)
The theoretical part of this bachelor´s thesis contains a description of the effect of reactive oxygen species on oxidative stress by mitochondria and the consequences of antibiotics use in cell line culturing. The cell line studied in this bachelor´s thesis is the mesenchymal stem cells (MSCs). The practical part describes the procedure of the experiment including cell culturing, passaging and cell labeling. The proposed experiment was repeated with sufficient number of repetitions. Finally, confocal microscopy images were processed in the MATLAB programming environment.
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Evaluation of glutathione content in plants as a marker of heavy metals environmental contamination
Borková, Marie ; Stoupalová, Michaela (referee) ; Opatřilová,, Radka (advisor)
Dependence of glutathione concentration on the amount of thallium in the plant was studied. Observed plant was maize (Zea mays) which was divided to two parts – root and overground. Two culture procedures were elaborated where seeds and young seedlings were cultivated in a solution of thallium of concentration 0, 1, 3, 5, 8, a 10 µmol/l. Extraction agents used during extraction were phosphate buffer and solution of ascorbic acid. Determination of glutathione was realized by capillary electrophoresis (CE) and high performance liquid chromatography (HPLC). Diode array detector (DAD) was used in both methods. Quantification of the thallium amount in the plant was done by method of inductively coupled plasma-atomic emission spectrometry (ICP-AES).
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The effect of air pollution on oxidative stress markers in newborns
Ambrož, Antonín ; Rössner, Pavel (advisor) ; Rubeš, Jiří (referee) ; Gábelová, Alena (referee)
In everyday life, humans are exposed to toxic substances of anthropogenic origin. These substances can also be found in the ambient air and their impact poses a long-term risk for human health. Respirable particulate matter (PM) of aerodynamic diameter < 2.5 µm (PM2.5) is intensively studied, along with carcinogenic polycyclic aromatic hydrocarbons (PAHs), bound to it, such as benzo[a]pyrene (B[a]P), a reference carcinogenic PAH. Owing to small size, PM2.5 can penetrate the human body primarily via the airways and represent an increased health risk compared to larger particles. The negative health impacts of anthropogenic PM2.5, generated e.g. by fossil fuel combustion, are linked with its small size, relatively large surface, as well as with PAHs and other substances adsorbed on PM surface. PAHs, generated by an incomplete combustion of organic matter, can enter organism either via ingestion of contaminated food, water or via inhalation of polluted air. PAHs affect organisms via genotoxic, mutagenic, carcinogenic, embryotoxic and other adverse effects. One of the common denominators of these effects is oxidative stress, which is also considered to be the main mechanism of action caused by PM in the human organism. Oxidative damage induced by reactive oxygen species (ROS) may affect any cellular...
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Redox signaling to chromatin
Plšková, Zuzana
Chromatin is a highly dynamic structure, which is constantly subjected to regulation in response to environmental conditions. This response is mediated by chromatin modifiers, such as histone acetyltransferases, which deposit histone post-translational modifications, resulting in changes in gene expression. One of such modifiers in plants is GENERAL CONTROL NON-REPRESSIBLE 5 (GCN5). Despite its importance in plant growth and development, it remains poorly understood how GCN5 is regulated. Emergine evidence points towards redox regulation of chromatin remodeling that can be executed via redox modifications of chromatin modifiers. To investigate possible redox regulation of GCN5, we used redox insensitive lines, carrying GCN5 with mutated cysteine to serine. Even though gcn5 mutants displayed enhanced susceptibility to paraquat-induced oxidative stress, the mutated lines phenocopied the wild type. We further probed the interactome of GCN5 to identify putative functional partners, whose association with GCN5 could be altered under oxidative stress conditions, or affected by its redox status. Mutating of a single cysteine residue in GCN5 did not result in significant changes of its interactome, suggesting that additional single and higher order mutants need to be explored.
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Pathophysiological and genetic factors affecting serum uric acid level.
Hasíková, Lenka ; Závada, Jakub (advisor) ; Hrnčíř, Zbyněk (referee) ; Horák, Pavel (referee)
Introduction: Serum uric acid level (SUA) depends on the balance between its production and excretion. SUA is associated with several transmembrane proteins responsible for reabsorption (mainly URAT1 and GLUT9) and secretion (ABCG2) on the apical and basolateral membranes of the proximal tubules in the kidney, and in the case of ABCG2, it also correlates with its significant excretion through the gastrointestinal tract. Gout is a metabolic disease caused by the deposition of urate crystals in the joints and tissues. Chronic hyperuricemia is a primary risk factor for the development of gout; however, gout patients usually have a lower SUA during an acute gout attack than in the intercritical periods. The exact mechanism of this phenomenon is unknown. It has been speculated that the systemic inflammatory response can explain this discrepancy. The aim of the study is to determine whether treatment with specific inhibitors of the proinflammatory cytokine TNF (TNFi) affects SUA in patients with systemic rheumatic disease (SRD), and whether changes in SUA correlate with changes in selected proinflammatory cytokines or with the biomarker of oxidative stress, allantoin. Another aim is to determine the frequency and effect of allelic variants in the ABCG2 urate transporter gene in patients with primary...
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The effect of yttrium and samarium oxide nanoparticles on plant development
Brandová, Zuzana ; Soudek, Petr (advisor) ; Petrová, Šárka (referee)
This thesis is based on hydroponic experiment conducted on Hordeum vulgare with nanoparticles of rare earth elements in concentration 2·10-4 mol/L and REEs chlorides in concentration 3·10-4 mol/L. The comparison of the the effect on their acumulation of another nanoparticles in the sloution and normal medium is included as well. In this case the hydroxyapatite NPs were chosen in the same concentration as yttrium and samarium NPs. The role of type and form of the elements on their acumulation and impact is also studied. Negative effect of REEs was proven by enzymatic analysis that detects reducting enzymes and determination of plant pigment levels by high performance liquid chromatography. Because both types of substances can reduce reactive oxygen radicals. Acumulation of REEs was determined by ICP spektrometry. Key words: nanoparicles, reare earth elements, reductive enzymes, plant pigments, acumulation, oxidative stress
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Heme sensor proteins as potential biomarkers of cellular and oxidative stress processes induced by ionizing radiation
Vávra, Jakub ; Martínková, Markéta (advisor) ; Souček, Pavel (referee) ; Tichý, Aleš (referee)
[IN CZECH] Ionizing radiation is a potential inducer of the oxidative stress processes in cells. As a result, reactive radicals are formed in the intracellular space modifying the essential biomolecules. Ionizing radiation has either direct (radiation sickness) or indirect (malignant processes) effect on the organism. Therefore, a fast determination of the dose is required when suspected irradiation of the organism occurs. However, a method routinely applicable, fast enough and at the same time suitable for dose estimation based on biomarkers has not been developed so far. The aim of this thesis is to describe new properties of the selected heme sensor proteins and discuss their potential importance for the cellular adaptation to oxidative stress conditions. Specifically, the thesis is focused on two eukaryotic proteins, heme regulated inhibitor (HRI) and transcription factor p53. The study of functional regulation as well as the conformational changes of these proteins induced by heme is greatly emphasized. Besides, the optimization of the key experimental methods was conducted. Specifically, Phos-tag electrophoresis was applied for the kinetics study of HRI wild type and its Gly202Ser mutant form, which is a characteristics of lung cancer development. Unsurprisingly, for both HRI forms studied,...
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Role of oxidative stress in male infertility.
Dolečková, Barbora ; Tlapáková, Tereza (advisor) ; Šanovec, Ondřej (referee)
Oxidative stress is a phenomenon caused by an excess of reactive oxygen species (ROS), or by insufficient activity of antioxidants, that reduces these ROS levels and thereby protect the organism from oxidative damage. ROS have two types of origin: endogenous, which includes leukocytes and immature sperm, and exogenous, which includes factors such as air pollution caused by heavy metals, smoking tobacco products, obesity and others. Low levels of ROS have a positive effect on the physiological functions of the organism, including the process of spermatogenesis, where ROS participates in the course of hyperactivation and capacitation. However, increased levels of ROS trigger a number of cellular pathologies, whether the loss of fluidity of biological membranes due to lipid peroxidation, deformation of enzymatic proteins or DNA fragmentation, which negatively affects individuals' infertility. Due to the significant positive correlation of ROS scavenging by antioxidants with improving sperm parameters of an infertile individual, antioxidant therapy has recently begun to be used as a possible successful component of male idiopathic infertility treatment.
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Induction of oxidative stress in skin keratinocyte cells
Martínková, Nikola ; Jirkovská, Anna (advisor) ; Hradiská Breiterová, Kateřina (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Student: Nikola Martínková Supervisor: PharmDr. Anna Jirkovská, Ph.D. Consultant: Mgr. Petr Jošt, Ph.D. Title of diploma thesis: Induction of oxidative stress in skin keratinocyte cells Keratinocytes are part of the epidermis and represent the majority of cells in the upper layer of human skin. When these cells are damaged by mutagenic substances, changes in the composition of the genetic material may occur, and even cell death may occur. Such substances that induce changes in DNA and have mutagenic potential include the chemical warfare agent sulfur mustard. The purpose of this study was to compare the HaCaT keratinocyte cell line with the experimentally derived 3HSM4 cell line in response to hydrogen peroxide-induced oxidative stress. The 3HSM4 cells were prepared by selecting resistant clones of HaCaT cells that were repeatedly exposed to the cytotoxic effects of sulfur mustard. The results of the comparison show that 3HSM4 cells proved to be more resistant to hydrogen peroxide. When glutathione (GSH) synthesis was blocked in both cell types, the cytotoxic effect of hydrogen peroxide was the same. Although 3HSM4 cells were shown to be more resistant in terms of cytotoxicity, they were found to have much...
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Microplastics and their effects on the metabolism of animal cells
Fiedlerová, Gabriela ; Cajthaml, Tomáš (advisor) ; Černý, Jan (referee)
Microplastics are tiny particles smaller than 5 mm in size found in various environments, including seas, rivers and soil. These particles can be intentionally produced as part of cosmetic products or formed by the breakdown of larger plastic objects. Microplastics represent a serious threat to the environment and animals and could eventually reach humans through the food chain. This thesis deals with a critical and synthetic overview of the literature related to the influence of microplastics on the metabolism of animal cells. The thesis also considers the experimental conditions under which the data were obtained. In particular, factors such as the tested microplastics' shape, size and ageing are critically evaluated as these aspects are often neglected in the literature. Current knowledge shows that microplastics accumulate in the bodies of animals and cause physical and metabolic damage, inflammation, disruption of energy metabolism, protein metabolism and amino acid and lipid metabolism. The most serious effects of microplastics can be considered to be the formation of oxygen radicals, neurotoxicity, disruption of gametogenesis and offspring development.
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