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Study of population specific alterations of breast cancer predisposition genes in Czech Republic.
Judasová, Kristýna ; Ševčík, Jan (advisor) ; Holá, Dana (referee)
Breast cancer is the most frequent malignant disease in the female population worldvide. About 10 % of all cases are of hereditary origin. The inactivation of tumor suppressor gene BRCA1 is the main genetic predisposing factor in breast cancer in the Czech Republic. Primarily, BRCA1 participates in DNA double strand break repair. Depending on cell cycle phase, the damage is repaired by homologous recombination or non-homologous end joining. Alternative splicing variants of BRCA1 are frequently detected during the genetic screening of high risk patients. The clinical significance of these variants is unknown. Understanding of the nature of breast cancer genetics is the critical factor for early diagnosis. Based on earlier studies from the Institute of Biochemistry and Experimental Oncology 1st Faculty of Medicine Charles University, two alternative splicing variants which were repeatedly detected in patients, were chosen for functional analysis. The aim of this work is to investigate the impact of alternative splicing variants BRCA1Δ5 and BRCA1Δ10 on DNA double strand breaks repair. Particular variants were over- expressed in the cells of model system. Activity of homologous recombination (HR) and non-homologous end joining (NHEJ) was scored by in vitro DNA repair assay. The cellular localization of...
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Diversity, systematics, and phylogeny of families Opetiidae and Platypezidae (Diptera)
Tkoč, Michal ; Prokop, Jakub (advisor) ; Ševčík, Jan (referee) ; Zatwarnicki, Tadeusz (referee)
5 ABSTRACT This doctoral thesis is focused on flat-footed fly families Opetiidae, Platypezidae and genus Microsania (Insecta: Diptera). The thesis consists of general introduction to the systematics, taxonomy, diversity, zoogeography, phylogeny, and biology of the flat-footed flies. Systematics and taxonomy are summarized based on historical as well as recent literature. Each important taxon is shortly introduced and information about its taxonomy, morphology, diversity, distribution, and biology is provided. The next two chapters deal with fossil species and research on species diversity. The following part is a summary and plan for future research. Final chapter consists of 9 published peer-reviewed articles (5 in impacted international journals and 4 in journals without impact factor). The first article is a molecular phylogeny study of relationships between genera of flat- footed flies. The results show Platypezidae consisting of two well-supported clades, the first with the subfamilies Melanderomyiinae + Callomyiinae and the second formed by subfamily Platypezinae. Genus Microsania was resolved as a separate lineage distant from Platypezidae which clustered with Opetiidae as its sister group, both together forming a sister group to Platypezidae. The genus Agathomyia proved to be paraphyletic....
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Preprocessing of sewage sludge for pyrolysis process
Ševčík, Jan ; Rusník,, Igor (referee) ; Bodík,, Igor (referee) ; Hlavínek, Petr (advisor)
The concept of sludge management at the level of sludge treating technology of sewage treatment plants, as well as at national, continental or world level, requires fundamental improvement in the interest of sustainable development. Sewage sludge production is continuously increasing. Sewage sludge contains a number of pollutants but also a significant renewable resource for material transformation and / or energy recovery. Sewage sludge is composed of an inorganic mineral part and organic components. Its organic part is a source of energy and nutrients, but also contains harmful pollutants for exapmple heavy metals. However, there are technologies that offer the possibility of neutralizing pollutants while using other sludge components in terms of recycling energy, nutrients and other attractive sludge components. The mentioned technologies include mainly thermal methods of sewage sludge treatment and one of them is pyrolysis. Thessis focuses on sewage sludge prepartation for pyrolysis process, which includes mixing with additives and peletization of prepared mixture for purposes of agricultural or energetical utilisation. Technology for material transformation and energy utilization of sewage sludge, in particular the mentioned pyrolysis or torrefaction, require an insignificant degree of detailed research in order to expand it in the field of sludge management. Indeed, it is not only research that will enable direct correct use of this technology in practice. The results of such research are the basis for the drafting of the relevant legislation, which will subsequently enable the deployment of pyrolysis technologies in practice.
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The functional in vitro analysis of the BRCA1alternative splicing variants
Ševčík, Jan ; Kleibl, Zdeněk (advisor) ; Stopka, Tomáš (referee) ; Macůrek, Libor (referee)
BACKGROUND: The inactivation of the tumor suppressor gene BRCA1 is a predisposing factor for a breast/ovarian cancer development. Formation of cancer-specific alternative splicing variants with aberrant biological properties can represent additional mechanism decreasing the overall BRCA1 activity in DNA double strand break (DDSB) repair. In this study, we analyzed BRCA1 alternative splicing variants BRCA114-15 and 17-19 ascertained previously during the screening of high-risk breast cancer individuals. METHODS: We established a stable MCF-7 cell line-based model system for an in vitro analysis of BRCA1 variants. Using this system, we analyzed the impact of BRCA114-15 and 17-19 variants on DNA repair kinetics using comet assay and confocal immunomicroscopy. The capacity of DNA repair was assessed directly by an in vitro NHEJ assay and indirectly by a mitomycin C sensitivity test. The proliferation activities were determined by a clonogenic assay and growth curves. RESULTS: Overexpression of BRCA114-15 and 17-19 increases the endogenous level of DNA damage, slows down the DDSB repair, and decelerates the initial phase of radiation-induced foci formation and prolongs their persistence. Moreover, BRCA114-15 and 17-19 differentially influence the activity of HR and NHEJ and sensitivity of MCF-7 cells to ionizing...
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Design and manufacture of bending fixture
Ševčík, Jan ; Trčka, Tomáš (referee) ; Zemčík, Oskar (advisor)
The work deals with the design and manufacture of bending fixture. It contains a comparsion with current bending fixtures on the market and a justification for the production of a given type. Then the work contains a proposal of the constuction of individual parts with the selection and description of technologies of their production and the process of their production. The work also contains complete drawing ducumentation of all parts of the bending fixture. The conclusion of the work contains the possibilities of design modifications and economic and technological evaluation.
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Analýza stavu ekologického zemědělství a bioprodukce ve vybraném regionu Zlínského kraje - okres KROMĚŘÍŽ
Ševčík, Jan
The diploma thesis „Analysis of the conditions of the ecological agriculture and bioproduction in the selected region Zlín, Kroměříž district deals with the contemporary conditions of the ecological agriculture and bioproduction in the Kroměříž district and with the availability of a range of goods for consumers. Principals and goals of the ecological agriculture, legislation, control mechanisms and production of the organic food are described in the literature review. History and development of the eco-agriculture in both the Czech Republic and the world are also described here. Next section focuses on the description of the Kroměříž district and its environmental conditions for agriculture as well as of the contemporary conditions of the eco-agriculture and production of the organic food. Last part focuses on the analysis of the companies in which the overall benefits for the region is found out through a questionnaire survey. The second questionnaire will be targeting directly on consumers of the organic food. The research findings and subsequent discussion are summarised in the conclusion.
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Sanitary technical installations and gas pipeline in the hotel
Ševčík, Jan ; Wierzbická, Helena (referee) ; Vaščáková, Alena (advisor)
Diploma thesis deals with design of sanitary and gas instalation in hotel building. It is hotel with four floors and basement. Theteoretical part is focused on grey water treatment systems and possibility of their use in the building. The Computational part and project includes design of sanitary and stormwater sewer, water supply system, gas installations a their service pipes to current utility lines.
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Technologies of reuse of wastewater in place of need
Podhrázský, Martin ; Ševčík, Jan (referee) ; Hlavínek, Petr (advisor)
Final thesis is focusing on possibilities hidden in the potential of wastewaters in sewers. The goal of this thesis is to present technologies and experiences with water reclamation and with heat recovery systems using wastewaters as a source of heat energy. The theoretical part is divided in two main topics. The first topic is focused on bringing important knowledge and experiences with designing and operating decentralized sewer mining systems, which are, due to less favourable environmental conditions, largely applicated and developed in countries such as Australia, California, etc. The second topic is focusing on heat recovery from sewage water and possibilities of heating and cooling buildings. A possibility of using heat energy from sewage for heating and cooling a swimming pool building located in Brno, Czech Republic was assessed in the last part of this thesis. As a source of wastewater was considered a sewer main located near the pool building. Payback periods and economical effectivity of investing into wastewater heat recovery system was compared with centralized heat supply system, which is a very common source of heat energy in Czech Republic.
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Clinical classification of sequence variants in non-coding regulatory regions in breast cancer susceptibility genes.
Bubáková, Eliška ; Ševčík, Jan (advisor) ; Vodička, Pavel (referee)
Inactivation of tumor supressor gene BRCA1 causes a life-long risk of breast carcinoma development. Genetic screenings of indicated individuals from high-risk families help to identify large number of sequence variants in known predisposing genes. Majority of discovered variants doesn't have clinical significance yet which causes a big problem for diagnostics. Some of these variants are found within regulatory non-coding regions of gene. A part of the clinical classification of variants is their functional characterization. The goal of this thesis was to create a model system for functional characterization of variants in non-coding regions and to verify its function. Model system was based on targeted gene manipulation by co-transfecting CRISPR-Cas9 construct and donor construct that contained a portion of BRCA1 gene sequence with analyzed modifications, into U2 OS cells. The cells have stably integrated DR-GFP system which allows the activity of homologous recombination (HR) to be determined. Monoallelic modifications were induced into U2 OS cells. These modifications were in a Kozak sequence region of BRCA1 gene. Expression level of BRCA1 mRNA was determined by qRT-PCR, which showed the same levels of mRNA in all cells with analyzed alterations. Next, expression level of BRCA1 protein was...
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