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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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Energetic use of wastewater sludge and microwave pyrolysis products
Šimek, Radim ; Ševčík, Jan (referee) ; Hlavínek, Petr (advisor)
The diploma thesis is focused on the energy utilization of sewage sludge before and after microwave pyrolysis process. The first part of the thesis deals with sludge management, treatment of sludge and its subsequent use or disposal. In the second part of the thesis prepared and modified samples of sludge from WWTP 1 and WWTP 2 are subjected to microwave pyrolysis process. Subsequently, samples are taken for analyzes to determine the total organic carbon, the specific surface area, the heavy metal content and the calorific values. The resulting data was processed in Microsoft Office Excel and presented at work. At the end of the thesis two case studies are then proposed for a specific design of the conceptual location of microwave pyrolysis for the sludge drying process and the drying process of the sewage sludge in the direct combustion boiler room.
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Accounting Directives for Company
Machačová, Hana ; Ševčík, Jan (referee) ; Pěta, Jan (advisor)
The bachelor thesis is focused on internal guidelines of the company ITAB Shop Concept CZ, a.s. The first part is devoted on theoretical background of internal directives, accounting and inventory valuation and legislative bases governing the formal and concept aspects of these documents.. The second part contains a company introduction and evaluation of the existing guidelines. The last part contains draft of internal guideline dealing with comprehensive inventory valuation and accounting of stocks.
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