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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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Nonlinear phenomena in electrokinetic chromatography
Dovhunová, Magda ; Dubský, Pavel (advisor) ; Koval, Dušan (referee) ; Ševčík, Juraj (referee)
Capillary electrophoresis often uses complexing agents since the interaction between the analyte and the complexing agent can result in achieving or improving the separation. Examples of such methods can be electrokinetic chromatography or affinity capillary electrophoresis (ACE). ACE is used to determine the complexing parameters. In case of chiral separation, this issue gets complicated, since the parameters of the two analytes (enantiomers) are not completely independent to one another. Therefore, a procedure has been proposed in this thesis, that should always be used to evaluate the complexing parameters of two enantiomers. Statistical evaluation of these parameters was assessed as well. This work also proposes a method that allows to determine the relative migration order of two enantiomers in two different complexing separation systems. The mathematical description of electrophoresis is based on continuity equations, that are inherently nonlinear. However, these equations can be linearized to obtain an approximate analytical solution. There was recently presented a generalized model, that enables inclusion of complete complexing equilibria in the theoretical description of electromigration. Thus, various phenomena, including nonlinear ones, associated with complexation can be predicted. This...
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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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Chromatograhic characterization of chiral and achiral separation systems
Kučerová, Gabriela ; Tesařová, Eva (advisor) ; Čabala, Radomír (referee) ; Ševčík, Juraj (referee)
Dissertation thesis is a 5-publications' collection concerning characterization and application potential of cyclodextrins, polysaccharides and macrocyclic antibiotics based chiral stationary phases. The effects of stationary phase and mobile phase are studied. This approach ensures the complex insight into separation systems studied. Systems with different nature of chiral selector were studied by HPLC. Namely, macrocyclic antibiotics and derivatized polysaccharides were used for experiments. Former ones provided better results for enantioseparation of non-coded amino acids than latter ones. Dynamic coating procedure was used for preparation of a new chiral stationary phase. Characterization of new cationic cyclodextrin based chiral stationary phase was performed. Linear free energy relationship method was used for characterization of two different separation systems, i.e. newly prepared stationary phase and commercially available stationary phase. Based on results obtained, newly prepared stationary phase showed better results for separation of different achiral groups of analysts. New stationary phase prepared by dynamic coating was compared with chromatographic system, in which the chiral selector was used as a mobile phase additive. The chiral selector used for the two different approaches was...
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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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Bonded hybrid joints in the automotive industry
Ševčík, Jan ; Hanáčková, Pavla (referee) ; Kubíček, Jaroslav (advisor)
This diploma thesis deals with bonded hybrid joints which are used for connecting parts of modern coachwork. Firstly, the thesis describes in general the process of production of the whole coachwork, from cutting of the blanks to the surface treatment including the analysis of the used materials. Subsequently, the problem of joining body parts through adhesives, but also by welding, soldering or other methods is discussed. Another part of this thesis is dedicated to an experiment in which the qualities of bonded hybrid joints were determined. Different ways of realizing the joint have been made which differed in the size of the glued surface, in the welding methods and in the process of their creation. The produced joints were tested for their strength and quality.
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The Development of Small Company
Fiedler, Viktor ; Śevčík, Jiří (referee) ; Śevčík, Jiří (referee) ; Koráb, Vojtěch (advisor)
This bachelor’s thesis presents possibilities and directions of the development of small company. It describes the structure of small company and the problematic of its development. It analyses present economical, financial and business situation inside and outside the company. In the second part author introduces some proposals for improving several parts of company.
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