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Recombinant expression of transcription factor TEAD1.
Řeháková, Jana ; Novák, Petr (advisor) ; Bláha, Jan (referee)
TEAD1 is one of the members of the TEAD transcription factor family. This transcription factor is very important, for example for development of organs. The structure of the entire TEAD1 transcription factor is not now yet known. Nevertheless, structure of two important domains are known. The TEA binding domain, which is important for the binding of the transcription factor TEAD1 to DNA, and the transactivation domain, to which coactivators bind. TEAD1 binds to DNA and upon coactivator bind it affects the transcription of various genes. Genes, that are affected by the transcription factor TEAD1 includes genes regulating proliferation, differentiation and apoptosis of cells. TEAD1 is also the target of the Hippo signalling pathway, which is active in adulthood and prevents abnormal growth of organs. Important for the activity of transcriptional factor TEAD1 are post-translation modifications, such as palmitoylation and phosphorylation. To discover the entire structure of the transcriptional factor TEAD1 and the way it interacts with DNA, the transcriptional factor TEAD1 was prepared recombinationally by expression in cells of Escherichia coli bacteria. Suitable conditions for production of the transcriptional factor TEAD1 were found and the cleavage of the histidine tag by thrombin was performed....
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Study of structure and interaction of human lymphocyte receptors
Bláha, Jan
Natural killer (NK) cells are an essential part of immune system, providing self-surveillance of virally infected, stress transformed or cancerous cells. NKR-P1 receptors and their ligands from clec2 gene family represent an alternate missing-self recognition system of NK cells based on interaction of highly related C-type lectin-like receptors. Human NKR-P1 has been described more than twenty years ago but still remains the sole human orthologue of this receptor family, particularly numerous in rodents. On binding to its cognate ligand LLT1, NKR-P1 can relay inhibitory or co-stimulatory signals. Although being interesting targets for their potential role in tumor immune evasion and autoimmunity, nature of their interaction is still unclear. To elucidate the architecture of their interaction, we developed a generally applicable method for recombinant expression of human NKR-P1 and LLT1 and their homologues based on transfection of HEK293S GnTI- cells. Further, we described a stabilizing mutation His176Cys, that enables for expression of highly stable and soluble LLT1. Finally, we have crystallized LLT1 and human NKR-P1 in different glycosylation states both as individuals and in complex. While both structures of LLT1 and NKR-P1 follow the classical C-type lectin-like superfamily fold, contrary to...
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The regulation of brown bullhead - \kur{Ameiurus nebulosus} (Lesueur, 1819)
BLÁHA, Jan
The presence of non-native fish species is one of the causes of loss of biodiversity or even degradation of entire ecosystems. The spread of non-native species is often of invasive nature, which is accompanied by a number of direct and indirect effects on the original organisms. These species are often characterized by greater aggressiveness, resistance, growth rate, early sexual maturity period or the presence of anti predatory mechanisms. The expansion of non-native species is mostly and significantly the result of human behaviour. This bachelor thesis is focused on the possibility of regulation of Brown Bullhead or its similar relative Black Bullhead. Brown Bullhead has been present in our country since 1890. The regulation was done by the means of traps. Different kinds of baits were also used in order to maximize the efficiency and selectivity. The data on the actual occurrence of these species in the Czech Republic has been obtained based on the mapping. Historical data need to be confirmed or disproved of and an assessment of its occurrence had to be made as accurately as possible. The thesis also describes other non-native fish species currently present in the waters of the Czech Republic that can affect water ecosystems. The conclusion also contains suggestions of some preventive measures that could be taken to eliminate the possibility of further spread of non-native species of fish in the Czech Republic.
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Production of IP camera parts on FDM 3D printer
Blaha, Jan ; Šeniglová, Hana (referee) ; Zemčík, Oskar (advisor)
Bachelor thesis dealing with production of IP camera body, using 3D printing method. This thesis contains two parts. The first one is research part and the second is design part. The first part summarizes the problems in the area of industrial cameras and information about 3D printing method. At the end of the research section, a suitable material is selected for printing on the FDM printer. The design part describes prototype design and production of the selected part. Design includes control calculations and technological design assessment. At the end of the bachelor thesis, there is presentation of technical-economic evaluation and recommendations for the future.
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Study of structure and interaction of human lymphocyte receptors
Bláha, Jan ; Vaněk, Ondřej (advisor) ; Šulc, Miroslav (referee) ; Obšil, Tomáš (referee)
Natural killer (NK) cells are an essential part of immune system, providing self-surveillance of virally infected, stress transformed or cancerous cells. NKR-P1 receptors and their ligands from clec2 gene family represent an alternate missing-self recognition system of NK cells based on interaction of highly related C-type lectin-like receptors. Human NKR-P1 has been described more than twenty years ago but still remains the sole human orthologue of this receptor family, particularly numerous in rodents. On binding to its cognate ligand LLT1, NKR-P1 can relay inhibitory or co-stimulatory signals. Although being interesting targets for their potential role in tumor immune evasion and autoimmunity, nature of their interaction is still unclear. To elucidate the architecture of their interaction, we developed a generally applicable method for recombinant expression of human NKR-P1 and LLT1 and their homologues based on transfection of HEK293S GnTI- cells. Further, we described a stabilizing mutation His176Cys, that enables for expression of highly stable and soluble LLT1. Finally, we have crystallized LLT1 and human NKR-P1 in different glycosylation states both as individuals and in complex. While both structures of LLT1 and NKR-P1 follow the classical C-type lectin-like superfamily fold, contrary to...
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Analysis of the weaknesses of IT process
Bláha, Jan ; Svatá, Vlasta (advisor) ; Říhová, Zora (referee)
The thesis focuses on research of IT process weakness. In theoretical part are described basic terms and also frameworks or best practise of process maturity rating. Selected practice ITIL and COBIT are then used for rating in practical part. For this purpose is choosen one of process which is outsourced by external contractor. Rating is made using free accessible tools, which are offered by selected practice and which offer rating process. Result is established according to consultation with people involved in project and their rating using mentioned tools.
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Regulatory mechanisms of fetoplacental vasculature in hypoxia
Kafka, Petr ; Cvachovec, Karel (advisor) ; Zoban, Petr (referee) ; Bláha, Jan (referee)
The placenta and the lungs are the only organs that perform a function of blood oxygenation and have a common characteristic - vasoconstriction in response to acute hypoxia. Thus the reaction is in an opposite manner than smooth muscle of systemic arteries does. In this study we focused on the mechanisms that regulate fetoplacental blood vessels in conditions associated with hypoxia. An experimental model of isolated dually perfused rat placenta was used. Our findings demonstrate an essential role of Rho kinase signaling pathway in the mechanism of this hypoxic fetoplacental vasoconstriction (HFPV). Chronic exposure to hypoxia causes a sustain elevation of vascular resistance on fetoplacental blood vessels. It was demonstrated in experiment with low-viscosity salt solution perfusion. Our current results show a significant increase of the fetoplacental vascular resistance with blood perfusion which better reflects in vivo conditions. An increase in fetoplacental vascular resistance may lead to placental hypoperfusion and consequent fetal undernutrition, which is considered one of the key causes of serious fetal and neonatal problems, especially intrauterine growth restriction. Diabetes mellitus is a well-known factor affecting fetal growth. Both chronic hypoxia and diabetes act on vessels partly...
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The impact of the cartographic method on students' map skills level
Havelková, Lenka ; Hanus, Martin (advisor) ; Bláha, Jan Daniel (referee)
The presented diploma thesis focuses on the map skills' problematic, which is currently becoming widespread in the Czech research environment. In contrast to the previous empirical studies, this thesis is devoted to the thematic map skills and their level of development among students of grammar schools and geographically oriented branches of university studies. Due to the significant differences between used cartographic methods of visualization on thematic maps the main aim of this diploma thesis is to identify and explain used cartographic method's influence on the level of these important skills. To achieve this aim, it was necessary to focus on different parts of the curriculum (intended, implemented, learned) and to use several scientific methods because this topic hasn't been well established yet and therefore only few theoretical and empirical studies dealt with this specific issue. Firstly, the review of literature focused on thematic cartography, map skills and on identifying their level of development among students was done. Due to the lack of specialized resources the model of thematic map skills was created. This specific model was further used during didactic test's designing and also for its results' interpretation. Similarly for the creation of appropriate didactic test -...
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