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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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Cartographic Works for Children and Youth: State of Art and Possibilities of GIS Utilization
Škardová, Marta ; Bláha, Jan Daniel (advisor) ; Hudeček, Tomáš (referee)
Abstract:: The aim of this paper is to present cartographic works for children and youth. The main object is the introduction to the problems of cartographic production for children and youth and the execution of value analysis in terms of cartography. The analysis focuses on the use of mapping language - character key and means of expresion, in this part is an efort to answer the question whether the cartographic works for children and youth are the real maps or rather an illustration. The assesment of selected product by their users - children continue the analasis. Another aim is to compare traditional methods of creating those works with the possibilities of modern computer cartography, three ways of making are demonstrated - by hand, using a graphic application and using cartographic application. Then the paper tries to assess the use of geographic and cartographic knowledge in products for children and youth, in this question the paper use knowledges gained in consultation with author from practise.
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Influence of crystalloid and colloid solutions on blood coagulation using the rotational thromboelastometry (ROTEM) method
Binterová, Silvie ; Durila, Miroslav (advisor) ; Vaněk, Tomáš (referee) ; Bláha, Jan (referee)
Fluid resuscitation with crystalloid and colloid solutions is a common treatment in perioperative medicine. However, a variety of unbalanced or balanced solutions is used in clinical practice and there is still a vivid debate going on regarding the selection of optimal fluid with minimal negative effect on coagulation. The goal of the dissertation was to investigate the adverse effect of balanced crystalloids and colloids on whole blood coagulation measured by method of rotational thromboelastometry. In the first phase of the work we had assessed the adverse effect of balanced crystalloid, hydroxyethyl starch and gelatin after dilution of blood with the solution in vitro. Parametrs of EXTEM and FIBTEM tests were evaluated by using rotational thromboelastometry. In the second phase of the work we evaluated the negative effect of infusion solution after dilution in vivo. We had obtained blood samples from 30 patients during knee arthroscopy before and after administration of 500 ml of crystalloid or hydroxyethyl starch or gelatin. Parametrs of EXTEM and FIBTEM tests were evaluated by using rotational thromboelastometry.In compliance with the results of the dissertation, hydroxyethyl starch has the most obvious negative effect on clot formation followed by gelatin and finally by crystalloids. Based on...
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Assessment of mental maps in GIS
Kynčlová, Martina ; Hudeček, Tomáš (advisor) ; Bláha, Jan Daniel (referee)
This work deals with the possibilities of assessments of mental maps using GIS tools. The main goal is to create a collection of methods to evaluate positioning and relative accuracy of mental maps. The method principles are based on methods of GIS. The emphasis is on maximum versatility and automation the assessment procedure. For all proposed methods there are further discussed the advantages and disadvantages. First part of this work deals with the theme of mental maps, followed by a brief introduction to GIS and their use in the field mental maps. The main part is devoted to the description and practical examples of evaluation methods. At the conclusion there is a discussion of the proposed methods and the appropriateness of use GIS at this issue in general. This work was created under the project No. 26609 called "Mental Maps: Object and Device for Assessment" supported by the Grant Agency of Charles University.
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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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