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Photoperiodic entraiment of circadian clock in suprachiasmatic nucleus
Parkanová, Daniela ; Vopálenský, Václav (referee) ; Sumová, Alena (advisor)
Most of physiological processes run in the organisms persistently, they begin in a definite rhythm again and again. The greatest attention is paid to the rhythms, whose period is equal to one day - they are called circadian rhythms. In case of mammals, these circadian rhythms are under control of the central circadian clock that resides in the suprachiasmatic nucleus, a part of the anterior hypothalamus. The mechanism of rhythm generation is based on interacting transcriptional-translational feedback loops that control expression of the clock genes in every single cell. Clock-controlled genes transmit these rhythms into the whole organism where they drive many physiological processes. Clock genes are expressed also in the peripheral oscillators (for example in liver, lungs, heart) and are under direct control of the central oscillator. Circadian clock needs to be entrained everyday to the external time to function precisely. The main entraining cue is the light part of the day. The length of the light part of the day, i.e. photoperiod, changes during the year rapidly in our latitudes and the central oscillator has to adapt to the changes all the time. The length of the photoperiod is encoded directly in the central oscillator by the transcriptional-translational relations among the clock genes and...
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Regulation of the expression of MHC class I molecules and other immunoactive molecules on tumor cells
Moravcová, Simona ; Vopálenský, Václav (referee) ; Reiniš, Milan (advisor)
4 Abstract Regulation of the expression of MHC class I molecules and other immunoactive molecules on tumor cells The aim of this master thesis project was to characterize the effects of IFNγ, TNFα and of the epigenetic agents 5-azacytidine (5AC) and 5-aza-2'-deoxycytidine (5AZAD) on the expression of molecules important for antigen presentation and modulation of the immune responses against tumors (MHC class I molecules and other immunoactive molecules CD54, CD80, B7-H1, B7-H2 and CD1d) on tumor cells. The experimental model used for this purpose were the HPV16-associated murine tumor cell lines, either MHC class I positive or negative. The goal was to determine the changes in surface expression of these molecules after treatment by FACS studies and also the expression at the mRNA level using qPCR. Expression of these proteins was also compared in the experiments in vitro on tumor cell lines and ex vivo on tumor cells explanted from animals. The most interesting result is the observation that 5AZAD increases the expression of B7-H1 which inhibits T cell mediated immune response and that 5AZAD and IFNγ act synergistically in the induction of the expression of MHC class I, CD54 and B7-H1 molecules. Klíčová slova: imunoeditace, protinádorová imunita, HVP16, MHC glykoproteiny I. třídy, B7-H1, IFNγ, TNFα,...
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Construction and characterization of recombinant adenylate cyclase toxoid of bacterium Bordetella pertussis carrying mycobacterial antigen TB7.7
Mikulecký, Pavel ; Vopálenský, Václav (referee) ; Staněk, Ondřej (advisor)
Bacterium Mycobacterium tuberculosis is an etiological agent of a deadly disease called tuberculosis that presents a global problem. According to The World Health Organization there are more than 2 billions people infected with latent tuberculosis all over the world. There is still need of specific, sensitive, quick and economic available method for identification of infected individuals. Currently in vitro blood tests are considered to be the best way of diagnosis. They are based on restimulation of specific T lymphocytes by mycobacterial antigens derived from virulent strains. There are several different approaches for enhancing of direct antigen delivery into antigen presenting cells and promising one is a genetically detoxified adenylate cyclase toxin (CyaA) of bacteria Bordetella pertussis. The main aim of the thesis includes construction and subsequent characterization of biological properties of CyaA protein carrying specific mycobacterial antigen TB7.7 in translocating domain. Here is shown that fusion protein CyaA-TB7.7 can form cation selective pores in target cell membranes and is able to deliver antigens into the cytosol of APC to be presented on surface with molecules MHC class II. Genetically detoxified CyaA- TB7.7 protein will be used to supplement current approaches such as also in vitro...
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Establishment and phenotyping of three mouse congenic strains.
Flachs, Petr ; Trachtulec, Zdeněk (advisor) ; Vopálenský, Václav (referee)
Mouse congenic strains are widely used as an important genetic tool for various purposes of research. Congenic strains are special inbred strains in which part of the genome of one mouse is transferred to another by backcrossing the donor mouse to the recipient strain with appropriate selection. The classical ten-generation breeding protocol is commonly utilized and adapted for different experimental needs. Our department is focused on the study of hybrid sterility between two related mouse strains M. m. musculus and M. m. domesticus. The first aim of this work was the utilization of congenics to stabilize three transgenic lines, which previously helped to identify first mammalian hybrid sterility gene Hst1 as Prdm9 (Mihola et al, 2009). Bacterial artificial chromosomes (BACs) present in transgenes encompassed several genes including Prdm9. The second goal was to look for some new phenotypes of our congenic strains, which may be causally associated with genes encoded by the BACs, as well as an additional research on the hybrid sterility phenomena. We established three new congenic strains on C57BL/6J (B6) background and found significant differences between transgenics versus their littermate controls in weights of testis, gonadal fat, body, liver, and heart. Powered by TCPDF (www.tcpdf.org)
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