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Charakter of the host cell iduced by vaccinia virus and inhibitory effects of the lipoic acid on vaccinia virus growth
Spišáková, Martina ; Mělková, Zora (advisor) ; Němečková, Šárka (referee) ; Pospíšek, Martin (referee)
Vaccinia virus is a typical member of the poxvirus family. It had been successfully used during the worldwide smallpox eradication campaign. Currently, it is used as vector for prophylactic, as well as experimental purposes. This PhD thesis stems from the findings previously published in our lab. It is partially focused on a further characterization of vaccinia virus effects on the type of host cell death. The results point to the activation of apoptosis during vaccinia virus infection, but it cannot be completed and the cell dies by necrosis. Our attempts to shift the necrotic type of cell death induced by vaccinia virus infection towards apoptosis using a pharmacological inhibition of activity of a key enzyme PARP (Poly-(ADP-ribose) polymerase) remained unsuccessful. The effects of vaccinia virus-encoded anti-apoptotic factors appear superior to the inhibition of this single enzyme. The second part of the thesis is focused on inhibitory effects of a redox-modulating compound, lipoic acid, on vaccinia virus infection. Our results demonstrated its inhibitory effects in cell lines of different embryonic origin. It appears that the lipoic acid inhibits vaccinia virus growth at the stage of late gene expression or possibly later, during virus morphogenesis. Lipoic acid could be potentially used as a supportive...
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Structure and function of IRES elements in yeast
Černý, Jiří ; Pospíšek, Martin (advisor) ; Fišer, Radovan (referee)
Translation initiation is an important step in control of gene expression. Cellular mRNAs are usually translated via classical cap-dependent pathway, but some of them can also be translated by an alternative pathway like IRES mediated one. In the first part of my work I studied a possibility of IRES dependent translation initiation of RRN3 and BAS1 mRNAs from yeast Saccharomyces cerevisiae. I inserted BAS1 and RRN3 5'UTR into the intercistronic spacer of pFGAL4h and its promotreless version and measured the activity of secondary reporters in enegineered yeast strains. My data indicates presence of cryptic promoter in tested regions. To confirm this results I first inserted the RRN3 5'UTR into the intercistronic region of another bicistronic plasmid - pRFh and measured expression of second cistron. The results were similar like in the case of pFGAL4h system. The presence of shorter RNAs transcribed from cryptic promoter within the intercistronic region was shown using real time PCR. My quest in the second part of this work was to prepare RNAs useful for next investigation of translation control of lariat capped RNAs produced by GIR1 ribozyme in a slime mold Didymium iridis. I have prepared a wide set of DNA molecules coding different versions of GIR1 in front of the firefly luciferase ORF which served as...
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Chemical modifications of polypropylene surfaces
Pospíšek, Marek ; Petrůj, Jaroslav (referee) ; Kučera, František (advisor)
Diploma thesis is focused on modification and characterization of polypropylene (PP) products surfaces by the chemical way. In theoretical part of thesis are summarized the current knowledge of this topic including characterization methods. Polypropylene surfaces were modified by acrylic monomers (acrylic acid, acrylic acid with 20 mol% content of sodium acrylate and acryl amide). Modified surfaces were characterized by infrared spectroscopy with ATR technique. The comparative parameter was carbonyl index determined from absorption peaks of functional methyl groups (2950 cm-1 ) and functional carbonyl groups (1700-1750 cm-1) of FTIR-ATR spectra. Determination of effect of initiator concentration on the modification process was done for acetophenone and benzophenone. There were determined the time dependence of carbonyl index for time from 0 to 600 s of mentioned monomers. In the same condition were modified test specimens designed for preparation of adhesive joints. Joints were bonded by cyanoacrylate adhesive that provide reaction with surface of polyacrylic acid. There were founded conditions of modification that provided strength of adhesive joints higher than strength of original PP. There were identified changes of surface roughness by using of confocal microscopy.
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Synthesis of Lanthanidocene Based Compounds
Pospíšek, Marek ; Zmrzlý, Martin (referee) ; Richtera, Lukáš (advisor)
The aim of this bachelor thesis is to summarize the issues of the polymerization catalytic systems based on lanthanidocene complexes. These complexes are subsequently divided by the structure into four basic groups. One part of this work deals with an overview of the most interesting results of the catalytic systems, which are then analyzed and assessed either in its entirety or point of view of catalytic activity, living mechanism of polymerisation and stereo or regioselectivity. The focus is also placed on type of monomers or the conditions of polymerization. In this part of thesis are also described general methods for their synthesis. Found analogies and generalization are commented from the point of view of the design of the catalytic systems for the purpose of choosing the stereo or regioselectivity, molar mass and its distribution. This gives significance to these complexes as catalytic systems for obtaining the required polymer properties. The acquired knowledge is used for the theoretical design of the catalytic system for controlled polymerization.
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Formation of 6-substitued 13H-1,5-benzodiazepino(2,3b) quinoxalines during the reaction of 4-deoxy-2,3-aldosdiuloses with o-phenylenediamine
Gabriel, Jiří ; Pospíšek, M. ; Palková, Z. ; Němeček, Jan ; Havlíček, Vladimír ; Volc, Jindřich ; Kubátová, Elena
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