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The estimation of oxidative DNA damage using the single-cell gel electrophoresis (commet assay).
Škubalová, Věra ; Hochmann, Jiří (advisor) ; Slyšková, Jana (referee)
The oxidative cell damage represents one of the most common type of damage, which concerns all cell components. It produces the oxidative modifications of lipid structure, proteins and DNA. We concentrated on the last one in this work. The oxidative changes result in modification of replication cell cycle, which are represented for example by carcinogenesis, mutagenesis or by cell aging. Cells are provided by repairing mechanism. Owing to this mechanism the cells are able to repair this damage and stay viable. The oxidative damage is induced by various factors, for example free oxygen radicals or UV radiation, which we used for producing oxidative damage of DNA in this work. It is possible to measure the oxidative damage by using different methods, the most common way is using the comet assay. It represents an important method used for detection of single and doublestrand breaks, for detection of alcali-labile sites, searching for DNA- DNA/DNAprotein cross-linking, oxidative and alkylate damage of DNA. Only a little number of cells are needed for the measurement, which is the most important advantage of this method. We also used the modification of comet assay. It means the application of the reparative enzymes, which are able to recognize the specific oxidative damage of DNA. We used three...
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The influence of the epigallokatechin gallate, elagic acid and curcumin on the cytotoxic effect of mutagens.
Tlučhořová, Drahomíra ; Hochmann, Jiří (advisor) ; Polívková, Zdeňka (referee)
Naturally occurring phenols epigallocatechin gallate (EGCG), ellagic acid and curcumin exhibit antioxidant, antimutagenic and anticarcinogenic activity in a lot of in vivo and in vitro studies. In our study we used comet assay test to determine if these compounds are capable to affect DNA damage of AA8 and HepG2 cells caused by 2-amino-3-methylimidazo [4,5-f] quinoline (IQ), N-nitroso-N-methylurea (MNU) and UVC radiation. Results should say if comet assay test is an appropriate method for antimutagenity tests. We were interested if compounds, which are able to moderate effects of mutagens, would also affect their cytotoxicity. Except MNU and UVC radiation we used also hydrogen peroxide as a reference mutagen. In comet assay curcumin was not able to protect against MNU induced DNA damage sufficiently. However, ellagic acid effectively reduced mutagenic effect of MNU. EGCG decreased the level of DNA breaks induced by IQ and UVC radiation and affected both DNA damage induction and repair of cells incubated with MNU. We chose only EGCG for cytotoxicity tests because it showed the greatest antimutagenity. EGCG increased the number of formed colonies of cells treated with hydrogen peroxide, likely to its antioxidant properties. However EGCG did not decrease cytotoxicity of MNU and UVC radiation and the...
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Pharmacological approach to the results of the estrogen and progesterone receptors in the breast tumours.
Bařinková, Vlasta ; Hochmann, Jiří (advisor) ; Bunček, Martin (referee)
In the part devoted to discussion of different authors about the importance of the progesterone receptor (PR) value we concentrated primarily to the works of foreign authors published in professional journals. We divided the opinions of this experts to such, which show that PR testing is needless and to the part about profitability of this testing. In the chapter about importance PR testing some authors critize, that some patients - negative in estrogen receptors (ER) (such of them that are simultaneously PRpositive) could be deprived in the possibility of hormonal therapy, because PR testing wouldn't be made. And in the addition, the other - (in this case) the less effective treatment would be given them preferentially. PR testing is important also for detection of the group of ER-positive tumors with PR negativity. At this group is the smaller possibility that the tumors would respond to the hormonal treatment (in contrast to ER+/PR+). In the part devoted to gene therapy we described first the information about learning from the textbooks. Futher we describe different modes of classification of the gene therapy of the breast cancer in the foreign literature. We modified their classification further according to our necessity to these subheads: (1) mutation compensation of differentiation genes,...
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