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The use of comet assay for measurement of DNA integrity in clinical and applied research
Bagryantseva, Yana ; Novotná, Božena (advisor) ; Rubeš, Jiří (referee) ; Štětina, Rudolf (referee)
Single cell gel electrophoresis or comet assay combined with enzymes of excision repair is a method for measuring DNA strand breaks and oxidative damage. Using this approach we analysed ineffective hematopoiesis in patients with low-risk MDS. Refractory anemia (RA) exhibited a higher DNA instability in bone marrow cells when compared to controls and the extent of DNA fragmentation correlated with cytopenia. No similar relationship was observed in RA with ring sideroblasts (RARS), although the levels of DNA breaks markedly exceeded even the values detected in RA. Both groups of patients also showed high levels of oxidative damage to DNA. However, there was no clear relationship to the levels of serum ferritin, cytopenia or associated inflammation. This suggested that the oxidative DNA damage per se is not responsible for extensive apoptosis in low-risk MDS. In any case, it undoubtedly contributes to genome instability and disease progression. The second part of thesis was aimed to the impact of air pollution and genetic polymorphisms on oxidative damage to DNA, lipids and proteins of city bus drivers and garagemen. Both groups exhibited a higher level of DNA breaks and oxidative damage to proteins than the controls, while an increased level of lipid peroxidation was detected only in bus drivers. The...

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