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Impact of habitat type and long-term in vitro cultivation on photosynthetic characteristics of snow algae and their resistance to stress induced by UV radiation
Zázvorková, Michaela ; Nedbalová, Linda (advisor) ; Kvíderová, Jana (referee)
Snow algae are psychrophilic microorganisms, that inhabit snow fields in mountains and polar regions, which creates colored snow in good conditions. Most species belong to order Chlamydomonadales (Chlorophyta) with complicated life cycles, containing flagellates and immobile stages (cysts). Extreme environment of snow is characteristed by low temperature, problems with availability of water and lack of nutrition. Depending on location and phase of life cycle, snow algae have to deal with excess or lack of solar radiation, the important component of which is also dangerous UV radiation. The light conditions differ substantially from open location above the forest level or polar regions to forest habitats. In the first part of this work I compared reactions of photosynthetic apparatus of snow algae strains from forest and forestless habitats to different intensity of radiation, then I dealt with assessing any changes related to long-term cultivation in laboratory. Based on measurement of rapid light curves on PAM fluorometer I have determined some characteristics of photosynthetic apparatus of individual strains (parameters α and Ik), which indicate adaptation to low or high light intensity. For some strains, it was possible to compare the results obtained at a three-year interval. In the second part...
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Genome stability of human induced pluripotent stem cells
ŘEHÁKOVÁ, Daniela
Human induced pluripotent stem cells (hiPSCs) play roles in both disease modeling and regenerative medicine. It is critical that the genomic integrity of the cells remains intact and that the DNA repair systems are fully functional. I focused on the detection of DNA double-strand breaks (DSBs) by phosphorylated histone H2AX (known as H2AX) and p53-binding protein 1 (53BP1) in multiple lines of hiPSCs, their source cells, human embryonic stem cells (hESCs) and cells differentiated from hiPSCs. I measured spontaneously occurring DSBs throughout the process of reprogramming and during long-term in vitro culture and differentiation process. To assess the variations in the functionality of the DNA repair system among the samples, the number of DSBs induced by gamma irradiation and the decrease over time was analyzed. The foci number was detected by fluorescence microscopy separately for the G1 and S/G2 cell cycle phases. Source cells contained a low number of non-replication-related foci, while this number increased after reprogramming into hiPSCs and then decreased again after long-term in vitro passaging or differentiation. The artificial induction of DSBs revealed that the repair mechanisms function well in the source cells and hiPSCs at low passages but fail to recognize a substantial proportion of DSBs at high passages. Observations suggest that cellular reprogramming increases the DSB number but that the repair mechanism functions well. However, with long-term hipSCs culture reparation capacity decreases.
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