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Orthohantaviruses in the reservoir and atypical hosts in the Czech Republic: spillover infection and indication of virus-specific tissue tropism
KAMIŠ, Jan
Aim of this study was to reveal the presence of hantaviruses in natural reservoir rodent hosts in selected urban areas in the Czech Republic. Hantavirus rodent hosts were trapped, sampled and tested for hantavirus RNA in different tissues. Universal and specific primers for amplification of the large and medium fragments of hantavirus genomic RNA were used. Phylogenetic relationships were based on the obtained nucleotide sequences. Four different hantaviruses were detected, including two species pathogenic (or potentially pathogenic) for humans, further suggesting a threat for public health. Moreover, inter-family spillover infections and hantavirus species-associated tissue tropism were recorded in rodent hosts.
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Hlodavci jako rezervoár hantavirů
KAMIŠ, Jan
This study reveals the presence of hantaviruses in free-living rodents and insectivores in urban areas in the Czech Republic. A large number of hantavirus reservoir hosts were tested for hantavirus RNA in different tissues, using universal and specific primers for amplification of the large and medium fragments of hantavirus genomic RNA. Phylogenetic relationships of obtained nucleotide sequences of hantaviruses were reconstructed. Four different species of hantaviruses were detected, including two species pathogenic (or potentially pathogenic) for humans, suggesting a threat for public health. Additionally, inter-family spillover infections and hantavirus species-associated tissue tropism were recorded in rodent hosts.
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Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies
Fornůsková, Daniela ; Zeman, Jiří (advisor) ; Hyánek, Josef (referee) ; Stiborová, Marie (referee)
Mgr. Daniela Fornuskova PhD thesis Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies ABSTRACT The mammalian organism fully depends on the oxidative phosphorylation system (OXPHOS) as the major energy (ATP) producer of the cell. Disturbances of OXPHOS may be caused by mutations in either mitochondrial DNA (mtDNA) or nuclear DNA (nDNA). One part of the thesis is focused on the role of early and late assembled nuclear-encoded structural subunits of cytochrome c oxidase (CcO) as well as Oxa1l, the human homologue of the yeast mitochondrial Oxa1 translocase, in the biogenesis and function of the human CcO complex using stable RNA interference of COX4, COX5A, COX6A1 and OXA1L, as well as expression of epitope-tagged Cox6a, Cox7a and Cox7b, in HEK (human embryonic kidney)- 293 cells. Our results indicate that, whereas nuclear- encoded CcO subunits Cox4 and Cox5a are required for the assembly of the functional CcO complex, the Cox6a subunit is required for the overall stability of the holoenzyme. In OXA1L knockdown HEK-293 cells, intriguingly, CcO activity and holoenzyme content were unaffected, although the inactivation of OXA1 in yeast was shown to cause complete absence of CcO activity. In addition, we compared OXPHOS protein deficiency patterns in mitochondria from skeletal...
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Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies
Fornůsková, Daniela ; Zeman, Jiří (advisor) ; Hyánek, Josef (referee) ; Stiborová, Marie (referee)
Mgr. Daniela Fornuskova PhD thesis Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies ABSTRACT The mammalian organism fully depends on the oxidative phosphorylation system (OXPHOS) as the major energy (ATP) producer of the cell. Disturbances of OXPHOS may be caused by mutations in either mitochondrial DNA (mtDNA) or nuclear DNA (nDNA). One part of the thesis is focused on the role of early and late assembled nuclear-encoded structural subunits of cytochrome c oxidase (CcO) as well as Oxa1l, the human homologue of the yeast mitochondrial Oxa1 translocase, in the biogenesis and function of the human CcO complex using stable RNA interference of COX4, COX5A, COX6A1 and OXA1L, as well as expression of epitope-tagged Cox6a, Cox7a and Cox7b, in HEK (human embryonic kidney)- 293 cells. Our results indicate that, whereas nuclear- encoded CcO subunits Cox4 and Cox5a are required for the assembly of the functional CcO complex, the Cox6a subunit is required for the overall stability of the holoenzyme. In OXA1L knockdown HEK-293 cells, intriguingly, CcO activity and holoenzyme content were unaffected, although the inactivation of OXA1 in yeast was shown to cause complete absence of CcO activity. In addition, we compared OXPHOS protein deficiency patterns in mitochondria from skeletal...
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