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The effect of air pollution on oxidative stress markers in newborns
Ambrož, Antonín ; Rössner, Pavel (advisor) ; Rubeš, Jiří (referee) ; Gábelová, Alena (referee)
In everyday life, humans are exposed to toxic substances of anthropogenic origin. These substances can also be found in the ambient air and their impact poses a long-term risk for human health. Respirable particulate matter (PM) of aerodynamic diameter < 2.5 µm (PM2.5) is intensively studied, along with carcinogenic polycyclic aromatic hydrocarbons (PAHs), bound to it, such as benzo[a]pyrene (B[a]P), a reference carcinogenic PAH. Owing to small size, PM2.5 can penetrate the human body primarily via the airways and represent an increased health risk compared to larger particles. The negative health impacts of anthropogenic PM2.5, generated e.g. by fossil fuel combustion, are linked with its small size, relatively large surface, as well as with PAHs and other substances adsorbed on PM surface. PAHs, generated by an incomplete combustion of organic matter, can enter organism either via ingestion of contaminated food, water or via inhalation of polluted air. PAHs affect organisms via genotoxic, mutagenic, carcinogenic, embryotoxic and other adverse effects. One of the common denominators of these effects is oxidative stress, which is also considered to be the main mechanism of action caused by PM in the human organism. Oxidative damage induced by reactive oxygen species (ROS) may affect any cellular...
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Role of chromosomal aberrations to evaluate genetic risk of exposure to carcinogens.
Rössnerová, Andrea ; Šrám, Radim (advisor) ; Rubeš, Jiří (referee) ; Kuglík, Petr (referee)
(in English) Air pollution is a serious worldwide problem associated with the risk of cancer. The negative effect of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs), including benzo[a]pyrene (B[a]P), on human health is analyzed using specific biomarkers. Among them biomarkers of early effect play an important role. This work summarizes the results of cytogenetic analyses performed by fluorescence in situ hybridization (FISH) (whole chromosome painting of chromosomes #1 and #4) and automated image analysis of micronuclei (MN). During the analyses a total set of 1304 samples was analyzed by the FISH method and 885 samples by the automated image analysis of MN. Studied groups including city policemen, garage men, bus drivers, administrative workers, mothers, newborns, healthy children and children with bronchial asthma and laboratory workers were from Prague, Ostrava and Ceske Budejovice. The locations significantly differed in levels of air pollutants and the type of air pollution. The exposure of participants of the study was assessed by personal and stationary monitoring. The impact of other factors including age, smoking or intake of vitamins was also evaluated in these studies. The results obtained by the FISH method in Prague showed the impact of seasonal variability of concentrations of...
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A study of aneuploidy in gametes and embryos
Diblík, Jan ; Macek, Milan (advisor) ; Forejt, Julius (referee) ; Rubeš, Jiří (referee)
The thesis deals with improvement and clinical application of molecular cytogenetic methods for reproductive genetics. These methods include both clinical investigations used for improvement of diagnostic and therapeutic care for infertile couples and experimental methods that can become the basis of new diagnostic tools. The thesis concentrates on the study of aneuploidies, because they constitute a major complication of human reproduction especially by means of assisted reproductive technologies. Aims The main practical aim was the introduction of fluorescence in situ hybridization (FISH) for evaluation of chromosomes in sperm, polar bodies and blastomeres for prefertilisation and preimplantation diagnosis of aneuploidies. The main scientific objective was the study of chromosome localization in nuclei of blastomeres, that are removed from human embryos for preimplantation diagnosis. The aim of this study was to find, whether the localization of chromosomes in relation to the nuclear center and periphery is ruled by the same rules as in other cell types in later stages of development and whether the localization is influenced by aneuploidy. Next aim was to search for peripheral localization of chromosome X in embryos with more than one copies of the chromosome X. This could be a manifestation of X...
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Modulation of sporadic colorectal cancer risk by polymorphisms and haplotypes of mismatch repair genes
Tulupova, Elena ; Šrám, Radim (advisor) ; Rubeš, Jiří (referee) ; Slabý, Ondřej (referee)
Sporadic colorectal cancer (CRC) is a common disease with complex aetiology and diverse molecular phenotypes. Failure of DNA repair systems is one of the leading determinants of cancer onset and development. The efficiency of these systems and susceptibility to cancer can be affected by genotype variations, including common single nucleotide polymorphisms (SNPs). In this work, an association between SNPs and haplotypes of DNA mismatch repair (MMR) genes, SNPs and their combinations in other DNA repair genes, and a risk of sporadic CRC was investigated in a hospital-based case-control study. As result of our study, certain MMR SNPs and haplotypes altered CRC risk, as demonstrated for the first time in the Czech population. Individual SNPs in DNA repair genes seem to have a limited effect on CRC risk, with possible modification by age or smoking. Several of the associations observed were site-specific, confirming the molecular heterogeneity of CRC. DNA repair capacity varies significantly between individuals, between different tissues of the same organism, and also between malignant and normal cells. To assess the background level of this variability, the association between SNPs in DNA repair genes and the individual DNA repair capacity in healthy individuals was investigated. Several polymorphisms...
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The effect of air pollution on oxidative stress markers in newborns
Ambrož, Antonín ; Rössner, Pavel (advisor) ; Rubeš, Jiří (referee) ; Gábelová, Alena (referee)
In everyday life, humans are exposed to toxic substances of anthropogenic origin. These substances can also be found in the ambient air and their impact poses a long-term risk for human health. Respirable particulate matter (PM) of aerodynamic diameter < 2.5 µm (PM2.5) is intensively studied, along with carcinogenic polycyclic aromatic hydrocarbons (PAHs), bound to it, such as benzo[a]pyrene (B[a]P), a reference carcinogenic PAH. Owing to small size, PM2.5 can penetrate the human body primarily via the airways and represent an increased health risk compared to larger particles. The negative health impacts of anthropogenic PM2.5, generated e.g. by fossil fuel combustion, are linked with its small size, relatively large surface, as well as with PAHs and other substances adsorbed on PM surface. PAHs, generated by an incomplete combustion of organic matter, can enter organism either via ingestion of contaminated food, water or via inhalation of polluted air. PAHs affect organisms via genotoxic, mutagenic, carcinogenic, embryotoxic and other adverse effects. One of the common denominators of these effects is oxidative stress, which is also considered to be the main mechanism of action caused by PM in the human organism. Oxidative damage induced by reactive oxygen species (ROS) may affect any cellular...
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Modulation of sporadic colorectal cancer risk by polymorphisms and haplotypes of mismatch repair genes
Tulupova, Elena ; Šrám, Radim (advisor) ; Rubeš, Jiří (referee) ; Slabý, Ondřej (referee)
Sporadic colorectal cancer (CRC) is a common disease with complex aetiology and diverse molecular phenotypes. Failure of DNA repair systems is one of the leading determinants of cancer onset and development. The efficiency of these systems and susceptibility to cancer can be affected by genotype variations, including common single nucleotide polymorphisms (SNPs). In this work, an association between SNPs and haplotypes of DNA mismatch repair (MMR) genes, SNPs and their combinations in other DNA repair genes, and a risk of sporadic CRC was investigated in a hospital-based case-control study. As result of our study, certain MMR SNPs and haplotypes altered CRC risk, as demonstrated for the first time in the Czech population. Individual SNPs in DNA repair genes seem to have a limited effect on CRC risk, with possible modification by age or smoking. Several of the associations observed were site-specific, confirming the molecular heterogeneity of CRC. DNA repair capacity varies significantly between individuals, between different tissues of the same organism, and also between malignant and normal cells. To assess the background level of this variability, the association between SNPs in DNA repair genes and the individual DNA repair capacity in healthy individuals was investigated. Several polymorphisms...
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The use of BEAS-2B cell line for micronucleus assay in genetic toxicology
Červená, Tereza ; Rössner, Pavel (advisor) ; Rubeš, Jiří (referee)
This thesis deals with the application of BEAS-2B cell line for micronucleus assay in genetic toxicology. It is divided into two main parts: a) theoretical introduction to the analysis and search for suitable models for testing the impact of air pollution and manufactured nanoparticles, b) practical part that describes the results of micronuclei induction by polycyclic aromatic hydrocarbons (PAHs), extractable organic matter (EOM) from diesel exhaust particles obtained from emissions of three types of fuel and engineered nanoparticles. BEAS-2B cell line is a nonmalignant human model of lung epithelium which seems to be suitable for micronucleus assay. This assay is commonly used for determining the genotoxicity of various substances to wide variety of cell cultures and also in human studies. In this thesis, the following substances were tested: benzo[a]pyrene, 3-nitrobenzanthrone and 1-nitropyrene as carcinogenic PAHs commonly found in polluted air; EOMs from exhaust particles of 100 % diesel fuel, a blend of diesel fuel and 30 % of biodiesel, 100 % biodiesel and two types of engineered nanoparticles (TiO2 and Ag). The cells were treated with the compounds for 28, 48 and 72 hours. The results confirm the suitability of BEAS-2B cell line as a model for testing the genotoxicity of substances under...
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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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Role of chromosomal aberrations to evaluate genetic risk of exposure to carcinogens.
Rössnerová, Andrea ; Šrám, Radim (advisor) ; Rubeš, Jiří (referee) ; Kuglík, Petr (referee)
(in English) Air pollution is a serious worldwide problem associated with the risk of cancer. The negative effect of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs), including benzo[a]pyrene (B[a]P), on human health is analyzed using specific biomarkers. Among them biomarkers of early effect play an important role. This work summarizes the results of cytogenetic analyses performed by fluorescence in situ hybridization (FISH) (whole chromosome painting of chromosomes #1 and #4) and automated image analysis of micronuclei (MN). During the analyses a total set of 1304 samples was analyzed by the FISH method and 885 samples by the automated image analysis of MN. Studied groups including city policemen, garage men, bus drivers, administrative workers, mothers, newborns, healthy children and children with bronchial asthma and laboratory workers were from Prague, Ostrava and Ceske Budejovice. The locations significantly differed in levels of air pollutants and the type of air pollution. The exposure of participants of the study was assessed by personal and stationary monitoring. The impact of other factors including age, smoking or intake of vitamins was also evaluated in these studies. The results obtained by the FISH method in Prague showed the impact of seasonal variability of concentrations of...
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A study of aneuploidy in gametes and embryos
Diblík, Jan ; Macek, Milan (advisor) ; Forejt, Julius (referee) ; Rubeš, Jiří (referee)
The thesis deals with improvement and clinical application of molecular cytogenetic methods for reproductive genetics. These methods include both clinical investigations used for improvement of diagnostic and therapeutic care for infertile couples and experimental methods that can become the basis of new diagnostic tools. The thesis concentrates on the study of aneuploidies, because they constitute a major complication of human reproduction especially by means of assisted reproductive technologies. Aims The main practical aim was the introduction of fluorescence in situ hybridization (FISH) for evaluation of chromosomes in sperm, polar bodies and blastomeres for prefertilisation and preimplantation diagnosis of aneuploidies. The main scientific objective was the study of chromosome localization in nuclei of blastomeres, that are removed from human embryos for preimplantation diagnosis. The aim of this study was to find, whether the localization of chromosomes in relation to the nuclear center and periphery is ruled by the same rules as in other cell types in later stages of development and whether the localization is influenced by aneuploidy. Next aim was to search for peripheral localization of chromosome X in embryos with more than one copies of the chromosome X. This could be a manifestation of X...
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