Aplikace metod molekulární dosimetrie(stanovení DNA aduktů,Comet assay,stanovení metabolických genotypů) pro určení genotoxicity komplexních směsí polyaromatických uhlovodíků a jejich nitroderivátů.Zhodnocení genet. poškození profesionální expozice PAU.

• MeSH
• biologické markery MeSH

• Konspekt
• Obecná genetika. Obecná cytogenetika. Evoluce
• NLK Obory
• biochemie
• embryologie a teratologie
• pracovní lékařství
• genetika, lékařská genetika
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

• Konspekt
• Biologické vědy
• NLK Obory
• biologie
• NLK Publikační typ
• učebnice vysokých škol

DNA adducts are markers of carcinogen exposure and of their biological effect; they have been shown to be related to mutagenesis, and therefore they could be a predictive biomarker of human cancer. The objective of this study was to assess if there is a relationship between vitamins A, C, and E, which are known to play a significant role as free radical scavengers and antioxidant agents, and biomarkers of genotoxicity and oxidative stress. Three hundred and fifty-six subjects from Czech Republic, Slovak Republic and Bulgaria, who completed a questionnaire on dietary information and had a measurement of plasma A, C, E vitamins, DNA adduct levels (benzo[a]pyrene (B[a]P) and bulky (DNA-Tot) DNA adducts) and oxidative damage (cyclic pyrimidopurinone N-1,N2 malondialdehyde-2 deoxyguanosine (M1dG) and 8-oxo-7,8-dihydro-2_deoxyguanosine (8-oxodG)) were analyzed. A significant inverse correlation was observed between plasma vitamin levels and both benzo[a]pyrene (B[a]P) and bulky DNA adducts. Vitamin A was also significantly inversely correlated with M1dG, a marker of oxidative damage. The associations were stronger in non-smokers than in smokers. Dietary intake of certain antioxidants such as vitamins is associated with reduced levels of markers of DNA damage (B[a]P and DNA-Tot) and oxidation (M1dG and 8-oxodG) measured in peripheral white blood cells. This could contribute to the protective role of such a dietary pattern on cancer risk. The protective effect of dietary vitamins is less evident in smokers.

• MeSH
• adukty DNA účinky léků   MeSH
• biologické markery analýza   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• oxidační stres účinky léků   MeSH
• průřezové studie MeSH
• průzkumy a dotazníky MeSH
• senioři MeSH
• vitaminy aplikace a dávkování   farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

• MeSH
• angiotensin konvertující enzym MeSH
• delece genu MeSH
• dýchání MeSH
• ergometrie statistika a číselné údaje   MeSH
• interpretace statistických dat MeSH
• kardiovaskulární systém MeSH
• lidé MeSH
• polymorfismus genetický MeSH
• spirometrie statistika a číselné údaje   MeSH
• sporty MeSH
• výkonnost MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Slovenská republika MeSH

The fluorescence in situ hybridization (FISH) technique with whole chromosome painting for chromosomes #1 and #4 was used to study the impact of air pollution containing higher concentrations of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) in three European cities, Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). In each site were followed an exposed group, who were police officers or bus drivers who work usually through busy streets for at least 8h, and a reference group, who spent more than 90% of their daily time indoors. In Prague, a significant increase was observed in percentage of aberrant cells (% AB.C.) in the police officers compared to the reference group (0.33+/-0.25 versus 0.24+/-0.18, p<0.05). In Kosice, the exposed group differed from reference in the endpoints F(G)/100 1.52+/-1.18 versus 1.12+/-1.30, p<0.05; % AB.C. 0.30+/-0.19 versus 0.21+/-0.20, p<0.05; t/1000 3.91+/-3.18 versus 2.84+/-3.10, p<0.05. In Sofia were followed two exposed groups: police officers and bus drivers. All FISH endpoints were significantly higher in police officers compared to reference group (F(G)/100 1.60+/-0.99 versus 0.82+/-0.79, p<0.01; % AB.C. 0.25+/-0.14 versus 0.13+/-0.13, p<0.01; t/1000 4.19+/-2.65 versus 2.13+/-2.05, p<0.05; rcp 1.46+/-1.07 versus 0.70+/-0.76, p<0.05). In bus drivers compared to reference there was an increase in % AB.C. (0.25+/-0.18 versus 0.13+/-0.13, p<0.05). This is the first study when FISH method was used to analyze the impact of environmental air pollution. According to the original hypothesis it is expected that the most important group of chemicals responsible for the biological activity of air pollution represent c-PAHs.

• MeSH
• biologické markery MeSH
• chromozomální aberace MeSH
• dospělí MeSH
• financování organizované MeSH
• hybridizace in situ fluorescenční metody   MeSH
• karcinogeny životního prostředí toxicita   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• monitorování životního prostředí MeSH
• policie MeSH
• polycyklické aromatické uhlovodíky toxicita   MeSH
• pracovní expozice MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH

Principal aims of this study were at first, to find a relevant human derived cell line to investigate the genotoxic potential of PAH-containing complex mixtures and second, to use this cell system for the analysis of DNA adduct forming activity of organic compounds bound onto PM10 particles. Particles were collected by high volume air samplers during summer and winter periods in three European cities (Prague, Kosice, and Sofia), representing different levels of air pollution. The genotoxic potential of extractable organic matter (EOM) was compared with the genotoxic potential of individual carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) as well as their artificial mixtures. Metabolically competent human hepatoma HepG2 cells, confluent cultures of human diploid lung fibroblasts (HEL), and the human monocytic leukemia cell line THP-1 were used as models. DNA adducts were analyzed by (32)P-postlabeling. The total DNA adduct levels induced in HepG2 cells after exposure to EOMs were higher than in HEL cells treated under the same conditions (15-190 versus 2-15adducts/10(8) nucleotides, in HepG2 and HEL cells, respectively). THP-1 cells exhibited the lowest DNA adduct forming activity induced by EOMs (1.5-3.7adducts/10(8) nucleotides). A direct correlation between total DNA adduct levels and c-PAH content in EOM was found for all EOMs in HepG2 cells incubated with 50microg EOM/ml (R=0.88; p=0.0192). This correlation was even slightly stronger when B[a]P content in EOMs and B[a]P-like adduct spots were analyzed (R=0.90; p=0.016). As THP-1 cells possess a limited metabolic capacity for most c-PAHs to form DNA reactive intermediates and are also more susceptible to toxic effects of PAHs and various EOM components, this cell line seemed to be an inappropriate system for genotoxicity studies of PAH-containing complex mixtures. The seasonal variability of genotoxic potential of extracts was stronger than variability among the three localities studied. In HepG2 cells, the highest DNA adduct levels were induced by EOM collected in Prague in the winter period, followed by Sofia and Kosice. However, in the summer sampling period, the order was quite opposite: Kosice>Sofia>Prague. When the EOM content per m(3) of air was taken into consideration in order to compare real exposures of humans to genotoxic compounds in all three localities, extracts from respirable dust particles collected in Sofia exhibited the highest genotoxicity regardless of the sampling period. The results indicate that most of DNA adducts detected in cells incubated with EOMs have their origin in low concentrations of c-PAHs representing 0.03-0.17% of EOM total mass. Finally, our results suggest that HepG2 cells have a metabolic capacity for PAHs similar to human hepatocytes and represent therefore the best in vitro model for investigating the genotoxic potential of complex mixtures containing PAHs among the three cell lines tested in this study.

• MeSH
• adukty DNA analýza   MeSH
• financování organizované MeSH
• karcinogeny životního prostředí toxicita   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• organické látky toxicita   MeSH
• pevné částice toxicita   MeSH
• polycyklické aromatické uhlovodíky metabolismus   toxicita   MeSH
• testy genotoxicity metody   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH

Acellular assay of calf thymus DNA+/-rat liver microsomal S9 fraction coupled with (32)P-postlabelling was used to study the genotoxic potential of organic compounds bound onto PM10 particles collected in three European cities-Prague (CZ), Kosice (SK) and Sofia (BG) during summer and winter periods. B[a]P alone induced DNA adduct levels ranging from 4.8 to 768 adducts/10(8) nucleotides in the concentration dependent manner. However, a mixture of 8 c-PAHs with equimolar doses of B[a]P induced 3.7-757 adducts/10(8) nucleotides, thus suggesting the inhibition of DNA adduct forming activity by interaction among various PAHs. Comparison of DNA adduct levels induced by various EOMs indicates higher variability among seasons than among localities. DNA adduct levels for Prague collection site varied from 19 to 166 adducts/10(8) nucleotides, for Kosice from 22 to 85 and for Sofia from 6 to 144 adducts/10(8) nucleotides. Bioactivation with S9 microsomal fraction caused 2- to 7-fold increase in DNA adduct levels compared to -S9 samples, suggesting a crucial role of indirectly acting genotoxic EOM components, such as PAHs. We have demonstrated for the first time a significant positive correlation between B[a]P content in EOMs and total DNA adduct levels detected in the EOM treated samples (R=0.83; p=0.04). These results suggest that B[a]P content in EOM is an important factor for the total genotoxic potential of EOM and/or B[a]P is a good indicator of the presence of other genotoxic compounds causing DNA adducts. Even stronger correlation between the content of genotoxic compounds in EOMs and total DNA adduct levels detected (R=0.94; p=0.005) was found when eight c-PAHs were taken into the consideration. Our findings support a hypothesis that a relatively limited number of EOM components is responsible for a major part of its genotoxicity detectable as DNA adducts by (32)P-postlabelling.

• MeSH
• financování organizované MeSH

• MeSH
• angiotensin konvertující enzym genetika   MeSH
• chronická obstrukční plicní nemoc enzymologie   epidemiologie   MeSH
• finanční podpora výzkumu jako téma MeSH
• genetická predispozice k nemoci epidemiologie   MeSH
• genetické testování metody   MeSH
• lidé MeSH
• plicní hypertenze enzymologie   epidemiologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

• MeSH
• dospělí MeSH
• finanční podpora výzkumu jako téma MeSH
• glutathiontransferasa genetika   MeSH
• lidé MeSH
• nádory plic diagnóza   genetika   MeSH
• nemalobuněčný karcinom plic diagnóza   genetika   MeSH
• obstrukční plicní nemoci diagnóza   genetika   MeSH
• oxidační stres imunologie   MeSH
• polymorfismus genetický MeSH
• rizikové faktory MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

Pozadie problému: Inzerčno (I)/delečný (D) polymorfizmus lokalizovaný v intróne 16 génu pre angiotenzín-konvertujúci enzým (ACE) je asociovaný so zvýšeným rizikom rozvoja ochorení kardiovaskulárneho systému. Avšak jeho vzťah k pľúcnej hypertenzii nie je jasný. Predošlé práce ukázali, že u pacientov s primárnou pľúcnou hypertenziou sa vyskytuje ACE DD genotyp častejšie ako genotyp s neprítomnosťou D alely. Hypotéza: Predpokladáme, že pacienti s chronickou obštrukčnou chorobou pľúc (CHOCHP), ktorí sú nositeľmi D alely, majú vyššie riziko rozvoja pľúcnej hypertenzie. Pacienti a metódy: Súbor tvorilo 40 pacientov hospitalizovaných s diagnózou CHOCHP, u ktorých bolo zrealizované funkčné vyšetrenie pľúc použitím bodypletyzmografie, tlak v pľúcnici bol určený echokardiograficky. Na stanovenie genotypu v l/D polymorfizme génu pre ACE bola použitá polymerázová retazová reakcia (PCR). Výsledky: Echokardiograficky stanovené tlaky (stredný a systolický) v arteria pulmonalis boli signifikantne vyššie v skupine pacientov s DD a ID génovým polymorfizmom pre ACE oproti pacientom s genotypom II (15,2 ±1,6 oproti 8,8 ± 0,8 mm Hg, p < 0,01; 32,9 ± 2,5 oproti 21,6 ± 2,3 mm Hg, p < 0,05). V multivariátiiej lineárnej regresnej analýze boli I/D génový polymorfizmus pre ACE a FEVl jedinými nezávislými prediktormi tiakov v pľúcnici. Záver: Naše predbežné výsledky ukázali, že I/D génový polymorfizmus pre ACE má u pacientov s CHOCHP významný vztah k riziku rozvoja pľúcnej hypertenzie. K hlbšiemu poznaniu genetického rizika rozvoja pľúcnej hypertenzie bude potrebné uskutočniť v budúcnosti ďalšie sledovania na väčších súboroch pacientov.

Background: The insertion (1)/deletion (D) polymorphism at intion 16 of angiotensin-converting enzyme (ACE) gene has been associated with an increased risk of cardiovascular diseases, however, its relationship to pulmonary hypertension is unclear. However, the ACE DD genotype is more prevalent in patients with primary pulmonary hypertension than the non-DD genotype. Hypothesis: In patients with chronic obstructive pulmonary disease (COPD), the carriers of D allelle are at higher risk of developing pulmonary hypertension. Patients and methods: In 40 consecutive patients with COPD, lung function was assessed using bodyplethysmography, pulmonary artery pressures were determined using echocardiography. The ACE gene I/D polymorphism was detected by polymerase chain reaction. Results: Mean and peak pulmonary artery pressures assessed by echocardiography were significantiy higher in the DD+ID group compared to the II group (15.2 ±1.6 versus 8.8 ± 0.8 mm Hg, p < 0.01; 32.9 ± 2.5 versus 21.6 ± 2.3 mm Hg, p < 0.05, respectively). The I/D ACE gene polymorphism and FEVl were the only independent predictors of pulmonary artery pressures in multiple linear regression analysis. Conclusion: Our preliminary data indicate that the l/D ACE gene polymorphism is linked to the risk of development of pulmonary hypertension in patients with COPD. Further studies in large groups of patients with COPD are needed to shed more light on this issue.

• MeSH
• chronická obstrukční plicní nemoc genetika   MeSH
• lidé MeSH
• plicní hypertenze genetika   MeSH
• polymorfismus genetický genetika   MeSH
• prospektivní studie MeSH
• renin-angiotensin systém genetika   MeSH
• Check Tag
• lidé MeSH