DNA damage can impair normal cellular functions and result in various pathophysiological processes including cardiovascular diseases and cancer. We compared the genotoxic potential of diverse DNA damaging agents, and focused on their effects on the DNA damage response (DDR) and cell fate in human lung cells BEAS-2B. Polycyclic aromatic hydrocarbons [PAHs; benzo[a]pyrene (B[a]P), 1-nitropyrene (1-NP)] induced DNA strand breaks and oxidative damage to DNA; anticancer drugs doxorubicin (DOX) and 5-bromo-2'-deoxyuridine (BrdU) were less effective. DOX triggered the most robust p53 signaling indicating activation of DDR, followed by cell cycle arrest in the G2/M phase, induction of apoptosis and senescence, possibly due to the severe and irreparable DNA lesions. BrdU not only activated p53, but also increased the percentage of G1-phased cells and caused a massive accumulation of senescent cells. In contrast, regardless the activation of p53, both PAHs did not substantially affect the cell cycle distribution or senescence. Finally, a small fraction of cells accumulated only in the G2/M phase and exhibited increased cell death after the prolonged incubation with B[a]P. Overall, we characterized differential responses to diverse DNA damaging agents resulting in specific cell fate and highlighted the key role of DNA lesion type and the p53 signaling persistence.

• Klíčová slova
• Anticancer drugs, Cell fate, DNA damage response, Genotoxicity, Polycyclic aromatic hydrocarbons,
• MeSH
• apoptóza MeSH
• bromodeoxyuridin farmakologie   MeSH
• DNA MeSH
• lidé MeSH
• nádorový supresorový protein p53 * metabolismus   MeSH
• plíce metabolismus   MeSH
• polycyklické aromatické uhlovodíky * toxicita   MeSH
• poškození DNA MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bromodeoxyuridin MeSH
• DNA MeSH
• nádorový supresorový protein p53 * MeSH
• polycyklické aromatické uhlovodíky * MeSH

TiO2 along with nano-TiO2 are commonly found in consumer products. In vivo studies have observed an accumulation of nano-TiO2 in macrophages. However, characteristics of nano-TiO2 determining toxicity remain unclear. In our study, the cytotoxic effects of 14 diverse nano-TiO2 on THP-1 macrophage-like cells were measured by 3 cytotoxicity assays (MTS, WST-1 and LDH). Total averaged cytotoxicity was calculated using principal component analysis. Characteristics of all 14 nano-TiO2 included hydrodynamic diameter, zeta potential, shape, polydispersity index (PDI) and concentration; moreover, crystal form, specific surface area and crystallite size were measured for 10 nano-TiO2.The variables affecting cytotoxicity were chosen using LASSO (least absolute shrinkage and selection operator). Except for concentration, PDI in media measured within 1 h after preparation of the nanomaterial dispersion was selected as a variable affecting cytotoxicity: stable dispersion resulted in higher cytotoxic effects. Crystallite size has been shown to have nonlinear effects (particles of sizes between 20 and 60 nm were cytotoxic while smaller and larger ones were not) and thus it has been excluded from LASSO. The shape (particles/fibre) and crystal form did not affect the cytotoxicity. PDI and the nonlinear effect of size could be an explanation for the inconsistencies of the cytotoxicity of nano-TiO2 in various studies.

• Klíčová slova
• Cytotoxicity, LASSO, Macrophages, Nano-TiO(2), Polydispersity,
• MeSH
• endotoxiny analýza   MeSH
• kultivační média MeSH
• lidé MeSH
• makrofágy účinky léků   MeSH
• nanočástice chemie   toxicita   MeSH
• povrchové vlastnosti MeSH
• THP-1 buňky MeSH
• titan chemie   toxicita   MeSH
• velikost částic MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• endotoxiny MeSH
• kultivační média MeSH
• titan MeSH
• titanium dioxide MeSH Prohlížeč

Smoking in pregnancy increases a woman's risk of preterm delivery resulting in serious neonatal health problems and chronic lifelong disabilities for the children (e.g., mental retardation, learning problems). To study the effects of tobacco smoke on the placental transcriptome, we performed gene expression profiling on placentas from women exposed to tobacco smoke in pregnancy (N = 12) and from those without significant exposure (N = 64). Gene expression profiles were determined by Illumina HumanRef-8 v2 Expression BeadChips with 18,216 gene probes. Microarray data were normalized by quantile method and filtered for a detection P-value <0.01. Differential gene expression was determined by moderated t-statistic. A linear model was fitted for each gene given a series of arrays using lmFit function. Multiple testing correction was performed using the Benjamini and Hochberg method. Abundant levels of transcripts were found for genes encoding placental hormones (CSH1, CSHL1), pregnancy-specific proteins (PSG3, PSG4, PAPPA), and hemoglobins (HBB, HBG, HBA). Comparative analysis of smokers vs nonsmokers revealed the differential expression of 241 genes (P < 0.05). In smoker cohort, we detected high up-regulation of xenobiotic genes (CYP1A1, CYP1B1, CYB5A, COX412), collagen genes (e.g., COL6A3, COL1A1, COL1A2), coagulation genes (F5, F13A1) as well as thrombosis-related genes (CD36, ADAMTS9, GAS6). In smokers, we identified deregulated genes that show tissue non-specific induction and may be considered as general biomarkers of tobacco smoke exposure. Further, we also found genes specifically deregulated in the exposed placentas. Functional annotation analysis suggested processes and pathways affected by tobacco smoke exposure that may represent molecular mechanisms of smoke-induced placental abnormalities.

• MeSH
• dospělí MeSH
• kohortové studie MeSH
• kouření genetika   metabolismus   MeSH
• lidé MeSH
• messenger RNA biosyntéza   genetika   MeSH
• novorozenec MeSH
• placenta metabolismus   fyziologie   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody   MeSH
• stanovení celkové genové exprese metody   MeSH
• těhotenství MeSH
• zpožděný efekt prenatální expozice genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• messenger RNA MeSH

(32)P-postlabelling and PAH-ELISA using the antiserum #29 were employed to analyze DNA adducts in venous and umbilical cord blood and the placenta of 79 mothers giving birth to 80 living babies in Prague (Czech Republic). Ambient air exposure was measured by stationary measurements of basic air pollutants (PM2.5, c-PAHs) during the entire pregnancy. Tobacco smoke exposure was assessed by questionnaire data and by plasma cotinine levels. The total DNA adduct levels in the lymphocytes of mothers and newborns were elevated by 30-40% (p<0.001) compared with the placenta. B[a]P-like DNA adduct (adduct with the identical chromatographic mobility on TLC as major BPDE derived DNA adduct) levels were elevated in the blood of mothers compared with the placenta and the blood of newborns (p<0.05 and p<0.01). In tobacco smoke-exposed mothers, higher DNA adduct levels in the blood of mothers and newborns compared with the placenta were found (p<0.001), whereas the total and B[a]P-like adduct levels were comparable in the blood of mothers and newborns. B[a]P-like adducts were elevated in the blood of mothers unexposed to tobacco smoke compared with that of corresponding newborns and the placenta (p<0.01). Total and B[a]P-like DNA adducts were increased in the placenta of tobacco smoke-exposed compared with unexposed mothers (p<0.001 and p<0.01). In lymphocytes of tobacco smoke-exposed mothers, the comparison of total adduct levels (1.18+/-0.67 vs. 0.92+/-0.28) and B[a]P-like DNA adducts (0.22+/-0.12 adducts/10(8) nucleotides vs. 0.15+/-0.06 adducts/10(8) nucleotides) with newborns indicated a 30-40% increase of adducts in mothers. Almost equal PAH-DNA adduct levels were detected by anti-BPDE-DNA ELISA in the placenta of tobacco smoke-exposed and -unexposed mothers. Our results suggest a protective effect of the placental barrier against the genotoxic effect of some tobacco smoke components between the circulation of mother and child. We found a correlation between adduct levels in the blood of mothers and newborns.

• MeSH
• 7,8-dihydro-7,8-dihydroxybenzo(a)pyren 9,10-oxid metabolismus   MeSH
• adukty DNA krev   metabolismus   MeSH
• biologické markery krev   MeSH
• dospělí MeSH
• ELISA MeSH
• fetální krev metabolismus   MeSH
• kotinin krev   MeSH
• kouření * MeSH
• látky znečišťující vzduch krev   MeSH
• lidé MeSH
• lymfocyty účinky léků   MeSH
• maternofetální výměna látek MeSH
• matka - expozice noxám * MeSH
• mladý dospělý MeSH
• novorozenec MeSH
• placenta účinky léků   MeSH
• plod krevní zásobení   MeSH
• polycyklické aromatické uhlovodíky krev   MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 7,8-dihydro-7,8-dihydroxybenzo(a)pyren 9,10-oxid MeSH
• adukty DNA MeSH
• benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA MeSH Prohlížeč
• biologické markery MeSH
• kotinin MeSH
• látky znečišťující vzduch MeSH
• polycyclic aromatic hydrocarbons-DNA adduct MeSH Prohlížeč
• polycyklické aromatické uhlovodíky MeSH

Oxidative damage to macromolecules may have numerous negative health consequences. We measured oxidative damage to DNA, proteins and lipids in 80 newborns and 79 mothers, analyzed the effect of mother's tobacco smoke exposure on oxidative stress, and assessed correlations between oxidative stress markers and bulky and PAH (polycyclic aromatic hydrocarbons)-specific DNA adducts. Mean levels (+/-S.D.) of 8-oxodeoxyguanosine (8-oxodG) per 10(5) dG in the placenta were 2.85+/-0.78; we did not see a difference between 8-oxodG levels in newborns born to mothers exposed and unexposed to tobacco smoke. Protein carbonyl levels, a marker of protein oxidation, were comparable in the umbilical cord and in maternal venous blood plasma (17.4+/-3.2 and 17.6+/-4.2nmol/ml plasma in newborns and mothers, respectively, p=0.66). Lipid peroxidation measured as levels of 15-F(2t)-isoprostane (15-F(2t)-IsoP) in plasma was significantly higher in newborns than in mothers (362+/-129 and 252+/-130pg/ml in newborns and mothers, respectively, p<0.001). We did not find any effect of tobacco smoke exposure on either biomarker in any group. Levels of both protein carbonyls and 15-F(2t)-IsoP in cord blood significantly correlated with those in maternal plasma (p<0.001). 8-oxodG levels positively correlated with plasma carbonyls in cord plasma, as well as with cotinine levels (marker of tobacco smoke exposure) in maternal plasma. 8-oxodG levels also correlated with bulky DNA adducts in lymphocyte DNA of newborns and mothers and with PAH-DNA adducts in the placenta. Our results showed higher lipid peroxidation in newborns than in mothers, close correlation of analyzed oxidative stress markers between newborns and mothers, and a relationship between oxidative stress and induction of DNA adducts.

• MeSH
• 8-hydroxy-2'-deoxyguanosin MeSH
• adukty DNA krev   MeSH
• biologické markery krev   MeSH
• deoxyguanosin analogy a deriváty   metabolismus   MeSH
• dospělí MeSH
• ELISA MeSH
• F2-isoprostany metabolismus   MeSH
• fetální krev chemie   MeSH
• karbonylace proteinů MeSH
• kotinin analýza   MeSH
• kouření * MeSH
• krevní proteiny analýza   MeSH
• látky znečišťující vzduch krev   MeSH
• lidé MeSH
• lymfocyty účinky léků   MeSH
• maternofetální výměna látek MeSH
• matka - expozice noxám * MeSH
• mladý dospělý MeSH
• novorozenec MeSH
• oxidace-redukce MeSH
• oxidační stres * MeSH
• peroxidace lipidů MeSH
• placenta metabolismus   MeSH
• polycyklické aromatické uhlovodíky krev   MeSH
• těhotenství MeSH
• vitamin A analýza   MeSH
• vitamin E analýza   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 5-F(2t)-isoprostane MeSH Prohlížeč
• 8-hydroxy-2'-deoxyguanosin MeSH
• adukty DNA MeSH
• biologické markery MeSH
• deoxyguanosin MeSH
• F2-isoprostany MeSH
• kotinin MeSH
• krevní proteiny MeSH
• látky znečišťující vzduch MeSH
• polycyclic aromatic hydrocarbons-DNA adduct MeSH Prohlížeč
• polycyklické aromatické uhlovodíky MeSH
• vitamin A MeSH
• vitamin E MeSH

Methylated chrysenes (MeChry) are important cigarette smoke constituents and 5-MeChry has been listed as possibly carcinogenic to humans. Although a major attention has been in past paid especially to mutagenic, tumor-initiating effects of MeChry, little is known about toxic effects of MeChry related to tumor promotion. As the position of methyl group has been repeatedly observed to determine genotoxic effects of MeChry, we examined both genotoxic and nongenotoxic effects of MeChry, using rat liver cell lines as experimental models. All six MeChry were relatively efficient aryl hydrocarbon receptor (AhR) agonists, with 3- and 6-MeChry being the most potent inducers of the AhR-mediated reporter gene activity. All six compounds disrupted contact inhibition in rat liver epithelial WB-F344 cells, a process previously reported to be AhR-dependent, suggesting that MeChry may interfere with cell cycle control in an AhR-dependent manner. In contrast, only 5- and 6-MeChry were found to acutely inhibit gap junctional intercellular communication (GJIC), another parameter correlating with tumor promoting effects of xenobiotics. Both 5- and 6-MeChry were efficient inducers of mRNA expression of enzymes involved in metabolic activation of polycyclic aromatic hydrocarbons, including cytochromes P450 1A1/1B1 and aldo-keto reductase 1C9. However, only 5-MeChry, and not 6-MeChry, induced significant formation of DNA adducts in rat liver epithelial cells, which corresponded with its ability to induce high accumulation of cells in S-phase. On the other hand, 5-MeChry induced neither apoptosis related to DNA damage nor phosphorylation of p53 tumor suppressor. Taken together, our results suggest that methyl group position may affect both genotoxic and nongenotoxic effects of MeChry, such as formation of DNA adducts and inhibition of GJIC. All MeChry showed a potency to disrupt cell proliferation control, while 5-MeChry was a single compound inducing DNA damage, disruption of cell cycle control and inhibition of GJIC in rat liver cells.

• MeSH
• adukty DNA metabolismus   MeSH
• buněčný cyklus účinky léků   MeSH
• chryseny toxicita   MeSH
• enzymová indukce účinky léků   MeSH
• epitelové buňky účinky léků   MeSH
• játra účinky léků   MeSH
• karcinogeny toxicita   MeSH
• krysa rodu Rattus MeSH
• mezerový spoj účinky léků   MeSH
• mezibuněčná komunikace účinky léků   MeSH
• receptory aromatických uhlovodíků fyziologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 5-methylchrysene MeSH Prohlížeč
• 6-methylchrysene MeSH Prohlížeč
• adukty DNA MeSH
• chryseny MeSH
• karcinogeny MeSH
• receptory aromatických uhlovodíků MeSH

The effect of exposure to organic compounds adsorbed onto respirable air particles (<2.5microm) on DNA adducts in lymphocytes was studied in a group of non-smoking policemen (N=109, aged 35+/-0.9 years) working in the downtown area of Prague and spending >8h daily outdoors. Personal exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) adsorbed on respirable particles was monitored in each subject for 48h before biological sampling. DNA adducts were analyzed by a (32)P-postlabelling assay, and total DNA adduct levels and B[a]P-like spots were determined. Further biomarkers included cotinine levels in urine to control for exposure to tobacco smoke, plasma levels of vitamins A, E and C and polymorphisms of metabolic genotypes (GSTM1, GSTP1, GSTT1, CYP 1A1-Msp I and Ile/Val, MTHFR, MS), DNA repair genotypes (XRCC1, hOGG1 and XPD exons 6 and 23) and the p53 gene (p53 Msp I and BstU I). All the biomarkers of exposure and effect were analyzed repeatedly during a period of one year at 2-3 month intervals (January, March, June, September 2004) to cover periods with high (winter) and low (summer) levels of air pollution. The highest personal exposure to c-PAHs was found in January (8.1+/-8.8ng/m(3)), while the other three sampling periods exhibited 3-4-fold lower c-PAH exposure. The total DNA adducts were only slightly elevated in January (2.08+/-1.60) compared to March (1.66+/-0.65), June (1.96+/-1.73) and September (1.77+/-1.77). B[a]P-like DNA adducts, however, were significantly higher in January than in the March and June sampling periods (0.26+/-0.14 vs. 0.19+/-0.12 and 0.22+/-0.13, respectively; p<0.0001 and p=0.017) indicating that c-PAH exposure probably plays a crucial role in DNA adduct formation in lymphocytes. No effect of individual metabololic or DNA repair genotypes on DNA adduct levels was observed. However, the combination of two genotypes encoding enzymes metabolizing c-PAHs - CYP 1A1 and GSTM1 - was associated with the levels of total and B[a]P-like DNA adducts under conditions of increased exposure to c-PAHs. Our study suggests that DNA adducts in the lymphocytes of subjects exposed to increased c-PAH levels are an appropriate biomarker of a biologically effective dose, directly indicating whether or not the extent of exposure to these compounds is related to an increased mutagenic and carcinogenic risk.

• MeSH
• adukty DNA analýza   MeSH
• benzopyren analýza   toxicita   MeSH
• biologické markery analýza   MeSH
• dospělí MeSH
• genotyp MeSH
• karcinogeny životního prostředí analýza   toxicita   MeSH
• látky znečišťující vzduch v pracovním prostředí analýza   toxicita   MeSH
• látky znečišťující vzduch analýza   toxicita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfocyty chemie   účinky léků   MeSH
• mutageny analýza   toxicita   MeSH
• policie * MeSH
• polycyklické aromatické uhlovodíky analýza   toxicita   MeSH
• polymorfismus genetický MeSH
• pracovní expozice * MeSH
• roční období MeSH
• znečištění ovzduší škodlivé účinky   analýza   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• adukty DNA MeSH
• benzo(a)pyrene-DNA adduct MeSH Prohlížeč
• benzopyren MeSH
• biologické markery MeSH
• karcinogeny životního prostředí MeSH
• látky znečišťující vzduch v pracovním prostředí MeSH
• látky znečišťující vzduch MeSH
• mutageny MeSH
• polycyclic aromatic hydrocarbons-DNA adduct MeSH Prohlížeč
• polycyklické aromatické uhlovodíky MeSH

BACKGROUND: During studies on the health of children aged 3 or 4.5 years in Teplice and Prachatice districts of the Czech Republic, we focused also on the extent of smoking in the families and exposure of children to environmental tobacco smoke. METHODS AND RESULTS: In 1128 questionnaires administered to mothers of children born in 1994-1998, 35.6% of mothers indicated that they smoked and 48.9% of fathers/partners (N = 1075) were smokers. Including other family members, there were 41.6% families without any smoker, 30.1% of families with one smoker and 24% families with two smokers (out of 1061 households). Urine samples of 523 pairs of mothers and children (aged 4.5 years) were assayed for cotinine using a RIA radioimmunoassay. Concentration of cotinine was higher than 500 ng cotinine/mg creatinine (the cut-off value for smoking) in 199 of 523 mothers (38%). Exposure of children to environmental tobacco smoke (cotinine levels over 20ng/mg creatinine) was detected in 48.2% of 523 children. There were more children with cotinine levels over 20 ng in Teplice (59.2% of 287 children) than in Prachatice district (34.7% of 236 children). CONCLUSIONS: Cotinine levels in child's urine were significantly positively associated with maternal cotinine levels as well as with smoking of mother and father, and were lower in children visiting kindergarten.

• MeSH
• kotinin moč   MeSH
• kouření epidemiologie   MeSH
• lidé MeSH
• předškolní dítě MeSH
• rodiče * MeSH
• znečištění tabákovým kouřem * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• kotinin MeSH
• znečištění tabákovým kouřem * MeSH