Search

Ambient air particulate matter (PM) represents a class of heterogeneous substances that form one component of air pollution. Oxidative stress has been implicated as an important action mechanism for PM on the human organism. Oxidative damage induced by reactive oxygen species (ROS) may affect any cellular macromolecule.The aim of our study was to investigate the impact of air pollution on oxidative DNA damage [8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG)] and lipid peroxidation [15-F2t-isoprostane (15-F2t-IsoP)] in the urine and blood from mothers and newborns from two localities with different levels of air pollution: Ceske Budejovice (CB), a locality with a clean air, and Karvina, a locality with high air pollution. The samples from normal deliveries (38–41 week+) of nonsmoking mothers and their newborns were collected in the summer and winter seasons.Higher PM2.5 concentrations were found in Karvina than in CB in the summer 2013 (mean±SD: 20.41±6.28 vs. 9.45±3.62μg/m3, P

Personal exposures to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) bound to airborne particulate matter 2.5 μm (PM2.5) were measured in the context of a large-scale molecular epidemiological study in order to identify the impacts of air pollution on human health. Sampling was carried out in three industrial cities in the Czech Republic: Ostrava, Karvina and Havirov. The city of Prague, exhibiting much lower industrial air pollution but a high level of traffic, served as a control. The first monitoring campaigns were held in winter and were repeated in the summer of 2009. The active personal monitors PV 1.7 for PM2.5-bound c-PAHs were used. Non-smoking city policemen from Prague, Karvina and Havirov, and office workers from Ostrava, participated in the study. All participants completed a personal questionnaire and a time-location-activity diary. The average personal winter exposure to c-PAHs (sum of the eight PAHs-benz[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[g,h,i]perylene, benzo[k]fluoranthene, chrysene, dibenz[a,h]anthracene and indeno[1,2,3-c,d]pyrene) was highest in Karvina, 39.1, followed by Ostrava at 15.1 and Prague at 4.3 ng/m(3). The winter levels were significantly higher than the summer values (P0.001): 4.3 in Karvina, 3.0 in Ostrava, 1.6 in Havirov and 1.0 ng/m(3) in Prague. The average personal benzo[a]pyrene winter/summer exposures were: 6.9/0.6 in Karvina, 2.5/0.4 in Ostrava, 0.8/0.1 in Prague and 0.2 ng/m(3) in summer in Havirov. In this study, we examined personal exposure to c-PAHs and tested it for associations with potential predictor variables collected from questionnaires, addressing life style factors and day-to-day activities. We found outdoor concentration, environmental tobacco smoke exposure, home heating fuel of coal, wood or gas, frequency of exhaust fan use, cooking and commuting by a car to be the main determinants of personal exposure.

Populations living in industrialised regions are at higher risk of a number of diseases and shortened life span. These negative effects are primarily brought about by damage to cells and macromolecules caused by environmental pollutants. In this study, we analysed the effect of exposure to benzo[a]pyrene, a particulate matter of aerodynamic diameter < 2.5 µm (PM2.5), and benzene on oxidative stress markers [including 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), 15-F(2t)-isoprostane (15-F2t-IsoP) and protein carbonyls] and cytogenetic parameters (stable and unstable chromosomal aberrations). The samples were collected from subjects living in the Ostrava region characterised by very high levels of air pollution and in Prague with comparatively lower concentrations of pollutants in three seasons (winter 2009, summer 2009 and winter 2010). Despite several-fold higher concentrations of air pollutants in the Ostrava region, the levels of stable aberrations (genomic frequency of translocations per 100 cells, percentage of aberrant cells and frequency of acentric fragments) were mostly comparable in both locations. The frequency of unstable aberrations measured as the number of micronuclei was unexpectedly significantly lower in the Ostrava region subjects in both seasons of 2009. Urinary excretion of 8-oxodG did not differ between locations in either season. Lipid peroxidation measured as levels of 15-F2t-IsoP in blood plasma was elevated in the Ostrava subjects sampled in 2009. Protein oxidation was higher in Prague samples collected in summer 2009. Multivariate analyses conducted separately in subjects from Prague and Ostrava showed a negative association between the frequency of micronuclei and concentrations of benzo[a]pyrene and PM2.5 in both regions. A positive relationship was observed between lipid peroxidation and air pollution; protein oxidation seems to be positively affected by PM2.5 in both regions.

A higher incidence of asthma is one of the serious problems confronting urban populations worldwide. The aim of the present study was to analyze the effect of age, gender, smoking, vitamin intake, genetic polymorphisms in genes related to the metabolic activation of polycyclic aromatic hydrocarbons (PAHs) and their detoxification and oxidative damage to DNA, lipids and proteins on the frequency of micronuclei (MN) in a group of 175 children (81 with bronchial asthma and 94 healthy controls) aged 6-15 years. The study group from the most polluted region of the Czech Republic, Ostrava, was followed in November 2008, when the mean concentration of benzo[a]pyrene (B[a]P) measured by stationary monitoring was 11.4±9.8ng/m(3). The results of cotinine analysis revealed active smoking in 15 children. The frequency of MN per 1000 binucleated cells (MN/1000 BNC), measured by automated image analysis, indicated a significant risk for smoking children with asthma in comparison with smoking control children (4.25±1.54 and 3.00±0.77, respectively, p

DNA integrity was investigated in the lymphocytes of 50 bus drivers, 20 garagemen and 50 controls using the comet assay with excision repair enzymes. In parallel, 8-oxo-7,8-dihydro-2′-deoxyguanosine and 15-F2t-isoprostane levels in the urine and protein carbonyl levels in the plasma were assessed as markers of oxidative damage to DNA, lipids and proteins. Exposure to carcinogenic polycyclic aromatic hydrocarbons (cPAHs) and volatile compounds was measured by personal samplers for 48 and 24 h, respectively, before the collection of biological specimens. Both exposed groups exhibited a higher levels of DNA instability and oxidative damage to biological macromolecules than the controls. The incidence of oxidized lesions in lymphocyte DNA, but not the urinary levels of 8-oxodG, correlated with exposure to benzene and triglycerides increased this damage. Oxidative damage to lipids and proteins was associated with exposure to cPAHs and the lipid peroxidation levels positively correlated with age and LDL cholesterol, and negatively with vitamin C. The carriers of at least one variant hOGG1 (Cys) allele tended to higher oxidative damage to lymphocyte DNA than those with the wild genotype, while XPD23 (Gln/Gln) homozygotes were more susceptible to the induction of DNA strand breaks. In contrast, GSTM1 null variant seemed to protect DNA integrity.

The aim of the present study was to investigate the seasonal variability of markers of oxidative damage to lipids (15-F2t-isoprostane, 15-F2t-IsoP) and proteins (protein carbonyl levels) in 50 bus drivers and 50 controls from Prague, Czech Republic, and to identify factors affecting oxidative stress markers. The samples were collected in three seasons with different levels of air pollution. The exposure to environmental pollutants (carcinogenic polycyclic aromatic hydrocarbons, c-PAHs, particulate matter, PM2.5 and PM10, and volatile organic compounds, VOC) was monitored by personal and/or stationary monitors. For the analysis of both markers, ELISA techniques were used. The median levels of individual markers in bus drivers versus controls were as follows: 15-F2t-IsoP (nmol/mmol creatinine): winter 2005, 0.81 versus 0.68 (p

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 spending8h 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; p0.0001 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.

The capital city of Prague is one of the most polluted localities of the Czech Republic. The effect of exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) adsorbed onto respirable air particles (2.5 microm) on chromosomal aberrations was studied in a group of city policemen (street patrol, aged 34+/-8 years) working in the downtown area of Prague and spending daily8h outdoors (N=61) in months of January and March 2004. Ambient air particles (PM10, PM2.5) and c-PAHs were monitored using Versatile Air Pollution Sampler (VAPS), and personal exposure was evaluated using personal samplers during working shift. Chromosomal aberrations were analyzed by fluorescent in situ hybridization (FISH) and conventional cytogenetic analysis. Urinary cotinine, plasma levels of vitamins A, E and C, folate, total cholesterol, HDL, LDL cholesterols and triglycerides were also analyzed as possible effect modifiers. During the sampling period the particulate air pollution monitored by VAPS was in January versus March as follows: PM10 55.6 microg/m3 versus 36.4 microg/m3, PM2.5 44.4 microg/m3 versus 24.8 microg/m3, c-PAHs 19.7 ng/m3 versus 3.6 ng/m3, and B[a]P 4.3 ng/m3 versus 0.8 ng/m3. Significant differences were observed for all FISH endpoints studied for the sampling in January and March (%AB.C.=0.27+/-0.18 versus 0.16+/-0.17, p0.001, F(G)/100=1.32+/-1.07 versus 0.85+/-0.95, p0.01, AB/1000 (aberrations/1000 cells)=4.27+/-3.09 versus 2.59+/-2.79, p0.001) while conventional cytogenetic analysis did not reveal any differences in the frequency of chromosomal aberrations. Factors associated with an increased level of translocations by FISH indicated the effect of age, cholesterol, LDL-cholesterol and vitamin C. We may conclude that FISH indicates that the city policemen in Prague represent a group of increased genotoxic risk. This is the first study reporting that translocations induced by c-PAHs in peripheral lymphocytes last only several weeks.

Exposure to ambient air pollution is associated with many diseases. Oxidative and nitrosative stress are believed to be two of the major sources of particulate matter (PM)-mediated adverse health effects. PM in ambient air arises from industry, local heating, and vehicle emissions and poses a serious problem mainly in large cities. In the present study we analyzed the level of oxidative and nitrosative stress among 50 bus drivers from Prague, Czech Republic, and 50 matching controls. We assessed simultaneously the levels of 15-F(2t)-isoprostane (15-F(2t)-IsoP) and 8-oxodeoxyguanosine (8-oxodG) in urine and protein carbonyl groups and 3-nitrotyrosine (NT) in blood plasma. For the analysis of all four markers we used ELISA techniques. We observed significantly increased levels of oxidative and nitrosative stress markers in bus drivers. The median levels (min, max) of individual markers in bus drivers versus controls were as follows: 8-oxodG: 7.79 (2.64-12.34)nmol/mmol versus 6.12 (0.70-11.38)nmol/mmol creatinine (p

Background: Benzo[a]pyrene and benzene are classified as proven human carcinogens.There are high concentrations of these pollutants in the eastern part of the Czech Republic. Aims: We measured pers...

Background and Aims: Ostrava Region in the Northern Moravia (Silesia) is the most polluted region in the Czech Republic by carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) as benzo[a]pyrene (...

Oxidative stress is believed to be one of the mechanisms of effects of air pollution to human health. We investigated levels of 8-oxodeoxyguanosine (8-oxodG), a marker of oxidative damage to DNA, in urine samples of 894 children from two districts in the Czech Republic: Teplice and Prachatice. We assessed the association between 8-oxodG levels and exposure to particulate matter of different size: ≤10 μm (PM10), ≤2.5 μm (PM2.5) and carcinogenic polycyclic aromatic hydrocarbons (c-PAHs); as well as between 8-oxodG levels and individual lifestyle, health and pregnancy outcomes. An ELISA technique was used for analysis of 8-oxodG levels. Median levels (range) of 8-oxodG in children from Teplice vs. Prachatice were as follows: 14.6 (3.1–326.5) nmol/mmol vs. 15.2 (3.0–180.8) nmol/mmol creatinine (p = 0.34). Levels of 8-oxodG were elevated in children exposed to environmental tobacco smoke (ETS) (p

The capital city of Prague is one of the most polluted localities of the Czech Republic. Therefore, the effect of exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) adsorbed onto respirable air particles ( 8 h outdoors (N = 53). Age- and sex-matched healthy volunteers spending > 90% daily time indoors were chosen as controls (N = 52). Ambient air particles (PM10, PM2.5) and c-PAHs were monitored using versatile air pollution sampler (VAPS), and personal exposure was evaluated using personal samplers during working shift. Chromosomal aberrations were analyzed by conventional cytogenetic analysis and fluorescent in situ hybridization (FISH). Urinary cotinine plasma levels of vitamins A, E and C, folate, total cholesterol, HDL, LDL cholesterols and triglycerides were also analyzed as possible effect modifiers. Genotypes CYP1A1*2A, CYP1A1*2C, GSTM1, GSTP1, GSTT1, EPHX1, NAT2, hOGG1, XRCC1, XPD, p53 BstI, p53 MspI, MTHFR677, and MS2656 were determined by PCR-based RFLP assays. The following levels of air pollution were recorded during the study period (mean from HiVol sampling): PM10 62.6g/m 3 , c-PAHs 24.7 ng/m 3 ,B [a]P 3.50 ng/m 3 . The conventional cytogenetic analysis did not reveal any differences between the group of policemen exposed to the ambient air pollution and the control group. The cytogenetic analysis by FISH analysis used the whole chromosome painting probes for chromosomes #1 and #4 (Cambio, UK). It detected a significant increase in all studied endpoints in the policemen compared to controls (% AB.C. = 0.33 ± 0.25 versus 0.24 ± 0.18, p < 0.05, FG/100 = 1.72 ± 1.57 versus 1.25 ± 1.11, p < 0.05, AB/1000 (aberrations/1000 cells) = 5.58 ± 4.62 versus 3.90 ± 3.06, p < 0.05). CYP1A1*2C (Ile/Ile), XPD 23 (Lys/Lys), and XPD 6 (CC) genotypes were associated with an increase of aberrant cells by conventional method. Factors associated with an increased level of translocations by FISH included age, smoking, B[a]P-like DNA adducts (corresponding to the exposure of c-PAHs), folate, polymorphisms of CYP1A1*2C, GSTP1, EPHX1, p53 MspI and MTHFR. Ambient air exposure to c-PAHs significantly increased FISH cytogenetic parameters in nonsmoking policemen. We may conclude that FISH indicates that the city policemen in Prague represent a group of increased genotoxic risk. This is the first study Abbreviations: PAHs, polycyclic aromatic hydrocarbons; c-PAHs, carcinogenic polycyclic aromatic hydrocarbons; B[a]P, benzo[a]pyrene; PM10, particulate matter < 10m; PM2.5, particulate matter < 2.5m; EXP, exposed group, policemen; CON, control group; CA, chromosomal aberrations; CCA, conventional cytogenetic analysis; FISH, fluorescence in situ hybridization; PBL, peripheral blood lymphocytes; PCR, polymerase

We analyzed the effect of exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) in ambient air on the plasma levels of p53 and p21 WAF1 proteins among city policemen, bus drivers and controls in three European cities: Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). p53 and p21 WAF1 proteins are key regulators of the cell cycle and are accepted as universal markers of genotoxic stress and DNA damage. In total 204 exposed subjects (100 smokers, 104 nonsmokers) and 152 controls (54 smokers, 98 nonsmokers) were analyzed. Personal exposure to c-PAHs was evaluated using personal samplers during the working shift. The levels of p53 and p21 WAF1 proteins were assessed by ELISA assay. There were no differences between the levels of either protein between exposed and controls, or smokers and nonsmokers, in any city. However, we observed significant differences in p53 plasma levels in all subjects regardless of the exposure status between the individual cities (median values: 5, 31, 234 pg/ml, p < 0.001, for Prague, Kosice and Sofia, respectively). The levels correspond to the differences in exposure levels to c-PAHs and benzo[a]pyrene (B[a]P) in the individual cities. A multiple linear regression analysis confirmed that c-PAHs exposure is a variable significantly affecting levels of both proteins in all locations. When all subjects were divided into the group exposed to below-median levels of c-PAHs and the group exposed to above-median levels of c-PAHs we found significantly higher p53, as well as p21 WAF1 levels in the above-median exposure group (p53, 167 pg/ml versus 25 pg/ml, p < 0.001; p21 WAF1 , 2690 pg/ml versus 2600 pg/ml, p < 0.05). Among all subjects p53 plasma levels were positively correlated with p21 WAF1 levels, exposure to B[a]P, c-PAHs and levels of total DNA adducts; for p21 WAF1 levels we observed the positive correlation with cotinine, c-PAHs exposure, total and B[a]P-like DNA adduct levels. In conclusion our results suggest that p53 and p21 WAF1 proteins plasma levels may be useful biomarkers of c-PAHs environmental exposure.

Organic compounds adsorbed to air particles (PM10) induced DNA adducts and embryotoxicity in in vitro and in vivo studies. Carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) induced 45-50% of all DNA adducts caused by organic matter bound to PM10. Placental bulky DNA adducts were affected by air pollution, smoking, genotypes and vitamin C. Higher DNA adduct levels were observed in nonsmoking mothers delivering children with intrauterine growth retardation (IUGR). An increased risk of IUGR was established in the pregnancy outcome study for mothers exposed to c-PAHs >15 ng/m during the first month of gestation. All these results indicate that c-PAHs represent a very important group of air pollutants.

The relationships between DNA adducts and birth weight and between birth weight and genetic polymorphisms of metabolic genotypes were studied using DNA from the children part of placental samples. The samples were collected in the districts of Teplice and Prachatice and in the city of Prague. DNA adducts were analyzed by 32 P-postlabeling in a total of 199 subjects, genetic polymorphisms in 1013 subjects. GSTM1, GSTP1, GSTT1, CYP1A1*2A, and CYP1A1*2C genotypes were determined for each subject. The level of DNA adducts was not correlated with birth weight in any group (N = 199), including that from the polluted district of Teplice (N = 90). Birth weight was significantly decreased by smoking, ETS and alleles of CYP1A1*2C. The risk of low birth weight and prematurity was significantly increased by genotypes of GSTM1 and CYP1A1*2C and the combination GSTM1+CYP1A1*2A. Genotypes are primarily effect modifiers, whose effect incorporates the effect of environmental factors. This means that in the future, the impact of air pollution on children’s health should be studied together with their genetic polymorphisms.

Vitamin C, E and A levels in maternal and cord blood sera were examined at delivery in two districts of the Czech Republic. Information on personal and social characteristics, health, ethnicity, and lifestyle was also collected. A highly significant correlation between ascorbate levels in maternal and cord blood was found. Vitamin C levels in cord blood were about 1.7 times those in maternal blood. This ratio was much higher for mothers deficient in vitamin C: it was about 3 for deficient nonsmokers and as high as 5 for deficient mothers who smoked cigarettes (p < 0.01). This finding may suggest a compensatory mechanism in fetuses that are endangered by oxidative stress. The mean maternal blood levels of vitamin A and E were higher than in fetal blood (both p < 0.0001). The mean fetal/maternal ratios were 0.7 for vitamin A and 0.2 for vitamin E levels; these ratios were considerably higher for mothers deficient in a particular vitamin as compared with those for well-nourished mothers. Ascorbate levels were associated with maternal education and smoking. Significantly decreased vitamin C levels were observed in Gypsy mothers and their babies; this may be attributed to unfavorable diet and smoking habits: about 78% of Gypsy mothers admitted smoking as compared with 31% of Czech mothers.

The effect of exposure to carcinogenic polycyclic aromatic hydrocarbons adsorbed onto respirable air particles (PM2.5, diameter 2.5 μm) on DNA adducts and chromosomal aberrations was repeatedly studied in Prague, Czech Republic, in groups of policemen working in the downtown area and in bus drivers. Personal exposure was evaluated using personal samplers during working shifts. DNA adducts were analyzed in lymphocytes by the (32)P-postlabeling assay and chromosomal aberrations were analyzed by conventional cytogenetic analysis and fluorescent in situ hybridization (FISH). The impact of environmental pollution on DNA adducts and chromosomal aberrations was studied in a total of 950 subjects. Our results suggest that the environmental exposure of nonsmokers to concentrations higher than 1 ng benzo[a]pyrene/m(3) represents a risk of DNA damage, as indicated by an increase in DNA adducts and the genomic frequency of translocations determined by FISH.