Possible relationship between respiratory diseases and urinary concentrations of polycyclic aromatic hydrocarbon metabolites - a pilot study.

This study investigates the potential relationship between exposure to polycyclic aromatic hydrocarbons (PAHs), specifically monohydroxylated metabolites (OH-PAHs), in urine, and the prevalence of respiratory diseases in 2-year-old children residing in two locations within the Czech Republic - Ceské Budejovice (control location) and the historically contaminated mining district of Most. Despite current air quality and lifestyle similarities between the two cities, our research aims to uncover potential long-term health effects, building upon previous data indicating distinctive patterns in the Most population. A total of 248 urine samples were analysed for the presence of 11 OH-PAHs. Employing liquid-liquid extraction with ethyl acetate and clean-up through dispersive solid-phase extraction, instrumental analysis was conducted using ultra-high performance liquid chromatography coupled with tandem mass spectrometry. The incidence of respiratory diseases was assessed through questionnaires administered by paediatricians. The concentrations of OH-PAHs were elevated in urine samples from 2-year-olds in Most compared to those from Ceské Budejovice. The incidence of respiratory diseases showed statistically significant higher levels of OH-PAHs in children from Most, together with a higher incidence of influenza. This association underlines the impact of environmental PAH exposure on children's respiratory health. It suggests that elevated urinary OH-PAH levels indicate an increased risk of developing respiratory diseases in the affected population. Further studies are needed to clarify the possible long-term health effects and to contribute to sound public health strategies.

Prenatal exposure to polycyclic aromatic hydrocarbons and growth parameters.

BACKGROUND AND OBJECTIVES: It has long been known that airborne polycyclic aromatic hydrocarbons (PAHs) can negatively affect pregnancy and birth outcomes, such as birth weight, fetal development, and placental growth factors. However, similar studies yield divergent results. Our goal was to estimate the amount of monohydroxylated PAH (OH-PAH) metabolites in the urine of pregnant women/mothers and their newborns in relation to birth outcomes, such as placenta weight, Apgar 5', and the growth parameters of children up to the age of two. METHODS: Two cohorts of children born in 2013 and 2014 during the summer and winter seasons in the Czech Republic in the cities Karviná (N = 144) and Ceské Budejovice (N = 198), which differ significantly in the level of air pollution, were studied. PAH exposure was assessed by the concentration of benzo[a]pyrene (B[a]P) in the air and the concentration of 11 OH-PAH metabolites in the urine of newborns and mothers. Growth parameters and birth outcomes were obtained from medical questionnaires after birth and from pediatric questionnaires during the following 24 months of the child's life. RESULTS: Concentrations of B[a]P were significantly higher in Karviná (p < 0.001). OH-PAH metabolites were significantly higher in the mothers' as well as in the newborns' urine in Karviná and during the winter season. Neonatal length was shorter in newborns in Karviná (p < 0.001), but this difference evened out during the next 3 to 24 months. Compared to Ceské Budejovice, newborns in Karviná showed significantly lower weight gain between birth and three months after delivery. The OH-PAH metabolites in mothers' or newborns' urine did not affect birth weight. The presence of seven OH-PAH (top 25% of values of concentrations higher than the median) metabolites in the newborns' urine is associated with decreased length of newborn. Nine OH-PAH metabolites decreased placenta weight, which was the most significant, while seven OH-PAH metabolites decreased Apgar 5'. CONCLUSION: We have shown a possible connection between higher concentration of OH-PAH metabolites in newborns' urine and decreased length, head circumference, placenta weight, and Apgar 5', but not birth weight.

The Impact of Cesarean and Vaginal Delivery on Results of Psychological Cognitive Test in 5 Year Old Children.

Background and objectives: The impact of cesarean and vaginal delivery on cognitive development was analyzed in 5 year old children. Materials and Methods: Two cohorts of 5 year old children born in the years 2013 and 2014 in Karvina (Northern Moravia) and Ceske Budejovice (Southern Bohemia) were studied for their cognitive development related to vaginal (n = 117) and cesarean types of delivery (n = 51). The Bender Visual Motor Gestalt Test (BG test) and the Raven Colored Progressive Matrices (RCPM test) were used as psychological tests. Results: In the comparison of vaginal delivery vs. cesarean section, the children delivered by cesarean section scored lower and, therefore, achieved poorer performance in cognitive tests compared to those born by vaginal delivery, as shown in the RCPM (p < 0.001) and in the BG test (p < 0.001). When mothers' education level was considered, the children whose mothers achieved a university degree scored higher in both the RCPM test (p < 0.001) and the BG test (p < 0.01) compared to the children of mothers with lower secondary education. When comparing mothers with a university degree to those with higher secondary education, there was a significant correlation between level of education and score achieved in the RCPM test (p < 0.001), but not in the BG test. Conclusions: According to our findings, the mode of delivery seems to have a significant influence on performance in psychological cognitive tests in 5 year old children in favor of those who were born by vaginal delivery. Since cesarean-born children scored notably below vaginally born children, it appears possible that cesarean delivery may have a convincingly adverse effect on children's further cognitive development.

Relation between personal exposure and outdoor concentrations of carcinogenic polycyclic aromatic hydrocarbons during smog episode.

OBJECTIVES: To our knowledge this is the first study measuring personal exposure to carcinogenic polycyclic aromatic hydrocarbons (cPAHs) bound to airborne particulate matter ≤ 2.5 µm (PM2.5) in periods of high air pollution (smog episode) in which citizen were tracked. METHODS: Measurements were performed in industrial regions of the Czech Republic: Ostrava, Karviná, Havírov. The city of Prague served as a control. Personal monitoring was conducted by active personal monitors for 48 hours. Non-smoking city policemen from Prague, Karviná and Havírov, office workers from Ostrava city and volunteers from Ostrava-Radvanice and Bartovice participated in the study (N = 214). RESULTS: The average personal exposure to benzo[a]pyrene (B[a]P) was highest in Ostrava (17.2 ng/m3), followed by Karviná, Havírov, Radvanice and Bartovice, and Prague (14.2, 12.0, 9.3, and 2.8 ng/m3, respectively). We tested for association between the personal exposure to cPAHs and various health-related factors extracted from the questionnaires, including lifestyle factors and day-to-day activities. CONCLUSIONS: Exposure to outdoor cPAHs, environmental tobacco smoke (ETS), commuting, and time spent indoors (in restaurants, workplace or home) were found to be the main determinants of the personal exposure. Daily cPAHs measurements in highly polluted areas are needed for evaluating the personal exposure and to avoid its underestimation resulting from stationary monitoring.

Oxidative stress in newborns by different modes of delivery.

OBJECTIVES: The aim of our study is to investigate the impact of the type of delivery - vaginal vs. cesarean section on oxidative damage determined as the lipid peroxidation (15-F2t-isoprostane (15-F2t-IsoP) in the cord blood of newborns and venous blood from mothers in 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. RESUTLS: In Karvina, the concentration of PM2.5 was higher than in CB in the summer 2013 (mean±SD: 20.41±6.28 vs. 9.45±3.62 µg/m3, p<0.001) and in the winter 2014 (mean±SD: 53.67±19.76 vs. 27.96±12.34 µg/m3, p<0.001). Similarly, the concentration of B[a]P was higher in Karvina than in CB in the summer 2013 (mean±SD: 1.16±0.91 vs. 0.16±0.26 ng/m3, p<0.001) and in the winter 2014 (5.36±3.64 vs. 1.45±1.19 ng/m3, p<0.001). Delivery procedures differed by the type of anesthesia; at the Cesarean section in CB was used general anesthesia in 73.8% vs. 20.8% in Karvina (p<0.001), epidural anesthesia in CB in 26.2% vs. 77.1% in Karvina (p<0.001), at vaginal delivery was local anesthesia used in CB in 58.9% vs. 14.1% in Karvina (p<0.001). In CB was oxidative stress higher after vaginal delivery (101.7±31.0 pg 15-F2t-isoP/ml plasma) vs. Cesarean section (83.9±26.9 pg 15-F2t-isoP/ml plasma, p<0.001), no difference between the type of delivery was observed in Karvina. CONCLUSION: No difference between the types of delivery was observed in mothers in CB as well as in Karvina. Oxidative stress in newborns in Karvina was significantly affected by the concentrations of PM2.5 and B[a]P in the polluted air.

Short-term impact of atmospheric pollution on fecundability.

BACKGROUND: Epidemiologic studies have reported associations between air pollution levels and semen characteristics, which might in turn affect a couple's ability to achieve a live birth. Our aim was to characterize short-term effects of atmospheric pollutants on fecundability (the month-specific probability of pregnancy among noncontracepting couples). METHODS: For a cohort of births between 1994 and 1999 in Teplice (Czech Republic), we averaged fine particulate matter (PM2.5), carcinogenic polycyclic aromatic hydrocarbons, ozone, nitrogen dioxide (NO2), and sulfur dioxide levels estimated from a central measurement site over the 60-day period before the end of the first month of unprotected intercourse. We estimated changes in the probability of occurrence of a pregnancy during the first month of unprotected intercourse associated with exposure, using binomial regression and adjusting for maternal behaviors and time trends. RESULTS: Among the 1,916 recruited couples, 486 (25%) conceived during the first month of unprotected intercourse. Each increase of 10 µg/m in PM2.5 levels was associated with an adjusted decrease in fecundability of 22% (95% confidence interval = 6%-35%). NO2 levels were also associated with decreased fecundability. There was no evidence of adverse effects with the other pollutants considered. Biases related to pregnancy planning or temporal trends in air pollution were unlikely to explain the observed associations. CONCLUSIONS: In this polluted area, we highlighted short-term decreases in a couple's ability to conceive in association with PM2.5 and NO2 levels assessed in a central monitoring station.

Analysis of biomarkers in a Czech population exposed to heavy air pollution. Part II: chromosomal aberrations and oxidative stress.

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 possible link between cognitive development in 5 years old children and prenatal oxidative stress.

BACKGROUND: To study the impact of oxidative damage associated with particulate matter< 2.5 µm (PM2.5) during prenatal period on the cognitive development in five years old children. METHODS: Two cohorts of children aged five years, born in the years 2013 and 2014, were studied for their cognitive development in the polluted district Karvina and the control district Ceske Budejovice. Exposure to PM2.5 in the ambient air was measured for each mother during the 3rd trimester of pregnancy. Oxidative damage was determined from the level of biomarkers at delivery in mothers´ and newborns´ urine as 8-oxo-7,8-dihydro-2´-deoxyguanosine (8-oxodG) and in plasma as 15-F2t-isoprostane levels (15-F2t-IsoP). The Bender Visual Motor Gestalt Test (BG test) and the Raven Colored Progressive Matrices (RCPM test) were used as psychological cognitive tests. RESULTS: Average concentrations of PM2.5 ± SD in the 3rd trimester of mothers´ pregnancies were 37.7 ± 14.7 µg/m3 and 17.1 ± 4.8 µg/m3 in Karvina and Ceske Budejovice, respectively (p < 0.001). The maternal level of 15-F2t-IsoP in plasma at the time of delivery was significantly associated with the results of the RCPM test (p < 0.05) and the BG test (p < 0.05) in five years old children. CONCLUSIONS: Lipid peroxidation in maternal plasma at the time of delivery has an adverse effect on the results of psychological cognitive tests in five years old children.

Factors affecting the frequency of micronuclei in asthmatic and healthy children from Ostrava.

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<0.05). Girls in the control group had 16% higher levels of MN in comparison with boys. Markers of oxidative damage to DNA, proteins and lipids were not associated with asthma in this study. Higher levels of MN were associated with increased levels of protein carbonyl groups. We conclude that smoking asthmatic children are at higher risk of DNA damage measured as the frequency of micronuclei in peripheral blood lymphocytes.

Micronuclei levels in mothers and their newborns from regions with different types of air pollution.

The aim of this study was to analyze genetic damage in human lymphocytes measured using automated image analysis of micronuclei (MN) in a group of 178 mothers and their newborns from two locations in the Czech Republic. The concentrations of benzo[a]pyrene (B[a]P), particulate matter of aerodynamic diameter <2.5 µm (PM2.5) and benzene were measured by stationary monitoring in the winter season of 2008/2009 in the capital city of Prague and in Ceske Budejovice, a regional city in a rural area. The 3-month mean concentration of B[a]P before delivery was lower in Prague in comparison with Ceske Budejovice: 1.9 ± 0.5ng/m³ vs. 3.2 ± 0.2ng/m³ (p<0.001). The opposite trend was found for PM2.5 and benzene: 27.0 ± 2.5µg/m³ and 2.5 ± 0.5µg/m³ vs. 24.5 ± 0.7µg/m³ and 2.1 ± 0.8µg/m³ (p<0.001) for Prague vs. Ceske Budejovice, respectively. The average age of the mothers was 31 years (range, 18-49 years). The frequencies of MN per 1000 binucleated cells were 8.35 ± 3.06 vs. 6.47 ± 2.35 (p<0.001) for mothers from Prague and Ceske Budejovice, respectively, and 2.17 ± 1.32 vs. 3.82 ± 2.43 (p<0.001) for newborns from Prague and Ceske Budejovice, respectively. Other factors, including vitamin intake, exposure to tobacco smoke, body mass index (BMI) before pregnancy, the education level of the mothers and the impact of the mothers' and fathers' ages were analyzed in our study. The results suggest that the different sensitivity of the study groups to various mixtures of carcinogenic pollutants could be affected by significant differences in lifestyle factors. Possible higher genetic damage was analyzed in newborns of smoking mothers, and the birth weight of this group was 7.4% lower (p<0.05) in comparison with the newborns of nonsmoking mothers. No impact of the age of the mothers or fathers on MN frequency in the newborns was observed.

Effect of Polycyclic Aromatic Hydrocarbons Exposure on Cognitive Development in 5 Years Old Children.

Objectives: To analyze the impact of polycyclic aromatic hydrocarbons (PAHs) in ambient air at the time of delivery and five years of age on cognitive development in five year old children. Materials and Methods: Two cohorts of children born in the years 2013 and 2014 from Karvina (Northern Moravia, n = 70) and Ceske Budejovice (Southern Bohemia, n = 99) were studied at the age of five years for their cognitive development related to the exposure to PAHs, determined in the ambient air as the concentration of benzo[a]pyrene (B[a]P) and OH-PAH (hydroxy-PAH) metabolites in urine of the newborns at the time of delivery. As psychological tests, the Bender Visual Motor Gestalt Test (BG test) and the Raven Colored Progressive Matrices (RCPM test) were used. Results: Concentrations of B[a]P in the third trimester of mother's pregnancies were 6.1 ± 4.53 ng/m3 in Karvina, and 1.19 ± 1.28 ng/m3 (p < 0.001) in Ceske Budejovice. Neither the outcome of the RCPM test nor the BG test differed between children in Karvina vs. Ceske Budejovice, or boys vs. girls. Cognitive development in five year old children was affected by the higher exposure to PM2.5 during the third trimester in girls in Karvina. Conclusions: We did not observe any significant effect of prenatal PAH exposure on psychological cognitive tests in five year old children.

Urinary 8-oxodeoxyguanosine levels in children exposed to air pollutants.

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:

Seasonal variability of oxidative stress markers in city bus drivers. Part II. Oxidative damage to lipids and proteins.

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<0.01); summer 2006, 0.62 versus 0.60 (p=0.90); winter 2006, 0.76 versus 0.51 (p<0.001); carbonyl levels (nmol/ml plasma): winter 2005, 14.1 versus 12.9 (p=0.001); summer 2006, 17.5 versus 16.6 (p=0.26); winter 2006, 13.5 versus 11.7 (p<0.001). Multivariate logistic regression identified PM levels measured by stationary monitors over a period 25-27 days before urine collection as a factor positively associated with lipid peroxidation, while protein oxidation levels correlated negatively with both c-PAHs and PM levels. In conclusion, markers of oxidative damage to lipids and proteins were increased in bus drivers in winter seasons, but not in summer. Lipid peroxidation was positively correlated with c-PAHs and PM exposure; protein oxidation correlated negatively and was highest in summer suggesting another factor(s) affecting protein carbonyl levels.

Seasonal variability of oxidative stress markers in city bus drivers. Part I. Oxidative damage to DNA.

We investigated the seasonal variability of 8-oxodeoxyguanosine (8-oxodG), a marker of oxidative damage to DNA, in urine of 50 bus drivers and 50 controls in Prague, Czech Republic, in three seasons with different levels of air pollution: winter 2005, summer 2006 and winter 2006. The exposure to environmental pollutants (carcinogenic polycyclic aromatic hydrocarbons, c-PAHs, particulate matter (PM), and volatile organic compounds (VOC)) was monitored by personal and/or stationary monitors. For the analysis of 8-oxodG levels, the ELISA technique was used. Bus drivers were exposed to significantly higher levels of c-PAHs in winter 2006, while in the other two seasons the exposure of controls was unexpectedly higher than that of bus drivers. We did not see any difference in VOC exposure between both groups in summer 2006 and in winter 2006; VOC were not monitored in winter 2005. 8-OxodG levels were higher in bus drivers than in controls in all seasons. The median levels of 8-oxodG (nmol/mmol creatinine) in bus drivers vs. controls were as follows: winter 2005: 7.79 vs. 6.12 (p=0.01); summer 2006: 6.91 vs. 5.11 (p<0.01); winter 2006: 5.73 vs. 3.94 (p<0.001). Multivariate logistic regression analysis identified PM2.5 and PM10 levels, measured by stationary monitors during a 3-day period before urine collection, as the only factors significantly affecting 8-oxodG levels, while the levels of c-PAHs had no significant influence.

Chromosomal aberration frequencies determined by conventional methods: Parallel increases over time in the region of a petrochemical industry and throughout the Czech Republic.

The rationale for cytogenetic monitoring to determine if safe maximum allowable concentrations (MAC) of genotoxic chemicals are being maintained in a workplace is that exposure levels that do not increase chromosomal aberration frequencies are without harmful effects. Such monitoring, widely used in occupational health programs in the Czech Republic (CR), includes workers exposed to 1,3-butadiene (BD) or other chemicals. Studies of BD exposed workers in the years 1992, 1993, 1994, 1998, and 2004 compared mean frequencies of cells carrying chromosomal aberrations (frequency of aberrant cells=%AB.C.) in exposed workers with those in non-exposed matched controls in the same plant or in other individuals living in the region of the same petrochemical industry. Workers potentially exposed to acrylonitrile at this site were also evaluated in 2000, along with another unexposed matched control group. The %AB.C. values of exposed workers and their controls were also compared with reference values determined for normal individuals (ages 20-59 years) throughout the CR. Substantial discrepancies were noted between subjects in the region of the petrochemical industry (exposed workers and controls) for the years 2000 and 2004 and the reference CR-wide normal values that had been determined during an earlier time period. The matched non-exposed controls at the petrochemical industry site showed a mean %AB.C. value of 1.56+/-1.23% (N=25) in 1998; this rose to a mean of 2.65+/-2.29% (N=33) in 2000. In 2004, values for non-exposed matched controls at the industry site were 2.64+/-1.75% for males (N=25) and 2.38+/-1.74% (N=26) for females. However, the earlier determined CR-wide %AB.C. mean reference values for normal individuals were 1.77+/-1.16% (N=1305) for the interval 1977-1988 and 1.45+/-1.17% (N=2140) for the interval 1991-1999. As both reference values are substantially lower than those determined in 2000 and 2004 for the non-exposed matched controls at the petrochemical industry site, an analysis of the CR-wide mean normal individual reference values for this same 2000-2004 period was conducted. Unexpectedly, it was found that this reference value too had risen to 1.95+/-1.36% (N=1045) and was comparable to the concurrent matched control values at the petrochemical industry site where the monitoring studies were conducted. This substantial increase in %AB.C. values in 2000 and 2004, therefore, has occurred throughout the CR and is probably unrelated to chemicals uniquely present at the petrochemical industry site.

Environmental exposure to carcinogenic polycyclic aromatic hydrocarbons--the interpretation of cytogenetic analysis by FISH.

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 daily >8h 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, p<0.001, F(G)/100=1.32+/-1.07 versus 0.85+/-0.95, p<0.01, AB/1000 (aberrations/1000 cells)=4.27+/-3.09 versus 2.59+/-2.79, p<0.001) 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.

Impact of air pollution and genotype variability on DNA damage in Prague policemen.

DNA integrity was analyzed in the lymphocytes of 65 non-smoking city policemen during January and September 2004 using the comet assay combined with excision repair enzymes. Information about inhalation exposure was obtained by (1) stationary monitoring of PM2.5 and carcinogenic polycyclic aromatic hydrocarbons (cPAHs) during the sampling periods and (2) personal exposure monitoring of cPAHs 48h before blood sampling. The data were completed by a lifestyle questionnaire. Regardless of the season of the year, policemen working outdoors (exposed group) exhibited higher levels of DNA damage than those working indoors (controls). Within the exposed group, the levels of both unspecified and oxidative DNA damage detected in January significantly exceeded those found in September. The controls did not show analogous inter-seasonal variability. The winter levels of oxidative DNA damage positively correlated with exposure to cPAHs, probably reflecting increased oxidative stress as a result of high concentrations of PM2.5. In comparison with the wild type genotype, the carriers of at least one mutated allele, CYP1A1*2C (Ile/Val), MTHFR 2656 or MS 2656, and the EPHX1-medium phenotype appeared to be more susceptible specifically to the induction of oxidative DNA damage, while the p53 MspI mutation predisposed the carrier to a higher incidence of both breaks and oxidative lesions in DNA. In contrast, GSTM1-null and vitamin C tended rather to protect DNA integrity.

Oxidative and nitrosative stress markers in bus drivers.

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<0.01); 15-F(2t)-IsoP: 0.81 (0.38-1.55)nmol/mmol versus 0.68 (0.39-1.79)nmol/mmol creatinine (p<0.01); carbonyl levels: 14.1 (11.8-19.0)nmol/ml versus 12.9 (9.8-16.6)nmol/ml plasma (p<0.001); NT: 694 (471-3228)nmol/l versus 537 (268-13833)nmol/l plasma (p<0.001). 15-F(2t)-IsoP levels correlated with vitamin E (R=0.23, p<0.05), vitamin C (R=-0.33, p<0.01) and cotinine (R=0.47, p<0.001) levels. Vitamin E levels also positively correlated with 8-oxodG (R=0.27, p=0.01) and protein carbonyl levels (R=0.32, p<0.001). Both oxidative and nitrosative stress markers positively correlated with PM2.5 and PM10 exposure. In conclusion, our study indicates that exposure to PM2.5 and PM10 results in increased oxidative and nitrosative stress.

Chromosomal aberrations in environmentally exposed population in relation to metabolic and DNA repair genes polymorphisms.

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 (<2.5mum) on chromosomal aberrations was studied in a group of policemen (males, aged 22-50 years) working in the downtown area of Prague and spending daily >8h 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 62.6microg/m(3), c-PAHs 24.7ng/m(3), B[a]P 3.50ng/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, F(G)/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 that has reported a relationship between DNA adducts (biomarker of exposure) and chromosomal aberrations by FISH (biomarker of effect).

Air pollution by carcinogenic PAHs and plasma levels of p53 and p21(WAF1) proteins.

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, 234pg/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, 167pg/ml versus 25pg/ml, p<0.001; p21(WAF1), 2690pg/ml versus 2600pg/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.