Urinary polycyclic aromatic hydrocarbons and monohydroxy metabolites as biomarkers of exposure in coke oven workers.

OBJECTIVES: To assess exposure to polycyclic aromatic hydrocarbons (PAHs) using 13 unmetabolised PAHs (U-PAHs) and 12 monohydroxy metabolites (OHPAHs) in urine, and to compare the utility of these biomarkers. METHODS: 55 male Polish coke oven workers collected urine spot samples after a workshift. U-PAHs (naphthalene, acenaphtylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[k]fluoranthene, benzo[b]fluoranthene, benzo[a]pyrene) were determined by automatic solid phase micro-extraction followed by gas chromatography/mass spectrometry (GC/MS). OHPAHs (1- and 2-hydroxynaphthalene, 2- and 9-hydroxyfluorene, 4-, 9-, 3-, 1- and 2-hydroxyphenanthrene, 1-hydroxypyrene, 6-hydroxychrysene, 3-hydroxybenzo[a]pyrene) were determined, after liquid/liquid extraction and derivatisation, by GC/MS. RESULTS: U-PAHs from naphthalene to chrysene were found in 100% of samples, and heavier U-PAHs in 7-22% of samples. OHPAHs up to 1-hydroxypyrene were found in 100% of samples, while 6-hydroxychrysene and 3-hydroxybenzo[a]pyrene were always below the quantification limit. Median naphthalene, phenanthrene, pyrene, chrysene and benzo[a]anthracene levels were 0.806, 0.721, 0.020, 0.032 and 0.035 microg/l, while hydroxynaphthalenes, hydroxyphenanthrenes and 1-hydroxypyrene levels were 81.1, 18.9 and 15.4 microg/l. For each chemical, the ratio between U-PAH and the corresponding OHPAH ranged from 1:26 to 1:1000. Significant correlations between logged values of U-PAHs and OHPAHs, between U-PAHs, and between OHPAHs were found, with Pearson's r ranging from 0.27 to 0.97. CONCLUSION: Current analytical techniques allow specific and simultaneous measurement of several urinary determinants of PAHs in humans. The results of these measurements support the use of U-PAHs as biomarkers of exposure and suggest the spectrum of chemicals to be investigated, including carcinogenic chrysene and benzo[a]anthracene, should be widened.

Non-smoking coke oven workers show an occupational PAH exposure-related increase in urinary mutagens.

We examined the urinary mutagenicity in the YG1024 Salmonella typhimurium strain in the presence of S9 mix, of 31 male non-smoking coke oven workers and an equal number of controls matched for gender and dietary habits. Occupational PAH exposure to the workers was assessed by means of the individual urinary post-shift excretion of 1-pyrenol (mean +/- S.D.: 5.41 +/- 6.06 micromole/mol creatinine). Eleven urinary extracts of workers (35.5%) were clearly mutagenic (with at least a doubling of the number of spontaneous revertants), against only two samples in the control group (6.5%) (chi2-test; chi2 = 7.883; P < 0.01). Moreover, the mean mutagenic activity level corrected for dilution/concentration of the urine was about three times higher in coke oven workers than in matched controls (mean +/- S.D. (range) 495 +/- 407 (89.7-1603) versus 186 +/- 113 (14.2-524) net revertants/mmol creatinine; Mann-Whitney U-test, z = 3.86, P < 0.001). Simple linear regression analysis showed that the coke workers' urinary mutagenic activity is associated with the PAH occupation-related urinary excretion of 1-pyrenol (r = 0.41, P = 0.0215). This study definitely demonstrates an occupation-related exposure of coke oven workers' bladder epithelium to mutagenic PAH metabolites. This factor, mainly in the case of high exposure studied here, may account for a higher bladder cancer risk in coke oven workers.

Association between urinary 1-hydroxypyrene and genotoxic effects in coke oven workers.

AIMS: To investigate whether current occupational exposure of coke oven workers to polycyclic aromatic hydrocarbons (PAHs) results in genotoxic effects measured in peripheral blood lymphocytes and whether these biomarkers are associated with the biomarkers of exposure. METHODS: Blood and urine samples were collected immediately after a shift at the end of a working week from 50 coke oven workers and 50 control workers not exposed to PAHs. Methods included: (1) biomarkers of exposure: urinary 1-hydroxypyrene (HpU), urinary mutagenicity by the plate Salmonella test with strains TA98 and YG1024 after metabolic activation, expressed as mutagenic rate (MR98 and MR1024, respectively), urinary cotinine; and (2) biomarkers of biological effects in peripheral blood lymphocytes (PBL): sister chromatid exchanges (SCE/cell), cells of high frequency of SCE (% HFC), micronuclei (MN/1000 cells), chromosomal aberrations (CA/100 cells), and DNA damage by the Comet assay. RESULTS: Occupational exposure to PAH resulted in significantly increased levels of HpU and mutagenic effect of urine. Median values of these biomarkers in coke oven workers were: 9.0 micromol/mol creatinine for HpU, 2.7 for MR98, and 8.2 for MR1024, compared to the controls: HpU = 0.6 micromol/mol creatinine, MR98 = 1.2, and MR1024 = 5.5. Occupational exposure caused significant induction of SCE, HFC, and MN in coke oven workers: median SCE = 5.9, HFC = 12.0%, MN = 6.0 compared to the controls: 3.9, 5.0%, and 3.0, respectively. No effect of occupational exposure was found in relation to CA and DNA damage measured with the Comet assay. HpU concentration was positively associated with SCE and HFC. The concentration of urinary 1-hydroxypyrene corresponding to a 5% probability of increased SCE was 1.0 micromol/mol creatinine. CONCLUSIONS: The occupational exposure to PAHs resulted in measurable biological effects (SCE, HFC, MN). In coke oven workers an increased level of SCE was not observed below the level of 1.0 micromol HpU/mol creatinine.

The effect of coal stoves and environmental tobacco smoke on the level of urinary 1-hydroxypyrene.

The objective of this study was to investigate a relationship between indoor air pollution from heating and cooking with coal-burning stoves and from environmental tobacco smoke (ETS), and the level of urinary 1-hydroxypyrene (1-OH-PY). 1-OH-PY was analysed in children living in three areas of Silesia, a province in Poland. Urine samples were collected in winter, (1) from children exposed to ETS and smoke resulting from indoor coal-burning and (2) from control children. Airborne particulates had been sampled by use of stationary samplers by the Regional Sanitary-Epidemiological Station, Katowice throughout 12 months prior to the urine sampling. The urinary level of 1-OH-PY tended to increase in children exposed to ETS, but the increase was not significant. The concentrations of 1-OH-PY in urine of passive smokers were significantly elevated only in Bytom where an index of smoking parents of the studied children was highest as compared to other areas. Exposure to polycyclic aromatic hydrocarbons (PAH) due to domestic heating and cooking with coal-burning stoves resulted in significantly increased levels of 1-OH-PY. The results of this study indicate that the uptake of PAH due to indoor air pollution strongly affected the level of 1-OH-PY and that the main source of PAH in indoor air was the household use of coal for heating and/or cooking. When the results associated with this kind of exposure were excluded, median 1-OH-PY levels from the three examined areas assumed a pattern more similar to that of the benzo(a)pyrene (BaP) concentrations in ambient air.

Evaluation of intra- and interindividual variation of urinary 1-hydroxypyrene, a biomarker of exposure to polycyclic aromatic hydrocarbons.

The study was performed on 30, 8-year-old children living in an industrial town of the Upper Silesia region. Morning urine samples were collected on 6 consecutive days. Intraindividual variation of urinary 1-hydroxypyrene concentrations, calculated as a coefficient of variance (CV), ranged from 14 to 109% whereas inter-individual variation ranged from 69 to 109%. Three-way analysis of variance disclosed a significant effect of sex, exposure to environmental tobacco smoke and day of examination on 1-hydroxypyrene concentrations not corrected for creatinine. The appropriate sample size for population studies and the minimum number of observations for the individual assessment of environmental exposure to PAHs calculated on the basis of inter- and intraindividual variability of 1-hydroxypyrene concentrations in urine amounted to 164 and 99, respectively. Urinary 1-hydroxypyrene may be considered a good indicator of environmental exposure to PAHs at the group level.

Exposure of coke-oven workers to polycyclic aromatic hydrocarbons based on biological monitoring results.

This study assessed the exposure of coke-oven workers to polycyclic aromatic hydrocarbons (PAH) based on two methods of biological monitoring (urinary mutagenicity by Ames test and urinary 1-hydroxypyrene) and concentrations of benzo(a)pyrene in the air collected by personal sampling. The coke-oven workers were classified into four job categories. There was no significant correlation among three methods applied. According to all methods, the workers most exposed to PAH were those working at the topside area of the coke plant, Coke-oven workers excreted significantly more mutagenic substances and 1-hydroxypyrene (HpU) in urine than the controls. Mutagenic substances in urine of coke-oven workers and the control group were detected only with TA98 after metabolic activation. Median HpU levels of coke-oven workers classified into three job categories (A, B, C) exceeded the biological exposure limit. The effect of smoking on urinary mutagenicity and urinary 1-hydroxypyrene was statistically significant.

Can we detect mutagenic activity of urinary sediment by the Ames test?

Using the Salmonella typhimurium strain TA98 we tested the mutagenicity of filtrate and sediment of urine collected from children and coke-oven workers living in the town of Dabrowa Górnicza. Mutagenic substances were detected in samples of urinary filtrate taken either from environmentally exposed children or from occupationally exposed coke-oven workers. The mutagenic effect was found only in acetone extracts of urinary filtrate in the presence of promutagenic activating fraction S9. Beta-glucuronidase/arylsulphatase treatment hydrolysed the conjugates contained urinary filtrate into compounds that were toxic towards tester strain. The mutagenic effect of urine should be tested only in urinary filtrate as we have never detected mutagenic substances in urinary sediment.

[Test for the effects of mutagenic toxic fungal metabolites originating from municipal landfill sites]. / Badanie efektu mutagennego metabolitów grzybów toksynotwórczych pochodzacych z wysypiska odpadów komunalnych.

The purpose of the study is to assess the mutagenic effect of mycotoxins produced by moulds growing on municipal landfill sites. Mutagenicity of toxic fungal metabolites was determined by the Salmonella plate incorporation assay with two strains of bacteria: TA98 and TA100, with and without metabolic activation. The results obtained indicate that there is a severe hazard caused by these mycotoxins detected main by TA98 with metabolic activation. The most mutagenic mixture of mycotoxins acting directly on both strains was produced by Aspergillus nidulans. The highest mutagenic effect detected by TA98 with metabolic activation was found in the mixture of mycotoxins produced by one of three isolated Aspergillus fumigatus varieties.

[Impact of different solvents on results of genotoxicity studies of airborne dust taken from work environments]. / Wplyw róznych rozpuszczalników na wyniki badan genotoksycznosci pylów wystepujacych w powietrzu na stanowiskach pracy.

Mutagenicity and toxicity of extracts of airborne particulate matters, sampled during electrolysis of aluminium, cooking of coal and low-temperature carbonization processes, were determined by the Salmonella plate incorporation assay with strains TA98 and TA100. Organic materials were extracted with benzene, cyclohexane or toluene from airborne particles by sonication. The results showed that: a) the extracts were mutagenic only to strain TA98 (without and after metabolic activation); b) there was a similar efficiency of applied solvents in extraction of mutagenic substances detectable with TA98, and c) the toluene extracts were most toxic towards test bacteria TA98.

[Myocardial infarction-related arrhythmia after rehabilitation treatment in a sanatorium]. / Zaburzenia rytmu serca u chorych po zawale serca poddanych rehabilitacji sanatoryjnej.

The study involved 100 patients (80 men and 20 women), aged between 27 and 69 years treated at the Department of Cardiology, Medical Academy in Lublin for the first myocardial infarction, and classified for a 4-week rehabilitation in sanatorium. A 24-hours ECG recording with Holter technique (Medilog 3000 manufactured by Oxford Medilog Timed) was performed twice. In the first record cardiac arrhythmias ( extrasystole both supra- and ventricular, ventricular tachycardia) were more frequent in comparison with the results of the second recording carried out after a 4-week rehabilitation.