Tobacco-smoke exposure indicators and urinary mutagenicity

In this study, the correlation of indicators of external (i.e. mean daily intake of condensate, nicotine, tobacco and tobacco proteins, and daily number of cigarettes smoked) and of internal tobacco-smoke exposure (i.e. urinary 1-pyrenol, nicotine and its metabolites and trans,trans-muconic acid) with urinary mutagenicity, detected on YG1024 Salmonella typhimurium strain with S9, were examined in 118 smokers.An increase in urinary mutagenicity was clearly significantly correlated with each external and internal indicators of exposure to tobacco smoke (correlation coefficient (r) ranging between 0.22 and 0.54, P<0.01), with a greater extent in the case of indicators of internal dose. In multiple regression analysis, among the indicators of external exposure, daily tobacco intake was the only variable significantly associated with urinary mutagenicity (t=2.47, P=0.015, with partial contribution to r²=5.15%). Instead, when all indicators of exposure (external and internal) were considered in the analysis, the influence of urinary 1-pyrenol on urinary mutagenicity was predominant, followed by those of urinary trans,trans-muconic acid and nicotine plus metabolites (t=4.63, 2.73 and 2.08, P<0.001, P=0.002 and 0.04, with partial contribution to r²=17.0, 6.66 and 3.96%, respectively), with no influence at all of external tobacco-smoke exposure indicators.In conclusion, our results show that indicators of internal dose are better correlated with formation of mutagens in urine of smokers. Among these, the best indicator was urinary 1-pyrenol and this result designates the combustion processes of tobacco as the determining step for the formation of urinary mutagens. However, as these biomarkers cannot be analysed the amount of daily tobacco intake represent the best valuable index of external (presumptive) exposure to tobacco-smoke genotoxins.     

Hemoglobin adducts and urinary mercapturic acids in rats as biological indicators of butadiene exposure

Binding of 1,2-epoxy-3-butene, the primary metabolite of butadiene, to hemoglobin (Hb) and excretion of its mercapturic acid in urine were studies as potential indicators of butadiene exposure. Four groups of Wistar rats were exposed to butadiene at 0, 250, 500 and 1000 ppm 6 h/day, 5 days/week, during 2 weeks. Blood was collected at the end of exposure and 17 days later for analysis of hemoglobin adducts and adduct stability. Urine was collected each day during exposure (afternoon samples) and in between exposures (morning samples). Adducts of 1,2-epoxy-3-butene to N-terminal valine in Hb were measured using the N-alkyl Edman procedure and GC/MS of the thiohydantoin derivatives. The corresponding mercapturic acid was analysed, after deacetylation, through derivatization with phthaldialdehyde and HPLC with fluorescence detection. The Hb adducts proved to be stable and are therefore useful for dosimetry of long-term exposure to butadiene. The adduct levels increased linearly with exposure dose up to 1000 ppm (3 nmol/g Hb at 1000 ppm). The increase with exposure dose of the mercapturic acid concentration in urine was also compatible with a linear does response up to 1000 ppm. The sensitivity of both analytical methods needs to be improved for their application to human samples.     

Determination of urinary hippuric acid and o-cresol levels as biological indicators of toluene exposure in shoe-workers and glue sniffers.

In this study, groups exposed to toluene either intentionally (glue sniffers) or unintentionally (shoe-workers) were compared. The groups were evaluated in terms of urinary levels of the toluene metabolites hippuric acid and o-cresol. Results were also compared with control values. Hippuric acid levels were determined by high performance liquid chromatography and o-cresol levels by gas liquid chromatography. The levels of hippuric acid and o-cresol were found to be statistically significantly higher in glue sniffers than in shoe-workers (p <0.001) or controls (p <0.001). In addition, the differences between the levels of urinary hippuric acid and o-cresol in the shoe-workers and in the controls were statistically significant (p <0.05 and p <0.001, respectively). These results suggest that extremely high levels of urinary hippuric acid and o-cresol indicate massive exposure to toluene.     

Urinary mutagenicity in nonsmokers following exposure to fresh diluted sidestream cigarette smoke

Ten healthy male and 10 healthy female 'never-smoking' subjects (ages 21-50) participated in a 5-day environmental room study to determine if an acute exposure to a high level of fresh diluted sidestream smoke (FDSS) would alter urinary mutagenicity. On Monday, Tuesday, Thursday and Friday, the 20 subjects sat in environmental rooms for 7.33h and were exposed to filtered and humidified air. On Wednesday, the 20 subjects were exposed in the environmental rooms for 7.33h to an average respirable suspended particle (RSP) concentration of 179 microg/m(3) of FDSS generated by machine smoking 1R4F Kentucky reference cigarettes. This level of FDSS is approximately three times the ETS level seen in the top 5% of US workplaces which allow smoking. A cumulative 7.33h air sample from each environmental room was collected and determined to be mutagenic by Ames Salmonella assay. Subjects' urinary mutagenicity was measured on Wednesday as compared with Tuesday or Thursday by assaying concentrates of 24h urine samples in Ames Salmonella bacterial strains TA98 and YG1024. Diet was strictly controlled on all study days, with broiled and pan-fried meat not served to minimize ingestion of mutagenic protein pyrolysis products. Although all the urinary mutagenicity values were within the range reported for minor changes in diet, the subjects experienced a small but statistically significant increase (p<0.05) in urinary mutagenicity in strain YG1024, but not in the less sensitive strain TA98 on the day of FDSS exposure.     

Human urinary and blood toxicokinetics of beryllium after accidental exposure

PurposeBeryllium is known to have adverse health effects and is classified as carcinogenic to humans. However, data on systemic beryllium exposure in humans are rare and especially human toxicokinetics are largely uncharted. As such, the first reported multi-annual course of blood and urine concentrations after a high exposure scenario provides important new insights.MethodsFor a medical follow-up biomonitoring samples were collected for 56 months from a male subject after an accidental and multi-faceted high exposure. Sampling started on day 2 post-exposure for urine and day 147 for blood. The samples were analyzed by inductively coupled mass spectrometry (ICP-MS) and plotted longitudinally as a function of time. Terminal half-lives were calculated assuming a first-order elimination process.Main findingsBoth matrices showed highly increased initial concentrations (about 100-fold), despite the 147-day delay in blood sampling, and a marked decline over time. In urine, a two-phase excretion process was suspected based on the longitudinal data. Calculations gave terminal half-lives of 117.5 days and 666.5 days for phases 1 and 2, respectively. Blood kinetics called for a terminal half-life of 103.5 days. Elimination kinetics in blood and urine were comparable, simultaneously gathered samples showed an excellent correlation (R² = 0.985).Principal conclusionsThe long-term follow-up after a high initial exposure to beryllium provides the first detailed insights into the elimination course of systemically available beryllium in humans. Conform kinetics of beryllium in urine and blood and the strong correlation between both parameters indicate high data validity and support the good representation of the current systemically available beryllium by urine and blood concentration in humans. The relatively long terminal half-lives in both matrices suggest a possible accumulation in humans in case of repeated exposures.     

Biochemical stress indicators of greater wax moth exposure to organophosphorus insecticides

Although acetylcholinesterase (AChE) is the primary target of organophosphorus insecticides (OPs), increasing evidence regarding their secondary effects suggests that OPs disturb homeostasis of insects by generating free radical intermediates that trigger lipid peroxidation. We therefore investigated alterations in lipid peroxidation product, malondialdehyde (MDA) content, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, in conjunction with AChE activity as biochemical stress indicators in greater wax moth, Galleria mellonella (L.) larvae for OPs methyl parathion (MP) and ethyl parathion (EP). The effects of MP and EP were first investigated by rearing the young larvae on an artificial diet containing 0.01, 0.1, 1, 10, and 100 ppm of each insecticide. Second, the mature larvae were injected with 0.05, 0.5, 5, 50, and 500 ng of insecticides for determining the changes in biochemical stress responses. The diet with lowest level of MP significantly decreased the activities of all measured enzymes, whereas it increased MDA content. However ALT and AST were significantly higher in the larvae reared with the diet with high levels of MP than in control larvae. All tested levels of MP resulted in a decrease in AChE activity. The lowest level of EP in diet (0.01 ppm) significantly increased ALT activity, whereas it reduced that of AChE. This insecticide at 0.1 ppm resulted in reduced AST activity, but 1 ppm in diet elevated AST activity and MDA content. EP at 0.1 ppm and higher levels in the diet reduced ALT activity. All dietary EP levels significantly decreased AChE activity. ALT, AST, and AChE were lower in larvae fed with the diet containing 100 ppm ethyl parathion compared with larvae on control diet. MP at 50 ng per larva increased ALT and AST activities from 35.42 +/- 0.74 and 26.34 +/- 0.83 to 203.57 +/- 1.09, and 122.90 +/- 1.21 U/g, respectively, when the mature larvae were injected. All injected doses of EP dramatically reduced both ALT and AST activities, but only the lowest and highest levels of this insecticide decreased AChE activity. The lowest level of this insecticide also significantly increased MDA content in larvae. High levels of both insecticides increased MDA content. We observed a significant higher increase in MDA content in the larvae reared with 10 ppm EP (102.16 +/- 1.57 nmol/g protein) than the control group (30.28 +/- 1.42 nmol/g protein). These results suggest that OPs caused the metabolic and synaptic dysfunctions in greater wax moth and alter its biochemical physiology in response to oxidative stress.     

Fluroxene mutagenicity

The commercially available volatile anesthetic fluroxene (2,2,2-trifluoroethyl vinyl ether) which contains the stabilizer N-phenyl-1-napthylamine, was tested for mutagenicity using four strains of S. typhimurium, TA1535, TA1537, TA98 and TA100, and one strain of E. coli, WP2. In addition, purified fluroxene; N-phenyl-1-napthylamine; trifluoroethanol, a major metabolite of fluoroxene; and urine from rats anesthetized with fluroxene were tested. Several procedures were utilized including exposure of bacteria to vapor in desiccators and in liquid suspension. Results indicate that fluroxene, but not its stabilizer, was mutagenic to strains TA1535, TA100 and WP2 only in liquid suspension and only in the presence of a rat-liver enzyme system. Trifluoroethanol and urine from fluroxene-treated rat were not mutagenic to any strain of bacteria. These findings indicate that fluroxene is a promutagen which requires preincubation before it is recognized. Further experiments were performed with enzymes prepared from mouse, hamster and human liver. Fluroxene was mutagenic only in the presence of enzymes prepared from Aroclor 1254 pretreated rodents. Since fluroxene was not mutagenic in the presence of enzymes prepared from three human livers, the significance of these findings to man are unclear.     

Occupational exposure to anaesthetic gases and urinary excretion of D-glucaric acid

In order to ascertain whether the urinary excretion of D-glucaric acid (DGA) might be a suitable biomarker of effect in monitoring workers exposed to anaesthetic gases, we measured DGA before and after an operating session (and, in some workers, before and after a 2-week vacation) in 229 workers of surgical units and in 229 controls. In the former, we also measured urinary levels of nitrous oxide (N₂O) and isofiurane after at least 4 h of exposure. For all subjects, information on age, smoking habits, daily intake of alcohol, coffee, and drugs, history of liver or kidney disease was collected. Study subjects were ranked according to: exposure (class 0: subjects not exposed; class 1: N₂     

In vivo mutagenicity of the carcinogenic 1-nitropyrene. A model of a potential asbestos exposure

The environmental carcinogen 1-nitropyrene was orally and intraperitoneally administered to rats in a single dose of 30 mmol/kg. Mutagenicity of excreted urine was tested in Salmonella typhimurium TA 98 and 100 strains. The mutagenic pattern of urine in case of oral exposure proved to be completely different as compared to the intraperitoneal administration. Frame-shift mutagen(s) was/were detected only after enzymatic deconjugation of sulphate or glucuronide metabolites within the first 24 h. Base-pair substitution-type mutagenicity was only detected in the urine samples collected after intraperitoneal treatment. Since environmental asbestos exposure involves carcinogenic effects of adsorbed polycyclic aromatic hydrocarbons, this animal model provides a useful tool for testing fiber-associated nitroarenes, in both mechanistic and risk assessment studies.     

Decrease in environmental pollution and changes in biological indicators in occupational exposure to lead

The Authors examine the variations of the levels of PbB and EPP in 39 workers at known lead exposure and evaluate the capacity of these parameters to follow the measured decreases of the environmental pollution. They conclude that the variations of the mean PbB values are well related to environmental pollution and that the diagram on the probability paper of median values and their corresponding standard deviations allows to calculate the number of exposed workers with biological values above a fixed limit.     

Fluroxene mutagenicity.

The commercially available volatile anesthetic fluroxene (2,2,2-trifluoroethyl vinyl ether) which contains the stabilizer N-phenyl-1-napthylamine, was tested for mutagenicity using four strains of S. typhimurium, TA1535, TA1537, TA98 and TA100, and one strain of E. coli, WP2. In addition, purified fluroxene; N-phenyl-1-napthylamine; trifluoroethanol, a major metabolite of fluoroxene; and urine from rats anesthetized with fluroxene were tested. Several procedures were utilized including exposure of bacteria to vapor in desiccators and in liquid suspension. Results indicate that fluroxene, but not its stabilizer, was mutagenic to strains TA1535, TA100 and WP2 only in liquid suspension and only in the presence of a rat-liver enzyme system. Trifluoroethanol and urine from fluroxene-treated rat were not mutagenic to any strain of bacteria. These findings indicate that fluroxene is a promutagen which requires preincubation before it is recognized. Further experiments were performed with enzymes prepared from mouse, hamster and human liver. Fluroxene was mutagenic only in the presence of enzymes prepared from Aroclor 1254 pretreated rodents. Since fluroxene was not mutagenic in the presence of enzymes prepared from three human livers, the significance of these findings to man are unclear.     

Antimutagenic effects of diethyldithiocarbamate towards maleic hydrazide- and N-nitrosodiethylamine-induced mutagenicity in theTradescantia mutagenicity assay

InTradescantia, clone 4430, diethyldithiocarbamate (DEDTC) markedly decreased the frequency of somatic mutations induced by maleic hydrazide (MH) and N-nitrosodiethylamine (NDEA). In contrast, DEDTC had no such effect on N-methyl-N-nitrosourea-induced mutagenesis. The putative degradation and conversion products of MH (maleic acid diamide, succinic acid, maleic acid, lactic acid and hydrazine) exhibited no mutagenic activity in theTradescantia mutagenicity assay.     

60 Hz magnetic field exposure and urinary 6-sulphatoxymelatonin levels in the rat

Four separate experiments were carried out to investigate the effect of extremely low frequency magnetic field (MF) exposure (60 Hz, 1 mT rms) on urinary 6-sulphatoxymelatonin (aMT6s) levels in Sprague-Dawley rats. In the first experiment, immature male rats maintained under a regular 12 h daily photoperiod (white fluorescent light) were exposed to a 20 h daily MF exposure for 6 weeks. The second experiment was similar to the first, except that the MF exposure was limited to 10 days. In the third experiment, adult male rats acclimated to a combination of continuous dim red light and regular 12 h daily photoperiod (white fluorescent) were subjected to a single 1 h exposure to intermittent MF (1 min on and 1 min off cycles), 2 h before fluorescent lights went off. The fourth experiment was similar to the third, except that the animals received 2 consecutive days of 20 h daily exposure to intermittent MF, beginning 1 h before the fluorescent lights went off each day. In all four experiments, the circadian profile of urinary aMT6s was examined before, during, and after the MF exposure. No significant effect of 1 mT MF on indoleamine metabolism was observed in any of the above experiments. However, in one of the experiments (no. 4), both the control and the MF groups showed a lower aMT6s level during the exposure days, when compared with that of pre- and post-exposure days, suggesting that the existence of possible effects with lower field strengths at the range of stray field cannot be ruled out. Bioelectromagnetics 19:172–180, 1998. © 1998 Wiley-Liss, Inc.     

Application of urinary mutagen testing to detect workplace hazardous exposure and bladder cancer

The objectives of this biochemical epidemiologic case-control study were to evaluate urinary mutagen testing for occupational exposure assessment, and for possible screening for bladder cancer in the workplace. Thirty-seven patients (19 bladder cancer cases and 18 controls) completed a questionnaire. Two urine samples, i.e. a work sample taken while at work, and a home sample, were requested from each patient. Twenty-six patients (17 cases and 9 controls) gave a total of 47 24-h urine samples for mutagenicity testing by the Ames test. A positive Ames test was found to be associated significantly with current occupation with hazardous exposure (odds ratio = 3.7, 95%CI 1.1–12.9), and non-significantly with bladder cancer (odds ratio = 1.8, 95%CI 0.5–7.1). Our results show that the urinary Ames test has the potential of being used as a surveillance for current workplace hazardous exposure (sensitivity = 52%, specificity = 77%, positive predictive value = 72%, negative predictive value = 59%, positive likelihood ratio = 2.3), but not as a screening test for bladder cancer cases (sensitivity = 42%, specificity = 71%, positive predictive value = 3%, negative predictive value = 98%, positive likelihood ratio = 1.5).