Chronic ethanol consumption causes structural and functional reorganization in the hippocampus and induces alterations in the gene expression of gamma-aminobutyric acid type A receptors (GABAARs). Distinct forced intermittent exposure models have been used previously to investigate changes in GABAAR expression, with contrasting results. Here, we used repeated cycles of a Chronic Intermittent Ethanol paradigm to examine the relationship between voluntary, dependence-associated ethanol consumption, and GABAAR gene expression in mouse hippocampus. Adult male C57BL/6J mice were exposed to four 16-h ethanol vapor (or air) cycles in inhalation chambers alternated with limited-access two-bottle choice between ethanol (15%) and water consumption. The mice exposed to ethanol vapor showed significant increases in ethanol consumption compared to their air-matched controls. GABAAR alpha4 and delta subunit gene expression were measured by qRT-PCR at different stages. There were significant changes in GABAAR delta subunit transcript levels at different time points in ethanol-vapor exposed mice, while the alpha4 subunit levels remained unchanged. Correlated concurrent blood ethanol concentrations suggested that GABAAR delta subunit mRNA levels fluctuate depending on ethanol intoxication, dependence, and withdrawal state. Using a vapor-based Chronic Intermittent Ethanol procedure with combined two-bottle choice consumption, we corroborated previous evidences showing that discontinuous ethanol exposure affects GABAAR delta subunit expression but we did not observe changes in alpha4 subunit. These findings indicate that hippocampal GABAAR delta subunit expression changes transiently over the course of a Chronic Intermittent Ethanol paradigm associated with voluntary intake, in response to ethanol-mediated disturbance of GABAergic neurotransmission.

Quantification of platinum in human urine is a valuable technique for assessing occupational and environmental exposure to platinum. We hereby present a simple and sensitive method to monitor urinary platinum in hospital workers exposed to platinum-based anticancer agents. This method consists in: i) the sample digestion with an ultrapure HNO3/HCl (5:1) mixture in a sonicator for 2h, and ii) the Inductively Coupled Plasma Mass Spectrometry (ICP–MS) platinum determination on the 1:10 diluted solutions. Sample pre-treatment is simpler and faster when compared to other published procedures, which are typically multistep and time-consuming and often require expensive equipments. The ICP–MS analysis of all pre-treated samples shows good selectivity, sensitivity, repeatability and recoveries. It is noteworthy that in the present case both standard addition mode and the direct platinum determination with reference to an external calibration curve give satisfactory results for urinary platinum, while the standard addition mode is typically required in the literature methods.

Analytical difficulties and lack of a biological exposure index and reference values have prevented using unmetabolized urinary benzene (UB) excretion as a biomarker of low-level environmental exposure. To explore what environmental factors beyond active smoking may contribute to environmental exposure to benzene, we monitored UB excretion in a non-smoking, non-occupationally exposed sample of the general population.

In several recent epidemiological studies blood lead levels (BLLs) even below the current CDC intervention level of 10 microg/dl have been associated with reduced neurocognitive capacities of children, with no clear evidence of a "safe" threshold. We analyzed the relationship between the BLLs and the neurocognitive capacities of 205 Sardinian students aged 11 to 15 years, using 2 tests of the Swedish Performance Evaluating System (SPES) and the full-scale Intelligence Quotient (IQ) derived from the Wechsler Intelligence Scale for Children (WISC). The studied population included 104 children (61 males and 43 females) living in Portoscuso, a town 2 Km far from a lead smelter (mean BLLs: 5.98 +/- 2.2; max 11.5 microg/dl), and 101 age-matched students (55 males and 46 females) living in Sant'Antioco, a town about 20 Km far from the same smelter (mean BLLs: 2.08 +/- 0.8; max 4.5 microg/dl). Subjects with BLLs above 4 microg/dl performed worse in the SPES tests and scored about 5.0 points less on the full-scale IQ compared to the students with lower BLLs. The adjusted regression coefficients derived from the multivariate analysis showed that higher BLLs were significantly associated with worse performances in the SPES tests and with reduced IQ (0.94 points for each microg/dl of BLL). This study confirms the potential neurotoxicity of low-levels of lead suggesting the need of lowering the actual CDC "limit of concern" for children to values lower than 4 microg/dl, improving at the same time the environmental primary prevention for limiting the lead exposure of subjects living near the lead smelter.

Introduction: Conflicting opinions exist about the reliability of biomarkers of low-level exposure to benzene. We compared the ability of the urinary excretion of trans, trans-muconic acid (t,t-MA), s-phenilmercapturic acid (s-PMA) and urinary benzene (U-Benz) to detect low level occupational and environmental exposure to benzene. Methods: We monitored airborne benzene by personal air sampling, and U-Benz, s-PMA, t, t-MA and cotinine (U-Cotinine) in spot urine samples, collected at 8 am and 8 pm, in 32 oil refinery workers and 65 subjects, randomly selected among the general population of urban and suburban Cagliari, Italy. Information on personal characteristics, diet and events during the sampling day was acquired through in person interviews. Results: The median concentration of airborne benzene was 25.2 mu g/m(3) in oil refinery workers, and 8.5 mu g/m(3) in the general population subgroup. U-Benz in morning and evening samples was significantly more elevated among oil refinery workers than the general population subgroup (p=0.012, and p=7.4x10(-7), respectively) and among current smokers compared to non-smokers (p=5.2x10(-8), and p=5.2x10(-5) respectively). Benzene biomarkers and their readings in the two sampling phases were well correlated to each other. The Spearman's correlation coefficient with airborne benzene was significant for U-Benz in the evening sample, while no correlation was seen with t, t-MA and s-PMA readings in either samplings. The two benzene metabolites were frequently below limit of detection (LOD), particularly among the general population study subjects (17-9% and 39%, for t, t-MA and s-PMA respectively). Morning U-Cotinine excretion showed a good correlation with U-Benz in the morning and in the evening sampling (p<0.001), and with s-PMA in the evening sample (p<0.001), but not with t, t-MA in either samplings. t, t-MA in the evening sample was the only biomarker showing a moderate inverse correlation with BMI (p<0.05). The multiple regression analysis adjusting by BMI and number of cigarettes smoked during the day confirmed the results of the univariate analysis. Discussion: Our results suggest that unmetabolized U-Benz would allow a more reliable biomonitoring of low-level exposure to benzene than s-PMA and t,t-MA.

Background: We monitored urinary benzene excretion to examine factors affecting benzene uptake in a sample of the general population living near a petrochemical plant.Methods: Our study population included 143 subjects: 33 petrochemical plant workers (W) with low level occupational benzene exposure; 30 residents in a small town 2 km from the plant (2kmR); 26 residents in a second small town located 2 to 4 km from the plant (4kmR); and 54 urban residents 25 km from the plant (25kmR). Exposure to benzene was evaluated by personal air sampling during one work-shift for the W group, and from 8.00 to 20:00 for general population subgroups, and by urinary benzene (BEN-U).Results: Median airborne benzene exposure was 25, 9, 7 and 6 mu g/m(3) benzene among the W, 2kmR, 4kmR, and 25kmR subgroups, respectively; the highest level was found among the workers, while there was no significant difference among the other groups. Median BEN-U was 2 to 14-fold higher in smokers compared to non-smokers; among non-smokers BEN-U was the highest in W (median 236 ng/L), and lower in the 2kmR (48 ng/L) and 4kmR (63 ng/L) subgroups than in the 25kmR (120 ng/L) subgroup. A multiple linear regression analysis, explaining up to 73% of BEN-U variability, confirmed that active smoking and airborne benzene most strongly affected BEN-U. Among the non-smoking, non-occupationally exposed study subjects, a positive association was found between BEN-U and the distance of residence from the plant. This association was explained by increased exposure to urban traffic emissions in the study group residing at a greater distance from the plant. Environmental tobacco smoke had a marginally positive role.Conclusion: Among factors affecting benzene uptake in non-occupationally exposed individuals, urban residence contributes to benzene exposure more than residing in close proximity to a petrochemical plant. (C) 2011 Elsevier Ltd. All rights reserved.

For the first time we report a rapid and sensitive LC–MS–MS method for quantification of rotenone, deguelin, and rotenolone in human serum. The analytical procedure involves extraction with ethyl acetate without further clean-up. The active ingredients were separated on a C8 reversed-phase column by isocratic elution. Eleven simultaneous transitions of precursor ions were monitored. Excellent selectivity and sensitivity enables quantification and identification of low levels of rotenoids (LOD 2 ng mL−1, LOQ 5 ng mL−1) in human serum.

BACKGROUND:There is conflicting epidemiological evidence concerning an increase in risk of non-Hodgkin's lymphoma (NHL) associated with elevated blood levels of persistent organochlorine (OC) pesticides and polychlorobiphenyls (PCBs). METHODS:We measured the concentration of 17 OC pesticides, including hexachlorobenzene (HCB), four lindane isomers (alpha-, beta-, gamma- and delta-hexachlorocyclohexane (HCH)), two chlordane species (heptachlor and oxy-chlordane), four cyclodiene insecticides (aldrin, dieldrin, endrin and mirex), six dichloro-diphenyl-trichloroethane (DDT) isomers and nine PCB congeners (PCBs 28, 52, 101, 118, 138, 153, 170, 180 and 194) in plasma samples of 377 subjects, including 174 NHL cases and 203 controls from France, Germany and Spain. The risk of NHL and its major subtypes associated with increasing blood levels of OC pesticides and PCBs was calculated using unconditional logistic regression. RESULTS:Risk of NHL, diffuse large B cell lymphoma (DLBCL) and chronic lymphatic leukaemia (CLL) did not increase with plasma levels of HCB, beta-HCH, p,p'-dichloro-diphenyl-dichloroethylene (DDE), or total and individual PCBs or their functional groups, in the overall study population. Substantial heterogeneity in DLBCL risk associated with immunotoxic PCBs (p = 0.03) existed between the Spanish subgroup (odds ratio (OR) for immunotoxic PCB plasma level above the median vs below the median was 0.7, 95% CI 0.3 to 1.6) and the French and German subgroups combined (OR 3.2, 95% CI 0.9 to 11.5). CONCLUSION:We did not find evidence of an association between NHL risk and plasma level of OC pesticides and PCBs.

In several recent studies blood lead levels below 100 microg/L have been associated with reduced neurocognitive capacities and neurobehavioural performances in children, with no clear evidence of the lowest "safe" level not associated to adverse effects on the CNS. We analyzed blood lead concentration and the results of 5 neurocognitive tests, two derived from the Swedish Performance Evaluating System (SPES) and three from the Wechsler Intelligence Scale for Children-Revised (WISC-R), in 139 Sardinian adolescents living in Portoscuso, a town 2 Km far from a lead smelter, and in 72 age-matched students living in S. Antioco, a town about 15 Km far from the same smelter. The blood lead concentrations were lower than 100 microg/l in almost subjects, but, in average, significantly higher particularly among males, in the Portoscuso group compared to controls. The regression coefficients derived from the multivariate stepwise analysis, adjusted for the student's age and gender and for the educational and socio-economic level of parents, showed that reduced performances in neurocognitive test were significantly associated to increasing blood lead concentrations. According to previous our surveys in the same area and to the recent scientific literature on this topic, the present study suggests the need to further low the blood lead levels, considered by the U.S. CDC in 1991 as potentially "safe" for the children's CNS, to a threshold possibly < 50 microg/L.

PURPOSE: Potential sources of exposure to polycyclic aromatic hydrocarbons (PAHs) and genetic polymorphisms were investigated in relation to their contribution to interindividual variation in baseline levels of urinary 1-hydroxypyrene (1-OHP) excretion in subjects without occupational exposure to PAHs.METHODS: Urinary excretion of 1-OHP was measured in 114 subjects, including 48 women and 66 men. Questionnaire information was collected on possible environmental and individual sources of PAH exposure. A subset of 70 individuals also was evaluated for a single-nucleotide polymorphism (Ex7 + 295C -> T) in the cytochrome P-450 1A2 (CYP1A2) gene, and 61 of these also were evaluated for the glutathione transferase T1 (GSTT1) gene polymorphism.RESULTS: 1-OHP values did not show a significant seasonal variability and were unaffected by age; education; body mass index; smoking status, including passive smoking; or the C -> T base substitution in position 295 of exon 7 of the CYP1A2 gene. After reciprocal adjustment with logistic regression, living in a heavily trafficked urban area (odds ratio, 4.9; 95% confidence interval, 1.0-24.9), and frequent intake of grilled meat (odds ratio, 6.9; 95% confidence interval, 1.1-43.5) were significant predictors of background urinary 1-OHP levels of 0.50 mu g/g creatinine or greater. Elevated risks also were associated with daily alcohol intake greater than 65 g and the nonnull GSTT1 genotype.CONCLUSION: Our study shows that exposure to urban traffic, dietary habits, and the nonnull GSTT1 genotype may contribute to interindividual variation in background levels of 1-OHP urinary excretion in subjects without occupational exposure to PAHs.

The effects of low-level lead exposure on children's intelligence quotient (IQ) were investigated in 64 Sardinian adolescents (13-16 years old). To estimate potential early adverse effects on the Central Nervous System (CNS) due to very low-level lead, the Intelligence Quotient (IQ) of adolescents with present blood lead concentrations (PbB) generally below 10 microg/dl was measured. We analyzed blood lead concentration and individual IQ of 32 Sardinian children living in Portoscuso, a town 2 Km far from a lead smelter, and of other 32 controls living in S. Antioco, a town about 15 Km far from the same smelter. The Wechsler Intelligence Scale for Children Revised (WISC-R) was administered. The relation between IQ and blood lead concentration was estimated by linear multivariate models adjusting for several potential confounders, such as the educational and socio-economic level of the parents. The blood lead concentration was in average significantly higher in the Portoscuso group compared to controls. The linear model applied to the total population studied (n 64) showed that the blood lead concentration was inversely and significantly associated with IQ, with an extrapolated decline of 1.29 points in total IQ for each microg/dl increase of lead blood concentration. According to the recent scientific literature on this topic, results of our pilot study suggest the need to further lower the blood lead concentration for children to a threshold significantly below 10 microg/dl, value till now considered "safe" for the children's CNS.

Several recent meta-analyses, although some inconsistencies between studies, provide evidence for subtle deficits in neurobehavioural tests being associated with average blood levels (PbB) between 350 and 600 microg/l and for significant correlations between impairments in some test results and increasing indices present andlor cumulative exposure to inorganic lead. In this study we assessed the relationship between blood lead level (PbB) and results of some neurobehavioural tests derived from the SPES battery and from the WAIS-R among a sample of 94 lead smelter workers (median PbB: 302 microg/l; range: 60-690 microg/l). The stepwise multiple regression analysis shows that, even after adjusting for age, education level, score in the vocabulary test and for alcohol and cigarette consumption, the rise of PbB above 300 microg/l was significantly associated with the rise in the number of errors and in the response time of the Symbol Digit test. No significant relationship between PbB and results of the other tests was observed after adjusting for the main confounding factors. Despite the small size of this study and the few number of neurobehavioural tests applied, our results are suggestive for suble potential neurotoxic effects of inorganic lead even at the present exposure levels found in the studied sample of smelter workers and support the ACGIH BEI and the SCOEL recommendation for a biological limit of 300 microg/l of PbB.

Objectives: To explore reproductive outcomes in relation to occupational exposure to DDT.Methods: We inquired into the reproductive history, including total number of children, sex distribution in the offspring, time-to-pregnancy, and number of spontaneous abortions and stillbirths, of the spouses of 105 men first exposed to DDT in a 1946-1950 anti-malarial campaign in Sardinia, Italy. The time-to-pregnancy in months at the first successful conception was estimated from population Registrars. Cumulative DDT exposure during the anti-malarial campaign was retrospectively estimated.Results: The stillbirth rate was elevated and the male/female ratio in the offspring was reversed among DDT-exposed workers, and particularly among DDT applicators, compared to the unexposed subjects. Among DDT applicators, the stillbirth rate increased and the male/female ratio decreased by the tertile of cumulative DDT exposure. The fecundity ratio among spouses of DDT applicators was 0.72 (95% CI, 0.41,1.21) compared to the unexposed. The average number of children and abortion rate were unaffected by DDT exposure.Conclusions: The low statistical power of our study does not allow definitive conclusions. However, the results prompt further in-depth research into adverse reproductive outcomes and reduced fertility among men heavily exposed to DDT. (C) 2004 Elsevier Inc. All rights reserved.