Dichlorodiphenyltrichloroethane (DDT), DDT metabolites and pregnancy outcomes.

Organochlorine pesticides (OCPs) are persistent endocrine disruptors. OCPs cross the placenta; this prenatal exposure has been associated with adverse pregnancy outcomes. We investigated associations between prenatal exposure to OCPs and gestational age and birth weight in 600 infants born between 1960 and 1963. The primary OCP was 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT), its primary metabolite, 1,1'-dichloro-2,2'-bis(p-chlorophenyl)ethylene (p,p'-DDE) and the contaminant, 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl)-ethane (o,p'-DDT). Regression analysis indicated that for each natural log unit increase in p,p'-DDT, birth weight increased by 274 g (95% CI: 122, 425) when controlling for p,p'-DDE and o,p'-DDT. At a given level of p,p'-DDT exposure, o,p'-DDT and p,p'-DDE were associated with decreased birth weight. p,p'-DDE was negatively associated with length of gestation, controlling for p,p'-DDT and o,p'-DDT. These findings suggest opposing associations between exposure to p,p'-DDT and p,p'-DDE and birth weight. We did not find evidence to support mediation by maternal thyroid hormone status nor that the association differed by sex.

Prenatal polychlorinated biphenyl exposure is associated with decreased gestational length but not birth weight: archived samples from the Child Health and Development Studies pregnancy cohort.

BACKGROUND: Polychlorinated biphenyls (PCBs), known endocrine disruptors, were banned in 1979 but persist in the environment. Previous studies are inconsistent regarding prenatal exposure to PCBs and pregnancy outcomes. We investigated associations between prenatal exposure to PCBs and gestational length and birth weight. METHODS: In a sample of 600 infants (born between 1960 and 1963) randomly selected from Child Health and Development Studies participants followed through adolescence we measured 11 PCB congeners in maternal post partum sera (within three days of delivery). Length of gestation was computed from the reported first day of the last menstrual period (LMP) and delivery date. Linear regression was used to estimate associations between PCB exposure and gestational age and birth weight, adjusting for potential confounders. PCBs were grouped according to hypothesized biological action (1b (sum of weak phenobarbital inducers), 2b (sum of limited dioxin activity), and 3 (sum of CYP1A and CYP2b inducers)) or degree of ortho- substitution (mono, di, tri). Secondary analyses examined associations between total PCB exposure and exposure to individual congeners. RESULTS: Each unit increase in mono-ortho substituted PCBs was associated with a 0.30 week decrease (95% confidence interval (CI) -0.59, -0.016), corresponding to a 2.1 (95% CI -4.13, -0.11) day decrease in length of gestation. Similar associations were estimated for di-ortho substituted PCBs, (1.4 day decrease; (95% CI -2.9, 0.1)) and group 3 PCBs (0.84 day decrease; (95% CI -1.8, 0.11). We found similar associations in congener specific analyses and for the sum of congeners. CONCLUSIONS: Our study provides new evidence that PCB exposure shortens length of gestation in humans. This may have public health implications for population exposures.

Distribution of polybrominated diphenyl ethers (PBDEs) and other persistent organic pollutants in human serum from Greece.

Human serum samples (n=61) were collected in Attika, Greece between June and October 2007 and analyzed for polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane (HBCD), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs). Thirty samples were collected from computer clerks of a large computer company in Athens working full-time with computers, and thirty-one from a control population in the Attika region with no computer use. Σ(5)PBDE concentrations (sum of tri- to hexa-BDEs) in all samples (n=61) ranged from 0.68 to 13.3 ng g(-1) lipid, with a median of 1.07 ng g(-1) lipid. These concentrations are on the lower end of those reported from other countries, probably reflecting lower usage of PBDE-containing products or lower exposures to these chemicals. Individual and Σ(5)PBDE concentrations did not statistically differ between the two groups of computer clerks and non-computer users (p>0.05), with the exception of BDE 153 (p=0.033). The predominant congener was BDE 153, followed by BDEs 47, 100, 99, 183, 154 and 28. HBCD was also detected in 70% of the samples. BDE 209 was detected in 8 out 61 samples (13%), with concentrations ranging from 1.18 to 19.1 ng g(-1) lipid, and a median of 2.94 ng g(-1) lipid. No age dependency was found for PBDEs. Σ(11)PCB (sum of PCBs 74, 99, 118, 138, 146, 153, 156, 170, 180, 183 and 187,) in all samples (n=61) ranged from 36 to 402 ng g(-1) lipid, with a median of 110 ng g(-1) lipid. pp'-DDE concentrations ranged from 53.8 to 1649 ng g(-1) lipid, with a median of 268 ng g(-1) lipid. This is the first study to report levels of PBDEs in a possibly occupationally exposed subset of the Greek population.

Short and medium chain length chlorinated paraffins in UK human milk fat.

Chlorinated paraffins (also called polychlorinated n-alkanes -- PCAs) are a class of industrial chemicals comprising chlorinated straight chain hydrocarbons. They have a wide range of applications and are now found in a range of environmental compartments. We analysed a total of 25 human milk-fat samples, donated by 18 individuals from the urban London and more rural Lancaster areas in the UK, for short chain PCAs (C(10)-C(13) sPCAs) and medium chain PCAs (C(14)-C(17) mPCAs), using gas chromatography-ECNI high-resolution mass spectrometry. Our study confirms that trace quantities of PCAs can reach human milk-fat. sPCAs were detected in all but four samples, while mPCAs were detected in all samples. The median sPCA concentration was 180 ng/g fat (range of 49 to 820 ng/g fat -- detected values only) and the median mPCA concentration was 21 ng/g fat (range of 6.2 to 320 ng/g fat). No differences were noted in ranges of observed values for either sPCAs or mPCAs between samples from London and Lancaster. Most samples also exhibited similar patterns of sPCAs and mPCAs. One sample exhibited a different pattern for sPCAs and mPCAs, an observation that may be related to differences in exposure or biological factors for this individual.

Inter-individual differences in the ability of human milk-fat extracts to enhance the genotoxic potential of the procarcinogen benzo[a]pyrene in MCF-7 breast cells.

Environmental factors are believed to play an important role in cancer aetiology. Whether environmental pollutants act in isolation or in combination within mixtures remains unclear. Four human milk-fat extracts (from resident U.K. women) were screened for levels of organochlorinated and brominated compounds prior to being tested (1-50 mg-equiv) for micronucleus (MN)-forming activity in MCF-7 cells. Using the cytokinesis-block micronucleus assay, micronuclei (MNi) were scored in 1000 binucleate cells per treatment. Cell viability (% plating efficiency) and immunohistochemical detection of p53 induction were also measured. The effects of treatment with 1 mg-equiv of extract in combination with benzo[a]pyrene (BP) were also examined. BP-DNA adducts were detected and quantified by 32P-postlabeling analysis. Dose-related increases in MNi independent of pollutant concentrations were induced by milk-fat extracts. All four extracts elevated the percentage of p53 positive cells, although not always in a dose-related fashion. Some combinations resulted in profound low-dose-induced increases in MNi and significant elevations in the percentage of p53 positive cells, which occurred without further reduction in cell viability or mitotic rate. When one particular extract was combined with BP, a 100-fold increase in BP-DNA adducts was detected as compared with the levels induced by BP alone; an effect not induced by other extracts. This adduct-enhancing extract was fractionated into 14 fractions that were subsequently tested (1 mg-equiv of original extract) in combination with 0.01 microM BP. Fraction 1, into which nonpolar pollutants mostly eluted, enhanced MN-forming activity with BP. Surprisingly, the more polar and less likely to contain fat-soluble pollutants fractions 5 and 8 also enhanced MN-forming activity. No identifiable pollutants were present in these fractions. The results suggest that different environmental pollutants present in human tissue may influence the susceptibility of target cells to initiating events.

Different levels of polybrominated diphenyl ethers (PBDEs) and chlorinated compounds in breast milk from two U.K. Regions.

Polybrominated diphenyl ether (PBDE) congeners are constituents of flame retardants, and there is growing concern regarding their persistence, bioaccumulation, and toxicity. We collected breast milk samples between late 2001 and early 2003 from 54 U.K.-resident mothers. Of these, 27 originated from southeast England (London), and the other 27 originated from northwest England (Lancaster). Analysis of milk-fat extracts by gas chromatography-mass spectrometry was performed to determine the levels of 15 PBDE congeners, 15 polychlorinated biphenyl (PCB) congeners, and other selected chlorinated compounds. PCB and organochlorine (OC) levels in southeast samples were consistently higher, and significant differences (p < 0.05) were observed. Sigma PBDE levels ranged from 0.3 to 69 ng/g lipid (geometric mean, 6.6 ng/g), and PBDE-47 was the most abundant congener. Sigma PCB levels ranged from 26 to 530 ng/g lipid (geometric mean, 150 ng/g) and were composed mainly of PCB-153 (26%), PCB-138 (20%), and PCB-180 (13%). OC levels for 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT) and its metabolites (Sigma DDX) ranged from 24 to 2,300 ng/g lipid (geometric mean, 160 ng/g);Sigma hexachlorocyclohexane levels ranged from 1.2 to 1,500 ng/g lipid (geometric mean, 16 ng/g). Using nuclear magnetic resonance-based metabonomics, samples (n = 7) containing the highest contaminant levels were compared with samples (n = 7) containing the lowest levels. Excellent separation along the first principal component implied that the chemical constituents of the two groups were significantly different. Although reasons for such differences remain obscure, lifestyle factors associated with a more heterogeneous London cohort could be responsible. Identifying primary routes of contaminant exposures and their biologic effects is of great importance. Key words: breast milk, flame retardants, gas chromatography-mass spectrometry, milk-fat extracts, organochlorines, PBDE-47, persistent contaminants, polybrominated diphenyl ethers, polychlorinated biphenyls, United Kingdom.

Low dose induction of micronuclei by lindane.

Environmental contaminants possessing hormonal activity have long been suspected of playing a role in cancer causation. What is unclear is whether such agents elicit their effects through genotoxic and/or epigenetic mechanisms. gamma-Hexachlorocyclohexane (gamma-HCH, lindane) was tested in the 10(-12)-10(-4) M range. Chromosomal damage in MCF-7 breast cells and PC-3 prostate cells was assessed using the cytokinesis block micronucleus assay. Micronuclei (MNi) were scored in 1000 binucleate cells per treatment. Cell viability and cell cycle kinetics were also assessed, along with immunocytochemical and quantitative gene expression analyses of CDKN1A (P21WAF1/CIP1), BCL-2 and BAX. Following 24 h treatment, lindane (10(-12)-10(-10) M) induced increases (up to 5-fold) in MNi in both cell lines. Increases in MNi occurred in the absence of DNA single-strand breaks or cytotoxicity and, compared with benzo[a]pyrene and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, at low concentrations. Lindane induced more MNi than the alpha or beta stereoisomers of HCH. Low dose lindane (10(-12)-10(-10) M) significantly elevated the percentage of MCF-7 cells staining positive for Bcl-2 and of PC-3 cells staining positive for Bax. Only high dose lindane (10(-4) M) disrupted cell cycle kinetics with increases in percentage of cells in G1 and decreases in percentage of cells in G2/M. Despite a comparable high dose lindane induction of cell cycle arrest, marked increases in expression of P21WAF1/CIP1 were observed only in MCF-7 cells, although in PC-3 cells a significant increase (P < 0.0005) in the percentage of cells staining positive for p21Waf1/Cip1 was seen. These results suggest that 'environmental' concentrations of lindane can induce a number of subtle alterations in breast and prostate cells in the absence of cytotoxicity.

Polybrominated diphenyl ethers (PBDEs) in grey seals during their first year of life--are they thyroid hormone endocrine disrupters?

Blubber biopsy and blood samples were collected from weaned grey seal pups and juveniles in 1998 and 1999. The relationship between the concentration of total PDBEs in the blubber and the circulating concentrations of serum proteins, minerals and thyroid hormones was investigated. Fifty-four post-weaned pups were sampled and 55 first year juveniles, of which 13 were recaptured post-weaned pups. The median concentration of total PBDEs in the pups' blubber was 170 and 460 ng g(-1) in the first-year animals. Total blubber PBDEs contributed significantly to the prediction of circulating thyroid hormone concentrations, after controlling for age and condition. Cholesterol and albumin concentrations were also positively related to blubber PBDEs. The results of this study suggest a link between thyroid hormones and exposure to PBDEs in grey seals during their first year of life. Such an association is not alone sufficient evidence for a cause and effect relationship, however it is in accordance with the hypothesis that these compounds are endocrine disrupters in grey seal pups.