Similarities in the Endocrine-Disrupting Potencies of Indoor Dust and Flame Retardants by Using Human Osteosarcoma (U2OS) Cell-Based Reporter Gene Assays

Indoor dust is a sink for many kinds of pollutants, including flame retardants (FRs), plasticizers, and their contaminants and degradation products. These pollutants can be migrated to indoor dust from household items such as televisions and computers. To reveal high-priority end points of and contaminant candidates in indoor dust, using CALUX reporter gene assays based on human osteosarcoma (U2OS) cell lines, we evaluated and characterized the endocrine-disrupting potencies of crude extracts of indoor dust collected from Japan (<i>n</i> = 8), the United States (<i>n</i> = 21), Vietnam (<i>n</i> = 10), the Philippines (<i>n</i> = 17), and Indonesia (<i>n</i> = 10) and for 23 selected FRs. The CALUX reporter gene assays used were specific for compounds interacting with the human androgen receptor (AR), estrogen receptor α (ERα), progesterone receptor (PR), glucocorticoid receptor (GR), and peroxisome proliferator-activated receptor γ2 (PPARγ2). Indoor dust extracts were agonistic to ERα, GR, and PPARγ2 and antagonistic against AR, PR, GR, and PPARγ2. In comparison, a majority of FRs was agonistic to ERα and PPARγ2 only, and some FRs demonstrated receptor-specific antagonism against all tested nuclear receptors. Hierarchical clustering clearly indicated that agonism of ERα and antagonism of AR and PR were common, frequently detected end points for indoor dust and tested FRs. Given our previous results regarding the concentrations of FRs in indoor dust and in light of our current results, candidate contributors to these effects include not only internationally controlled brominated FRs but also alternatives such as some phosphorus-containing FRs. In the context of indoor pollution, high-frequency effects of FRs such as agonism of ERα and antagonism of AR and PR are candidate high-priority end points for further investigation

Peregrine falcon egg pollutants

<p>The role of the peregrine falcon (<i>Falco peregrinus</i>) as a bioindicator for the accumulation of organohalogen compounds and other persistent organic pollutants has been established in field studies. Biometric indices for damage caused by pollutants such as the shell thickness and the shell index were determined and the egg contents were analyzed for various pollutants by gas chromatography/mass spectrometry. A wide range of chemically activated luciferase expression (CALUX®) bioassays were performed on subsamples of the eggs.</p> <p>The following organohalogen compounds were found in the eggs of 2009: dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethane, heptachlor epoxide, hexachlorobenzene, dieldrin, hexachlorocyclohexane, polychlorobiphenyls and polychlorodibenzodioxins and polychlorodibenzofurans, polybromodiphenyl ethers, hexabromocyclododecane, tetrabromobisphenyl A, perfluoro compounds, and mercury. The DDT metabolite, dichlorodiphenyldichloroethene (DDE) (11,800 ng/g dry matter), was found to be the most highly concentrated egg contaminant followed by 2,2′,4,4′,5,5′-hexachlorobiphenyl (3800 ng/g). After a past general drop in pollution level, most egg contaminants presently plateau at levels that may still exceed limit values in foods of animal origin (DDE) or even toxicological thresholds (polychlorobiphenyls + polychlorodibenzodioxins, polybromodiphenyl ethers, perfluorooctanesulfonate, methylmercury).</p> <p>Accumulation of DDE could be shown in peregrine falcon eggs from the uplands of Southwest Germany with elevations up to 1500 m, presumably due to its global distribution and its cold condensation in higher altitudes. In contrast, the concentration of polychlorobiphenyls in falcon eggs decreases with elevation, indicating that these pollutants originate mainly from conurbations and local industrial sites.</p> <p>Significant negative correlations were found between both shell index and thickness and the concentration of Hg. A deleterious effect is also evident from a no-observed-adverse-effect level of 120 ng MeHg per gram egg determined by other authors in chronic feeding studies with ibises, which resulted in decreased egg productivity and male homosexual nesting and courtship behavior. The average Hg concentration in the peregrine falcon eggs from 2009 is almost four times higher than this level. MeHg accounted for 82.5% of the Hg present in the eggs of 2009 and 2010.</p> <p>The cell test DR CALUX® for screening of dioxin-like activities can be used to detect not only the 29 regulated dioxin-like substances but also many other persistent organic pollutants with dioxin-like potencies, such as mixed halogenated dioxins/biphenyls. In our case, the results of bioanalytical screening methods showed no additional effect of other compounds with dioxin-like activity.</p> <p>Ninety-three out of 177 analytes sought could be detected in the eggs. Chlorinated paraffins, organotin compounds, some pesticides that are still in use, and phthalic esters with the exception of traces of diethylhexyl phthalate could be excluded. All pollutants found in the eggs belong to substance classes banned by the Stockholm and Minamata Conventions.</p

Identification of Major Dioxin-Like Compounds and Androgen Receptor Antagonist in Acid-Treated Tissue Extracts of High Trophic-Level Animals

We evaluated the applicability of combining <i>in vitro</i> bioassays with instrument analyses to identify potential endocrine disrupting pollutants in sulfuric acid-treated extracts of liver and/or blubber of high trophic-level animals. Dioxin-like and androgen receptor (AR) antagonistic activities were observed in Baikal seals, common cormorants, raccoon dogs, and finless porpoises by using a panel of rat and human cell-based chemical-activated luciferase gene expression (CALUX) reporter gene bioassays. On the other hand, no activity was detected in estrogen receptor α (ERα)-, glucocorticoid receptor (GR)-, progesterone receptor (PR)-, and peroxisome proliferator-activated receptor γ2 (PPARγ2)-CALUX assays with the sample amount applied. All individual samples (<i>n</i> = 66) showed dioxin-like activity, with values ranging from 21 to 5500 pg CALUX-2,3,7,8-tetrachlorodibenzo-<i>p</i>-dioxin equivalent (TEQ)/g-lipid. Because dioxins are expected to be strong contributors to CALUX-TEQs, the median theoretical contribution of dioxins calculated from the result of chemical analysis to the experimental CALUX-TEQs was estimated to explain up to 130% for all the tested samples (<i>n</i> = 54). Baikal seal extracts (<i>n</i> = 31), but not other extracts, induced AR antagonistic activities that were 8–150 μg CALUX-flutamide equivalent (FluEQ)/g-lipid. <i>p,p′</i>-DDE was identified as an important causative compound for the activity, and its median theoretical contribution to the experimental CALUX-FluEQs was 59% for the tested Baikal seal tissues (<i>n</i> = 25). Our results demonstrate that combining <i>in vitro</i> CALUX assays with instrument analysis is useful for identifying persistent organic pollutant-like compounds in the tissue of wild animals on the basis of <i>in vitro</i> endocrine disruption toxicity