Revised relative potency values for PCDDs, PCDFs, and non-ortho-substituted PCBs for the optimized H4IIE-luc in vitro bioassay

While the World Health Organization 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalency factors are useful estimates of relative potencies of mixtures when conducting risk assessments, they are not useful when comparing the results of bioassays such as the H4IIE-luc to concentrations of TCDD equivalents calculated from instrumental analyses. Since there are thousands of dioxin-like compounds (DLCs), one use of screening assays is to determine if all of the aryl hydrocarbon receptor (AhR) active DLCs in a mixture have been accounted for in instrumental analyses. For this purpose, bioassay-specific relative potency (ReP) values are needed. RePs of 21 polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls that exhibit effects mediated through the AhR were determined by use of the H4IIE-luc assay. Different values of RePs are derived, depending on the statistical, curve-fitting methods used to derive them from the dose–response relationships. Here, we discuss the various methods for deriving RePs from in vitro data and their assumptions and effects on values of RePs. Full dose–response curves of 2,3,7,8-TCDD and other representative DLCs were used to estimate effective concentrations at multiple points (e.g., EC20-50-80), which were then used to estimate ReP of each DLC to 2,3,7,8-TCDD.     

The levels of PAHs and aryl hydrocarbon receptor effects in sediments of Taihu Lake,China

A total of 16 priority polycyclic aromatic hydrocarbons (PAHs) in sediment samples from Taihu Lake were analyzed by instruments, and sediment extracts were assayed for aryl hydrocarbon receptor (AhR)-mediated ethoxyresorufin-o-deethylase (EROD) induction using a rat hepatoma cell line (H4IIE). The cause–effect relationship between the observed EROD activity and chemical concentrations of PAHs was examined. Our results showed that sediment extracts could induce significant AhR effects, and the bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents of raw extracts (TEQ_bios) ranged from 2.7 to 39.8 pg g^?1 dw. Chemical analysis showed that 16 PAHs were all detected in all samples, and their total concentrations (Σ₁₆PAHs) ranged from 179.8 to 1,669.4 ng g^?1 dw. The abundance of sedimentary PAHs in the three regions (Meiliang Bay, Gonghu Bay, and Xukou Bay) showed a decreasing trend from the inflow region to the outflow region. Chemical analysis-derived TEQs (TEQ_cals) contributed by PAHs ranged from 1.6 to 20.7 pg g^?1 dw. The mean contribution rates (CRs) of PAHs to TEQ_bios were 48.9 %. In Meiliang Bay, EROD effects of 60 % samples were caused by PAHs whose CRs were more than 60 %, while in most sampling sites of Gonghu Bay and Xukou Bay, the CRs of PAHs to TEQ_bios were basically below 40 %. In addition, preliminary ecological risk assessment found that PAHs in sediments have very low ecological impact based on the chemical data of PAHs, while the sediments might pose an unacceptable risk to aquatic organisms and their predators based on the data of TEQ_bio. These findings showed that EROD effects of sediment extracts from Taihu Lake were also caused by other compounds, such as dioxins, polychlorinated biphenyls, etc., together.     

Tetrachlorodibenzo-p-dioxins and tetrachlorodibenzofurans in Atlantic coast striped bass and in selected Hudson River fish,waterfowl and sediments

In striped bass samples from the lower Hudson River and its estuary 2,3,7,8-tetrachlorodibenzo-$\text{p}$-dioxin (2,3,7,8-TCDD) was found at concentrations from 16 to 120 pg/g (ppt). Striped bass from two other locations (Rhode Island coastal waters and Chesapeake Bay, Maryland) had <5 ppt, 2,3,7,8-TCDD. The contaminant, 2,3,7,8-tetrachlorodibenzofuran (2,3,7,8-TCDF), was found in striped bass from all three locations with concentrations varying from 6 ppt in Chesapeake Bay to 78 ppt in the Hudson River. Results from a limited number of non-migratory fish (carp and goldfish) and sediments suggest that the upper Hudson River is not a source for 2,3,7,8-TCDD/2,3,7,8-TCDF contamination of striped bass.     

Inhibition of ethoxyresorufin-O-deethylase (EROD) activity in mixtures of 2,3,7,8-tetrachlorodibenzo-p-dioxin and polychlorinated biphenyls

Induction of ethoxyresorufin-O-deethylase (EROD) activity and porphyrin accumulation shows different structure-activity relationships for different polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Interactions between the two responses can strongly influence the induction and activity of EROD. The results support the conclusion that there are nonadditive interactions between nondioxin-like PCBs and dioxin-like compounds. The interaction between EROD activity and the porphyrin biosynthesis makes the prerequisite of additivity in the toxic equivalency factor concept for environmental mixtures highly spurious. Inhibition of EROD activity caused by non-dioxin like compounds could have a significant impact on the value of EROD activity as a biomarker in the present methods of risk assessment for these compounds.     

Transcriptional profiles induced by the Aryl Hydrocarbon Receptor agonists 2,3,7,8-tetrachlorodibenzo-p-dioxin, 2,3,7,8-tetrachlorodibenzofuran and 2,3,4,7,8-pentachlorodibenzofuran in primary rat hepatocytes

Toxicogenomics was used to examine mRNA expression profiles obtained from primary rat hepatocytes treated for 24 h with 0.01 or 1.0 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 0.02 or 2.0 nM 2,3,4,7,8-pentachlorodibenzofuran (2,3,4,7,8-PeCDF) and 0.1 or 10 nM 2,3,7,8-tetrachlorodibenzofuran (2,3,7,8-TCDF). The concentrations of 2,3,4,7,8-PeCDF and 2,3,7,8-TCDF were chosen to be equivalent to 2,3,7,8-TCDD’s concentration based on the toxic equivalency factor/toxic equivalent (TEF/TEQ) method for estimating biological potency. 2,3,7,8-TCDD at 1.0 nM altered the expression of 533 genes; 2,3,4,7,8-PeCDF at 2.0 nM altered 182 genes, and 2,3,7,8-TCDF at 10 nM altered 154 genes. Of these, 57 genes were affected by all three congeners. Agglomerative hierarchical clustering revealed distinct congener-dependent gene subclusters. Principal components analyses of the microarray data revealed that these congeners cluster independently of one another. Data presented here demonstrate that equivalent TEQ concentrations of 2,3,7,8-TCDD, 2,3,4,7,8-PeCDF and 2,3,7,8-TCDF, while altering the expression of a small battery of genes in common, also produce substantial congener specific alterations in gene expression.     

Concentrations and compositions of organochlorine contaminants in sediments, soils, crustaceans, fishes and birds collected from Lake Tai, Hangzhou Bay and Shanghai city region, China

Contamination by persistent organochlorines (OCs), such as DDTs, hexachlorocyclohexane isomers (HCHs), chlordane compounds (CHLs), hexachlorobenzene (HCB) and polychlorinated biphenyls (PCBs) were examined in sediments, soils, fishes, crustaceans, birds, and aquaculture feed from Lake Tai, Hangzhou Bay, and in the vicinity of Shanghai city in China during 2000 and 2001. OCs were detected in all samples analyzed, and DDT and its metabolites were the predominant contaminants in most sediments, soils and biota. Concentrations of p,p'-DDT and ratio of p,p'-DDT to SigmaDDTs were significantly higher in marine fishes than those in freshwater fishes. While the use of DDTs has been officially banned in China since 1983, these results indicate a recent input of technical DDTs into the marine environment around Hangzhou Bay. Comparison of organochlorine concentrations in fishes collected from Lake Tai and Hangzhou Bay suggests the presence of local sources of HCHs, chlordanes and PCBs at Lake Tai. Higher proportions of penta- and hexa-PCB congeners in fishes at Lake Tai may suggest the use of highly chlorinated PCB product, such as PCB(5), around this lake. To our knowledge, this is a first comprehensive study to examine the present status of organochlorine contamination in various environmental media, such as sediments, soils and wildlife, in China.     

AhR-active compounds in sediments of the Haihe and Dagu Rivers, China

Total concentrations of compounds that can cause activation of the aryl hydrocarbon receptor (AhR) in extracts of river sediments from various locations in the Haihe and Dagu Rivers, Tianjin, China were determined by use of the in vitro H4IIE-luc cell line. AhR-active compounds were isolated from sediments by Soxhlet extraction, and the crude extracts were fractionated using a Florisil column into three Fractions. The response of samples was compared to the TCDD standard and expressed as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TCDD-EQs). Significant dioxin-like activity was observed in each sample. The TCDD-EQs in crude extracts of sediments (SCEs), as determined with the bioassay were 2-4 times greater than the sum of the TCDD-EQs of the eluent from fractions separated with a Florisil column. The results also showed that Fractions 2 and 3 contained most of the AhR-mediated activity. The results obtained by using the bioassay were compared with those of previous measurements of the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) and the structurally-similar AhR-active polychlorinated biphenyls (PCBs). It was determined that sediments from the Dagu River contained greater concentrations of TCDD-EQ than did sediments from the Haihe River except at Jingangqiao (location R3), which is associated with industrial activities in the adjacent densely populated area. The concentrations of TCDD-EQs, based on the EC20 and the relative potency ranges, of SCEs ranged from 330 to 930 and 1200 to 13,900 pg TCDD-EQ g(-1) dry wt in Haihe and Dagu Rivers, respectively.     

Simple and rapid yeast reporter bioassay for dioxin screening: evaluation of the dioxin-like compounds in industrial and municipal waste incineration plants

The CROMIS AhR kit, a simple and rapid yeast bioassay kit, was developed and used to detect dioxins and dioxin-like compounds in 20 gas and solid samples collected from refuse incineration plants in Japan. The World Health Organization toxic equivalent (WHO-TEQ) values of the samples were also calculated using high-resolution gas chromatography/high--resolution mass spectrometry. The WHO-TEQ values of the samples varied greatly, ranging from 0.0021–6.3 ng/g to 0.00013–16 ng/m³N (normal cubic meter) in the solid and gas samples, respectively. 2,3,4,7,8-Pentachlorodibenzofuran (23478-PeCDF) and 1,2,3,7,8-pentachlorodibenzo-p-dioxin (12378-PeCDD) were the major contributors to the samples’ WHO-TEQ values. The yeast in the bioassay responded to these congeners, and the EC₅₀ values of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2378-TeCDD), 12378-PeCDD, and 2,3,4,7,8-PeCDF were 490, 560, and 590 nM, respectively. The incinerator samples were subjected to the bioassay to obtain 2378-TeCDD equivalent values (CROMIS-TEQ values). The CROMIS-TEQ values of the solid and gas samples ranged from 0.0019 to 5.64 ng/g and from 0.14 to 20 ng/m³N, respectively. The CROMIS-TEQ and WHO-TEQ values displayed good correlations (r ²?=?0.94 and 0.95 in the solid and gas samples, respectively), except for those of the samples with low dioxin concentrations (below the Japanese emission standards). Therefore, the CROMIS AhR kit is a useful tool for the initial screening of samples containing dioxin-like compounds.     

Characterization of trace organic contaminants in marine sediment from Yeongil Bay, Korea: 2. Dioxin-like and estrogenic activities

This study employed mechanism-specific in vitro bioassays to help characterize the occurrence and distribution of dioxin-like and estrogenic contaminants in sediment from Yeongil Bay, Korea. Approximately 85% of the sediments tested induced significant dioxin-like activity in the H4IIE-luc bioassay, while approximately 50% induced significant estrogenic activity in the MVLN bioassay. Instrumentally-derived estimates of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 17beta-estradiol equivalents tended to underestimate the magnitude of response observed in the bioassays, suggesting that compounds detected by chemical analysis did not account for all the activity associated with Yeongil Bay sediments, or that non-additive interactions were occurring. The greatest dioxin-like and estrogenic activity was associated with the mid-polarity Florisil fractions (F2) expected to contain polycyclic aromatic hydrocarbons (PAHs) as well as chlorinated dioxins and furans. As in previous studies of Korean coastal sediment, more polar fractions (F3) generated more modest responses both in terms of magnitude and the number of samples responding.     

Aryl hydrocarbon receptor (AhR) inducers and estrogen receptor (ER) activities in surface sediments of Three Gorges Reservoir,China evaluated with in vitro cell bioassays

Two types of biological tests were employed for monitoring the toxicological profile of sediment cores in the Three Gorges Reservoir (TGR), China. In the present study, sediments collected in June 2010 from TGR were analyzed for estrogen receptor (ER)- and aryl hydrocarbon receptor (AhR)-mediated activities. The estrogenic activity was assessed using a rapid yeast estrogen bioassay, based on the expression of a green fluorescent reporter protein. Weak anti-estrogenic activity was detected in sediments from an area close to the dam of the reservoir, and weak estrogenic activities ranging from 0.3 to 1 ng 17β-estradiol (E2) equivalents (EQ) g^?1 dry weight sediment (dw) were detected in sediments from the Wanzhou to Guojiaba areas. In the upstream areas Wanzhou and Wushan, sediments demonstrated additive effects in co-administration of 1 nM E2 in the yeast test system, while sediments from the downstream Badong and Guojiaba areas showed estrogenic activities which seemed to be more than additive (synergistic activity). There was an increasing tendency in estrogenic activity from upstream of TGR to downstream, while this tendency terminated and converted into anti-estrogenic activity in the area close to the dam. The AhR activity was detected employing rat hepatoma cell line (H4IIE). EROD activities were found homogenously distributed in sediments in TGR ranging from 200 to 311 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) EQ g^?1 dw for total AhR agonists and from 45 to 76 pg TCDD EQ g^?1 dw for more persistent AhR agonists. The known AhR agonists polycyclic aromatic hydrocarbon, polychlorinated biphenyl, and PCDD/F only explained up to 8 % of the more persistent AhR agonist activity in the samples, which suggests that unidentified AhR-active compounds represented a great proportion of the TCDD EQ in sediments from TGR. These findings of estrogenic potential and dioxin-like activity in TGR sediments provide possible weight-of-evidence of potential ecotoxicological causes for the declines in fish populations which have been observed during the past decades in TGR.     

Distribution of nutrients,heavy metals,and PAHs affected by sediment dredging in the Wujin'gang River basin flowing into Meiliang Bay of Lake Taihu

This paper's survey of the pollution of the Wujin'gang River is important because it is one of the main rivers flowing into Meiliang Bay of Lake Taihu in eastern China. Trace metals (TMs) in this paper are described according to their pollution index (P _i). Cluster analysis and correlation analysis are utilized for group sites and to assess co-contamination. Toxicity effect analysis was conducted using individual sediment quality guideline quotients (SQGQs) and mean SQGQs. The results showed that sediment from the Wujin'gang River basin was affected by nutrients, heavy metals, and polycyclic aromatic hydrocarbons (PAHs), which are an essential contamination source for both Meiliang Bay and Zhushan Bay of Lake Taihu. The discharge of TMs has significant correlations to total nitrogen (TN) and total phosphorus (TP); however, no significant correlations were observed between the content of PAHs and TMs. Toxicity effect results show that sediment in the Wujin'gang River basin threatens sediment-dwelling organisms. The harmful effect was mainly caused by heavy metals especially Cd, Cr, Ni, and Cu. Sediment dredging is an effective way to control pollution from internal rivers especially for the pollution of TN and heavy metals in the Wujin'gang River basin.     

Distribution of PCDD/Fs and dioxin-like PCBs in sediment and plants from a contaminated salt marsh (Tejo estuary,Portugal)

Concentrations and profiles of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) were investigated in sediment and plants collected from a salt marsh in the Tejo estuary, Portugal. The highest PCDD/F and dl-PCB concentrations were detected in uncolonized sediments, averaging 325.25?±?57.55 pg g^?1 dry weight (dw) and 8,146.33?±?2,142.14 pg g^?1 dw, respectively. The plants Sarcocornia perennis and Halimione portulacoides growing in PCDD/F and dl-PCB contaminated sediments accumulated contaminants in roots, stems, and leaves. It was observed that PCDD/F and dl-PCB concentrations in roots were significantly lower in comparison with stems and leaves. In general, concentration of ΣPCDD/Fs and Σdl-PCBs in H. portulacoides tissues were found to be twofold higher than those in S. perennis, indicating a difference in the accumulation capability of both species. Furthermore, congener profiles changed between sediments and plant tissues, reflecting a selective accumulation of low chlorinated PCDD/Fs and non-ortho dl-PCBs in plants.     

Patterns and sources of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in surficial sediments of Lakes Erie and Ontario

This study determines spatial trends and congener patterns of 2378-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in surficial sediments of Lakes Erie and Ontario. Sediments are enriched in 2378-PCDFs in Lake Ontario, and the PCDD/F concentrations increased from shallow near-shore sediments towards deep-water depositional zone sediments. In Lake Erie, sediments were dominated by octachlorodibenzo-p-dioxin, and the highest PCDD/F concentrations were observed in the western basin and the southern shoreline of the central basin with a decrease towards the eastern basin and the northern shoreline of the central basin. Principal components analysis revealed that chemical manufacture and disposal of chemical waste along the Niagara River has been a major PCDD/F source to Lake Ontario; while PCDD/Fs in Lake Erie are from multiple sources including industrial sources along the Detroit River, major tributaries along the southern shoreline of the lake, and atmospherically-derived material from the upper lakes and connecting channels.     

Longer-term and short-term variability in pollution of fluvial sediments by dioxin-like and endocrine disruptive compounds

Changes in pollutant loads in relatively dynamic river sediments, which contain very complex mixtures of compounds, can play a crucial role in the fate and effects of pollutants in fluvial ecosystems. The contamination of sediments by bioactive substances can be sensitively assessed by in vitro bioassays. This is the first study that characterizes detailed short- and long-term changes in concentrations of contaminants with several modes of action in river sediments. One-year long monthly study described seasonal and spatial variability of contamination of sediments in a representative industrialized area by dioxin-like and endocrine disruptive chemicals. There were significant seasonal changes in both antiandrogenic and androgenic as well as dioxin-like potential of river sediments, while there were no general seasonal trends in estrogenicity. Aryl hydrocarbon receptor-dependent potency (dioxin-like potency) expressed as biological TCDD-equivalents (BIOTEQ) was in the range of 0.5–17.7 ng/g, dry mass (dm). The greatest BIOTEQ levels in sediments were observed during winter, particularly at locations downstream of the industrial area. Estrogenicity expressed as estradiol equivalents (EEQ) was in the range of 0.02–3.8 ng/g, dm. Antiandrogenicity was detected in all samples, while androgenic potency in the range of 0.7–16.8 ng/g, dm dihydrotestosterone equivalents (DHT-EQ) was found in only 30 % of samples, most often during autumn, when antiandrogenicity was the least. PAHs were predominant contaminants among analyzed pollutants, responsible, on average, for 13–21 % of BIOTEQ. Longer-term changes in concentrations of BIOTEQ corresponded to seasonal fluctuations, whereas for EEQ, the inter-annual changes at some locations were greater than seasonal variability during 1 year. The inter- as well as intra-annual variability in concentrations of both BIOTEQ and EEQ at individual sites was greater in spring than in autumn which was related to hydrological conditions in the river. This study stresses the importance of river hydrology and its seasonal variations in the design of effective sampling campaigns, as well as in the interpretation of any monitoring results.     

Analysis of snake tissue and snake eggs for 2,3,7,8-tetrachlorodibenzo-p-dioxin via fused silica GC combined with atmospheric pressure ionization MS

Several snake tissue samples and one set of snake eggs were collected and analyzed for 2,3,7,8-tetrachlorodibenzo-$\text{p}$-dioxin. The isomer-specific analysis was performed using fused silica GC combined with atmospheric pressure ionization MS. The results ranged from a low of 38 parts-per-trillion (ppt) to high levels over 500 ppt.     

Persistent organic pollutants in Detroit River suspended sediments: polychlorinated dibenzo-p-dioxins and dibenzofurans,dioxin-like polychlorinated biphenyls and polychlorinated naphthalenes

Suspended sediments from the Detroit River were collected in 1999 and 2000 using sediment traps at sites ranging from western Lake Erie to southern Lake St. Clair and analyzed to determine the spatial distributions of contaminants including polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/PCDFs), dioxin-like PCBs (DLPCBs) and polychlorinated naphthalenes (PCNs). Concentrations of all three contaminant classes were clearly elevated at sites in the lower reaches of the river in the Trenton Channel. The potential influence of the Trenton Channel as a source of contamination to western Lake Erie was further evidenced by PCDD/PCDF homologue profiles, which indicated a contribution from chemical manufacturing in addition to the normal background combustion profile. Toxic equivalents (TEQs) for PCDDs/PCDFs generally exceeded those for DLPCBs; combined total TEQs in July 2000 for these two compound classes ranged from 2.30 pg/g in southern Lake St. Clair to 306 pg/g at a station just downstream of the outflow of Monguagon Creek in the Trenton Channel. The spatial distribution of PCN contamination was similar to that of PCDDs/PCDFs and DLPCBs, with the highest level of total PCNs (8200 ng/g) detected at a site in the Trenton Channel near Elizabeth Park; TEQs for PCNs in the Trenton Channel ranged from 73 to 3300 pg/g. The data indicate that PCNs represent a significant contribution to dioxin-like biological activity in Detroit River suspended sediments.     

Endocrine disruption of water and sediment extracts in a non-radioactive dot blot/RNAse protection-assay using isolated hepatocytes of rainbow trout

Goal, Scope and Background In order to evaluate the estrogenic activity of sediments and XAD water extracts of selected sites of the catchment area of the River Neckar, a river system in Southern Germany, an integrative assessment approach was used to assess the ecological hazard potential of endocrine-disrupting compounds in sediment and water. Methods The approach is based on estrogen receptor-mediated vitellogenin synthesis induced in isolated hepatocytes of rainbow trout and quantified in a non-radioactive dot blot/RNAse protection-assay in parallel to comprehensive chemical analyses of estrogenic substances. Results and Discussion Numerous investigated extracts revealed an estrogen activity comparable to that of the positive control (1 nM 17?-estradiol corresponding to 270 ng/L in the test medium). Based on a concentration factor of 30 in the extracts and a recovery of XAD resins of approximately 80 %, 17?-estradiol equivalent concentrations between 20 and 26.7 ng/L could be calculated downstream of a sewage treatment plant (< 0.1 ng/L for a reference site). A comparison of the bioassay-derived Bio-TEQs (toxicity equivalents) and the Chem-TEQs revealed a high correlation with a Pearson coefficient of 0.85, indicating that the same ranking of the samples could be obtained with respect to the endocrine disrupting potential with both chemical and bioanalytical analysis. However, the TEQ concentrations computed from chemical analyses were significantly lower than the bioassay-derived TEQ concentrations. In fact, in none of the samples, more than 14 % of the vitellogenin-inducing potency could be attributed to the substances (steroids, alkylphenols, bisphenol A, diethylstilbestrol) analyzed. A comparison of the endocrine disrupting potential of sediments extracted by the solvents acetone and methanol revealed lower biological effects for acetone-extracted samples. Possible reasons may be a masking of endocrine effects in acetone extracts by cytotoxicity, a low extraction efficiency of the solvent acetone, or anti-estrogen potencies of some extracted sediment compounds. Using a mass balance approach, the contribution of the compounds analyzed chemically (Chem-TEQs) to the total endocrine activity (Bio-TEQs) was calculated. Based on the very low detection limits, particularly of the steroids with their high TEF factors, results revealed that a calculation of the Chem-TEQs is associated with considerable scale inaccuracy: Whereas only 7-15 % of the biological effectiveness (Bio-TEQs) could be explained by endocrine substances identified above the detection limits, the assumption of concentrations slightly below the given detection limits would result in a significant over estimation (137-197 %) of the Bio-TEQs. Even the interassay variation of the dot blot assay with different fish donors for primary hepatocyte (factor 2 - 2.5) is relatively low, when compared to the large range of the Chem-TEQ concentrations (factor 20) obtained when applying different modes of calculation. Conclusions and Outlook Overall, only a minor portion of the endocrine activity detected by bioassays could be linked to compounds identified by chemical analysis. In vitro assays for assessment of endocrine activities are useful as sensitive integrating methods that provide quantitative estimates of the total activity of particular receptor-mediated responses. Although discrepancies may also result from different bioanalytical approaches, it is overall likely that bioanalytical and not chemical analytical approaches give the correct estimate of endocrine disrupting potencies in environmental samples. As a conclusion, assessment of endocrine disruption based on chemical analysis alone does not appear sufficient and further research into the spectrum of substances with potential endocrine activity as well as into additive or even synergistic effects in complex environmental samples is urgently needed.     

Cell bioassays for detection of aryl hydrocarbon (AhR) and estrogen receptor (ER) mediated activity in environmental samples

In vitro cell bioassays are useful techniques for the determination of receptor-mediated activities in environmental samples containing complex mixtures of contaminants. The cell bioassays determine contamination by pollutants that act through specific modes of action. This article presents strategies for the evaluation of aryl hydrocarbon receptor (AhR)-(hereafter referred as dioxin-like) or estrogen receptor (ER)-mediated activities of potential endocrine disrupting compounds (EDCs) in complex environmental mixtures. Extracts from various types of environmental or food matrices can be tested by this technique to evaluate their 2,3,7,8-tetrachlorodibenzop-dioxin equivalents (TCDD-EQs) or estrogenic equivalents (E₂-EQs) and to identify contaminated samples that need further investigation using resource-intensive instrumental analyses. Fractionation of sample extracts exhibiting significant activities, and subsequent reanalysis with the bioassays can identify important classes of contaminants that are responsible for the observed activity. Effect-directed chemical analysis is performed only for the active fractions to determine the responsible compounds. Mass-balance estimates of all major compounds contributing to the observed effects can be calculated to determine if all of the activity has been identified, and to assess the potential for interactions such as synergism or antagonism among contaminants present in the complex mixtures. The bioassay approach is an efficient (fast and cost effective) screening system to identify the samples of interest and to provide basic information for further analysis and risk evaluation.     

Polychlorinated naphthalenes in human adipose tissue from New York, USA

Polychlorinated naphthalenes (PCNs) are persistent, bioaccumulative, and toxic contaminants. Prior to this study, the occurrence of PCNs in human adipose tissues from the USA has not been analyzed. Here, we have measured concentrations of PCNs in human adipose tissue samples collected in New York City during 2003-2005. Concentrations of PCNs were in the range of 61-2500 pg/g lipid wt. in males and 21-910 pg/g lipid wt. in females. PCN congeners 52/60 (1,2,3,5,7/1,2,4,6,7) and 66/67 (1,2,3,4,6,7/1,2,3,5,6,7) were predominant, collectively accounting for 66% of the total PCN concentrations. Concentrations of PCNs in human adipose tissues were 2-3 orders of magnitude lower than the previously reported concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs). Concentrations of PCNs were not correlated with PCB concentrations. The contribution of PCNs to dioxin-like toxic equivalents (TEQs) in human adipose tissues was estimated to be <1% of the polychlorinated dibenzo-p-dioxin/dibenzofuran (PCDD/F)-TEQs.     

Aryl hydrocarbon receptor-mediated activity of atmospheric particulate matter from an urban and a rural site in Switzerland

Atmospheric particulate matter (PM) is an air-suspended mixture of solid and liquid particles that vary in size, shape, and chemical composition. Long-term exposure to elevated concentrations of fine atmospheric particles is considered to pose a health threat to humans and animals. In this context, it has been hypothesized that toxic chemicals such as polycyclic aromatic hydrocarbons (PAHs) play an important role. Some PAHs are known to be carcinogenic and it has been shown that carcinogenic effects of PAHs are mediated by the aryl hydrocarbon receptor (AhR). In this study, PM1 was collected at a rural and an urban traffic site during an intense winter smog period, in which concentration of PM1 often exceeded 50 μg m^?3. We applied an in vitro reporter gene assay (DR-CALUX) to detect and quantify PM1-associated chemicals that induce AhR-mediated gene expression. This activity was expressed as CALUX equivalents of 2,3,7,8-tetrachlorodibenzodioxin (PM-TCDD-CEQs). In addition, concentrations of PAHs in the PM1 extracts were determined using gas chromatography/high-resolution mass spectrometry. Concentrations of PM-TCDD-CEQs ranged from 10 to 85 pg m^?3 and from 19 to 87 pg m^?3 at the urban and rural site, respectively. By the use of known relative potency factors, the measured concentration of a PAH was converted into a PAH-TCDD-CEQ concentration. ΣPAH-TCDD-CEQ and PM-TCDD-CEQ were highly correlated at both sites (r² = 0.90 and 0.69). The calculated ΣPAH-TCDD-CEQs explain between 2% and 20% of the measured PM-TCDD-CEQs. Benzo[k]fluoranthene was the most important PAH causing approximately 60% of the total ΣPAH-TCDD-CEQ activity. In contrast to NO, CO, PM10, and PM1, the concentration of PM-TCDD-CEQs showed no significant difference between the two sites. No indications were found that road traffic emissions caused elevated concentrations of PM-TCDD-CEQs at the urban traffic site.