Parent Polychlorinated Biphenyls Research Articles

Chiral Polychlorinated Biphenyls Are Biotransformed Enantioselectively by Mammalian Cytochrome P-450 Isozymes to Form Hydroxylated Metabolites

In vitro incubations of purified rat cytochrome P-450 (CYP) 2B1 and human CYP 2B6 were performed to determine if CYP isozymes biotransform polychlorinated biphenyls (PCBs) enantioselectively. Enantioselective metabolism of chiral PCBs 45, 84, 91, 95, 132, and 136 and production of hydroxylated PCB metabolites (OH-PCBs) were observed, while no changes in PCB 183 atropisomer composition were observed for either isozyme. Enantiomer fractions (EFs) of parent PCBs, individually incubated as racemates at 25 ng/mL initial concentration, with rat CYP 2B1 ranged from 0.353 to 0.822. Enantioselectivity was also observed for PCBs 45 (EF = 0.437) and 132 (EF = 0.537) incubated at that concentration with human CYP 2B6. Both atropisomers of chiral PCBs appeared to be biotransformed simultaneously by rat CYP 2B1, except for (+)-PCB 132, but at different rates. Hydroxylated PCBs were identified using gas chromatography-high resolution mass spectrometry for all chiral PCBs enantioselectively transformed by CYPs. These metabolites did not correspond to any commercially available authentic standards, supporting the hypothesis that many unidentified OH-PCBs detected in wildlife may have arisen from in vivo biotransformation of chiral PCBs. A rough estimate suggested that more than half of the total congener metabolized by rat CYP 2B1 was converted to OH-PCBs. Similar concentration decreases were observed for congeners incubated with human CYP 2B6, but less OH-PCBs were formed. Formation of OH-PCBs via an enantioselective OH insertion mechanism was suggested, and may be a source of the unidentified OH-PCBs currently found in the environment.

Elimination Half-Lives of Polychlorinated Biphenyl Congeners in Children

The elimination kinetics of polychlorinated biphenyls (PCBs) in humans is difficult to assess in observational studies, because PCB exposure is never completely abolished. In a community with high dietary PCB exposures from whale blubber, we examined two groups of children with increased body burdens from breast-feeding. Follow-up was from ages 4.5 to 7.5 years (99 subjects) and 7 to 14 years (101 subjects). The calculations were performed by the use of structural equation models, with adjustment for body weight and dietary blubber intake as the main source of postnatal exposure. As a likely result of background exposures, apparent elimination half-lives were unexpectedly long when based on results from all cohort members. Subjects with exposures above the median and in the highest quartile showed half-lives of about 3-4 years for CB-138 and 4.5-5.5 years for CB-105 and CB-118; 6.5-7.5 years for CB-156, CB-170, and CB-187; and 7-9 years for CB-153 and CB-180. The longest half-lives correspond to elimination of the parent PCB solely with a daily fat excretion rate of 1-2 g, whereas shorter half-lives assume metabolic break-down.

Prediction of the health effects of polychlorinated biphenyls (PCBs) and their metabolites using quantitative structure–activity relationship (QSAR)

Polychlorinated biphenyls (PCBs) are a group of 209 persistent environmental contaminants that are slightly different but structurally related. PCBs are known to induce a variety of health effects and often have been toxicologically tested as complex commercial mixtures (Aroclors) but environmental exposure occurs separately to a small number of specific congeners. Recently, the Third National Report on Human Exposures to Environmental Chemicals, an assessment of exposure data of the National Health and Nutrition Examination Survey (NHANES), identified 35 individual PCB congeners in the U.S. population. These types of findings necessitate the toxicity evaluation of individual congeners but adequate toxicity data for most individual PCB congeners are not available. Due to this, a quantitative structure–activity relationship (QSAR) approach was used to assess the potential mutagenesis and carcinogenesis of individual congeners and their possible metabolites. The predictions were analyzed to define the underlying generalizations between the parent PCBs, their metabolites, and some important toxicological endpoints. This analysis reveals that (1) mono and di-chlorinated PCBs and their metabolites can be potential mutagens; (2) PCB benzoquinone metabolites could be carcinogenic but the weight of evidence is poor. These results support the hypothesis that environmental exposure to some PCBs and/or their metabolites could produce mutagenicity and/or carcinogenicity. Hence, these data should be considered as priority toxicological testing data needs. As with all computational toxicology analytical findings, these conclusions must yield to empirical data as they become available.

Hydroxy-PCBs, Methoxy-PCBs and Hydroxy-Methoxy-PCBs: Metabolites of Polychlorinated Biphenyls Formed In Vitro by Tobacco Cells

While the metabolism of polychlorinated biphenyls (PCBs) in plant cells is a rarely studied field, hydroxy-PCBs have been detected in several studies involving the use of various plant species. The ability of the tobacco (Nicotiana tabacum) callus culture WSC-38 to metabolize six dichlorobiphenyls under aseptic conditions was studied, and the resulting PCB metabolites were analyzed. WSC-38 cultures were cultivated with individual dichlorinated PCB congeners. The metabolites were identified based on mass spectra characteristics after gas chromatography separation. In addition, metabolites of PCB 9 (2,5-dichlorobiphenyl) were identified by comparing their retention characteristics with the available standards. In most cases at least two hydroxy-PCBs were produced from each parent PCB. Methoxy-PCBs and hydroxy-methoxy-PCBs were other groups of metabolites produced. To the best of our knowledge, ours is the first report to determine the presence of methoxy- and hydroxy-methoxy-metabolites of PCBs in plants. The role of the O-methyltransferases (OMTs) in the methylation of hydroxy-PCBs is discussed. As methoxy-metabolites of acetophenone were found among our samples, we posit that the OMTs responsible for the methylation of these compounds are also involved in the metabolism of PCBs in cultures of WSC-38.

Predatory Bird Species Show Different Patterns of Hydroxylated Polychlorinated Biphenyls (HO-PCBs) and Polychlorinated Biphenyls (PCBs)

Hydroxylated metabolites of polychlorinated biphenyls (HO-PCBs) have previously been associated with endocrine disrupting effects. Since metabolic capacity may differ among species, we investigated the levels and profiles of HO-PCBs and PCBs in livers of four predatory bird species from Belgium. Maximum concentrations for sum HO-PCBs were found in the common buzzard (Buteo buteo) up to 13 700 pg/g wet weight (ww). The most prominent HO-PCB congener in all bird species was 4-HO-CB 187 (up to 6420 pg/g ww in buzzard liver), followed by 4-HO-CB148 in the buzzard (up to 1820 pg/g ww), sparrowhawk (Accipiter nisus), and grey heron (Ardea cinerea), and by 3'-HO-CB138 in long eared owl (Asio otus) and in one grey heron (up to 985 pg/g ww and 3450 pg/g ww, respectively). The mean profile of the grey heron differed from the other species with 3'-HO-CB138 and 4-HO-CB163 contributing more to the sum HO-PCBs. This indicates that aquatic and terrestrial predatory bird species may show differences in their HO-PCBs profiles. Variation in the diet and species-specific accumulation and metabolism of PCBs are probably the most important causes for these differences. Correlations between HO-PCBs and their parent PCBs were only found significant for buzzards.

Monofluorinated analogues of polychlorinated biphenyls (F-PCBs): Synthesis using the Suzuki-coupling, characterization, specific properties and intended use

An improved method, a palladium-catalyzed cross-coupling reaction the so-called Suzuki-coupling, was used to synthesize 19 monofluorinated analogues (F-PCBs) of polychlorinated biphenyls (PCBs), including analogues of PCBs 28, 52 and 81. The coupling of chlorinated and/or fluorinated isomers of aryl boronic acids with bromo- and/or iodobenzenes gave the desired F-PCB congeners in good to excellent yields. The self-coupling product of the aryl boronic acids was the major impurity, but this limitation could be minimized. The influence of ortho-substitution by chlorine on the reaction time was studied in detail. The F-PCBs were characterized by means of 1H and 19F NMR and mass spectrometry (MS). F-PCBs show remarkably similar characteristics in many aspects to those of the corresponding non-fluorinated parent PCBs. Since fluorine is generally resistant to degradation, once incorporated into a substrate it functions as an indelible label that can be used to monitor structures and dynamics. F-PCBs may prove their usefulness as markers and tracers in mechanistic and toxicological studies. The aim of further research will be to test the suitability of fluorine as a sensitive probe in a broad range of applications, including synthesis, analytical, mechanistic and toxicological studies.

Differential Accumulation of Polychlorinated Biphenyl Congeners in the Terrestrial Food Web of the Kalamazoo River Superfund Site, Michigan

A series of field studies was conducted to determine the bioaccumulation of polychlorinated biphenyl (PCB) congeners in the terrestrial food web of the Kalamazoo River flood plain. Samples included colocated soils, native plants likely to be consumed by wildlife, several taxa of terrestrial invertebrates, small mammals, passerine bird eggs, nestlings, and adults, and great horned owl plasma and eggs. Mean concentrations of total PCBs in samples from the former Trowbridge impoundment were 6.5 mg/kg dry weight for soils and 0.023, 0.13, 1.3, 1.3, 1.6, and 8.2 mg/kg wet weight for plants, small herbivorous mammals, depurated earthworms, shrews, great horned owl eggs, and house wren eggs, respectively. Historical data from the Kalamazoo River have reported Aroclor-equivalent total PCB concentrations in the terrestrial food web; however, the degree of environmental weathering of the parent PCB mixtures was unknown. In this study, earthworms and composite samples of coleoptera exhibited PCB congener patterns that were similar to patterns in colocated soils. However, in plants, less chlorinated PCBs (e.g., mono-, di-, tri-, and tetrachlorinated biphenyls) were predominant, and in small mammals, there was a notable enrichment of PCBs 153, 180, 138, 118, and 99. In general, concentrations of PCBs were lower in most biota than in soil from the Kalamazoo River Area of Concern (KRAOC) although there was a modest biomagnification of PCBs from lower trophic level biota to highertrophic levels. As a consequence of environmental weathering of PCBs in the terrestrial food web of the KRAOC, the relative potency of the PCBs (expressed as mg TEQs/kg PCBs) decreased from soil to most biota. While there was a general trend, as expected, in which concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) increased with total PCBs, this relationship was rather poor (R2 = 0.13). Taken together, these data suggest that the differential accumulation of PCB congeners in the terrestrial food web can be explained by congener-specific differences in bioavailability from soil, exposure pathways, and metabolic potential of each of the food web components.

Developmental exposure to 4-hydroxy-2,3,3',4',5-pentachlorobiphenyl (4-OH-CB107): long-term effects on brain development, behavior, and brain stem auditory evoked potentials in rats.

In the present study the developmental neurotoxic effects of the PCB metabolite 4-OH-2,3,3',4',5-pentachlorobiphenyl (4-OH-CB107) were compared with effects caused by a mixture of parent polychlorinated biphenyl (PCB) congeners (Aroclor 1254). Pregnant female Wistar rats were exposed to 0.5 or 5 mg 4-OH-CB107, or 25 mg Aroclor 1254 per kg body weight from gestation days 10 to 16. Plasma thyroid hormone levels were significantly decreased in the offspring of all treatment groups at postnatal day 4 (PND 4). Behavioral experiments using an open field paradigm revealed an impaired habituation in male offspring of all treatment groups at PND 130. Passive avoidance experiments indicated significant influences on the time course of step-down latencies across trials in exposed male rats. Catalepsy induced by haloperidol showed increases in latencies to movement onset in female offspring exposed to 0.5 mg 4-OH-CB107 compared to Aroclor 1254 treated offspring at PND 168-175. Male offspring exposed to 4-OH-CB107 or Aroclor 1254 showed decreases in latencies compared to control animals. Brain stem auditory evoked potentials (BAEPs) measured at PND 300-310 showed significant increases in auditory thresholds in the low frequency range between Aroclor 1254 and 4-OH-CB107 (5 mg/kg bw) treated animals. Measurements of neurotransmitter levels revealed effects of Aroclor 154 exposure on both the dopaminergic and the serotonergic systems, whereas 4-OH-CB107 exposure affected dopaminergic and noradrenergic systems, with slight but not significant effects on the serotonergic system. These results indicate that 4-OH-CB107 is able to induce long-term effects on behavior and neurodevelopment. The observed effects for 4-OH-CB107 are similar to, but in some aspects different from, the effects observed after Aroclor 1254 exposure.

Polychlorinated biphenyls (PCBs) exert thyroid hormone-like effects in the fetal rat brain but do not bind to thyroid hormone receptors.

Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants routinely found in human and animal tissues. Developmental exposure to PCBs is associated with neuropsychologic deficits, which may be related to effects on thyroid hormone (TH) signaling in the developing brain. However, PCBs may interfere with TH signaling solely by reducing circulating levels of TH, or they may exert direct effects on TH receptors (TRs). Therefore, we tested whether maternal exposure to a commercial PCB mixture, Aroclor 1254 (A1254), exerts effects in the fetal brain by one or both of these mechanisms. Dams were dosed daily with 0, 1, or 4 mg/kg A1254 from gestational day 6 (GD6) until they were sacrificed on GD16. A1254 significantly reduced circulating levels of triiodothyronine (T3) and thyroxine (T4) in pregnant rats but increased the expression of several TH-responsive genes in the fetal cortex, including neuroendocrine-specific protein A (NSP-A), RC3/neurogranin, and Oct-1. These findings are consistent with a direct action of PCBs on TRs. However, we did not identify parent PCB congeners or metabolites that bound to rat TRs isolated from hepatic nuclei. These findings indicate that PCBs can interfere with TH signaling in the fetal brain by direct actions on the fetus rather than by producing maternal hypothyroidism.

Hydroxylated and methylsulfone-containing metabolites of polychlorinated biphenyls in the plasma and blubber of bowhead whales (Balaena mysticetus).

Bowhead whale (Balaena mysticetus) blubber (n = 20) and plasma (n = 19) samples were collected during the 1997 to 2000 Inuit subsistence harvests in Barrow, Alaska, USA, to quantify the concentrations of methylsulfone (MeSO2)-containing and hydroxylated (OH) polychlorinated biphenyl (PCB) metabolites in this cetacean. The distribution of MeSO2-PCBs in blubber was dominated by 4-MeSO2-substituted congeners, the most abundant being 4-MeSO2-CB-70, 3'-MeSO2-CB-132, and 4-MeSO2-CB-64. Mean (+/- 1 standard error) sum (sigma) MeSO2-PCBs concentrations in blubber were low (6.23 +/- 0.81 ng g(-1) lipid normalized) compared to concentrations previously reported in other marine mammals. However, similar ratios of MeSO2-PCB metabolites to parent PCB congeners among marine mammals suggest that cytochrome P450 2B-like biotransformation and other necessary enzyme-mediated processes and mechanisms that influence the formation and clearance of MeSO2-PCBs exist in the bowhead whale. Pentachlorophenol was the most abundant halogenated phenolic compound quantified in bowhead plasma (1.55 +/- 0.19 ng g(-1) wet wt). Despite indirect evidence for arene epoxidation of the biphenyl moiety inferred from MeSO2-PCB formation, sumOH-PCB concentrations in bowhead plasma were low (1.52 +/- 0.31 ng g(-1) wet wt) compared to humans and marine mammals and were comprised of only two detectable OH-PCB congeners (4'-OH-CB-130 and 4-OH-CB-187). Further research is required to elucidate the toxicokinetics and distribution of OH-PCBs in this cetacean.

Increased [ [formula omitted]]Phorbol Ester Binding in Rat Cerebellar Granule Cells and Inhibition of [formula omitted] Buffering in Rat Cerebellum by Hydroxylated Polychlorinated Biphenyls

Our previous structure–activity relationship (SAR) studies indicated that the effects of polychlorinated biphenyls (PCBs) on neuronal Ca 2+ homeostasis and protein kinase C (PKC) translocation were associated with the extent of coplanarity. Chlorine substitutions at ortho position on the biphenyl, which increase the non-coplanarity, are characteristic of the most active congeners in vitro. In the present study, we investigated the effects of selected hydroxylated PCBs, which are major PCB metabolites identified in mammals, on the same measures where PCBs had differential effects based on structural configuration. These measures include PKC translocation as determined by [ 3H ]phorbol ester ([ 3H ]PDBu) binding in cerebellar granule cells, and Ca 2+ sequestration as determined by 45Ca 2+ uptake by microsomes isolated from adult rat cerebellum. All the selected hydroxy-PCBs with ortho-chlorine substitutions increased [ 3H ]PDBu binding in a concentration-dependent manner and the order of potency as determined by E 50 (concentration that increases control activity by 50%) is 2′,4′,6′-trichloro-4-biphenylol (32 ± 4 μM), 2′,5′-dichloro-4-biphenylol (70 ± 9 μM), 2,2′,4′,5,5′-pentachloro-4-biphenylol (80 ± 7 μM) and 2,2′,5′-trichloro-4-biphenylol (93 ± 14 μM). All the selected hydroxy-PCBs inhibited microsomal 45Ca 2+ uptake to a different extent. Among the hydroxy-PCBs selected, 2′,4′,6′-trichloro-4-biphenylol is the most active in increasing [ 3H ]PDBu binding as well as inhibiting microsomal 45Ca 2+ uptake. 3,5-Dichloro-4-biphenylol and 3,4′,5-trichloro-4-biphenylol did not increase [ 3H ]PDBu binding, but inhibited microsomal 45Ca 2+ uptake. This effect was not related to ionization of these two hydroxy-PCBs. Hydroxylated PCBs seemed to be as active as parent PCBs in vitro. These studies indicate that PCB metabolites such as hydroxy-PCBs might contribute significantly to the neurotoxic responses of PCBs.

Influence of the consumption of fatty Baltic Sea fish on plasma levels of halogenated environmental contaminants in Latvian and Swedish men.

We examined the influence of widely varied consumption of fatty fish from the Baltic Sea and of age on plasma concentrations of polychlorinated biphenyls (PCBs), polychlorobiphenylols (OH-PCBs), 2, 2-bis(4-chlorophenyl)-1,1,1-trichloroethane (4,4'-DDT), 2, 2-bis(4-chlorophenyl)-1,1-dichloroethane (4,4'-DDE), 2,2',4, 4'-tetrabromodiphenyl ether (BDE-47), hexachlorobenzene (HCB), and pentachlorophenol (PCP) in Latvian and Swedish men. Both age and fish consumption were significantly correlated with the concentrations of [sigman]PCB, [sigman]OH-PCB, 4,4'-DDE, 4,4'-DDT, and HCB. In the case of BDE-47, no significant relationship with age was observed, and fish consumption had the largest relative effect on plasma concentrations of this contaminant. This relationship may be a result of exposure to BDE-47 having been more recent than that of PCBs and DDE, or because the half-life of BDE-47 may be shorter than that of PCB and DDE. Latvian men demonstrated higher plasma levels of DDE and DDT but lower levels of [sigman]PCB and PCP than did Swedish men. The corresponding levels of HCB and BDE-47 were similar in both countries. The Spearman's rank correlation coefficient obtained by comparing the level of the metabolite 4-hydroxy-2,3,3',4',5-pentachlorobiphenyl (4-OH-CB107) to the combined levels of its parent compounds, 2,3,3',4, 4'-pentachlorobiphenyl (CB-105) and 2,3',4,4',5-pentachlorobiphenyl (CB-118), was higher than the median correlation coefficient obtained upon comparing the level of this metabolite to all other possible combinations of two PCB levels. No other increased correlation between metabolite and parent PCB concentration was observed.

Biological activity associated with noncoplanar polychlorinated biphenyls after microbial dechlorination of aroclor 1242® and aroclor 1254®

AbstractBioremediation of polychlorinated biphenyls (PCBs) byanaerobic microbial dechlorination occurring naturally in the subsurface and in engineered systems results in mixtures of lower‐chlorinated, primarily ortho‐substituted biphenyls. The purpose of this study was to determine whether this process of bacterial dechlorination results in a mixture that differs in biological activity from that of the parent PCB mixture. Two biological assays sensitive to the action of ortho‐substituted PCBs were employed: insulin release by RINm5F cells, and superoxide anion (O2) production by rat neutrophils. The PCB mixtures Aroclor 1242® and Aroclor 1254® were incubated for nine months with microbes from PCB‐contaminated sites (Silver Lake, MA, USA, or River Raisin, MI, USA), and the products of dechlorination were then extracted. Exposure of RINm5F cells to dechlorinated Aroclor 1242 or 1254 product mixtures caused an increase in insulin release similar to the hormone release from cells exposed to non‐dechlorinated Aroclors. When tested alone, several of the major products identified in the dechlorination mixture (i.e., 2,2′,4,4′‐tetrachlorobiphenyl, 2,2′,4‐trichlorobiphenyl [TCB], 2,3′,4‐TCB, 2,3′,5‐TCB, and 2,2′‐dichlorobiphenyl) caused an increase in insulin release. In studies using neutrophils isolated from rat peritoneum, the amount of O2 produced on exposure to product mixtures resulting from dechlorination of Aroclor 1242 was not different from the amount produced in nondechlorinated controls. The product mixture resulting from Aroclor 1254 dechlorination by organisms from River Raisin increased generation of O2− relative to the parent Aroclor. Taken together, these results suggest that anaerobic dechlorination of Aroclor mixtures of PCBs does not reduce the biological activities associated with lightly chlorinated and ortho‐substituted PCBs. This observation has implications for the usefulness of PCB bioremediation efforts that involve only anaerobic dechlorination.

BIOLOGICAL ACTIVITY ASSOCIATED WITH NONCOPLANAR POLYCHLORINATED BIPHENYLS AFTER MICROBIAL DECHLORINATION OF AROCLOR 1242® AND AROCLOR 1254®

Bioremediation of polychlorinated biphenyls (PCBs) byanaerobic microbial dechlorination occurring naturally in the subsurface and in engineered systems results in mixtures of lower-chlorinated, primarily ortho-substituted biphenyls. The purpose of this study was to determine whether this process of bacterial dechlorination results in a mixture that differs in biological activity from that of the parent PCB mixture. Two biological assays sensitive to the action of ortho-substituted PCBs were employed: insulin release by RINm5F cells, and superoxide anion (O2) production by rat neutrophils. The PCB mixtures Aroclor 1242® and Aroclor 1254® were incubated for nine months with microbes from PCB-contaminated sites (Silver Lake, MA, USA, or River Raisin, MI, USA), and the products of dechlorination were then extracted. Exposure of RINm5F cells to dechlorinated Aroclor 1242 or 1254 product mixtures caused an increase in insulin release similar to the hormone release from cells exposed to non-dechlorinated Aroclors. When tested alone, several of the major products identified in the dechlorination mixture (i.e., 2,2′,4,4′-tetrachlorobiphenyl, 2,2′,4-trichlorobiphenyl [TCB], 2,3′,4-TCB, 2,3′,5-TCB, and 2,2′-dichlorobiphenyl) caused an increase in insulin release. In studies using neutrophils isolated from rat peritoneum, the amount of O2 produced on exposure to product mixtures resulting from dechlorination of Aroclor 1242 was not different from the amount produced in nondechlorinated controls. The product mixture resulting from Aroclor 1254 dechlorination by organisms from River Raisin increased generation of O2− relative to the parent Aroclor. Taken together, these results suggest that anaerobic dechlorination of Aroclor mixtures of PCBs does not reduce the biological activities associated with lightly chlorinated and ortho-substituted PCBs. This observation has implications for the usefulness of PCB bioremediation efforts that involve only anaerobic dechlorination.

Assessment of PCBs and hydroxylated PCBs as potential xenoestrogens: In vitro studies based on MCF-7 cell proliferation and induction of vitellogenin in primary culture of rainbow trout hepatocytes.

In the present study, four structurally diverse polychlorinated biphenyls (PCBs) were chosen from a set of 20 PCBs selected to represent the 154 tetra- through hepta-chlorinated biphenyls. The purpose was to determine estrogenic activities of the chosen PCBs and five of their hydroxylated derivatives (OH-PCBs). A human breast cancer cell line (MCF-7) and primary cultures of rainbow trout (Oncorhyncus mykiss) hepatocytes were used to determine estrogenic effects. The PCBs 2,2',4,6,6'-pentachlorobiphenyl (104) and 2,2',3, 4', 5,6,6'-heptachlorobiphenyl (188), and the hydroxylated PCBs 2,2', 4',6'-tetrachloro-4-biphenylol (4'-50), 2',4', 6'-trichloro-4-biphenylol (4'-30), 2',3,5, 5'-tetrachloro-4-biphenylol (4'-72), 2',3,3',5', 6'-pentachloro-4-biphenylol (4'-112), and 2',3,4',5, 6'-pentachloro-4-biphenylol (4'-121) significantly increased MCF-7 cell proliferation. The coaddition of hydroxytamoxifen, an estrogen antagonist, inhibited increased cell proliferation. The activity of the hydroxylated PCBs 4'-50 and 4'-30 was significantly higher at all nominal concentrations tested as compared to the corresponding PCB, viz., PCB 104. The hydroxylated PCBs 4'-50, 4'-30, 4'-72 and 4'-112 induced vitellogenin synthesis in rainbow trout hepatocytes. Significant differences were found in the MCF-7 system between the parent PCB and its hydroxylated derivative, viz., for 4'-50/4'-30 and 104, and in the rainbow trout hepatocyte assay between 4'-112 and 112, respectively. No activity was observed for PCB 58 in any of the two assays in the present study. Even though cells from two different species (human and fish) are used in the present study, the results obtained by the two methods agree fairly well. In both studies the hydroxylated PCBs were more active than the PCBs, and 4'-30 was the most active compound second only to 17beta-estradiol. http://link.springer-ny. com/link/service/journals/00244/bibs/37n2p145.html

Examination of the estrogenicity of 2,4,6,2',6'-pentachlorobiphenyl (PCB 104), its hydroxylated metabolite 2,4,6,2',6'-pentachloro-4-biphenylol (HO-PCB 104), and a further chlorinated derivative, 2,4,6,2',4',6'-hexachlorobiphenyl (PCB 155).

Several studies have reported that polychlorinated biphenyls (PCBs) exhibit estrogenic activity; however, it is not clear if these responses are associated with the polychlorinated hydrocarbon or its hydroxylated metabolite. In order to further test this hypothesis, a battery of in vitro and in vivo assays were used to investigate the estrogenic and antiestrogenic activities of 2,4,6,2',6'-pentachlorobiphenyl (PCB 104), its para-hydroxylated derivative 2,4,6,2',6'-pentachloro-4-biphenylol (HO-PCB 104), and its para-chlorinated derivative 2,4,6,2',4',6'-hexachlorobiphenyl (PCB 155). PCB 104 was found to 1) compete with tritiated 17beta-estradiol (E2) for binding to the mouse uterine estrogen receptor (ER); 2) induce gene expression in MCF-7 human breast cancer cells transiently transfected with the Gal4-human ER chimeric construct (Gal4-HEGO) and the Gal4-regulated luciferase reporter gene (17m5-G-Luc); and 3) increase MCF-7 cell proliferation in a dose-dependent manner. HO-PCB 104 exhibited greater estrogenic activity than PCB 104 in the in vitro assays examined. However, gas chromatographic-mass spectrophotometric analysis of extracts prepared from MCF-7 cells incubated with PCB 104 failed to detect the presence of the expected major metabolite HO-PCB 104. The estrogenic activity of the para-chlorinated derivative, PCB 155, was minimal compared to PCB 104 and HO-PCB 104, but it did exhibit significant antiestrogenic activity following co-treatment with 1 nM E2. Co-treatment of PCB 104 with 1 nM E2 had no effect on reporter gene expression compared to E2 alone, while 10 microM HO-PCB 104 exhibited additivity with 1 nM E2. At a dose of 202 mg/kg,PCB 104 increased uterine wet weight in ovariectomized CD-1 mice and induced vaginal epithelial cell cornification at 202, 16, and 1.7 mg/kg in a dose-dependent manner. These studies demonstrate that in addition to the hydroxylated metabolites, selected parent PCB congeners may also exhibit estrogenic and antiestrogenic activities.

Characterization of hepatic microsomal cytochrome P-450 from rats treated with methylsulphonyl metabolites of polychlorinated biphenyl congeners

The inducing potency of 3-methylsulphonyl(MeSO 2)-2,2′,4′,5,5′-pentachlorobiphenyl (pentaCB), which was one of the major MeSO 2 metabolites of polychlorinated biphenyls (PCBs) present in seal blubber[1], on the hepatic drug metabolizing enzyme activities was examined in comparison with that of the parent compound and phenobarbital (PB). The inducing fashion of the above enzymes and changes in the contents of PB-inducible P-450 forms by 2,3′,4′,5-tetrachlorobiphenyl (tetraCB) (IU-70), 2,2′,3′,4′,5-pentaCB (IU-87), 2,2′,4′,5,5′-pentaCB (IU-101) and 2,2′,3′,4′,5,5′-hexachlorobiphenyl (hexaCB) (IU-141), and their MeSO 2 metabolites were investigated in rats. Administration at various doses (0.2-1.0 μmol/kg) of 3-MeSO 2-2,2′,4′,5,5′-pentaCB produced nearly dose-related increases in the hepatic concentration of this methyl sulphone, in the contents of cytochromes P-450 and b 5, and in activities of aminopyrine N-demethylase, 7-ethoxycoumarin O-deethylase and benzo[ a]pyrene hydroxylase of liver microsomes. Major PB-inducible forms, CYP2B1, CYP2B2, CYP3A2 and CYP2C6 were induced with four PCBs (342 μmol/kg) and their 3-MeSO 2 metabolites (0.5–10 μmol/kg), indicating that 3-MeSO 2 metabolites were strong PB-type inducers of hepatic drug-metabolizing enzymes. 3-MeSO 2-2,2′,4′,5,5′-pentaCB was an especially strong inducer. On the other hand, four PB-inducible forms of cytochrome P-450 were not induced with the 4-MeSO 2 isomers. The relation between liver concentrations of the corresponding 3-MeSO 2 derivatives and induction of four PB-inducible forms of cytochrome P-450 after administration of four PCBs and their 3-MeSO 2 derivatives further confirmed that the 3-MeSO 2 metabolites played an important role in the induction which parent PCB congeners caused on the hepatic drug-metabolizing enzyme system.

Carcinogenicity of Black Rock Harbor Sediment to the Eastern Oyster and Trophic Transfer of Black Rock Harbor Carcinogens from the Blue Mussel to the Winter Flounder

The eastern oyster (Crassostrea virginica) developed neoplastic disorders when experimentally exposed both in the laboratory and field to chemically contaminated sediment from Black Rock Harbor (BRH), Bridgeport, Connecticut. Neoplasia was observed in oysters after 30 and 60 days of continuous exposure in a laboratory flow-through system to a 20 mg/L suspension of BRH sediment plus postexposure periods of 0, 30, or 60 days. Composite tumor incidence was 13.6% (49 neoplasms in 40, n = 295) for both exposures. Tumor occurrence was highest in the renal excretory epithelium, followed in order by gill, gonad, gastrointestinal, heart, and embryonic neural tissue. Regression of experimental neoplasia was not observed when the stimulus was discontinued. In field experiments, gill neoplasms developed in oysters deployed in cages for 30 days at BRH and 36 days at a BRH dredge material disposal area in Central Long Island Sound, and kidney and gastrointestinal neoplasms developed in caged oysters deployed 40 days in Quincy Bay, Boston Harbor. Oysters exposed to BRH sediment in the laboratory and in the field accumulated high concentrations of polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), and chlorinated pesticides. Chemical analyses demonstrated high concentrations of PCBs, PAHs, chlorinated pesticides, and heavy metals in BRH sediment. Known genotoxic carcinogens, co-carcinogens, and tumor promoters were present as contaminants. The uptake of parent PAH and PCBs from BRH sediment observed in oysters also occurs in blue mussels (Mytilus edulis). Winter flounder fed BRH-contaminated blue mussels contained xenobiotic chemicals analyzed in mussels. The flounder developed renal and pancreatic neoplasms and hepatotoxic neoplastic precursor lesions, demonstrating trophic transfer of sediment-bound carcinogens up the food chain.

Carcinogenicity of Black Rock Harbor sediment to the eastern oyster and trophic transfer of Black Rock Harbor carcinogens from the blue mussel to the winter flounder.

The eastern oyster (Crassostrea virginica) developed neoplastic disorders when experimentally exposed both in the laboratory and field to chemically contaminated sediment from Black Rock Harbor (BRH), Bridgeport, Connecticut. Neoplasia was observed in oysters after 30 and 60 days of continuous exposure in a laboratory flow-through system to a 20 mg/L suspension of BRH sediment plus postexposure periods of 0, 30, or 60 days. Composite tumor incidence was 13.6% (49 neoplasms in 40, n = 295) for both exposures. Tumor occurrence was highest in the renal excretory epithelium, followed in order by gill, gonad, gastrointestinal, heart, and embryonic neural tissue. Regression of experimental neoplasia was not observed when the stimulus was discontinued. In field experiments, gill neoplasms developed in oysters deployed in cages for 30 days at BRH and 36 days at a BRH dredge material disposal area in Central Long Island Sound, and kidney and gastrointestinal neoplasms developed in caged oysters deployed 40 days in Quincy Bay, Boston Harbor. Oysters exposed to BRH sediment in the laboratory and in the field accumulated high concentrations of polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), and chlorinated pesticides. Chemical analyses demonstrated high concentrations of PCBs, PAHs, chlorinated pesticides, and heavy metals in BRH sediment. Known genotoxic carcinogens, co-carcinogens, and tumor promoters were present as contaminants. The uptake of parent PAH and PCBs from BRH sediment observed in oysters also occurs in blue mussels (Mytilus edulis). Winter flounder fed BRH-contaminated blue mussels contained xenobiotic chemicals analyzed in mussels. The flounder developed renal and pancreatic neoplasms and hepatotoxic neoplastic precursor lesions, demonstrating trophic transfer of sediment-bound carcinogens up the food chain.

Polychlorinated biphenyl retention time standards obtained by chemical dechlorination of polychlorinated biphenyl isomers

Polychlorinated biphenyls (PCBs) were obtained by the partial catalytic dechlorination of individual PCBs having up to six chlorine atomes. The dechlorination was accomplished with sodium borohydride and nickel boride catalysis generated in situ. All of the expected PCB congeners were observed when decholorinating PCBs with four or less chlorine atoms. The mixtures were separated by gas chromatography on a methyl silicone capillary column. Identification of individual congeners was made by comparison of retention indices (RI) to literature, experimental and estimated values and by cross comparison of peaks generated from different parent PCBs. Various methods of estimating RIs were evaluated. The effects of varying concentration and reaction time was studied along with the relative reactivity of chlorines in different positions. The partially dechlorinated PCB mixtures are particularly useful as retention time standards for PCBs with less than four chlorine atoms.