Polychlorinated biphenyls (PCBs) are ubiquitious lipophilic environmental pollutants. At least some of the PCB congeners and mixtures are hepatic tumor promoters. The mechanisms are not fully understood and might be multifactorial Besides being the most abundant congener in the environment, 2,2,4,4,5,5-hexachlorobiphenyl (PCB-153), has been previously shown to increase hepatocyte proliferation 48h after exposure in rats. The goal of this study was to determine whether hepatic Kupffer cells are important in the promoting activity of PCBs. The hypothesis of this study was that modulation of Kupffer cell activity by PCBs may contribute to PCB-induced liver tumor promotion. The experimental approach consisted on three in vivo models (tumor promotion model and two short term exposure models) and one in vitro model. In the tumor promotion model, glycine inactivation of Kupffer cells did not significantly influence the promoting activity of PCB-77 (3,3,4,4-tetrachlorobiphenyl) or PCB-153. For the short term exposure model, we investigated the effect of Kupffer cell inactivation by glycine and the effect of Kupffer cell depletion on PCB-153s impact on hepatocyte proliferation. The oil used as a vehicle had no significant effect on any of the end points considered. Inhibition of Kupffer cells with glycine or the absence of Kupffer cells did not affect cell proliferation or NF-B activation after PCB treatment compared to the control. In vitro, PCB-153 increased DNA binding activity of NF-B in Kupffer cells but did not significantly increase the TNF- concentration in the medium. In conclusion, PCB-153 increased the number of preneoplastic foci per liver in the casein group but had no significant effect on cell proliferation, and Kupffer cells do not seem to play a role in hepatocyte proliferation.

Diseases, such as cardiovascular disease (CVD), are linked to chronic low levels of inflammation. This inflamed state is the product of risk factors including exposure to environmental pollutants, such as polychlorinated biphenyls (PCBs), which are correlated with increased risk for CVD and diabetes. In response to this health risk, our research addresses the mechanisms by which coplanar PCBs elicit an inflammatory response and the mitigation of PCB-induced inflammation through dietary intervention using docosahexaenoic acid (DHA), an omega-3 lipid. Investigators from the University of Kentucky Engineering Department are developing remediation technologies that detoxify PCBs through dechlorination. We studied the cellular toxicity of coplanar PCB 77 remediation products in primary vascular endothelial cells. The dechlorination products elicited different toxicological responses, which were less than the parent compound and contributed to the overall inflammatory response. The presence of PCB 77 at any concentration was sufficient to promote an inflammatory response, which was attenuated with complete dechlorination. PCB 77 is a good model for coplanar PCB-induced toxicity, but in environmental and human samples, coplanar PCB 126 is detected more frequently. Using different doses of PCB 126, we determined that acute exposure to 5 μmol PCB 126/kg mouse was sufficient to produce an inflammatory response without inducing a toxic wasting phenotype. PCB-induced inflammation was attenuated in vitro by DHA-derived neuroprostanes. Applying this information, we fed mice a DHA-enriched diet and exposed them to PCB 126. Liver and adipose lipid profiles confirm an increase in omega-3 fatty acid composition and DHA metabolites, and changes in gene expression indicate a heightened anti-oxidant response in the presence of PCB-induced inflammation. These data provide an overview of the in vivo response to a PCB-induced inflammation after DHA dietary feeding. We have demonstrated that PCB-induced endothelial dysfunction is propagated through lipid domains called caveolae. Caveolae are also signaling domains for toll-like receptor 4 (TLR4), and receptor for lipopolysaccharide (LPS). Similar to PCBs, TLR4 signaling is inhibited by DHA. We compared the caveolae-associated signaling response after exposure to coplanar PCB 126 or LPS. The domain localization of caveolae was altered by both PCB 126 and LPS. Our study determined that PCB 126-induced inflammation was not inhibited by a TLR4-specific inhibitor, but caveolae-based signaling was critical to both PCB 126- and LPS-induced inflammation. Environmental pollutants, such as coplanar PCBs, are risk factors in the development of chronic diseases. Here we investigate possible signaling pathways associated with environmental toxicity and apply potential dietary interventions with omega-3 lipids.

This dissertation combines laboratory and field experiments to investigate the mechanisms of atropisomer enrichment for chiral polychlorinated biphenyls (PCBs) and their metabolites in organisms. Stereoselective biotransformation and bioaccumulation were identified as two major reasons for the different environmental fate of PCB atropisomers. Other affecting factors, such as presence of nanoparticles and changes in feeding ecology of organisms, also affect the fate of chiral contaminants. In vitro incubations of rat cytochrome P-450 2B1 (CYP2B1) isozyme with chiral PCBs indicated that different biotransformation kinetics and competition among PCB congeners or between atropisomers were two main factors affecting atropisomer enrichment. Different interactions between chiral PCB congeners or atropisomers with rat CYP2B1 may occur at the molecular level. Non-racemic meta-hydroxylated-PCBs (5-OH-PCBs) were the major metabolites. CYP-mediated stereoselective formation of dihydroxylated PCBs from OH-PCBs was observed. Gold nanoparticles affected biotransformation activity of rat CYP2B1 and changed PCB atropisomeric composition, directly by electrostatic interaction, or indirectly by changes to the surrounding ionic strength. Thus, stereoselective metabolism of chiral PCBs and OH-PCBs by CYPs is a major mechanism for atropisomer enrichment of PCBs and their metabolites in the environment, with the degree of enrichment dependent, at least in part, on charged nanoparticles and stereoselective interference of atropisomers with each other at the enzyme level. The atropisomer compositions of chiral PCBs were measured in the marine biota of Cumberland Sound (Canada) and Svalbard (Norway). High trophic level organisms, including harp seal, beluga, and narwhal reported for the first time, had species-specific atropisomer signatures, likely due to a combination of in vivo biotransformation and trophic transfer. PCB chiral signatures in Greenland sharks supported the hypothesis that some of these PCB atropisomer compositions shifted over time and space, possibly due to a change in feeding ecology. To our knowledge, this is the first report to investigate temporal trends of PCB atropisomer signatures in Arctic biota.

Persistent organic pollutants (POPs) have been contaminating the marine environment since the mid 2Oth Century and continue to do so today. The polychlorinated biphenyls (PCBs) are of particular concern, since they are found at high concentrations in marine mammals throughout the Northern Hemisphere, and have been associated with endocrine disruption, reproductive impairment, immunotoxicity, and outbreaks of disease. In this study, samples were obtained from free-ranging harbour seal (Phoca vituilna) pups, which were live-captured in British Columbia, Canada, and Washington State, USA, in order to assess adverse health effects associated with POPs on the immune system of these mammals. PCBs were the most abundant of the 31 POPs measured in seals, and represented the greatest toxicological concern on the basis of established reference values for laboratory rodents and aquatic wildlife consumers. Seal immune function was assessed using traditional measures of immunotoxicity, including hematology, innate immune function, and adaptive immune function, and related to PCB concentrations while carefully controlling for confounding factors such as age, sex and condition. PCB concentrations negatively correlated to phagocytosis, T lymphocyte proliferative responses, (thymosin-a1-induced) lymphocyte signalling, and lymphocyte counts, and positively associated with the respiratory burst of phagocytes and aryl hydrocarbon receptor (AhR) expression of white blood cells, suggesting chemical-associated immunotoxicity. Parallel experiments, in which harbour seal white blood cells were exposed in vitro to 2,3,7,8-tetrachlorodibenzo-p-dioxin, a potent immunotoxicant, further supported the hypothesis that the observed reduction in immunocompetence in free-ranging seals may be due to an AhR-mediated mechanism of immunotoxicity. Principal component analysis (PCA) of immunological endpoints combined evidence of PCB-associated effects on the immune system as a whole. However, PCA also identified a difference between the immunological profiles of urban seals and those from remote sites, consistent with elevated pathogen abundance due to biological pollution at urban sites. In a second approach, PCB concentrations were related to concentrations of vitamin A and expression levels of its receptor, the retinoic acid receptor alpha (RARa), which are known to be sensitive to PCB exposure. More contaminated seals were characterised by lower vitamin A concentrations in circulation and in the blubber, as well as higher RARa expression in the blubber. AhR expression and concentrations of thymosin-a1 (a thymic hormone important in lymphocyte development and immune function) did not relate to contaminants when their levels were investigated in seal tissues (blubber, skin), contrasting their sensitivity in white blood cells. These results implied that PCB-associated toxicity may be reflected if a biomarker represents a primary lesion, but might not be extrapolated among tissues in all cases. Secondly, although blubber represents the primary site of PCB storage, it is not necessarily the site of the highest toxicity. In summary, significant evidence of immunotoxicity and disruption of immune function-related biomarkers has been provided in a healthy group of free-ranging marine mammals which contributes to the weight of evidence that environmental contaminants may render marine mammal populations more vulnerable to disease through immunotoxicity. Immune function measures in free-ranging harbour seal pups exposed to contaminants in vivo, in combination with harbour seal white blood cells in vitro, furthermore suggested that immunotoxicity may take place through an AhR-mediated mechanism of action. An unexpected finding was the evidence of a second, independent, impact on the immune system of seals, consisting of biological pollution. The combination of both chemical and biological pollution, that would imply both diminished immune responses and increased pathogen loads, may represent the largest threat to the health of marine mammals in many parts of the world.

The fate and bioaccumulation of a contaminant entering the marine environment through wastewater outfalls depends on the contaminant’s persistence and affinity for particles. The physical characteristics of the receiving environment, e.g. current velocity, sedimentary processes, and the availability of organic carbon are also important. However, these latter effects are not usually evaluated quantitatively. This thesis investigates the near-field accumulation in sediment and biota of particle-reactive polybrominated diphenyl ethers (PBDEs) entering coastal waters via two municipal outfalls: one discharging into a high energy, low sedimentation environment near Victoria, B.C., Canada; the other into a low energy, high sedimentation environment near Vancouver, B.C. We used 210Pb profiles in sediment box cores together with an advection-diffusion model to determine surface mixing and sedimentation rates, and to model the depositional history of PBDEs at these sites. A particularly important finding of this study is that the very high energy environment to the southeast of the Victoria outfall accumulates PBDEs despite not having net sediment accumulation. Although the discharge of PBDEs was much lower from the Victoria outfall than from Vancouver, some sediment PBDE concentrations were higher near Victoria. Most PBDEs were dispersed beyond the near-field at both sites, but a greater proportion was captured in the sediment near the Vancouver outfall where rapid burial was facilitated by inorganic sediment supplied from the nearby Fraser River. Clearly, treating wastewater to the same level, regardless of local oceanographic conditions, will not result in a uniform environmental footprint. Total PBDE concentrations in benthic invertebrate communities were higher near Vancouver than Victoria, despite lower concentrations in sediments, and correlated with organic carbon normalized sediment concentrations. Principal Components Analysis suggested uptake of individual PBDE congeners was determined by sediment properties (TOC, grain size), whereas PCB congener uptake was governed by physico-chemical properties (octanol-water partitioning coefficient). Our results suggest that sediment quality guidelines for PBDEs and likely PCBs may be more relevant if corrected to TOC content in sediment. In addition, where enhanced wastewater treatment increases the ratio of PBDEs to particulate organic carbon in effluent, nearfield benthic invertebrates may face increased PBDE accumulation. This underlines the need for source control of persistent organic contaminants, which cannot be broken down by conventional wastewater treatment. / Graduate

Ligand I- along with nonionic surfactant, Triton X-100 or anionic surfactant, sodium dodecyl sulfate (SDS) were applied as soil washing agents to desorb Cd from both naturally and artificially contaminated soils. After seven consecutive washings, up to 90% of Cd was desorbed from both soils. Triton X-100 with I- showed a higher capacity to desorbing Cd than did SDS with I-. The increase of ligand concentration was a critical factor for increasing leaching capacity. Without the ligand, surfactant alone could not desorb Cd effectively from either soil. After seven consecutive washings, a sequential extraction experiment was performed for soil residuals to define the soil fraction of Cd removed by the washing agent. Among the washing agents, only Triton X-100/I- could remove Cd from the exchangeable fraction of both soils. / Various ligands including I-, SCN-, and I-/SCN- in combination with Triton X-100 were tested for their efficacy in desorbing heavy metals such as Cd, Zn, Cu, and Pb from a field contaminated soil. Cadmium was preferentially desorbed by Triton X-100/I- whereas Zn and Cu were preferentially desorbed by Triton X-100/SCN-. The mixture of I- and SCN- with Triton X-100 desorbed the most Cd and Cu, but not for Zn, as I- inhibited Zn desorption. Sequential extraction experiments after seven washings showed that metals held in the exchangeable fraction can be desorbed only by a combination of ligand and surfactant. / Nonionic surfactants having different alkyl chain lengths in combination with ligand I- were tested for the desorption of Cd and PCBs from soil. Cadmium desorption was increased at the lower surfactant concentration and higher ligand concentration. The increase in the hydrophilic alkyl chain length of the surfactant adversely affected Cd desorption. Up to 100% of PCBs were successfully removed by most of surfactant-ligand combinations and the desorption was less dependent, compared to heavy metals, on the concentration of washing agents and length of alkyl chain. The linear relationships between number of washings and Cd desorption, and between alkyl chain length and Cd desorption was defined. (Abstract shortened by UMI.)

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning). Uppsala : Sveriges lantbruksuniv., 2003. / Härtill 4 uppsatser.

Thesis (M.Sc.) (Hons.) -- University of Western Sydney, 2002. / A thesis submitted to the University of Western Sydney in fulfillment of the requirements for admission to the [degree of] Masters of Science (Honours). Bibliography : leaves 156-163.