Micro-distribution of arsenic in toenail clippings using laser ablation inductively coupled plasma mass spectrometry: implications for biomonitoring.

Toenails are a common monitoring tool for arsenic exposure, but the risk of external contamination of toenails has cast doubt on its usefulness. The main objective of this study is to investigate the micro-distribution of arsenic through the dorsoventral plane of nail clippings to understand endogenous vs exogenous sources. We used laser-ablation inductively coupled plasma mass spectrometry to measure arsenic through a dorsoventral cross-section of the nail plate collected from reference (N = 17) and exposed individuals (N = 35). Our main results showed (1) bulk toenail concentrations measured using ICP-MS in this study ranged from 0.54 to 4.35 µg/g; (2) there was a double-hump pattern in arsenic concentrations, i.e., dorsal and ventral layers had higher arsenic than the inner layer; (3) the double-hump was more pronounced in the exposed group (ventral: 6.25 µg/g; inner: 0.75 µg/g; dorsal: 0.95 µg/g) than the reference group (ventral: 0.58 µg/g; inner: 0.15 µg/g; dorsal: 0.29 µg/g) on average; (4) the distribution was, in part, associated with different binding affinity of nail layers (i.e., ventral > dorsal > inner); (5) most individuals in the higher exposure group showed > 25% contamination in ventral and dorsal nail layers; and (6) there were no statistically significant correlations between LA-ICP-MS arsenic with either bulk toenail arsenic or urine arsenic from the same individuals. Our results on micro-distribution and binding affinity provide insight into the impact of external contamination on arsenic concentrations and show how LA-ICP-MS can access the protected inner nail layer to provide a more accurate result.

Variation in biomarker levels of metals, persistent organic pollutants, and omega-3 fatty acids in association with genetic polymorphisms among Inuit in Nunavik, Canada.

BACKGROUND: Biomarker measures of contaminant exposure and nutrient status can help increase understanding of the risks and benefits associated with the consumption of traditional foods by Inuit. While gene-environment and gene-nutrient interactions may help explain variations in biomarker measures, the role of genetic polymorphisms is largely understudied especially for vulnerable sub-populations. OBJECTIVE: The aim of this study was to characterize the relationship between single nucleotide polymorphisms (SNPs) in key genes and blood concentrations of environmental chemicals and nutrients among Inuit. METHODS: Blood samples from 665 individuals who participated in the Qanuippitaa Survey (Nunavik, Canada) in 2004 were analyzed for toxicants and nutrients. DNA was extracted and 140 SNPs in classes relevant to the toxicokinetics and/or toxicodynamics of the target contaminants and nutrients, and/or are involved in cardiovascular health and lipid metabolism were genotyped using the Sequenom iPLEX Gold platform. RESULTS: Geometric means (µg/L) of mercury (Hg), cadmium (Cd), lead (Pb), DDE, PCB-153, and selenium (Se) were 11.1, 2.8, 39.9, 2.9, 1.1 and 301.2, respectively. Red blood cell membrane levels of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were 5.1%/total fatty acid (TFA) and 1.3%/TFA respectively. Out of 106 SNPs which met our inclusion criteria, biomarker levels for Hg, Cd, Pb, DDE, PCB-153, DHA, and EPA differed (p < 0.05) by genotype for 20, 13, 12, 19, 21, 9 and 8 SNPs, respectively. Following Bonferroni correction (p < 0.0005), only 9 SNPs remained significant (rs2274976 in MTHFR, rs174602 in FADS2, rs7115739 and rs74771917 in FADS3, rs713041 in GPX4, rs2306283 and rs4149056 in SLCO1B1, rs1885301 in ABCC2/MRP2, and rs4244285 in CYP2C19; 5 associated with Hg, 2 with Pb, 2 with DDE, 4 with PCB-153, 1 with DHA). CONCLUSIONS: The findings suggest that polymorphisms in environmentally-responsive genes can influence biomarker levels of key toxicants and nutrients. While there are no immediate clinical or public health implications of these findings, we believe that such gene-environment and gene-nutrient studies provide a foundation that will inform and provide direction to future studies.

Associations of health status and diabetes among First Nations Peoples living on-reserve in Canada.

OBJECTIVE: Our objective is to describe self-reported health status, prevalence of diabetes and obesity and their associations in participants from the First Nations Food, Nutrition and Environment Study (FNFNES) in order to identify possible correlates of health in First Nations adults. METHODS: FNFNES is a participatory study with First Nations Peoples living on reserve lands south of the 60th parallel. Health and diabetes were self-reported, and prevalence of obesity was evaluated. Socio-demographic and lifestyle factors and traditional food (TF) activities were investigated for associations with health parameters. RESULTS: High prevalence rates of overweight/obesity (78-91%) and diabetes (19% age-standardized prevalence) were found. Smoking rates were high and physical activity was low. In multivariable analyses, obesity was associated with region, income source, age, gender, smoking and self-reported health; diabetes and lesser self-reported health were associated with obesity and lower education. Diabetes was strongly associated with lesser self-reported health and weakly associated with being a smoker. CONCLUSION: We have identified possible correlates of health in this population that can help to better understand the underlying concerns and identify solutions for First Nations and their partners. We urge governments and First Nations to address the systemic problems identified with a holistic ecosystem approach that takes into consideration the financial and physical access to food, particularly TF, and the facilitation of improved health behaviour. New mechanisms co-developed with First Nations leadership should focus on supporting sustainable, culturally safe and healthy lifestyles and closing the gaps in nutrition and food insecurity.

RéSUMé: OBJECTIFS: Décrire l'état de santé autodéclaré, les prévalences de diabète et d'obésité et leurs associations chez les participants de l'Étude sur l'alimentation, la nutrition et l'environnement des Premières Nations (EANEPN) afin d'identifier d'éventuelles associations avec la santé chez les adultes des Premières Nations. MéTHODE: L'EANEPN est une étude participative entreprise avec les Peuples des Premières Nations vivant dans les réserves au sud du 60e parallèle. L'état de santé et le diabète ont été autodéclarés et la prévalence de l'obésité a été évaluée. Les facteurs sociodémographiques et de style de vie et les activités d'apports en aliments traditionnels (AT) ont été étudiés pour leurs associations avec les paramètres de santé. RéSULTATS: Une prévalence élevée de surpoids/obésité (78­91%) et de diabète (19% ­ taux normalisé selon l'âge) a été observée. Le taux de tabagisme était élevé et l'activité physique était faible. Dans les analyses multivariées, l'obésité était associée avec la région, la source de revenu, l'âge, le sexe, le tabagisme et l'état de santé autodéclaré; le diabète et l'état de santé faible autodéclarés étaient positivement associés à l'obésité et à moins d'années d'éducation. Le diabète était fortement associé à un faible état de santé autodéclaré et légèrement avec le tabagisme. CONCLUSION: Nous avons identifié des corrélats de santé potentiels dans cette population qui peuvent nous aider à mieux comprendre les préoccupations sous-jacentes. Nous exhortons les gouvernements et les Premières Nations à s'attaquer de concert aux problèmes systémiques identifiés avec une approche écosystémique holistique qui prend en considération l'accès financier et physique à la nourriture, en particulier les AT, et qui facilite une amélioration dans les comportements en santé. De nouveaux mécanismes élaborés conjointement avec les dirigeants des Premières Nations devraient viser à soutenir des modes de vie durables, culturellement sécuritaires et sains et à combler les lacunes en matière de nutrition et d'insécurité alimentaire.

The Gut Microbial Community Structure of the North American River Otter (Lontra canadensis) in the Alberta Oil Sands Region in Canada: Relationship with Local Environmental Variables and Metal Body Burden.

The Alberta Oil Sands Region in Canada is home to one of the largest oil bitumen deposits in the world. The North American river otter (Lontra canadensis) is a top predator with a small home range and is sensitive to disturbances; it has been designated as a sentinel species for the potential impacts of the natural resource exploitation on freshwater ecosystems in the Alberta Oil Sands Region. With an increasing interest in noninvasive biomarkers, recent studies suggest that gut microbiota can be used as a potential biomarker of early biological effects on aquatic wildlife. The goal of the present study was to determine the river otter gut microbial structure related to environmental variables characterizing mining activities and metal body burden. We obtained 18 trapped animals from and surrounding the surface mineable area of the Alberta Oil Sands Region. The gut microbial community structure was characterized using high-throughput sequencing of 16S rRNA gene amplicon analyses. Trace metal concentrations in the liver were measured by inductively coupled plasma-mass spectrometry. Our study revealed that the gut bacteria of river otters in the Alberta Oil Sands Region clustered in 4 groups dominated by Peptostreptococcaceae, Carnobacteriaceae, Enterobacteriaceae, Clostridiaceae, and Nostocaceae. We show that arsenic, barium, rubidium, liver-body weight ratio, and δ15 N were associated with each cluster. When comparing affected versus less affected sites, we show that river otter gut bacterial community and structure are significantly related to trophic level of the river otter but not to Alberta Oil Sands Region mining activities. Our study reveals that the gut bacterial dynamics can provide insights into the diet and habitat use of river otters but that more work is needed to use it as a pollution biomarker. Environ Toxicol Chem 2020;39:2516-2526. © 2020 SETAC.

Geospatial analysis of the patterns of chemical exposures among biota in the Canadian Oil Sands Region.

Understanding the patterns of chemical exposure among biota across a landscape is challenging due to the spatial heterogeneity and complexity of the sources, pathways, and fate of the different chemicals. While spatially-driven relationships between contaminant sources and biota body burdens of a single chemical are commonly modelled, there has been little effort on modelling chemical mixtures across multiple wildlife species in the Canadian Oil Sands region. In this study, we used spatial principal components analysis (sPCA) to assess spatial patterns of the body burdens of 22 metals and Potentially Toxic Elements (PTEs) in 492 individual wildlife, including fur-bearing mammals, colonial waterbirds, and amphibians collected from the Canadian Oil Sands region in Canada. Spatial analysis and mapping both indicate that some of the complex exposures in the studied biota are distributed randomly across a landscape, which suggests background or non-point source exposures. In contrast, the pattern of exposure for seven metals and PTEs, including mercury, vanadium, lead, rubidium, lithium, strontium, and barium, exhibited a clustered pattern to the east of the open-pit mining area and in regions downstream of oil sands development which indicates point-source input. This analysis demonstrated useful methods for integrating monitoring datasets and identifying sources and potential drivers of exposure to chemical mixtures in biota across a landscape. These results can be used to support an adaptive monitoring program by identifying regions needing additional monitoring, health impact assessments, and possible intervention strategies.

Health risk assessment of arsenic exposure among the residents in Ndilǫ, Dettah, and Yellowknife, Northwest Territories, Canada.

There are concerns in Yellowknife, Northwest Territories, Canada, about arsenic exposure due to past mining operations, particularly the former Giant Mine. The objective of this study was to characterize the risk of arsenic exposure and associated risk factors among the local residents. Arsenic (As) and its species were quantified in urine (n = 1966) using inductively coupled mass spectrometry. Children in the study were found to have significantly higher (p < 0.05) urinary inorganic-related As (uiAs) concentrations than children in the general Canadian population, as well as adults in the study. Additionally, uiAs concentrations in children, particularly those above the 95th percentile, are above the Biomonitoring Equivalents (BE) levels that are associated with dermal effects, vascular problems and cancer risks. Multiple linear regression results showed that market seafood (fish and shellfish) and rice consumption frequency were significantly positively associated with uiAs. Specific to children, drinking lake water was positively associated with uiAs. Specific to adults, consumption of local mushrooms and berries were significantly positively associated with uiAs while there was a significant negative association with age, smoking and recreational water activities. The risk factors identified in this research can be used for public health education to lower arsenic intake. Overall, these results support the need for an ongoing monitoring program.

A metabolomics study on effects of polyaromatic compounds in oil sand extracts on the respiratory, hepatic and nervous systems using three human cell lines.

Polyaromatic compounds (PACs) are by-products of combustion and are the major pollutants from the oil and gas industry. However, the mechanism of PACs induced toxicity still remains elusive. The aim of this study was to elucidate the effects of a typical mixture of PACs found in oil sand extract (OSE) on the respiratory, hepatic and nervous systems in humans using in vitro cell culture models followed by non-targeted metabolomics analysis. OSE collected from Alberta, Canada was fractionated into PAC and alkane fractions, and their effects after 24 h exposure on the cell viability measured by MTT assay in three human cell lines (A549, HepG2, and SK-N-SH) were studied. The PAC fractions showed significant dose-dependent cytotoxicity. A549 cells showed the highest sensitivity to OSE extracts, followed by SK-N-SH and HepG2. In contrast, the alkane fractions showed no effects on cell viability. The three human cell lines were further exposed with the PACs at 10% and 20% lethal concentration for 24 h. Metabolomics analysis of the cell extracts indicated that PACs treatments showed different disruptions on possible metabolic pathways on the three cell lines. PACs altered the sex steroid hormone metabolism and regulated the levels of leukotrienes metabolites in all three cell types. The amino acids L-cysteine, L-glutamine, L-tyrosine that are known to cause respiratory effects were significantly up-regulated in A549 cells. The PACs treated HepG2 cells showed down-regulation in metabolites responsible for the inflammatory mediation. Treatment of the differentiated SK-N-SH cells showed up-regulated metabolites involved with butanoate, fatty acid, and pyrimidine metabolism. Leukotriene metabolites were found to be significantly increased in all PACs treated cells. In conclusion, our results showed that PACs in OSE can alter the metabolism of the human lung, liver and neuronal cells and may induce toxicity in multiple target organs.

The Use of Geographic Information Systems for Spatial Ecological Risk Assessments: An Example from the Athabasca Oil Sands Area in Canada.

There is an acknowledged need in ecotoxicology for methods that integrate spatial analyses in risk assessment. This has resulted in the emergence of landscape ecotoxicology, a subdiscipline of ecotoxicology. However, landscape ecotoxicology has yet to become common practice in risk assessment due to the underdevelopment of techniques and a lack of standardized methods. In the present study, we demonstrate how geographic information systems (GISs) can serve as a standardized platform to integrate data, assess spatial patterns of ecotoxicological data for multiple species, and assess relationships between chemical mixture exposures and effects on biota for landscape ecotoxicological risks assessment. We use data collected under the Joint Oil Sands Monitoring Program in the Athabasca Oil Sands Region in Alberta, Canada. This dataset is composed of concentrations of contaminants including metals and polycyclic aromatic compounds, and health endpoints measured in 1100 biological samples, including tree swallows, amphibians, gull and tern eggs, plants, and mammals. We present 3 examples using a GIS as a platform and geospatial analysis to: 1) integrate data and assess spatial patterns of contaminant exposure in the region, 2) assess spatial patterns of exposures to complex mixtures, and 3) examine patterns of exposures and responses across the landscape. We summarize the methods used in the present study into a workflow for ease of use. The GIS methods allow researchers to identify hot spots of contamination, use georeferenced monitoring data to derive quantitative exposure-response relationships, and assess complex exposures with more realism. Environ Toxicol Chem 2019;38:2797-2810. © 2019 SETAC.

Seafood consumption patterns, their nutritional benefits and associated sociodemographic and lifestyle factors among First Nations in British Columbia, Canada.

OBJECTIVE: To describe seafood consumption patterns in First Nations (FN) in British Columbia (BC) and examine lifestyle characteristics associated with seafood consumption; to identify the top ten most consumed seafood species and their contributions to EPA and DHA intake; and to estimate dietary exposure to methylmercury, polychlorinated biphenyls and dichlorodiphenyldichloroethylene. DESIGN: Dietary and lifestyle data from the First Nations Food Nutrition and Environment Study, a cross-sectional study of 1103 FN living in twenty-one communities across eight ecozones in BC, Canada, were analysed. Seafood consumption was estimated using a traditional FFQ. Seafood samples were analysed for contaminant contents. RESULTS: Seafood consumption patterns varied significantly across BC ecozones reflecting geographical diversity of seafood species. The top ten most consumed species represented 64 % of total seafood consumption by weight and contributed 69 % to the total EPA+DHA intake. Mean EPA+DHA intake was 660·5 mg/d in males, 404·3 mg/d in females; and 28 % of FN met the Recommended Intake (RI) of 500 mg/d. Salmon was the most preferred species. Seafood consumption was associated with higher fruit and vegetable consumption, lower smoking rate and increased physical activity. Dietary exposure to selected contaminants from seafood was negligible. CONCLUSIONS: In FN in BC, seafood continues to be an essential part of the contemporary diet. Seafood contributed significantly to reaching the RI for EPA+DHA and was associated with a healthier lifestyle. Given numerous health benefits, seafood should be promoted in FN. Efforts towards sustainability of fishing should be directed to maintain and improve access to fisheries for FN.

Neurotoxicity of alkylated polycyclic aromatic compounds in human neuroblastoma cells.

Polycyclic aromatic compounds (PAC) are ubiquitous environmental pollutants originating from incomplete combustion processes. While the toxicity of parent PAC such as benzo[a]pyrene (BaP) is well characterized, effects of other alkyl-PAC dibenzothiophene (DBT) and retene (Ret) are not well established. The aim of this study was to examine the underlying relative neurotoxic mechanisms attributed to BaP (parent PAH), DBT and Ret (alkyl-PACs) using human neuroblastoma SK-N-SH cells. The lethal concentrations (LC10 and LC20) were found at approximately 10 µM and 40 µM, respectively after 24-h exposure of SK-N-SH cells. It was hypothesized that PAC trigger reactive oxygen species (ROS) production, leading to activation of apoptotic signaling pathways. Differentiated neuronal cells were treated with three compounds at (0.5-40 µM) for 24 h. There was a significant concentration-dependent increase in levels of ROS, even at sub-lethal levels of 1 µM Ret. The mitochondrial membrane potential (MMP) was significantly decreased. Real-time RT-PCR results showed up-regulation of pro-apoptotic genes and down-regulation of antioxidative genes expression in BaP-, DBT-, and Ret-treated SK-N-SH cells. Cytochrome c protein levels and lipid peroxidation (LPO) were also significantly elevated in a concentration-related manner. Data demonstrated that BaP-, DBT-, or Ret-induced neuronal cell damage involved oxidative stress generation through mitochondria-mediated apoptosis pathway. Alkyl-PAC also exhibited higher potency in ROS induction and reduction of MMP than parent PAC. These findings may be important for environmental risk assessment attributed to exposure to PAC.

Acrylamide-induced neurotoxicity in primary astrocytes and microglia: Roles of the Nrf2-ARE and NF-κB pathways.

Acrylamide (AA) is a common food contaminant formed during food heat processing that has neurotoxic effects. We hypothesize that AA induces oxidative stress in astrocytes and microglia, leading to neurotoxicity. Oxidative status, translocation of Nrf2 and NF-κB, and related down-stream targets were measured in primary astrocytes and microglia obtained from BALB/c mice. The results showed that AA increased reactive oxygen species (ROS) formation and reduced glutathione levels, causing successive events associated with oxidative stress, including 4-hydroxynonenal and 8-hydroxy-2-deoxyguanosine adduct formation, in both cell types. Both Nrf2 and NF-κB pathways were activated, but Nrf2 and its downstream antioxidative genes acted at earlier stages in both cell types before NF-κB activation. After NF-κB activation, related cytokines, including IL-6, TNF-α, G-CSF, and IL-1ß, were released and cell viability decreased. Greater ROS generation, faster glutathione reduction, and increased oxidative adduct formation were observed in microglia compared with astrocytes. Moreover, Nrf2/NF-κB and its downstream genes were up-regulated much faster and to greater degrees in microglia than astrocytes. These results clarify the roles of the Nrf2 and NF-κB pathways in AA-induced neurotoxicity. These cellular responses may provide new insights for the development of adverse outcome pathway approaches for risk assessments of AA.

Association between fish consumption, dietary omega-3 fatty acids and persistent organic pollutants intake, and type 2 diabetes in 18 First Nations in Ontario, Canada.

BACKGROUND: First Nations (FNs) populations in Canada experience a disproportionally higher rate of obesity and type 2 diabetes (T2D) compared to the general population. Recent data suggest that a high consumption of fish may help prevent T2D. On the other hand, fish might also be a potential source of environmental contaminants which could potentially be a risk factor for T2D. OBJECTIVE: To investigate the potential associations between self-reported T2D and consumption of locally-harvested fish, dietary long-chain omega-3 fatty acids (n-3FAs) and persistent organic pollutants intake among adult FNs living on reserve in Ontario. DESIGN: Data from the First Nations Food Nutrition and Environment Study, which included a cross-sectional study of 1429 Ontario FNs adults living in 18 communities across 4 ecozones in 2012 were analyzed. Social and lifestyle data were collected using household interviews. The consumption of locally-harvested fish was estimated using a traditional food frequency questionnaire along with portion size information obtained from 24hr recalls. Fish samples were analyzed for the presence of contaminants including dichlorodiphenyldichloroethylene (DDE) and polychlorinated biphenyls (PCBs). Dietary intakes of DDE and PCBs were estimated using community-specific levels of DDE/PCBs in fish species. Multiple logistic regression models adjusted for potential covariates including age, gender, body mass index, physical activity, total energy intake, smoking, and education were developed. RESULTS: The prevalence of T2D in Ontario FNs was 24.4%. A significant positive association between fish consumption of one portion per week and more and T2D compared to no fish consumption was found (OR=2.5 (95% CI: 1.38-4.58). Dietary DDE and PCBs intake was positively associated with T2D (OR=1.09 (95%CI: 1.05-1.75) for DDE and OR=1.07 (95%CI: 1.004-1.27) for PCBs) per unit increase in DDE/PCBs while n-3-FAs intake, adjusted for DDE/PCBs intake, showed an inverse effect against T2D among older individuals (OR=0.86 (95% CI: 0.46-0.99). CONCLUSION: Our results support previous findings that exposure to DDE and PCBs may increase the risk of T2D. Elevated levels of contaminants in fish may counteract with potentially beneficial effects of n-3FAs from fish consumption. However, the overall health benefits of high consumption of fish with a high n-3 FAs content may outweigh the adverse effect of contaminants.

Occurrence, sources and human exposure assessment of SCCPs in indoor dust of northeast China.

Short-chain chlorinated paraffins (SCCPs) are widely used chemicals in household products and might cause adverse human health effects. However, limited information is available on the occurrence of SCCPs in indoor environments and their exposure risks on humans. In this study the concentrations, profiles and human exposure of SCCPs in indoor dust from five different indoor environments, including commercial stores, residential apartments, dormitories, offices and laboratories were characterized. The SCCPs levels ranged from 10.1 to 173.0 µg/g, with the median and mean concentration of 47.2 and 53.6 µg/g, respectively. No significant difference was found on concentrations among the five microenvironments. The most abundant compounds in indoor dust samples were homologues of C13 group, Cl7 group and N20 (N is the total number of C and Cl) group. In the five microenvironments, commercial stores were more frequently exposed to shorter carbon chained and higher chlorinated homologues. Three potential sources for SCCPs were identified by the multiple linear regression of factor score model and correspondence analysis. The major sources of SCCPs in indoor dust were technical mixtures of CP-42 (42% chlorine, w/w) and CP-52 b (52% chlorine, w/w). The total daily exposure doses and hazard quotients (HQ) were calculated by the human exposure models, and they were all below the reference doses and threshold values, respectively. Monte Carlo simulation was applied to predict the human exposure risk of SCCPs. Infants and toddlers were at risk of SCCPs based on predicted HQ values, which were exceeded the threshold for neoplastic effects in the worst case. Our results on the occurrences, sources and human exposures of SCCPs will be useful to provide a better understanding of SCCPs behaviors in indoor environment in China, and to support environmental risk evaluation and regulation of SCCPs in the world.

Effect of acrylamide-induced neurotoxicity in a primary astrocytes/microglial co-culture model.

Acrylamide (AA), is a common food contaminant generated by heat processing. Astrocytes and microglia are the two major glial cell types in the brain that play pivotal but different roles in maintaining optimal brain function. The objective of this study is to investigate the neurotoxicity of AA, using a primary astrocytes/microglia co-culture model. Co-cultural cells obtained from Balb/c mice were cultured and treated with 0-1.0mM AA for 24-96h. Cell viability, reactive oxygen species (ROS) generation, oxidative end produces formation and glutathione (GSH) levels were measured. The expression of nuclear-E2-related factor 2(Nrf2), and nuclear factor kappa-beta (NF-κB) and selected down-stream genes were measured. Results showed that AA treatment led toa dose-dependent toxicity. Oxidative stress was induced as indicated by an increase of ROS, a decrease of GSH levels, and an increase in the formation of 4-hydroxynonenal-adduct and 8-hydroxy-2-deoxyguanosine-adduct. Both Nrf2 and NF-κB pathway contributed to the initiation of oxidative stress but the timing of two factors was different. Nrf2 and its related downstream genes were activated earlier than that in NF-κB pathway. In conclusion, AA-induced neurotoxicity attribute to oxidative stress via Nrf2 and NF-κB pathway. Moreover, the co-culture cell model was proven to be a viable model to study AA neurotoxicity.

A Comparative Assessment of Arsenic Risks and the Nutritional Benefits of Fish Consumption in Kuwait: Arsenic Versus Omega 3-Fatty Acids.

Inorganic and organic forms of arsenic (As), as well as omega-3 fatty acids were measured in 578 fish/seafood samples that belong to 15 species of commonly consumed seafood in Kuwait. Arsenic speciation data, with the toxicological profile of inorganic arsenic (iAs) and fish consumption rates were applied in a probabilistic risk assessment to estimate the risk from exposure to iAs. The nutritional benefits of omega-3-fatty acid levels in various species of fish were taken into consideration. Results showed that the mean daily intake of iAs through fish consumption among the Kuwaiti population was 0.058 µg/kg/day, and the 95th percentile was 0.15 µg/kg/day. Although the mean intake level did not exceed the incremental lifetime cancer risk (ILCR) at 1 × 10-4, the 95th percentile of iAs intake showed an ILCR of 2.7 × 10-4. Kuwaiti children (aged 6-12 years) were found to have a higher mean intake of iAs at 0.10 µg/kg/day with 68% of children in this category, exceeding the risk specific dose associated with an ILCR of 1 × 10-4. The fish species, hammor (grouper; Epinephelus coioides), is the top contributor to iAs intake, and tuna is the major source of omega 3-fatty acids for the Kuwaiti population.

Air pollution, neighbourhood and maternal-level factors modify the effect of smoking on birth weight: a multilevel analysis in British Columbia, Canada.

BACKGROUND: Maternal smoking during pregnancy negatively impacts fetal growth, but the effect is not homogenous across the population. We sought to determine how the relationship between cigarette use and fetal growth is modified by the social and physical environment. METHODS: Birth records with covariates were obtained from the BC Perinatal Database Registry (N = 232,291). Maternal smoking status was self-reported as the number of cigarettes smoked per day usually at the first prenatal care visit. Census dissemination areas (DAs) were used as neighbourhood-level units and linked to individual births using residential postal codes to assign exposure to particulate air pollution (PM2.5) and neighbourhood-level attributes such as socioeconomic status (SES), proportion of post-secondary education, immigrant density and living in a rural place. Random coefficient models were used with cigarettes/day modeled with a random slope to estimate its between-DA variability and test cross-level interactions with the neighbourhood-level variables on continuous birth weight. RESULTS: A significant negative and non-linear association was found between maternal smoking and birth weight. There was significant between-DA intercept variability in birth weight as well as between-DA slope variability of maternal smoking on birth weight of which 68 and 30 % respectively was explained with the inclusion of DA-level variables and their cross-level interactions. High DA-level SES had a strong positive association with birth weight but the effect was moderated with increased cigarettes/day. Conversely, heavy smokers showed the largest increases in birth weight with rising neighbourhood education levels. Increased levels of PM2.5 and immigrant density were negatively associated with birth weight, but showed positive interactions with increased levels of smoking. Older maternal age and suspected drug or alcohol use both had negative interactions with increased levels of maternal smoking. CONCLUSION: Maternal smoking had a negative and non-linear dose-response association with birth weight which was highly variable between neighbourhoods and evidence of effect modification with neighbourhood-level factors. These results suggest that focusing exclusively on individual behaviours may have limited success in improving outcomes without addressing the contextual influences at the neighbourhood-level. Further studies are needed to corroborate our findings and to understand how neighbourhood-level attributes interact with smoking to affect birth outcomes.

Methylmercury alters glutathione homeostasis by inhibiting glutaredoxin 1 and enhancing glutathione biosynthesis in cultured human astrocytoma cells.

Methylmercury (MeHg) is a neurotoxin that binds strongly to thiol residues on protein and low molecular weight molecules like reduced glutathione (GSH). The mechanism of its effects on GSH homeostasis particularly at environmentally relevant low doses is not fully known. We hypothesized that exposure to MeHg would lead to a depletion of reduced glutathione (GSH) and an accumulation of glutathione disulfide (GSSG) leading to alterations in S-glutathionylation of proteins. Our results showed exposure to low concentrations of MeHg (1µM) did not significantly alter GSH levels but increased GSSG levels by ∼12-fold. This effect was associated with a significant increase in total cellular glutathione content and a decrease in GSH/GSSG. Immunoblot analyses revealed that proteins involved in glutathione synthesis were upregulated accounting for the increase in cellular glutathione. This was associated an increase in cellular Nrf2 protein levels which is required to induce the expression of antioxidant genes in response to cellular stress. Intriguingly, we noted that a key enzyme involved in reversing protein S-glutathionylation and maintaining glutathione homeostasis, glutaredoxin-1 (Grx1), was inhibited by ∼50%. MeHg treatment also increased the S-glutathionylation of a high molecular weight protein. This observation is consistent with the inhibition of Grx1 and elevated H2O2 production however; contrary to our original hypothesis we found few S-glutathionylated proteins in the astrocytoma cells. Collectively, MeHg affects multiple arms of glutathione homeostasis ranging from pool management to protein S-glutathionylation and Grx1 activity.

Increased F3-Isoprostanes in the Canadian Inuit Population Could Be Cardioprotective by Limiting F2-Isoprostane Production.

CONTEXT: F3-isoprostanes (F3-IsoPs), derived from peroxidation of eicosapentaenoic acid (C20:5n-3), could be cardioprotective by limiting production of F2-isoprostanes (F2-IsoPs), a cardiovascular disease risk factor. OBJECTIVE: The objective of the study was to determine whether the n-3-polyunsaturated (PUFA)-rich Inuit diet is associated with a lower plasma ratio of F2-IsoPs to F3-IsoPs. DESIGN: This was a cross-sectional observational study. SETTING: The study was conducted in 36 Canadian Arctic Inuit communities. PARTICIPANTS: Participants included a random subset (n = 233) of Inuit adults taken from a population-based survey. MAIN OUTCOME MEASURES: Plasma F2-IsoPs and F3-IsoPs, cardiometabolic risk factors (blood lipids, C-reactive protein, blood pressure, fasting glucose) and markers of dietary exposure (erythrocyte n-3 and n-6 PUFA, blood levels of Se, mercury, polychlorinated biphenyls) were measured. RESULTS: Inuit aged 40 years old and older vs younger Inuit showed higher concentrations of plasma F3-IsoPs and erythrocyte n-3 PUFA and lower plasma F2-IsoPs concentrations despite having higher blood lipids, fasting glucose, systolic blood pressure, and percentage body fat. Plasma F3-IsoPs were not associated with any cardiometabolic measures. When subjects were categorized into tertiles according to total n-3 PUFA erythrocyte concentrations, F3-IsoPs increased with increasing tertiles, whereas the F2-IsoP to F3-IsoP ratio was lowest at the highest n-3 tertile. The F2-IsoP to F3-IsoP ratio was significantly predicted by C20:5n-3 (ß= -.365, P = .002); C20:4n-6:C20:5n-3 (ß = .056, P = .006), blood mercury (ß = -.812, P =.015), blood Se (ß = -1.95, P = .015), and smoking (ß = .745, P = .025). CONCLUSIONS: Plasma F3-IsoPs were not associated with cardiometabolic risk factors previously seen with F2-IsoPs. Higher n-3 fatty acid status was associated with lower plasma F2-IsoPs and higher plasma F3-IsoPs, which provides partial explanation to the cardioprotective effects of the n-3 PUFA-rich Inuit diet.

Superoxide produced in the matrix of mitochondria enhances methylmercury toxicity in human neuroblastoma cells.

The mechanism of intracellular metabolism of methylmercury (MeHg) is not fully known. It has been shown that superoxide (O2(-)), the proximal reactive oxygen species (ROS) generated by mitochondria, is responsible for MeHg demethylation. Here, we investigated the impact of different mitochondrial respiratory inhibitors, namely rotenone and antimycin A, on the O2(-)mediated degradation of MeHg in human neuroblastoma cells SH-K-SN. We also utilized paraquat (PQ) which generates O2(-) in the mitochondrial matrix. We found that the cleavage of the carbon-metal bond in MeHg was highly dependent on the topology of O2(-) production by mitochondria. Both rotenone and PQ, which increase O2(-) in the mitochondrial matrix at a dose-dependent manner, enhanced the conversion of MeHg to inorganic mercury (iHg). Surprisingly, antimycin A, which prompts emission of O2(-) into the intermembrane space, did not have the same effect even though antimycin A induced a dose dependent increase in O2(-) emission. Rotenone and PQ also enhanced the toxicity of sub-toxic doses (0.1 µM) MeHg which correlated with the accumulation of iHg in mitochondria and depletion of mitochondrial protein thiols. Taken together, our results demonstrate that MeHg degradation is mediated by mitochondrial O2(-), specifically within the matrix of mitochondria when O2(-) is in adequate supply. Our results also show that O2(-) amplifies MeHg toxicity specifically through its conversion to iHg and subsequent interaction with protein cysteine thiols (R-SH). The implications of our findings in mercury neurotoxicity are discussed herein.

Superoxide anion radical (O2(-)) degrades methylmercury to inorganic mercury in human astrocytoma cell line (CCF-STTG1).

Methylmercury (MeHg) is a global pollutant that is affecting the health of millions of people worldwide. However, the mechanism of MeHg toxicity still remains somewhat elusive and there is no treatment. It has been known for some time that MeHg can be progressively converted to inorganic mercury (iHg) in various tissues including the brain. Recent work has suggested that cleavage of the carbon-metal bond in MeHg in a biological environment is facilitated by reactive oxygen species (ROS). However, the oxyradical species that actually mediates this process has not been identified. Here, we provide evidence that superoxide anion radical (O2(-)) can convert MeHg to iHg. The calculated second-order rate constant for the degradation of 1µM MeHg by O2(-) generated by xanthine/xanthine oxidase was calculated to be 2×10(5)M(-1)s(-1). We were also able to show that this bioconversion can proceed in intact CCF-STTG1 human astrocytoma cells exposed to paraquat (PQ), a O2(-) generating viologen. Notably, exposure of cells to increasing amounts of PQ led to a dose dependent increase in both MeHg and iHg. Indeed, a 24h exposure to 500µM PQ induced a ∼13-fold and ∼18-fold increase in intracellular MeHg and iHg respectively. These effects were inhibited by superoxide dismutase mimetic MnTBAP. In addition, we also observed that a 24h exposure to a biologically relevant concentration of MeHg (1µM) did not induce cell death, oxidative stress, or even changes in cellular O2(-) and H2O2. However, co-exposure to PQ enhanced MeHg toxicity which was associated with a robust increase in cell death and oxidative stress. Collectively our results show that O2(-) can bioconvert MeHg to iHg in vitro and in intact cells exposed to conditions that simulate high intracellular O2(-) production. In addition, we show for the first time that O2(-) mediated degradation of MeHg to iHg enhances the toxicity of MeHg by facilitating an accumulation of both MeHg and iHg in the intracellular environment.