Long-term Low-level Exposure Research Articles

Transcriptome-centric approach to the derivation of adverse outcome pathway networks of vascular dysfunction after long-term low-level exposure of human endothelial cells to dibutyl phthalate

Dibutyl phthalate (DBP) is an endocrine disruptor that adversely affects reproduction; however, evidence suggests it can also impact other systems, including vascular function. The mechanisms underlying DBP-induced vascular dysfunction, particularly after long-term low-level exposure of endothelial cells to this phthalate, remain largely unknown. To address this gap, we used experimentally derived data on differentially expressed genes (DEGs) obtained after 12 weeks of exposure of human vascular endothelial cells EA.hy926 to the concentrations of DBP to which humans are routinely exposed (10−9 M, 10−8 M, and 10−7 M) and various computational tools and manual data curation to build the first adverse outcome pathway (AOP) network relevant to DBP-induced vascular toxicity. DEGs were used to infer transcription factors (molecular initiating events) and molecular functions and biological processes (key events, KEs) using the Enrichr database. The AOP-helpFinder 2.0, an artificial intelligence-based web tool, was used to link genes and KEs and assign confidence scores to co-occurred terms. We constructed the AOP networks using Cytoscape and then manually arranged KEs to depict the flow of mechanistic information across different levels of network organization. An AOP network was created for each DBP concentration, revealing several distinct high-confidence subnetworks that could be involved in DBP-induced vascular toxicity: the insulin-like growth factor subnetwork for 10−7 M DBP, the CXCL8-dependent chemokine subnetwork for 10−8 M DBP, and the fatty acid subnetwork for 10−9 M DBP. We also developed an AOP network providing a mechanistic insight into the dose-dependent effects of DBP in endothelial cells leading to vascular dysfunction. In summary, we present novel putative AOP networks describing the mechanistic flow of information involved in DBP-induced vascular dysfunction in a long-term low-level exposure scenario.

Long-term ambient air pollution and coronary atherosclerosis: Results from the Swedish SCAPIS study

Background and aimsDespite firm evidence for an association between long-term ambient air pollution exposure and cardiovascular morbidity and mortality, results from epidemiological studies on the association between air pollution exposure and atherosclerosis have not been consistent. We investigated associations between long-term low-level air pollution exposure and coronary atherosclerosis. MethodsWe performed a cross-sectional analysis in the large Swedish CArdioPulmonary bioImaging Study (SCAPIS, n = 30 154), a random general population sample. Concentrations of total and locally emitted particulate matter <2.5 μm (PM2.5), <10 μm (PM10), and nitrogen oxides (NOx) at the residential address were modelled using high-resolution dispersion models. We estimated associations between air pollution exposures and segment involvement score (SIS), coronary artery calcification score (CACS), number of non-calcified plaques (NCP), and number of significant stenoses, using ordinal regression models extensively adjusted for potential confounders. ResultsMedian 10-year average PM2.5 exposure was 6.2 μg/m3 (range 3.5–13.4 μg/m3). 51 % of participants were women and 51 % were never-smokers. None of the assessed pollutants were associated with a higher SIS or CACS. Exposure to PM2.5 was associated with NCP (adjusted OR 1.34, 95 % CI 1.13, 1.58, per 2.05 μg/m3). Associations with significant stenoses were inconsistent. ConclusionsIn this large, middle-aged general population sample with low exposure levels, air pollution was not associated with measures of total burden of coronary atherosclerosis. However, PM2.5 appeared to be associated with a higher prevalence of non-calcified plaques. The results suggest that increased risk of early-stage atherosclerosis or rupture, but not increased total atherosclerotic burden, may be a pathway for long-term air pollution effects on cardiovascular disease.

Protective effect of the extract from Aronia melanocarpa L. berries against cadmium-induced oxidative stress in the kidney: A study in an in vivo experimental model

The destructive impact of cadmium (Cd) on the oxidative/antioxidative status of the kidney, as well as the possible beneficial effect of co-administration of a 0.1% aqueous extract from Aronia melanocarpa L. berries (AM), were studied in a rat model of low-level and moderate general population exposure to this heavy metal (1 and 5 mg Cd/kg feed, respectively, for up to 24 months). Total antioxidative status (TAS) of the kidney and the main indices of the enzymatic (superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase) and non-enzymatic (reduced glutathione and thioredoxin) antioxidative barrier were assessed. The total oxidative status (TOS) and concentrations of hydrogen peroxide, xanthine oxidase, myeloperoxidase, and oxidized glutathione were measured as markers of oxidative status. The oxidative stress index (OSI) was calculated (TOS/TAS) to estimate the intensity of oxidative stress in the kidney. The exposure to Cd, dose- and duration-dependently, weakened the enzymatic and non-enzymatic antioxidative potential of the kidney leading to a decrease in its TAS, as well as enhanced oxidative status of this organ resulting in the development of oxidative stress. The administration of AM during the low-level and moderate intoxication with Cd significantly protected from this xenobiotic-induced disruption of the oxidative/antioxidative balance and development of oxidative stress in the kidney. In summary, even low-level long-term exposure to Cd may result in the occurrence of oxidative stress in the kidney, whereas supplementation with chokeberry products may improve the oxidative/antioxidative balance preventing oxidative stress in this organ.

Australia’s Ongoing Challenge of Legacy Asbestos in the Built Environment: A Review of Contemporary Asbestos Exposure Risks

Asbestos remains ubiquitous in the Australian built environment. Of the 13 million tonnes of asbestos products installed in earlier decades, an estimated 50% remain in situ today. Because of the extensive past use of asbestos, and the increasing age of these products, the potential for exposure to asbestos fibres in both indoor and outdoor environments remains high, even while the actual asbestos exposure levels are mostly very low. Sources of these exposures include disturbance of in situ asbestos-containing materials (ACMs), for example during renovations or following disaster events such as fires, cyclones and floods. Our understanding of the risk of asbestos-related disease arising from long-term low-level or background exposure, however, is poor. We provide the most up-to-date review of asbestos exposure risks currently affecting different groups of the Australian population and the settings in which this can manifest. From this, a need for low-level asbestos monitoring has emerged, and further research is required to address whether current exposure monitoring approaches are adequate. In addition, we make the case for proactive asbestos removal to reduce the risk of ongoing asbestos contamination and exposure due to deteriorating, disturbed or damaged ACMs, while improving long-term building sustainability, as well as the sustainability of limited resources.

Air pollution, alcohol consumption, and the risk of elevated liver enzyme levels: a cross-sectional study in the UK Biobank.

Evidences on the association between exposure to air pollution and liver enzymes was scarce in low pollution area. We aimed to investigate the association between air pollution and liver enzyme levels and further explore whether alcohol intake influence this association. This cross-sectional study included 425,773 participants aged 37 to 73years from the UK Biobank. Land Use Regression was applied to assess levels of PM2.5, PM10, NO2, and NOx. Levels of liver enzymes including AST, ALT, GGT, and ALP were determined by enzymatic rate method. Long-term low-level exposure to PM2.5 (per 5-μg/m3 increase) was significantly associated with AST (0.596% increase, 95% CI, 0.414 to 0.778%), ALT (0.311% increase, 0.031 to 0.593%), and GGT (1.552% increase, 1.172 to 1.933%); The results were similar for PM10; NOX and NO2 were only significantly correlated with AST and GGT Significant modification effects by alcohol consumption were found (P-interaction < 0.05). The effects of pollutants on AST, ALT, and GGT levels gradually increased along with the weekly alcohol drinking frequency. In conclusion, long-term low-level air pollutants exposure was associated with elevated liver enzyme levels. And alcohol intake may exacerbate the effect of air pollution on liver enzymes.

Global gene expression analysis reveals novel transcription factors associated with long-term low-level exposure of EA.hy926 human endothelial cells to bisphenol A

Bisphenol A (BPA) is an endocrine disruptor that binds to estrogen receptors (ER); however, studies have shown that the ER pathway was not always the primary molecular mechanism of BPA's action in cells and that gene transcription could be altered by different exposure times and doses. Here, we sought to understand the correlation between the BPA-responsive genes that have associated biological functions and the transcription factors (TFs) involved in their regulation by repeatedly exposing human endothelial cells EA.hy926 to three nanomolar concentrations of BPA (10−9 M, 10−8 M, and 10−7 M) for 14 weeks, after which changes in global gene expression were determined by RNA sequencing. Cytoscape plug-in iRegulon was used to infer TFs involved in the control of BPA-deregulated genes. The results show a minimal overlap in deregulated genes between three concentrations of BPA, with 10−9 M BPA having the highest number of deregulated genes. TF analysis suggests that all three concentrations of BPA were active in the absence of an ER-mediated pathway. A unique set of TFs (NES≥4) has been identified for each BPA concentration, including the NFκB family and CEBPB for 10−9 M BPA, MEF family, AHR/ARNT, and ZBTB33 for 10−8 M BPA, and IRF1-7 and OVOL1/OVOL2 for 10−7 M BPA, whereas STAT1/STAT2 were common TFs for 10−9 M and 10−7 M BPA. Overall, our data suggest that long-term low-level exposure of EA.hy926 cells to BPA leads to concentration-specific changes in gene expression that are not controlled by the ER-mediated signaling but rather by other mechanisms.

Genistein induces endocrine resistance in human breast cancer by suppressing H3K27 trimethylation.

Genistein (GE), the most important phytoestrogen in diet, is known to behave as a partial agonist of estrogen receptor α and shows a proliferative effect on the growth of breast cancer cell lines. Recent research has reported that long-term consumption of low doses of GE results in hormone-independent growth phenotypes of MCF-7 tumors, with increased HER2. Overexpression of HER2 has been associated with endocrine resistance in human breast cancer, but whether long-term low-level GE-induced HER2 expression is the cause of endocrine resistance remains to be determined. Short-term and long-term treatments with GE may have different effects on HER2 expression. We found that low doses of GE had estrogen-like effects and inhibited HER2 expression after short-term exposure in estrogen receptor-positive breast cancers cells. However, in contrast to short-term exposure, long-term exposure induced an increase in HER2 expression, which led to endocrine resistance. During long-term low-level exposure, the continuous activation of ERK1/2-phosphorylated EZH2 at Ser21 resulted in a decrease of lysine 27 trimethylation. As H3K27me3 levels decreased, the expression of interleukin-6 (IL-6) and IL-8 increased, and HER2 levels gradually increased, forming a feedback loop of ERK1/2/EZH2/IL-6 and IL-8/HER2. We identified a novel pathway by which EZH2 phosphorylation contributed to long-term low-level GE-induced HER2 overexpression and provided new insight for long-term low-level GE-induced acquired endocrine resistance. For breast cancer patients, long-term low-level use of soy supplements has potential health risks, and monitoring dietary exposure to GE is advisable when patients are treated with tamoxifen.

Breast Cancer Incidence in Relation to Long-Term Low-Level Exposure to Air Pollution in the ELAPSE Pooled Cohort.

Established risk factors for breast cancer include genetic disposition, reproductive factors, hormone therapy, and lifestyle-related factors such as alcohol consumption, physical inactivity, smoking, and obesity. More recently a role of environmental exposures, including air pollution, has also been suggested. The aim of this study, was to investigate the relationship between long-term air pollution exposure and breast cancer incidence. We conducted a pooled analysis among six European cohorts (n = 199,719) on the association between long-term residential levels of ambient nitrogen dioxide (NO2), fine particles (PM2.5), black carbon (BC), and ozone in the warm season (O3) and breast cancer incidence in women. The selected cohorts represented the lower range of air pollutant concentrations in Europe. We applied Cox proportional hazards models adjusting for potential confounders at the individual and area-level. During 3,592,885 person-years of follow-up, we observed a total of 9,659 incident breast cancer cases. The results of the fully adjusted linear analyses showed a HR (95% confidence interval) of 1.03 (1.00-1.06) per 10 μg/m³ NO2, 1.06 (1.01-1.11) per 5 μg/m³ PM2.5, 1.03 (0.99-1.06) per 0.5 10-5 m-1 BC, and 0.98 (0.94-1.01) per 10 μg/m³ O3. The effect estimates were most pronounced in the group of middle-aged women (50-54 years) and among never smokers. The results were in support of an association between especially PM2.5 and breast cancer. The findings of this study suggest a role of exposure to NO2, PM2.5, and BC in development of breast cancer.

Impact of In Vitro Long-Term Low-Level DEHP Exposure on Gene Expression Profile in Human Granulosa Cells.

Here, we applied a model of long-term exposure of human granulosa cells to low environmentally relevant levels of di(2-ethylhexyl) phthalate (DEHP). This approach provides more relevant data regarding the impact of DEHP on the function of human granulosa cells. The immortalized human granulosa cells HGrC1 were exposed to 50 nM and 250 nM DEHP for four weeks. The cells were collected every week to analyze the basal granulosa cells’ functions. A portion of the DEHP-exposed cells was stimulated with forskolin (FOR) for 48 h. Steroidogenesis was investigated using ELISA, whereas DNBQ sequencing and RT-qPCR were used to analyze gene expression. The results show that steroidogenesis was not affected by DEHP exposure. RNAsequencing shows that DEHP caused week- and concentration-specific changes in various genes and functions in HGrC1. Sulfotransferase family 1A member 3 (SULT1A3) and 4 (SULT1A4), which are involved in catecholamine metabolism, were the most prominent genes affected by DEHP under both the basal and FOR-stimulated conditions in all four weeks of exposure. This study showed, for the first time, that SULT1A3 and SULT1A4 are expressed in human granulosa cells, are regulated by FOR, and are affected by low-level DEHP exposure. These data provide new insight into the relationship between DEHP, SULT1A3, and SULT1A4 in human granulosa cells.

Long-Term Effects of Low-Level Blast Exposure and High-Caliber Weapons Use in Military Special Operators.

Chronic low-level blast exposure has been linked with neurological alterations and traumatic brain injury (TBI) biomarkers. Impaired smooth-pursuit eye movements (SPEM) are often associated with TBI. The purpose of this study was to determine whether long-term operators of low-level blast exposure or high-caliber weapons use displayed oculomotor behaviors that differed from controls. Twenty-six members of an elite military unit performed a computerized oculomotor testing task using an eye tracker and completed a concussion assessment questionnaire. The participants were split into a blast exposure group and control group. The blast exposure group had a history of exposure to low-level blasts or high-caliber weapon use. The results revealed significant differences in SPEM, saccades, and fixations between the blast exposure group and control group. The blast exposure group’s eye movements were slower, stopped at more frequent points when following a target, traveled further from the target in terms of both speed and direction, and showed higher rates of variation and inefficiency. Poor oculomotor behavior correlated with a higher symptom severity on the concussion assessment questionnaire. Military special operators exposed to long-term low-level blasts or high-caliber weapons usage displayed an impaired oculomotor behavior in comparison to controls. These findings further our understanding of the impact of long-term low-level blast exposure on the oculomotor behavior of military special operators and may inform practical implications for military training.

Molecular Fates of Organometallic Mercury in Human Brain.

Mercury is ubiquitous in the environment, with rising levels due to pollution and climate change being a current global concern. Many mercury compounds are notorious for their toxicity, with the potential of organometallic mercury compounds for devastating effects on the structures and functions of the central nervous system being of particular concern. Chronic exposure of human populations to low levels of methylmercury compounds occurs through consumption of fish and other seafood, although the health consequences, if any, from this exposure remain controversial. We have used high energy resolution fluorescence detected X-ray absorption spectroscopy to determine the speciation of mercury and selenium in human brain tissue. We show that the molecular fate of mercury differs dramatically between individuals who suffered acute organometallic mercury exposure (poisoning) and individuals with chronic low-level exposure from a diet rich in marine fish. For long-term low-level methylmercury exposure from fish consumption, mercury speciation in brain tissue shows methylmercury coordinated to an aliphatic thiolate, resembling the coordination environment observed in marine fish. In marked contrast, for short-term high-level exposure, we observe the presence of biologically less available mercuric selenide deposits, confirmed by X-ray fluorescence imaging, as well as mercury(II)-bis-thiolate complexes, which may be signatures of severe poisoning in humans. These differences between low-level and high-level exposures challenge the relevance of studies involving acute exposure as a proxy for low-level chronic exposure.

Mercury pollution in China: implications on the implementation of the Minamata Convention.

Mercury (Hg) is a toxic metal released into the environment through human activities and natural processes. Human activities have profoundly increased the amount of Hg in the atmosphere and altered its global cycling since the Industrial Revolution. Gaseous elemental Hg is the predominant form of Hg in the atmosphere, which can undergo long-range transport and atmospheric deposition into the aquatic systems. Hg deposition elevates the methylmercury (MeHg) level in fish through bioaccumulation and biomagnification, which poses a serious human health risk. Acute poisoning of MeHg can result in Minamata disease, while low-level long-term exposure in pregnant women can reduce the intelligence quotient of infants. After five sessions of intergovernmental negotiation, the Minamata Convention on mercury entered into force in August 2017 to protect human health and the environment from Hg pollution. Currently China contributes the largest quantity of Hg production, consumption, and emission globally. However, the status of Hg pollution in the environment in China and its associated health risk remains relatively unknown, which hinders the development of implementation plans of the Minamata Convention. In this paper, we provide a comprehensive review on the atmospheric release of Hg, distribution of air Hg concentration, human exposure to MeHg and health impacts caused by Hg pollution in China. Ongoing improvement of air pollution control measures is expected to further decrease anthropogenic Hg emissions in China. Air Hg concentrations in China are higher than the background values in the Northern Hemisphere, with spatial distribution largely influenced by anthropogenic emissions. Long-term observations of GEM in China show a decline in recent years. The net Hg transport outflow from China in 2013 is estimated to be 511 t year-1, and ∼60% of such outflow is caused by natural surface Hg emissions. Hg concentrations in fish and rice in China are relatively low and therefore the associated risks of human Hg exposure are low. Future research needs and recommendations for the implementation of the Minamata Convention are also discussed in this paper.

YAC128 mouse model of Huntington disease is protected against subtle chronic manganese (Mn)-induced behavioral and neuropathological changes

Manganese (Mn) is an essential micronutrient but excessive levels induce neurotoxic effects. Increasing evidence suggests a deficit of bioavailable Mn in Huntington disease (HD), an inherited neurodegenerative disease characterized by motor and cognitive disturbances. Previous studies have shown rescue of some molecular HD phenotypes by acute Mn exposure. This study simultaneously examined the potential for chronic Mn exposure to attenuate HD behavioral phenotypes, and for the HD genotype to offer protection against detrimental effects of chronic Mn exposure. In two independent studies a chronic Mn exposure paradigm was implemented in the YAC128 mouse model of HD and behavior was assessed at several timepoints. Study 1 exposed WT and YAC128 mice to twice weekly subcutaneous injections of 0, 5, 15, or 50 mg/kg MnCl[2] tetrahydrate from 12 to 32 weeks of age. A promising protective effect against motor coordination decline in 5 mg/kg MnCl[2] tetrahydrate-treated YAC128 mice was detected. Study 2 thus exposed WT and YAC128 mice to either 0 or 5 mg/kg MnCl[2] tetrahydrate from 12 to 52 weeks of age (with a partial randomized treatment crossover at 31 weeks). The same protective effect was not observed under these conditions at higher statistical power. We report subtle toxicological changes in exploratory behavior and total activity induced by chronic Mn exposure in WT mice only, despite similar total increases in brain Mn in WT and YAC128 mice. Further, chronic Mn treatment resulted in a 10–12 % decrease in striatal NeuN positive cell density in WT mice but not YAC128 mice, despite vehicle cell counts already being reduced compared to WT mice as expected for the HD genotype. The subtle changes observed in specific outcome measures, but not others, following long-term low-level Mn exposure in WT mice delineate the neurobehavioral and neuropathological effects at the threshold of chronic Mn toxicity. We conclude that these chronic low-dose Mn exposures do not significantly rescue behavioral HD phenotypes, but YAC2128 mice are protected against the subtle Mn-induced behavioral changes and decreased striatal neuron density observed in Mn-exposed WT mice.

Evaluation of the Role of KIM-1 in Detecting Early Nephrotoxicity in Lead-Exposed Workers.

Lead nephropathy usually starts silent. This study aimed to evaluate using kidney injury molecule 1 (KIM-1) as an early nephrotoxicity predictor of long-term low-level occupational lead exposure. History, examination, and laboratory investigations including: blood lead, urinary KIM-1, serum uric acid, creatinine, urea, sodium, potassium, serum albumin, and urine analysis were done on 35 lead-exposed workers and a matched control group. Higher blood lead levels were found among the exposed group compared to the control one. No statistically significant difference was found regarding renal failure manifestations or standard renal functions (uric acid, blood urea, and creatinine). Urinary KIM-1 was statistically significantly increased among the exposed group. Renal adverse effects were associated to lead fumes exposure. KIM-1 can be used as biomarker for detecting early renal affection among lead-exposed workers.

Investigating the impact of long term exposure to chemical agents on the chromosomal radiosensitivity using human lymphoblastoid GM1899A cells

This study aimed to investigate the impact of chronic low-level exposure to chemical carcinogens with different modes of action on the cellular response to ionising radiation. Human lymphoblastoid GM1899A cells were cultured in the presence of 4-nitroquinoline N-oxide (4NQO), N-nitroso-N-methylurea (MNU) and hydrogen peroxide (H2O2) for up to 6 months at the highest non-(geno)toxic concentration identified in pilot experiments. Acute challenge doses of 1 Gy X-rays were given and chromosome damage (dicentrics, acentric fragments, micronuclei, chromatid gaps/breaks) was scored. Chronic exposure to 20 ng/ml 4NQO, 0.25 μg/ml MNU or 10 μM H2O2 hardly induced dicentrics and did not significantly alter the yield of X-ray-induced dicentrics. Significant levels of acentric fragments were induced by all chemicals, which did not change during long-term exposure. Fragment data in combined treatment samples compared to single treatments were consistent with an additive effect of chemical and radiation exposure. Low level exposure to 4NQO induced micronuclei, the yields of which did not change throughout the 6 month exposure period. As for fragments, micronuclei yields for combined treatments were consistent with an additive effect of chemical and radiation. These results suggest that cellular radiation responses are not affected by long-term low-level chemical exposure.

Long-term low-level ambient air pollution exposure and risk of lung cancer – A pooled analysis of 7 European cohorts

Background/aimAmbient air pollution has been associated with lung cancer, but the shape of the exposure-response function - especially at low exposure levels - is not well described. The aim of this study was to address the relationship between long-term low-level air pollution exposure and lung cancer incidence. MethodsThe “Effects of Low-level Air Pollution: a Study in Europe” (ELAPSE) collaboration pools seven cohorts from across Europe. We developed hybrid models combining air pollution monitoring, land use data, satellite observations, and dispersion model estimates for nitrogen dioxide (NO2), fine particulate matter (PM2.5), black carbon (BC), and ozone (O3) to assign exposure to cohort participants’ residential addresses in 100 m by 100 m grids. We applied stratified Cox proportional hazards models, adjusting for potential confounders (age, sex, calendar year, marital status, smoking, body mass index, employment status, and neighborhood-level socio-economic status). We fitted linear models, linear models in subsets, Shape-Constrained Health Impact Functions (SCHIF), and natural cubic spline models to assess the shape of the association between air pollution and lung cancer at concentrations below existing standards and guidelines. ResultsThe analyses included 307,550 cohort participants. During a mean follow-up of 18.1 years, 3956 incident lung cancer cases occurred. Median (Q1, Q3) annual (2010) exposure levels of NO2, PM2.5, BC and O3 (warm season) were 24.2 µg/m3 (19.5, 29.7), 15.4 µg/m3 (12.8, 17.3), 1.6 10−5m−1 (1.3, 1.8), and 86.6 µg/m3 (78.5, 92.9), respectively. We observed a higher risk for lung cancer with higher exposure to PM2.5 (HR: 1.13, 95% CI: 1.05, 1.23 per 5 µg/m3). This association was robust to adjustment for other pollutants. The SCHIF, spline and subset analyses suggested a linear or supra-linear association with no evidence of a threshold. In subset analyses, risk estimates were clearly elevated for the subset of subjects with exposure below the EU limit value of 25 µg/m3. We did not observe associations between NO2, BC or O3 and lung cancer incidence. ConclusionsLong-term ambient PM2.5 exposure is associated with lung cancer incidence even at concentrations below current EU limit values and possibly WHO Air Quality Guidelines.

Urinary phthalate concentrations in the slovenian population: An attempt to exposure assessment of family units

Phthalates are widespread contaminants with differing chemical characteristics, which largely determine their product applications, and they can leach into the environment. Due to their endocrine disruptive properties at long-term low-level exposure, they propose a health threat to people that has been associated with several adverse health effects such as: decreased male fertility and impacts on neurological development. People are exposed to different phthalates on a daily basis. Accordingly, this study aims to determine urinary concentrations of seven phthalate metabolites in Slovenian mothers (n = 155), fathers (n = 77), and children (n = 155) within the European project DEMOCOPHES and to identify potential sources of exposure using questionnaire data on sociodemographic characteristics. Furthermore, the appropriateness of two adjustment methods (creatinine and specific gravity) has been evaluated. First morning urine samples were obtained from one urban and one rural location in 2011. Samples were analysed with Ultra Performance Liquid Chromatography Tandem Mass Spectrometry according to the COPHES SOP protocol by VITO NV laboratory in Belgium. All investigated metabolites were detected in all populations. Children's urinary concentrations exceeded those of adults for most metabolites. We observed variations in concentrations depending on sociodemographic and geographic characteristics, such as food and product sources (e.g. plastic packaging, tins, personal care products, PVC) as well as lifestyle and habits (e.g. living space, time spent outside). We observed geographic and sociodemographic differences in our populations that could be confirmed for the three populations separately and for family units. Concentrations are significantly higher at the rural sampling location as well as in households with a lower level of education. We found both the urinary concentrations and the intake doses to be within the European range as presented in the literature. Between creatinine and specific gravity, we found specific gravity the more appropriate option for phthalates. To our knowledge, this is the first study investigating exposure to phthalates in the Slovenian population while considering the common exposure of family units.

P.169 The effect of CDNF on N171-82Q mouse model of Huntington`s disease

Manganese (Mn) is an essential micronutrient but excessive levels induce neurotoxic effects. Increasing evidence suggests a deficit of bioavailable Mn in Huntington disease (HD), an inherited neurodegenerative disease characterized by motor and cognitive disturbances. Previous studies have shown rescue of some molecular HD phenotypes by acute Mn exposure. This study simultaneously examined the potential for chronic Mn exposure to attenuate HD behavioral phenotypes, and for the HD genotype to offer protection against detrimental effects of chronic Mn exposure. In two independent studies a chronic Mn exposure paradigm was implemented in the YAC128 mouse model of HD and behavior was assessed at several timepoints. Study 1 exposed WT and YAC128 mice to twice weekly subcutaneous injections of 0, 5, 15, or 50 mg/kg MnCl[2] tetrahydrate from 12 to 32 weeks of age. A promising protective effect against motor coordination decline in 5 mg/kg MnCl[2] tetrahydrate-treated YAC128 mice was detected. Study 2 thus exposed WT and YAC128 mice to either 0 or 5 mg/kg MnCl[2] tetrahydrate from 12 to 52 weeks of age (with a partial randomized treatment crossover at 31 weeks). The same protective effect was not observed under these conditions at higher statistical power. We report subtle toxicological changes in exploratory behavior and total activity induced by chronic Mn exposure in WT mice only, despite similar total increases in brain Mn in WT and YAC128 mice. Further, chronic Mn treatment resulted in a 10–12 % decrease in striatal NeuN positive cell density in WT mice but not YAC128 mice, despite vehicle cell counts already being reduced compared to WT mice as expected for the HD genotype. The subtle changes observed in specific outcome measures, but not others, following long-term low-level Mn exposure in WT mice delineate the neurobehavioral and neuropathological effects at the threshold of chronic Mn toxicity. We conclude that these chronic low-dose Mn exposures do not significantly rescue behavioral HD phenotypes, but YAC2128 mice are protected against the subtle Mn-induced behavioral changes and decreased striatal neuron density observed in Mn-exposed WT mice.

The progressive alteration of urine metabolomic profiles of rats following long-term and low-dose exposure to permethrin

Background: Permethrin is a type of widely used pyrethroid pesticide. Although acute toxicity of permethrin has been well-characterised, the non-acute toxicity of permethrin upon long-term exposure at low dose has been seldom studied yet. The current study investigates the time-course change of the metabolomic profiles of urine following the low level long-term exposure of permethrin and identified biomarkers of the chronic toxicity of permethrin.Methods: Male Wistar rats were administrated orally with permethrin (75 mg/kg body weight/day, 1/20 LD50) daily for consecutive 90 days. The urine samples from day 30, day 60, and day 90 after the first dosing were collected and analysed by 1H NMR spectrometry. Serum biochemical analysis was also carried out.Results: Permethrin caused significant changes in the urine metabolites such as taurine, creatinine, acetate, lactate, dimethylamine, dimethylglycine, and trimethylamine-N-oxide. These biological markers indicated prominent kidney and liver toxicity induced by permethrin. However, there was no change in serum biochemical parameters for the toxicity, indicating that metabolomic approach was much more sensitive in detecting the chronic toxicity.Conclusion: The time-course alteration of metabolomic profiles of the urine based on 1H NMR reflects the progressive development of the chronic toxicity with the long-term low-level exposure of permethrin.

Low-level ambient air pollution exposure and risk of lung cancer – a pooled analysis of 7 European cohorts

PDS 68: Outdoor air pollution, mortality and morbidity, Exhibition Hall (PDS), Ground floor, August 26, 2019, 10:30 AM - 12:00 PM Background/Aim: Ambient air pollution has been associated with lung cancer but the shape of the exposure-response function - especially at lower exposure levels - is not well described. The aim of this study was to address the relationship between long-term low-level air pollution exposure and lung cancer incidence. Methods: The “Effects of low level air pollution: A study in Europe” (ELAPSE) collaboration pools 7 cohorts from across Europe. We developed hybrid models combining monitoring and land use data, satellite observations, and dispersion model estimates for nitrogen dioxide (NO2), fine particulate matter (PM2.5), black carbon (BC), and ozone (O3) to assign exposure to participants’ residential addresses. We analyzed the pooled data by stratified Cox proportional hazards models adjusting for potential confounders. We developed linear and natural cubic spline models, analyses based on subsets and thresholds across the exposure distribution, and multi-pollutant models to disentangle potential inter-dependencies between pollutants. Results: A total of 307,550 persons were included in the analyses. During a mean follow-up of 18.1 years, 3,956 incident lung cancer cases occurred. Median (5–95%) exposure levels of NO2, PM2.5, BC and O3 (warm season) were 24.2 µg/m3 (12.8–39.5), 15.4 µg/m3 (8.7–19.4), 1.6 10-5/m (0.7–2.1), and 86.6 µg/m3 (70.4–92.9), respectively. We observed a higher risk for lung cancer with higher exposure to PM2.5 (HR 1.13, 95% 1.05-1.23 per 5 µg/m3) after adjustment for covariates. The association was similar across subsets of exposure-levels with no sign of a threshold below which no association was evident. The effect estimate was also robust to the inclusion of co-occurring pollutants. We did not find significantly positive associations between NO2, BC or O3 and lung cancer incidence. Conclusions: Our results indicate that long-term ambient PM2.5 exposure may contribute to lung cancer incidence even at concentrations lower than current EU limit values and WHO Air Quality Guidelines.