Human biomonitoring (HBM) monitors levels of environmental pollutants in human samples, which often is a topic of concern for residents near industrially contaminated sites (ICSs). Around an ICS area in Menen (Belgium), including a (former) municipal waste incinerator and a metal recovery plant, increasing environmental concentrations of dioxins and polychlorinated biphenyls (PCBs) were observed, causing growing concern among residents and authorities. The local community succeeded in convincing the responsible authorities to investigate the problem and offer research funding. Persistent organic pollutants (POPs) were measured in two consecutive HBM studies (2002–2006 and 2010–2011), in the context of the Flemish Environment and Health Study (FLEHS), as well as in soil and locally produced food. Meanwhile, local authorities discouraged consumption of locally produced food in a delineated area of higher exposure risk. Ultimately, HBM and environmental data enabled tailored dietary recommendations. This article demonstrates the usefulness of HBM in documenting the body burdens of residents near the ICS, identifying exposure routes, evaluating remediating actions and providing information for tailored policy strategies aiding to further exposure reduction. It also highlights the role of the local stakeholders as an example of community-based participatory research and how such an approach can create societal support for research and policy.

Citation for published version (APA): Vandermarken, T., Boonen, I., Van Langenhove, K., Goeyens, L., Denison, M. S., Croes, K., & Elskens, M. (2016). In vitro reporter gene activity of PCDD/Fs and dl-PCBs in wood ash from domestic heating systems (Belgium) using the H1L7.5c1 CALUX Bioassay. In Dioxin 2016, 36th International Symposium on Halogenated Persistent Organic Pollutants (Vol. 78, pp. 1107-1110). [Vol. 78] Organohalogen Compounds.

Sex hormone levels and timing of sexual maturation are considered important markers for health status of adolescents in puberty, and previous research suggests they might be influenced by metal exposure. In three campaigns of the Flemish Environment and Health Study (FLEHS I 2002-2006; FLEHS II 2007-2011 and FLEHS III 2012-2015), data were collected on internal exposure to metals (Cd, Cu, Pb, Cr, Mn, Tl, Ni, Sb, Hg, As and As species) and sexual maturation in 2671 14-15years old adolescents. All metals were measured in blood and/or urine, except total- and methylmercury which were measured in hair samples. Sex hormone levels were measured in blood serum of adolescent males of the cohorts of FLEHS I and FLESH II. The use of a uniform methodology in successive campaigns allows to confirm associations between exposure and health in different cohorts and over time. Furthermore, mathematical and statistical density correction methods using creatinine or specific gravity were tested for urinary markers. Significant associations between sex hormones and maturity markers were observed in the FLEHS I and II campaigns, when both were assessed together. Regardless of the applied correction method, creatinine correction systematically introduced bias due to associations of creatinine with sex hormones and maturation markers, especially in adolescent males, while this is not the case for specific gravity. A series of exposure-response associations were found, but several involving Cd, Pb, As, Tl and Cu persisted in different FLEHS campaigns. The effects of Pb and Cu on luteinizing hormone, (free) testosterone, (free) oestradiol and maturation support a xenoestrogenic agonistic action on the feedback of oestradiol to the hypothalamus-pituitary-gonadal axis. Our results suggest that specific care should be taken when selecting urine density correction for investigating associations with hormonal and maturation markers in adolescent males. Furthermore, the possibility of xenoestrogenic effects of certain metals in environmentally exposed adolescents warrants further investigation.

When compared to the European guidelines, PM10 (particulate matter up to 10-μm size) concentrations in Algeria are often exceeding the maximum limits, and in general, no information exists on the compounds bound on its surface. The objective of this study was to measure the dioxin-like activity of polychlorinated dibenzodioxines and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (PCBs) in the PM10 fraction at the Sour El Ghozlane cement plant in Algeria. PM10 samples (n = 23) were taken between 24 March and 15 April 2013, using a medium volume sampler and 47-mm PTFE filters. The 24-h samples were dried to determine the PM10 content and afterward extracted, cleaned up, and analyzed with the dioxin-responsive element–chemical-activated luciferase gene expression (DRE-CALUX) bioassay. Our results showed that the measured bioanalytical equivalents (BEQs) were similar to those in other international industrial sites worldwide. The PCDD/Fs and dioxin-like PCBs (dl-PCBs) were positively correlated (rho = 0.6, p = 0.002), indicating that they have similar sources. Furthermore, samples from March showed higher PCDD/F and dl-PCB BEQs and humidity but lower temperatures compared to samples from April, while there was no difference in the PM10 concentrations between the two months. These results reveal that PM10 alone is not a good proxy and that meteorological conditions are an important factor in assessing dioxin-like pollution in the atmosphere. It seems that, at present, there is no health hazard through direct airborne human exposure to dioxin-like pollutants in PM10 from this site. However, it is important to monitor these POPs for a longer period of time and also to gain more insight in their distribution between the particulate and gas phase in relation to meteorological conditions.

Background: Placental mitochondrial DNA (mtDNA) content can be indicative of oxidative damage to the placenta during fetal development and is responsive to external stressors. In utero exposure to environmental pollutants that may influence placental mtDNA needs further exploration.Objectives: We evaluated if placental mtDNA content is altered by environmental pollution in newborns and identified pollutants independently associated to alterations in placental mtDNA content.Methods: mtDNA content was measured in placental tissue of 233 newborns. Four perfluoroalkyl compounds and nine organochlorine compounds were quantified in cord blood plasma samples and six toxic metals in whole cord blood. We first applied a LASSO (least absolute shrinkage and selection operator) penalized regression model to identify independent associations between environmental pollutants and placental mtDNA content, without penalization of several covariates. Then adjusted estimates were obtained using an ordinary least squares (OLS) regression model evaluating the pollutants' association with placental mtDNA content, adjusted for several covariates.Results: Based on LASSO penalized regression, oxychlordane, p,p'-dichlorodiphenyldichloroethylene, beta-hexachlorocyclohexane, perfluorononanoic acid, arsenic, cadmium and thallium were identified to be independently associated with placental mtDNA content. The OLS model showed a higher placental mtDNA content of 2.71% (95% CI: 0.3 to 5.2%; p = 0.03) and 1.41% (0.1 to 2.8%, p = 0.04) for a 25% concentration increase of respectively cord blood beta-hexachlorocyclohexane and arsenic. For a 25% concentration increase of cord blood thallium, a 4.88% lower placental mtDNA content (95% CI: -9.1 to -0.5%, p = 0.03) was observed.Conclusion: In a multi-pollutant approach, low fetal exposure levels of environmental organic and inorganic pollutants might compromise placental mitochondrial function as exemplified in this study by alterations in mtDNA content.

Polycyclic aromatic hydrocarbons (PAHs) are of interest to human biomonitoring studies due to their carcinogenic potential. Traditionally metabolites of these compounds, like 1-hydroxypyrene, are monitored in urine, but recent methods allow the determination of the parent compounds in urine, which give additional information regarding sources and toxicity of PAHs. In order to assess the feasibility of incorporating these methods in a human biomonitoring study, the 16 USEPA parent PAHs were determined in 20 urine samples. These samples were obtained from 10 boys and 10 girls aged 14–16 years, participating in the third Flemish Environment and Health Study (Flanders, Belgium).Of these 16 parent PAHs, nine could be determined in more than 95% of the samples and three (including benzo(a)pyrene) in more than 50%. Several correlations were found between different PAHs, but not between pyrene and its metabolite 1-hydroxypyrene. Diagnostic PAH ratios in urine and air samples pointed towards combustion sources and are in line with the ratios in environmental samples. Benzo(a)pyrene, naphthalene and fluorene have the highest carcinogenic potential in our cohort, when using toxic equivalency factors. Some associations between PAH congeners and determinants of exposure were found, while fluorene and acenaphthylene were positively associated with thyroid hormone levels and benzo(a)pyrene showed a positive correlation with DNA damage by comet assay. These results confirm that parent PAHs in urine are useful as biomarkers of exposure in biomonitoring studies.

Early life exposure to endocrine disrupting chemicals (EDCs) has been frequently associated with impaired perinatal growth, an important risk factor for later onset of metabolic disorders.We analyzed whether the cord blood transcriptome showed early indications of alterations in metabolic processes in 195 human samples in relation to cord blood levels of dichlorodiphenyldichloroethylene (p,p'-DDE), polychlorinated biphenyl-153 (PCB-153), perfluorooctanoic acid (PFOA), and perfluorooctane sulfonate (PFOS).Overall, 39 metabolically relevant transcription factors were significantly enriched (31 by p,p'-DDE, 10 by PCB-153, 8 by PFOA, and 2 by PFOS). These included the glucocorticoid receptor (p,p'-DDE and PCB-153) and the progesterone receptor (PFOA and PFOS). The 'insulin receptor signaling', 'acute phase response signaling', 'Interleukin(IL)-6 signaling', and 'prolactin signaling' pathways were significantly enriched in relation to p,p'-DDE.Transcriptional changes at birth suggest a role for specific metabolic targets as a link between prenatal EDC exposure and metabolic disorders later in life. (C) 2016 Elsevier Inc. All rights reserved.

Background: Endocrine disrupting chemicals represent a broad class of compounds, are widespread in the environment and can pose severe health effects.Objectives: The objective of this study was to investigate and compare the overall estrogen and androgen activating potential of PM10 air samples at an urban, rural and industrial location in Flanders, using a human in vitro cell bioassay.Methods: PM10 samples were collected on glass fiber filters every six days between April 2013 and January 2014 using a high-volume sampler. Extraction was executed with a hexane/acetone mixture before analysis using a recombinant estrogen- or androgen responsive human carcinoma cell line. Results were expressed as bioanalytical equivalents (BEQs) per cubic meter of air.Results: High fluctuations in estrogenic activity were observed during the entire sampling period, with median BEQs of 32.1, 35.9 and 31.1 fg E2-Eq m(-3) in the industrial, urban and rural background area, respectively. Estrogenic activity was measured in 70% of the samples, while no androgenic activity was observed in any of the samples. The estrogenic activity in the industrial area was positively correlated with the airborne concentration of the sum of the non-carcinogenic PAH5 pyrene and fluoranthene (rho=0.48; p < 0.01) and the sum of the carcinogenic PAH5 (rho=0.36; p=0.05).Conclusions: This study showed that no androgenic activity was present in PM10 and that although the median estrogenic activity was rather low and comparable in the three locations, high fluctuations in estrogenic response exist over time. While atmospheric PAHs contributed to the observed estrogenic response, especially in the industrial area, the chemicals responsible for the majority of estrogenic activity remain to be identified. (C) 2016 Elsevier Inc. All rights reserved.

Endocrine disrupting chemicals represent a broad class of compounds, are widespread in the environment and can pose severe health effects. The objective of this study was to investigate the overall estrogen activating potential of PM10 air samples at an urban location with high traffic incidence in Flanders, using a human in vitro cell bioassay.PM10 samples (n = 36) were collected on glass fiber filters every six days between April 2013 and January 2014 using a high-volume sampler. Extraction was executed with a hexane/acetone mixture before analysis using a recombinant estrogen-responsive human ovarian carcinoma (BG1Luc4E2) cell line. In addition, several samples and procedural blanks were extracted with ultra-pure ethanol or acetonitrile to compare extraction efficiencies. Results were expressed as bioanalytical equivalents (BEQs) in femtogram 17 beta-estradiol equivalent (fg E2-Eq) per cubic meter of air.High fluctuations in estrogenic activity were observed during the entire sampling period, with mean and median BEQs of 50.7 and 35.9 fg E2-Eq m(-3), respectively. Estrogenic activity was measured in more than 70% of the samples and several sample extracts showed both high BEQs and high cytotoxicity, which could not be related to black carbon, PM10 or heavy metal concentrations. At this moment, it remains unclear which substances cause this toxicity, but comparison of results obtained with different extraction solvents indicated that acetone/hexane extracts contained more compounds that were cytotoxic and suppressive of responses than those extracted using ultra-pure ethanol. Although more research is needed, the use of a more polar extraction solvent seems to be advisable. (C) 2015 Elsevier Ltd. All rights reserved.

To follow time trends in exposure to environmental chemicals, three successive campaigns of the Flemish Environment and Health Study (FLEHS) have recruited and sampled in total 5825 participants between 2002 and 2014. Cord samples from newborns, urine and blood samples from 14 to 15 years old adolescents and from adults between 50 and 65 years old were analysed in geographical representative samples of the Flemish population. The data of the different campaigns were considered per age group and per biomarker after adjustment for predefined covariates to take into account differences in characteristics of the study populations over time. Geometric means were calculated. Multiple linear regression was used to evaluate time trends. The concentration of serum biomarkers for persistent organic pollutants (POPs), such as marker polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (p,p'-DDE), the major metabolite of dichlorodiphenyltrichloroethane (DDT), and hexachlorobenzene (HCB) expressed per g lipid, decreased significantly with time. The levels of DDE in all age groups and those of PCBs in cord and adolescent serum samples were almost halved in a time period of ten years. HCB levels were reduced by a factor of 4 in adolescents and in adults. Mean serum concentrations of the more recently regulated perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were significantly lower in cord samples of 2013 compared to samples of 2007. The decline was more pronounced for PFOS than for PFOA. In the same period, mean metabolite levels of di-2-ethylhexyl phthalate (DEHP) and of di-n-butyl phthalate (DBP) decreased significantly in urine samples of adolescents with sharper declines for DEHP than for DBP. Cadmium and lead levels in cord and adolescent blood samples were significantly lower in the recent campaigns than 10 years before. Also the mean urinary cadmium level in adults was 35% lower compared to adult samples of 2002. Such favourable trends were not observed for arsenic and thallium measured in cord blood. Similar, the concentrations of 1-hydroxypyrene, a marker for exposure to polycyclic aromatic hydrocarbons (PAHs), was not lower in urine from adolescents sampled in 2013 compared to 2003. In contrast, concentrations of t,t'-muconic acid, a marker of benzene exposure, showed clearly reduced levels. The FLEHS program shows that concentrations of well-regulated chemicals especially traditional POPs and cadmium and lead are decreasing in the population of Flanders. Response to regulatory measures seems to happen rapid, since concentrations in humans of specific regulated perfluorinated compounds and phthalates were significantly reduced in five years time. Biomarker concentrations for arsenic, thallium, and polyaromatic hydrocarbons are not decreasing in this time span and further follow up is warranted.

Rapid, easy and reliable quantification of amino acids is crucial in research on plant amino acid metabolism and nutritional improvement of crops via enrichment of essential amino acids. A recently reported analysis method, based on solid phase extraction (SPE), derivatization with methyl chloroformate and gas chromatography-mass spectrometry was optimized and tested on three-week-old Arabidopsis thaliana leaf tissues. Optimization of the SPE cleanup yielded recovery rates of minimum 95% for all amino acids (except arginine). Variations in accuracy and precision did not exceed 12.5%, except for cysteine, histidine and tryptophane, which were excluded from analysis. Quantification of overlapping peaks for isoleucine/threonine and proline/asparagine was possible by selection of two specific fragment ions for each amino acid. Of the 16 selected amino acids, 14 were quantified successfully in at least 75% of the samples, while methionine and tyrosine were only quantifiable in 6% and 42%, respectively. A case study on the aspartate super pathway confirmed the applicability of the optimized method on wild type and genetically modified plants: external supplementation of methionine or lysine yielded a 146-fold or 27-fold increase in the respective absolute amino acid levels compared with the control treatment. Induced expression of dhdps-r1 (a mutated lysine biosynthesis gene encoding a feedback insensitive enzyme) caused an 83-fold increase in absolute lysine levels.

Historical persistent organic pollutants (POPs) are banned from Antarctica under international treaty; but contemporary-use POPs can enter as additives within polymer and textile products. Over their useful lives these products may release additives in-situ. Indeed, we observed 226 and 109 ng/g dry weight (dw) of the total concentrations of α-, β- and γ-hexabromocyclododecane (HBCD) in indoor dust from McMurdo Station (U.S.) and Scott Station (New Zealand), respectively. Sewage sludge collected from wastewater treatment facilities at these stations exhibited ∑HBCD of 45 and 69 ng/g dw, respectively. Contaminants originally within the bases may exit to the local outdoor environment via wastewaters. Near McMurdo, maximum ∑HBCD levels in surficial marine sediments and aquatic biota (invertebrates and fish) were 2350 ng/g (total organic carbon basis) and 554 ng/g lipid weight, respectively. Levels declined with distance from McMurdo. Our results illustrate that Antarctic research stations serve as local HBCD sources to the pristine Antarctic environment.

Arsenic (As) is a compound naturally present in the environment. Recent studies have shown detectable levels of As in rice, rice food products, and apple juice. As is a well known carcinogen and is related to multiple health effects, for example dermal and kidney toxicity. Recent epidemiological studies have shown that As exposure is also a risk factor for cardiovascular diseases. We evaluated the exposure to As in a sample of 408 adolescents that were recruited from the general population in Flanders as part of the third Flemish Environment and Health Study (FLEHS3). We tested whether the current levels of As exposure were associated with oxidative stress biomarkers or biomarkers for DNA damage. In this study, the individual exposure to As was estimated by measuring total As in whole blood and different toxic As metabolites in urine: As III, As V, mono-methylarsonous acid (MMA), dimethylarsinous acid (DMA), and the non-toxic, organic compound arsenobetaine. The sum of the toxic compounds was defined as toxic relevant As (TRA). DNA damage was measured using the alkaline comet assay and the micronucleus test in whole blood samples, and by an enzyme-linked immunosorbent assay (ELISA, Gentaur) quantifying 8-hydroxy-2-deoxyguanosine (8-OHdG) in urine. Multiple linear regression models were used to estimate the change of effect for an increase of the exposure from P25 to P75 (P25-P75). All models were adjusted for gender, age, smoking status and statistically significant (p<0.05) covariates. The exposure biomarker As III was categorised due to the large number values below the quantification limit (LOQ). The effect estimate was given for the group with the highest As III exposure versus the group with values below LOQ. Multiple regression analysis showed significant positive associations of urinary inorganic As III (iAsIII) and MMA with DNA breaks measured by the comet assay (iAsIII: β=0.333, 95% CI=0.061-0.605; MMA: β=0.090, 95% CI=0.014-0.166). Urinary iAs levels were positively associated with concentrations of 8-OHdG (iAsIII: OR=1.231, 95% CI=1.107-1.369; iAsV: OR=1.074, 95% CI=1.014-1.137; MMA: OR=1.154, 95% CI=1.105-1.205; DMA: OR=1.099, 95% CI: 1.041-1.160; TRA: OR=1.131, 95% CI= 1.073-1.192). No statistically significant associations were found with the micronucleus test. Although 82% of the adolescents had a TRA concentration below the Biomonitoring Equivalents for non-carcinogenic effects (≤6.4 µg/l), the data obtained in this study indicate that chronic exposure of Flemish adolescents to very low levels of As, compared to studies in e.g. Bangladesh, and both in whole blood (total As) or urine (TRA), is associated with oxidative stress and DNA damage. The studies of the Flemish Center of Expertise on Environment and Health were commissioned, financed and steered by the Ministry of the Flemish Community (Department of Economics, Science and Innovation; Flemish Agency for Care and Health; and Department of Environment, Nature and Energy).

The Flemish Centre for Environment and Health started with human biomonitoring in 2002 (FLEHS I: 2002-2006). The main goal of the second human biomonitoring cycle (FLEHS II: 2007-2011), was to determine mean values for a large number of pollutants in a representative sample of the general Flemish population. Values for Cd and Pb were updated, and a group of previously undetermined metals and metalloids (As, Mn, Cu and Tl) were included in some of the age groups. In this human biomonitoring program, three different age groups of the general Flemish population were monitored: 255 newborns and their mothers, 210 adolescents aged 14-15, and 204 adults between 20 and 40 years old. Trace elements were determined in cord blood and maternal blood of the mothers, in blood and urine of adolescents and in urine of adults. Determinants of life-style and personal factors were taken into account. The levels of trace elements in cord blood and maternal blood were for most elements at the lower end of the range found in literature. For Pb, As and Tl, a strong correlation (respectively r=0.43, 0.55 and 0.33; p<0.05) was found between levels in cord blood (respectively 8.6, 0.54 and 0.017 μg/L) and maternal blood (11.1, 0.64 and 0.028 μg/L), indicating that they are transported via the placenta from mother to fetus. The levels found in the adolescents and adults were compared with results from international biomonitoring studies, and were found to be in the same ranges. With the exception of Pb, all trace elements increased with increasing age group population. Finally, the results also showed that the levels of Cd and Pb in blood for this campaign (e.g. for Pb 8.6 and 14.8 μg/L in neonates and adolescents respectively) were lower compared to the first campaign (e.g. for Pb 14.7 and 21.7 μg/L in neonates and adolescents respectively), indicating a decrease over time. However, differences in sampling strategies might partially explain this observed trend.