In vitro-to-in vivo correlation of the skin penetration, liver clearance and hepatotoxicity of caffeine.

This work illustrates the use of Physiologically-Based Toxicokinetic (PBTK) modelling for the healthy Caucasian population in in vitro-to-in vivo correlation of kinetic measures of caffeine skin penetration and liver clearance (based on literature experiments), as well as dose metrics of caffeine-induced measured HepaRG toxicity. We applied a simple correlation factor to quantify the in vitro and in vivo differences in the amount of caffeine permeated through the skin and concentration-time profiles of caffeine in the liver. We developed a multi-scale computational approach by linking the PBTK model with a Virtual Cell-Based Assay to relate an external oral and dermal dose with the measured in vitro HepaRG cell viability. The results revealed higher in vivo skin permeation profiles than those determined in vitro using identical exposure conditions. Liver clearance of caffeine derived from in vitro metabolism rates was found to be much slower than the optimised in vivo clearance with respect to caffeine plasma concentrations. Finally, HepaRG cell viability was shown to remain almost unchanged for external caffeine doses of 5-400 mg for both oral and dermal absorption routes. We modelled single exposure to caffeine only.

Micro-organic pollutants and biological response of mussels in marinas and ship building/breaking yards in Turkey.

Concentrations of PAHs, PCBs and OCPs in sediments and mussels (caged and/or native) were determined at 16 stations in six major sites of coastal Turkey. The biological effects of pollution were evaluated using sediment toxicity tests and enzyme activity assays. EROD, PROD, GST, AChE, CaE, and GR activities were evaluated using the digestive glands of mussels. The total PAH concentrations in the sediments varied between nd and 79,674 ng g(-1) dw, while the total OCP concentrations were in the range of nd to 53.7 ng g(-1) dw. The total PAH concentrations in mussels varied between 22.3 and 37.4 ng g(-1) ww. The average concentrations of total PCBs in mussels were 2795 pg g(-1) ww in the shipyard, 797 pg g(-1) ww in Marina 2 and 53 pg g(-1) ww in Marina 1 stations. The results of whole-sediment toxicity tests showed a strong correlation between toxicity test results and pollutant concentrations. Selected cytosolic enzyme activities in digestive glands differed significantly depending on localities. These differences in enzyme activities were mainly related to the different pollutant levels of the sampling sites. The micro-organic contaminant profile patterns, toxicity tests and biomarker studies showed that shipyards and shipbreaking yards are the major potential sources of organic pollution in coastal areas.

Use of passive samplers in pollution monitoring: a numerical approach for marinas.

Triolein-containing semipermeable membrane devices (SPMDs) and butyl rubber (BR) based sorbents were employed as passive samplers in 14 coastal stations of Turkey including shipyards and marinas to characterize time-integrated levels of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) and their relationship to potential pollution sources. Passive samplers of SPMDs and BR sorbents were deployed for 30days in the spring of 2012. The maximum concentrations of total PAH and PCB compounds sequestered by SPMDs were 3338 ng g(-1) SPMD and 4247 pg g(-1) SPMD. (END)-I and DDT-related compounds were dominant OCP compounds for most of the sites in passive samplers. Total PAH concentrations in SPMDs were found 1.2 to 8 times higher than the concentrations in BRs. However, BR sorbents were able to sample some PAHs which could not be sampled by SPMDs. The concentrations of PCBs and OCPs in BRs were similar or higher than SPMDs. SPMD-data were used to estimate the average ambient water concentrations of the contaminants. Two existing theoretical approaches have been used to derive the concentrations of hydrophobic pollutants in the ambient waters. The results were found very similar and range from 7318 to 183864 pg L(-1) for PAHs, from 2 to 186 pg L(-1) for PCBs, and from 98 to 848 pg L(-1) for OCPs. Furthermore, a simple numerical model was designed to estimate the boat-related water concentrations in marinas by using the seawater data supplied by SPMDs. The model was mainly built on the water concentration and the capacities of a particular marina and then applied to two sites in the second marina. A good correlation was found between the model outputs and SPMD-water data.

The acute effects of daily nicotine intake on heart rate--a toxicokinetic and toxicodynamic modelling study.

Joint physiologically-based toxicokinetic and toxicodynamic (PBTK/TD) modelling was applied to simulate concentration-time profiles of nicotine, a well-known stimulant, in the human body following single and repeated dosing. Both kinetic and dynamic models were first calibrated by using in vivo literature data for the Caucasian population. The models were then used to estimate the blood and liver concentrations of nicotine in terms of the Area Under Curve (AUC) and the peak concentration (Cmax) for selected exposure scenarios based on inhalation (cigarette smoking), oral intake (nicotine lozenges) and dermal absorption (nicotine patches). The model simulations indicated that whereas frequent cigarette smoking gives rise to high AUC and Cmax in blood, the use of nicotine-rich dermal patches leads to high AUC and Cmax in the liver. Venous blood concentrations were used to estimate one of the most common acute effects, mean heart rate, both at rest and during exercise. These estimations showed that cigarette smoking causes a high peak heart rate, whereas dermal absorption causes a high mean heart rate over 48h. This study illustrates the potential of using PBTK/TD modelling in the safety assessment of nicotine-containing products.

Screening of dioxin-like compounds by complementary evaluation strategy utilising ELISA, micro-EROD, and HRGC-HRMS in soil and sediments from Montevideo, Uruguay.

Polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDF) are persistent, toxic, and bioaccumulate in the environment. Due to their high analytical costs, these compounds are hardly regulated and mostly not monitored in the Third World. To overcome this, bioassays have been proposed as low-cost alternative methods. Two of the most established bioanalytical tools, the dioxin antibody-based enzyme-linked immunosorbent assay ELISA and the micro-EROD bioassay are evaluated and compared to high resolution gas chromatography and high resolution mass spectrometry (HRGC/HRMS) analytical methodology. The methods were tested using thirteen soils and sediment samples selected from diverse sites in Montevideo, Uruguay. The WHO2005 total toxic equivalent (WHO2005-TEQ) of soils ranged from 2.4 to 2212 (ng WHO2005-TEQ/kg dry sample) and from 0.14 to 9.4 (ng WHO2005-TEQ/kg dry sample) in sediments. This study shows significant contamination related to dioxin-like compounds, particularly in sites where uncontrolled burnings were carried out. ELISA and micro-EROD bioassay correlated well with HRGC/HRMS, R Spearman 0.773 and 0.913, respectively and were highly correlated to each other, R Spearman 0.879. Preliminary threshold values of bioassay toxic equivalents of 330 (ng/kg dry sample) for the micro-EROD bioassay and 220 (ng/kg dry sample) for ELISA are proposed.

Application of physiologically-based toxicokinetic modelling in oral-to-dermal extrapolation of threshold doses of cosmetic ingredients.

The application of physiologically based toxicokinetic (PBTK) modelling in route-to-route (RtR) extrapolation of three cosmetic ingredients: coumarin, hydroquinone and caffeine is shown in this study. In particular, the oral no-observed-adverse-effect-level (NOAEL) doses of these chemicals are extrapolated to their corresponding dermal values by comparing the internal concentrations resulting from oral and dermal exposure scenarios. The PBTK model structure has been constructed to give a good simulation performance of biochemical processes within the human body. The model parameters are calibrated based on oral and dermal experimental data for the Caucasian population available in the literature. Particular attention is given to modelling the absorption stage (skin and gastrointestinal tract) in the form of several sub-compartments. This gives better model prediction results when compared to those of a PBTK model with a simpler structure of the absorption barrier. In addition, the role of quantitative structure-property relationships (QSPRs) in predicting skin penetration is evaluated for the three substances with a view to incorporating QSPR-predicted penetration parameters in the PBTK model when experimental values are lacking. Finally, PBTK modelling is used, first to extrapolate oral NOAEL doses derived from rat studies to humans, and then to simulate internal systemic/liver concentrations - Area Under Curve (AUC) and peak concentration - resulting from specified dermal and oral exposure conditions. Based on these simulations, AUC-based dermal thresholds for the three case study compounds are derived and compared with the experimentally obtained oral threshold (NOAEL) values.

Comparative evaluation of ELISA kit and HPLC DAD for the determination of chlorpyrifos ethyl residues in water and sediments.

Enzyme linked immunosorbent assay (ELISA) kit is a versatile, cheap and relatively available tool that can be used in remote areas. In this study, performance of ELISA kit was evaluated in terms of accuracy, recovery, precision, sensitivity, cross reactivity and matrix interference for pesticide residue determination in water and sediment samples. This method was compared with high performance liquid chromatography (HPLC) which is not a commonly available analytical technique for chlorpyrifos ethyl residue analysis in developing countries. The ELISA kit had limits of detection (LOD) of 0.37 µg L(-1) and 0.42 µg Kg(-1) dry weight (dw), for chlorpyrifos ethyl in water and sediment samples, respectively using deionized water and a control sediment sample. Mean percentage recoveries and coefficients of variation (CV) for ELISA kit varied from 96.0±5.8% to 108.0±3.4% for water and sediment samples. Comparison between ELISA and HPLC analysis results using water and sediment samples from Lake Naivasha showed no significant difference in results (p≤0.05). Strong correlations (r2=0.9878 water samples and r1=0.9670, p<0.0001 for sediment samples, n=48) were reported between the methods for the two samples analyzed. Bland-Altman bias plot analysis showed that the two methods were in agreement within 95% confidence interval of limits -2.9 to 3.8 and -2.2 to 3.6 for water and sediment, respectively. Given the high sensitivity reported and the obtained acceptable limits of coefficient of variation and percentage recovery, ELISA appears to be a suitable rapid analytical tool in analysis of chlorpyrifos ethyl in water and sediment samples. Results demonstrate comparability to HPLC and could complement conventional tools in regular monitoring program particularly in developing countries. This will hasten results delivery for ecological risk assessment and timely execution of mitigation measures.

Water concentrations of PAH, PCB and OCP by using semipermeable membrane devices and sediments.

Water concentrations of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were estimated from semipermeable membrane devices (SPMDs) and from sediment pollutant concentrations. SPMDs were deployed in the Istanbul Strait and Marmara Sea and retrieved after 7 and 21 days. Performance reference compounds (PRCs) were used to determine the site-specific sampling rates of the compounds. Water concentrations (C(w)) of the analyzed compounds estimated by using two different calculation methods for SPMDs were found similar. C(w) of total PAHs estimated from SPMDs (C(w-spmd)) were found between 13 and 79 ng L⁻¹ and between 7.0 and 68 ng L⁻¹ for 7 and 21 days of deployments respectively. Water concentrations of PCBs using sediment data was found as between 0.001 and 11.0 ng L⁻¹. The highest value of C(w-spmd) for two deployments were 2.8 ng L⁻¹ for OCPs. C(w) estimated from sediment concentrations were generally higher than those estimated from SPMDs.

Association between chemical pattern in breast milk and congenital cryptorchidism: modelling of complex human exposures.

During the past four decades, there has been an increase in the incidence rate of male reproductive disorders in some, but not all, Western countries. The observed increase in the prevalence of male reproductive disorders is suspected to be ascribable to environmental factors as the increase has been too rapid to be explained by genetics alone. To study the association between complex chemical exposures of humans and congenital cryptorchidism, the most common malformation of the male genitalia, we measured 121 environmental chemicals with suspected or known endocrine disrupting properties in 130 breast milk samples from Danish and Finnish mothers. Half the newborns were healthy controls, whereas the other half was boys with congenital cryptorchidism. The measured chemicals included polychlorinated biphenyls (PCBs), polybrominated diphenyl-ethers, dioxins (OCDD/PCDFs), phthalates, polybrominated biphenyls and organochlorine pesticides. Computational analysis of the data was performed using logistic regression and three multivariate machine learning classifiers. Furthermore, we performed systems biology analysis to explore the chemical influence on a molecular level. After correction for multiple testing, exposure to nine chemicals was significantly different between the cases and controls in the Danish cohort, but not in the Finnish cohort. The multivariate analysis indicated that Danish samples exhibited a stronger correlation between chemical exposure patterns in breast milk and cryptorchidism than Finnish samples. Moreover, PCBs were indicated as having a protective effect within the Danish cohort, which was supported by molecular data recovered through systems biology. Our results lend further support to the hypothesis that the mixture of environmental chemicals may contribute to observed adverse trends in male reproductive health.

The Yangtze-Hydro Project: a Chinese-German environmental program.

Water of good quality is one of the basic needs of human life. Worldwide, great efforts are being undertaken for an assured water supply. In this respect, one of the largest water technology projects worldwide is the Yangtze Three Gorges Dam in China. There is a need for extensive scientific and technical understanding of the challenges arising from this large hydrological engineering project. German and Chinese groups from various scientific fields are collaborating to provide knowledge for the sustainable management of the reservoir. In this project description, the Yangtze Three Gorges Dam Project, its goals and challenges, are described in brief, and the contributions of the German research projects are presented.

Assessment of PCDD/F, PCB, OCP and BPA dietary exposure of non-breast-fed European infants.

The aim of this study is to obtain data on the exposure of non-breast-fed infants to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), polychlorinated biphenyls (PCB), organochlorine pesticides (OCP), and bisphenol A (BPA) and its chlorinated derivatives through consumption of commercial infant foods with largest shares of the market in 22 European Union countries. The E-Screen bioassay was employed to assess the oestrogenicity of the baby foods and the ethoxyresorufin-O-deethylase (EROD) induction was measured to determine the levels of PCDD/F and PCB. Consequently, the highest total effective xenoestrogen burden (TEXB) of 73.60 pM Eeq g(-1) was found in the soy-based formula and the EROD bioassay was always below the limit of quantification (LOQ) (3.5 pg g(-1)). Overall, the estimated dietary exposure to BPA via commercial baby foods was lower than the tolerable daily intake (TDI) of 50 µg kg(-1) body weight (bw). Furthermore, the findings indicated that the dietary exposure of 0-9-month-old infants through the products investigated here does not exceed the maximum TDI of 4 pg WHO-TEQ (toxic equivalents) kg(-1) bw. However, exposure to more than 2 pg WHO-TEQ kg(-1) bw day(-1) might occur for 0-4-month-old infants consuming 'starting' hypoallergenic formula. Moreover, analysis of OCP indicated that the dietary exposure of non-breast-fed infants was not harmful. Considering the importance of early development and the vulnerability of infants and children, it is essential to determine their dietary exposure to contaminants in order to decide which efforts of risk reduction should receive highest priority.

Bioaccumulation depuration kinetics and effects of benzo(a)anthracene on Mytilus galloprovincialis.

Polycyclic aromatic hydrocarbons (PAHs) are important environmental pollutants due to their persistence and bioaccumulation potential both in organisms and in sediments. In this study, bioaccumulation and depuration experiments were performed employing local Mediterranean mussel species, Mytilus galloprovincialis, with two biomarkers: filtration rate and lysosomal stability (neutral red retention) assay. Benzo(a)anthracene (BaA) was chosen as the model PAH compound due to its common presence in several matrices in the marine environment. Bioconcentration Factors (BCFs) for the mussels exposed to different BaA concentrations were calculated from both kinetic rate constants and from the experimental data and found between 27-3184 and 16-2745, respectively. Experimental QSAR values were found comparable to the previous QSAR models created for BaA. The effect of BaA exposure on the mussels showed a more clear dose-response relationship according to the results of lysosomal stability compared to the filtration rate.

Assessment of dietary exposure to PCDD/F and dioxin-like PCB in infant formulae available on the EU market.

In infant period, human milk and infant formulae are the major sources of exposure to dioxins (polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (PCB). Since in many parts of Europe the mothers are increasingly reluctant to breastfeed their babies, the main objective of the present study is to assess the level of exposure of non-breast-fed infants during their early development. Consequently, the concentration levels of PCDD/F and dioxin-like PCB were determined in industrial infant formulae including "starting" (aged 0-4months) and "follow-on" (after 4months) products of milk formula, soy formula and hypoallergenic infant formula, available on the EU market. Furthermore, dietary exposure to dioxin of exclusively formula-fed infants was assessed at each month from 0 to 9months. The results suggest that dietary exposure to dioxins via formulae is higher when consuming "starting" than "follow-on" infant formulae on a body-weight basis. Estimated dietary exposure after 4months of age was always below the lowest range of the Tolerable Daily Intake (TDI) of 1pg WHO-TEQkg(-1)bwd(-1) and Provisional Tolerable Monthly Intake (PTMI) of 70pg WHO-TEQkg(-1)bwmonth(-1). The highest estimated cumulative dietary exposure to PCDD/F and dioxin-like PCB was obtained considering infants of 0-1months fed with the "starting" hypoallergenic infant formula (2.8pg WHO-TEQkg(-1)bwd(-1) and 84pg WHO-TQkg(-1)bwmonth(-1)). However, these elevated levels are much lower than those observed in some studies in breast-fed infants. The study can be used for further risk assessments in regard to infant exposure.

Design and preparation of market baskets of European Union commercial baby foods for the assessment of infant exposure to food chemicals and to their effects.

The assessment of acute and chronic dietary exposure to contaminants in baby foods is needed to ensure healthy infant growth. Monthly European Union market baskets of commercial baby foods were designed for the first 9 months of life by the 'babyfood' study group of the CASCADE Network of Excellence for the specific purpose of assessing exposure to potentially toxic substances in infants fed with such foods. The present paper reports the different steps that led to the preparation of monthly pooled samples of commercial baby foods ('Infant formulae and follow-on formulae' and 'Other baby foods') that may constitute the extreme case of the diet for an infant who would not be breast fed at all. Several market baskets were generated for an 'average European Union infant' and for infants of four selected countries (Italy, Sweden, Spain, and the Slovakia), fed with either milk infant formulae, soy infant formulae or hypoallergenic infant formulae and weaned (at the fifth month) with commercial baby foods and beverages available on the European Union market. Market share data for 2007 for baby foods were used to design the baskets. Holding companies and the name of all their products were identified. Monthly diets for European Union infants were elaborated in terms of food categories (e.g. infant cereals) of typologies of products (e.g. infant cereals without gluten) and of a specific product. The number of baskets generated was 30 for 'Infant formulae and follow-on formulae' (including 62 products) and 13 for 'Other baby foods' (including 35 products). These market baskets were designed to be used for the determination of certain contaminants and nutrients in the diet of European Union infants and for the assessment of their effects on infant health.

Country-specific chemical signatures of persistent environmental compounds in breast milk.

Recent reports have confirmed a worldwide increasing trend of testicular cancer incidence, and a conspicuously high prevalence of this disease and other male reproductive disorders, including cryptorchidism and hypospadias, in Denmark. In contrast, Finland, a similarly industrialized Nordic country, exhibits much lower incidences of these disorders. The reasons behind the observed trends are unexplained, but environmental endocrine disrupting chemicals (EDCs) that affect foetal testis development are probably involved. Levels of persistent chemicals in breast milk can be considered a proxy for exposure of the foetus to such agents. Therefore, we undertook a comprehensive ecological study of 121 EDCs, including the persistent compounds dioxins, polychlorinated biphenyls (PCBs), pesticides and flame retardants, and non-persistent phthalates, in 68 breast milk samples from Denmark and Finland to compare exposure of mothers to this environmental mixture of EDCs. Using sophisticated, bioinformatic tools in our analysis, we reveal, for the first time, distinct country-specific chemical signatures of EDCs with Danes having generally higher exposure than Finns to persistent bioaccumulative chemicals, whereas there was no country-specific pattern with regard to the non-persistent phthalates. Importantly, EDC levels, including some dioxins, PCBs and some pesticides (hexachlorobenzene and dieldrin) were significantly higher in Denmark than in Finland. As these classes of EDCs have been implicated in testicular cancer or in adversely affecting development of the foetal testis in humans and animals, our findings reinforce the view that environmental exposure to EDCs may explain some of the temporal and between-country differences in incidence of male reproductive disorders.

PCDD/F and PCB in spruce forests of the Alps.

PCDD/F and PCB concentrations in remote mountainous spruce stands of the Central European Alps show strong geographic variation. Independent of the matrix (0.5 year old needles, humus or mineral soil), the highest pollutant levels were always found at the lateral zones of the mountain range. High levels coincided with strong precipitation, particularly along the northern margin of the study region. The most volatile PCB congener propagated farther into the colder, drier central Alps than the heavier species. Matrices with different accumulation history (needles and humus) repeatedly reflected different spatial emission patterns. Consistent with its much longer exposure, pollutant levels in humus exceeded those of needles by up to two orders of magnitude. Needle contamination varied with altitude but the vertical trends were highly variable between transsects and changed between years, too.

Vertical distribution of organochlorine pesticides in humus along Alpine altitudinal profiles in relation to ambiental parameters.

In forest soils along vertical profiles located in different parts of the Alps, concentrations of persistent organic pollutants (POPs), namely organochlorine pesticides (OCPs) like dichlorodiphenyltrichloroethanes (DDTs), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCH), heptachlor, aldrin, dieldrin and mirex, were measured. Though local characteristics of the sites are influenced by numerous factors like orographic and meteorological parameters, forest stand characteristics and humus parameters, we ascertained a marked vertical increase of concentrations of some organochlorine compounds in the soil. On the basis of climatological values of each site, we found that the contamination increase with altitude can be ascribed to a certain 'cold condensation effect'. In addition, the perennial atmospheric deposition of POPs is controlled by precipitation. Other key parameters explaining the accumulation of POPs are the soil organic carbon stocks, the turnover times, the re-volatilisation and degradation processes, which vary with altitude.

Novel sampling methods for atmospheric semi-volatile organic compounds (SOCs) in a high altitude alpine environment.

High- and low-volume active air samplers as well as bulk deposition samplers were developed to sample atmospheric SOCs under the adverse conditions of a mountain environment. Active sampling employed separate filters for different European source regions. Filters were switched depending on daily trajectory forecasts, whose accuracy was evaluated post hoc. The sampling continued on three alpine summits over five periods of four months. The prevailing trajectories varied stronger between sampling periods than between stations. The sampling equipment (active and bulk deposition) proved dependable for operation in a mountain environment, with idle times being mainly due to non-routine manipulations and connectivity.

Long-term air monitoring of organochlorine pesticides using Semi Permeable Membrane Devices (SPMDs) in the Alps.

Atmospheric sampling of organochlorine pesticides (OCPs) was conducted using Semi Permeable Membrane Devices (SPMDs) deployed in the Alps at different altitudinal transects for two consecutive exposure periods of half a year and a third simultaneous year-long period. Along all the altitude profiles, the sequestered amounts of OCPs increased in general with altitude. SPMDs were still working as kinetic samplers after half a year for the majority of the OCPs. However, compounds with the lowest octanol-air partition coefficient (K(oa)), reached equilibrium within six months. This change in the SPMD uptake was determined for the temperature gradient along the altitude profile influencing K(oa), OCPs availability in the gaseous phase, and SPMD performance. In sum, it seems two effects are working in parallel along the altitude profiles: the change in SPMD performance and the different availability of OCPs along the altitudinal transects determined by their compound properties and concentrations in air.

A comparison of Alpine emissions to forest soil and spruce needle loads for persistent organic pollutants (POPs).

The project MONARPOP analysed the concentrations of semivolatile organic compounds (SVOCs) in two important sink compartments, needles of Norway spruce (Picea abies [L.] Karst.) and forest soil from 40 remote Alpine forest sites in Austria, Germany, Italy, Slovenia and Switzerland. In the present study the load of PCDD/F, PCB, PBDE, PAH, HCB, HCH and DDT in the Alps calculated on the basis of measured data are compared with their estimated emissions in the Alpine region. It comes out that the masses of the studied pollutants stored in the forests are higher than the corresponding emissions in the Alpine area indicating that the Alps are a sink for POPs advected from surrounding areas. It is assumed that local emissions of PCDD/F and PAH deriving from biomass burning are probably underestimated and that the pool of these pollutants in the forests represents the accumulation over some decades.