An integrative risk assessment approach for persistent chemicals: A case study on dioxins,furans and dioxin-like PCBs in France

For persistent chemicals slowly eliminated from the body, the accumulated concentration (body burden), rather than the daily exposure, is considered the proper starting point for the risk assessment. This work introduces an integrative approach for persistent chemical risk assessment by means of a dynamic body burden approach. To reach this goal a Kinetic Dietary Exposure Model (KDEM) was extended with the long term time trend in the exposure (historic exposure) and the comparison of bioaccumulation with body burden references for toxicity. The usefulness of the model was illustrated on the dietary exposure to PolyChlorinatedDibenzo-p-Dioxins (PCDDs), PolyChlorinatedDibenzoFurans (PCDFs) and PolyChlorinated Biphenyls (PCBs) in France. Firstly the dietary exposure to these compounds was determined in 2009 and combined with its long term time trend. In order to take differences between the kinetics of PCDD/F and dl-PCBs into account, three groups of congeners were considered i.e. PCDD/Fs, PCB 126 and remaining dl-PCBs. The body burden was compared with reference body burdens corresponding to reproductive, hepatic and thyroid toxicity. In the case of thyroid toxicity this comparison indicated that in 2009 the probability of the body burden to exceed its reference ranged from 2.8% (95% CI: 1.5–4.9%) up to 3.9% (95% CI: 2.7–7.1%) (18–29 vs. 60–79 year olds). Notwithstanding the decreasing long-term time trend of the dietary dioxin exposure in France, this probability still is expected to be 1.5% (95% CI: 0.3–2.5%) in 2030 in 60–79 olds. In the case of reproductive toxicity the probability of the 2009 body burden to exceed its reference ranged from 3.1% (95% CI: 1.4–5.0%) (18–29 year olds) to 3.5% (95% CI: 2.2–5.2%) (30–44 year olds). In 2030 this probability is negligible in 18–29 year olds, however small though significant in 30–44 year olds (0.7%, 95% CI: 0–1.6%). In the case of hepatic toxicity the probability in 2009 even in 60–79 year olds already was negligible. In conclusion this approach indicates that in France dioxin levels in food form a declining, though still present, future health risk with respect to thyroid and reproductive toxicity.     

Molecular interactions of dioxins and DLCs with the ketosteroid receptors: an in silico risk assessment approach

Dioxins and dioxin-like compounds (DLCs) are the ones with poor water solubility and low volatility, resistant to physical, chemical and biological processes, persistent in the environment even under extreme conditions. Due to lipophilic nature, they get adhered to the fatty material and concentrate through biomagnification and bioaccumulation, thereby easily getting incorporated into food chains, paving the way to endocrine disruption via modulation of various human receptors. This in turn leads to certain adverse health effects. In the present study, a total of 100 dioxins and DLCs were taken and their binding pattern was assessed with the ketosteroid receptors, i.e. androgen (hAR), glucocorticoid (hGR), progesterone (hPR) and mineralocorticoid (hMR) in comparison to the corresponding natural steroids and a known endocrine disrupting xenobiotic, Bisphenol A (BPA). Most of the DLCs, particularly those bearing hydroxyl (-OH) group showed considerable affinities with ketosteroid receptors. On comparing D scores of all the dioxins and DLCs against all four receptors, compound 8-hydroxy-3,4-dichlorodibenzofuran(8-OH-DCDF) exhibited least D score of -9.549?kcal mol^?1 against hAR. 3,8-Dihydroxy-2-chlorodibenzofuran(3,8-DiOH-CDF), 4′-hydroxy-2,3,4,5-tetrachlorobiphenyl (4′-OH-TCB) and 4-hydroxy-2,2′,5′-trichlorobiphenyl(4-OH-TCB) also showed comparable molecular interactions with the ketosteroid receptors. These interactions mainly include H-bonding, π–π stacking, hydrophobic, polar and van der Waals’ interactions. In contrast, BPA and some natural ligands tested in this study showed lower binding affinities with these receptors than certain DLCs reported herein, i.e. certain DLCs might be more toxic than the proven toxic agent, BPA. Such studies play a pivotal role in the risk assessment of exposure to dioxins and DLCs on human health.     

A molecular epidemiological approach to health risk assessment of urban air pollution

The ambient air of urban centres is polluted with potentially toxic chemicals mostly arising from the combustion or fuels used for transport. Among these compounds, benzene raises particular concern due to its haematoxicity and leukaemogenic risks. Although limits of benzene in air have been established in the European Union (5 microg/m(3)), individual exposure levels--and therefore risk estimates--cannot merely be extrapolated from environmental concentrations. Molecular epidemiology can facilitate health risk assessment by investigating the relationship between exposure to environmental pollutants and quantification of biomarkers that lie on the pathway of carcinogenesis upstream of clinical disease. We review the available for biomarker studies regarding health risks linked to environmental benzene exposure, and make some suggestions for future work.     

Trace element risk assessment: essentiality vs. toxicity

Risk assessment of essential trace elements examines high intakes resulting in toxicity and low intakes resulting in nutritional deficiencies. This paper analyzes the risk assessments carried out by several U.S. governmental and private organizations for eight essential trace elements: chromium, copper, iodine, iron, manganese, molybdenum, selenium, and zinc. The compatibility of the toxicity values with the nutritionally essential values is examined, in light of recently derived values, termed Dietary Reference Intakes, set by the U.S. Food and Nutrition Board of the Institute of Medicine. The results show that although there are differences in the values set by the different organizations, increased coordination has resulted in values that are more compatible than revealed in past evaluations.     

A combined approach to drug metabolism and toxicity assessment.

The challenge of predicting the metabolism or toxicity of a drug in humans has been approached using in vivo animal models, in vitro systems, high throughput genomics and proteomics methods, and, more recently, computational approaches. Understanding the complexity of biological systems requires a broader perspective rather than focusing on just one method in isolation for prediction. Multiple methods may therefore be necessary and combined for a more accurate prediction. In the field of drug metabolism and toxicology, we have seen the growth, in recent years, of computational quantitative structure-activity relationships (QSARs), as well as empirical data from microarrays. In the current study we have further developed a novel computational approach, MetaDrug, that 1) predicts metabolites for molecules based on their chemical structure, 2) predicts the activity of the original compound and its metabolites with various absorption, distribution, metabolism, excretion, and toxicity models, 3) incorporates the predictions with human cell signaling and metabolic pathways and networks, and 4) integrates networks and metabolites, with relevant toxicogenomic or other high throughput data. We have demonstrated the utility of such an approach using recently published data from in vitro metabolism and microarray studies for aprepitant, 2(S)-((3,5-bis(trifluoromethyl)benzyl)-oxy)-3(S)phenyl-4-((3-oxo-1,2,4-triazol-5-yl)methyl)morpholine (L-742694), trovofloxacin, 4-hydroxytamoxifen, and artemisinin and other artemisinin analogs to show the predicted interactions with cytochromes P450, pregnane X receptor, and P-glycoprotein, and the metabolites and the networks of genes that are affected. As a comparison, we used a second computational approach, MetaCore, to generate statistically significant gene networks with the available expression data. These case studies demonstrate the combination of QSARs and systems biology methods.     

Trichloroethylene: mechanisms of renal toxicity and renal cancer and relevance to risk assessment.

1,1,2-Trichloroethylene (TCE) is an important solvent that is widespread in the environment. We have reviewed carcinogenicity data from seven bioassays with regard to renal injury and renal tumors. We report a consistent but low incidence of renal tubule carcinoma in male rats. Epidemiology studies on workers exposed to TCE (and other chlorinated solvents) indicate a weak association between high-level exposure and renal cancer. There appears to be a threshold below which no renal injury or carcinogenicity is expected to arise. TCE is not acutely nephrotoxic to rats or mice, but subchronic exposure to rats produces a small increase in urinary markers of renal injury. Following chronic exposure, pathological changes (toxic nephrosis and a high incidence of cytomegaly and karyomegaly) were observed. The basis for the chronic renal injury probably involves bioactivation of TCE. Based on the classification by E. A. Lock and G. C. Hard (2004, Crit. Rev. Toxicol. 34, 211-299) of chemicals that induce renal tubule tumors, we found no clear evidence to place TCE in category 1 or 2 (chemicals that directly or indirectly interact with renal DNA), category 4 (direct cytotoxicity and sustained tubule cell regeneration), category 5 (indirect cytotoxicity and sustained tubule cell regeneration associated with alpha2u-globulin accumulation), or category 6 (exacerbation of spontaneous chronic progressive nephropathy). TCE is best placed in category 3, chemicals that undergo conjugation with GSH and subsequent enzymatic activation to a reactive species. The implication for human risk assessment is that TCE should not automatically be judged by linear default methods; benchmark methodology could be used.     

Simulated impact of a fish based shift in the population n--3 fatty acids intake on exposure to dioxins and dioxin-like compounds.

Due to the favourable health effects of LC n-3 PUFAs, marine products have been recognised as a food group of special importance in the human diet. However, seafood is susceptible to contamination by lipophilic organic pollutants. The objective of this study was to evaluate intake levels of PCDDs, PCDFs and dioxin-like PCBs, by a probabilistic Monte Carlo procedure, in relation to the recommendation on LC n-3 PUFAs given by Belgian Federal Health Council. Regarding the recommendation, two scenarios were developed differing in LC n-3 PUFAs intake: a 0.3 E% and a 0.46 E% scenario. Total exposure to dioxins and dioxin-like substances in the 0.3 E% LC n-3 PUFAs scenario ranges from 2.31 pg TEQ/kg bw/day at the 5th percentile, over 4.37 pg TEQ/kgbw/day at the 50th percentile to 8.41 pg TEQ/kgbw/day at the 95th percentile. In the 0.46 E% LC n-3 PUFAs scenario, 5, 50 and 95th percentile are exposed to 2.74, 5.52 and 9.98 pg TEQ/kgbw/day, respectively. Therefore, if the recommended LC n-3 PUFAs intake would be based on fish consumption as the only extra source, the majority of the study population would exceed the proposed health based guidance values for dioxins and dioxin-like substances.     

A new approach for the assessment of acrylamide toxicity using a green paramecium.

Exposure to acrylamide induces neurotoxic effects in humans. In addition, it induces genotoxic, reproductive and carcinogenic effects in laboratory animals. However, no convenient bioassay system for assessing acrylamide toxicity to animal and plant cells has been proposed to date. The present study aims to evaluate acrylamide toxicity to a green paramecium, Paramecium bursaria, bearing many endosymbiotic algae, because some chemicals are highly toxic to paramecia or microalgae, and some protozoa are already used for evaluation of environmental contaminations. Results showed that high acrylamide concentrations (> or = 1500 mg/l) have a lethal effect on P. bursaria. Although low acrylamide concentrations (< or = 150 mg/l) induced less change on the paramecium growth, the number of endosymbiotically growing algal cells drastically decreased. The acrylamide concentration required to induce a 50% decrease in the cell number (IC(50)) was determined to be 7.8 mg/l for endosymbiotic algae, indicating that the algal sensitivity to acrylamide was 7 and 15 times higher than that of Syrian hamster embryo (SHE) cells and the host cells, respectively. Here, we propose the use of P. bursaria being a convenient and sensitive bioindicator as a new approach for the assessment of acrylamide toxicity.     

Factors affecting toxicity and efficacy of polymeric nanomedicines

Nanomedicine is the application of nanotechnology to medicine. The purpose of this article is to review common characteristics of polymeric nanomedicines with respect to passive targeting. We consider several biodegradable polymeric nanomedicines that are between 1 and 100 nm in size, and discuss the impact of this technology on efficacy, pharmacokinetics, toxicity and targeting. The degree of toxicity of polymeric nanomedicines is strongly influenced by the biological conditions of the local environment, which influence the rate of degradation or release of polymeric nanomedicines. The dissemination of polymeric nanomedicines in vivo depends on the capillary network, which can provide differential access to normal and tumor cells. The accumulation of nanomedicines in the microlymphatics depends upon retention time in the blood and extracellular compartments, as well as the type of capillary endothelium surrounding specific tissues. Finally, the toxicity or efficacy of intact nanomedicines is also dependent upon tissue type, i.e., non-endocrine or endocrine tissue, spleen, or lymphatics, as well as tumor type.     

The effects of limiting nutrient to algal toxicity assessment: A theoretical approach

The effects of limiting nutrient to algal toxicity tests are discussed. It has been shown, through theoretical derivation, that EC₅₀ values will vary with changes of nutrient concentration even if the effects of limiting nutrient and limiting toxicant on algal growth are independent. The variation in EC₅₀'s is governed by the degree of correlation between two variables: the tolerance and the activation level. The proposed theory shows a potential application for making a comparison of results obtained from different laboratories or methods and for applying laboratory results to field conditions.     

Factors modulating the epithelial response to toxicants in tracheobronchial airways

As one of the principal interfaces between the organism and the environment, the respiratory system is a target for a wide variety of toxicants and carcinogens. The cellular and architectural complexity of the respiratory system appears to play a major role in defining the focal nature of the pulmonary response to environmental stressors. This review will address the biological factors that modulate the response of one of the major target compartments within the respiratory system, the tracheobronchial airway tree. Individual airway segments respond uniquely to toxic stress and this response involves not only the target cell population, e.g. epithelium, but also other components of the airway wall suggesting a trophic interaction within all components of the airway wall in maintaining steady state and responding to injury. A number of biological factors modulate the nature of the response, including: (1) metabolic potential at specific sites for activation and detoxification; (2) the nature of the local inflammatory response; (3) age of the organism at the time of exposure; (4) gender of the exposed organism; (5) history of previous exposure; and (6) species and strain of the organism exposed.     

Factors affecting the soil arsenic bioavailability,accumulation in rice and risk to human health: a review

Arsenic (As), a class one carcinogen, reflects a disastrous environmental threat due to its presence in each and every compartment of the environment. The high toxicity of As is notably present in its inorganic forms. Irrigation with As contaminated groundwater in rice fields increases As concentration in topsoil and its bioavailability for rice crops. However, most of the As in paddy field topsoils is present as As(III) form, which is predominant in rice grain. According to the OECD-FAO, rice is the second most extensively cultivated cereal throughout the world. This cereal is a staple food for a large number of populations in most of the developing countries in sub-Saharan Africa, Latin America, South and South-east Asia. Rice consumption is one of the major causes of chronic As diseases including cancer for Asian populations. Thus, this review provides an overview concerning the conditions involved in soil that leads to As entrance into rice crops, phytotoxicity and metabolism of As in rice plants. Moreover, the investigations of the As uptake in raw rice grain are compiled, and the As biotransfer into the human diet is focused. The As uptake by rice crop represents an important pathway of As exposure in countries with high rice and rice-based food consumption because of its high (more than the hygienic level) As levels found in edible plant part for livestock and humans.     

Toxicokinetics as a key to the integrated toxicity risk assessment based primarily on non-animal approaches

Toxicokinetics (TK) is the endpoint that informs about the penetration into and fate within the body of a toxic substance, including the possible emergence of metabolites. Traditionally, the data needed to understand those phenomena have been obtained in vivo. Currently, with a drive towards non-animal testing approaches, TK has been identified as a key element to integrate the results from in silico, in vitro and already available in vivo studies. TK is needed to estimate the range of target organ doses that can be expected from realistic human external exposure scenarios. This information is crucial for determining the dose/concentration range that should be used for in vitro testing. Vice versa, TK is necessary to convert the in vitro results, generated at tissue/cell or sub-cellular level, into dose response or potency information relating to the entire target organism, i.e. the human body (in vitro–in vivo extrapolation, IVIVE). Physiologically based toxicokinetic modelling (PBTK) is currently regarded as the most adequate approach to simulate human TK and extrapolate between in vitro and in vivo contexts. The fact that PBTK models are mechanism-based which allows them to be ‘generic’ to a certain extent (various extrapolations possible) has been critical for their success so far. The need for high-quality in vitro and in silico data on absorption, distribution, metabolism as well as excretion (ADME) as input for PBTK models to predict human dose–response curves is currently a bottleneck for integrative risk assessment.     

An approach to risk assessment for TiO2

Titanium dioxide (TiO2) is a poorly soluble, low-toxicity (PSLT) particle. Fine TiO2 (<2.5 microm) has been shown to produce lung tumors in rats exposed to 250 mg/m3, and ultrafine TiO2 (< 0.1 microm diameter) has been shown to produce lung tumors in rats at 10 mg/m3. We have evaluated the rat dose-response data and conducted a quantitative risk assessment for TiO2. Preliminary conclusions are: (1) Fine and ultrafine TiO2 and other PSLT particles show a consistent dose-response relationship when dose is expressed as particle surface area; (2) the mechanism of TiO2 tumor induction in rats appears to be a secondary genotoxic mechanism associated with persistent inflammation; and (3) the inflammatory response shows evidence of a nonzero threshold. Risk estimates for TiO2 depend on both the dosimetric approach and the statistical model that is used. Using 7 different dose-response models in the U.S. Environmental Protection Agency (EPA) benchmark dose software, the maximum likelihood estimate (MLE) rat lung dose associated with a 1 per 1000 excess risk ranges from 0.0076 to 0.28 m2/g-lung of particle surface area, with 95% lower confidence limits (LCL) of 0.0059 and 0.042, respectively. Using the ICRP particle deposition and clearance model, estimated human occupational exposures yielding equivalent lung burdens range from approximately 1 to 40 mg/m3 (MLE) for fine TiO2, with 95% LCL approximately 0.7-6 mg/m3. Estimates using an interstitial sequestration lung model are about one-half as large. Bayesian model averaging techniques are now being explored as a method for combining the various estimates into a single estimate, with a confidence interval expressing model uncertainty.     

Coumarin metabolism, toxicity and carcinogenicity: relevance for human risk assessment.

The metabolism, toxicity and results of tests for carcinogenicity have been reviewed with respect to the safety for humans of coumarin present in foodstuffs and from fragrance use in cosmetic products. Coumarin is a natural product which exhibits marked species differences in both metabolism and toxicity. The majority of tests for mutagenic and genotoxic potential suggest that coumarin is not a genotoxic agent. The target organs for toxicity and carcinogenicity in the rat and mouse are primarily the liver and lung. Moreover, the dose-response relationships for coumarin-induced toxicity and carcinogenicity are non-linear, with tumour formation only being observed at high doses which are associated with hepatic and pulmonary toxicity. Other species, including the Syrian hamster, are seemingly resistant to coumarin-induced toxicity. There are marked differences in coumarin metabolism between susceptible rodent species and other species including humans. It appears that the 7-hydroxylation pathway of coumarin metabolism, the major pathway in most human subjects but only a minor pathway in the rat and mouse, is a detoxification pathway. In contrast, the major route of coumarin metabolism in the rat and mouse is by a 3,4-epoxidation pathway resulting in the formation of toxic metabolites. The maximum daily human exposure to coumarin from dietary sources for a 60-kg consumer has been estimated to be 0.02 mg/kg/day. From fragrance use in cosmetic products, coumarin exposure has been estimated to be 0.04 mg/kg/day. The total daily human exposure from dietary sources together with fragrance use in cosmetic products is thus 0.06 mg/kg/day. No adverse effects of coumarin have been reported in susceptible species in response to doses which are more than 100 times the maximum human daily intake. The mechanism of coumarin-induced tumour formation in rodents is associated with metabolism-mediated, toxicity and it is concluded that exposure to coumarin from food and/or cosmetic products poses no health risk to humans.     

A non-traditional approach to risk assessment of respiratory exposure to outdoor air pollutants.

The aim of this study was to verify a non-traditional assessment of respiratory exposure to outdoor air pollutants in industrial areas. The technique involved environmental biological monitoring' using domestic rabbits in the neighborhood of a mercury-producing plant. Rabbits were exposed whole-body to mercury emissions for 6 months in special cages near the plant. Control rabbits were kept using the same schedule outside the polluted area. Potential toxicity was assessed by: (a) measurement of Hg-accumulation in lungs and other body tissues by atomic absorption spectroscopy (AAS); (b) scanning electron microscopy (SEM) of the tracheal surface; and (c) measurement of toxic effects on pulmonary alveolar macrophages (PAM) by a rossette assay (Fc immunoglobulin binding). We found increased Hg concentrations in the lungs, kidneys, liver, heart, brain and bone; changes in the mucosal relief of the trachea, and depression of PAM Fc receptor (FcR) activity to IgG. A 6-month chamber exposure of Wistar rats to the aerosol created from solid particles of the mercury-producing plant revealed the increase of Hg-content in the same body tissues except the brain, and, less intense morphologic changes on the tracheal relief. Biomonitoring using environmental exposure of domestic rabbits might be useful in screening for possible health hazards to the respiratory system from complex outdoor aerosols.     

The integrated project ReProTect: a novel approach in reproductive toxicity hazard assessment

Validated alternative test methods are urgently needed for safety testing of drugs, chemicals and cosmetics. Whereas some animal tests for topical toxicity have been successfully replaced by alternative methods, systemic toxicity testing requires new test strategies in order to achieve an adequate safety level for the consumer. Substantial numbers of animals are required for the current in vivo assays for drugs, chemicals and cosmetics and a broad range of pioneering alternative methods were already developed. These prerequisites motivate the development of a tiered testing strategy based on alternative tests for reproductive toxicity hazard. In the Integrated Project ReProTect, a consortium set up by the European Centre for the Validation of Alternative Methods (ECVAM) takes the lead to manage the development of a testing strategy in the area of reproductive toxicity. The reproductive cycle can be broken down into well-defined sub-elements, namely male and female fertility, implantation and pre/postnatal development. In this project, in vitro, in silico and sensor technologies will be developed, leading to testing strategies, that shall be implemented and disseminated.     

Relay bioavailability and toxicity of isometamidium residues: a model for human risk assessment

The bioavailability and potential toxicity of the residues of the antitrypanosomal drug isometamidium (ISMM) in bovine tissues were investigated in male Sprague-Dawley rats. They were allowed to feed for 7 and 21 days on a standard diet, to which were added lyophilized tissues from a calf treated im with a combination of 45 mg 14C-ISMM and 73 mg unlabelled ISMM (1 mg/kg bodyweight). Cumulative excretion of radioactivity of the residues in feces of the rats was on average 90% of the dose. No radioactivity was detectable in the tissues examined, including the kidney, liver, spleen, muscle, stomach and small intestine. No clinical effects were seen in any of the rats, and both gross and histopathological examinations did not reveal any lesions. In rats given 14C-ISMM (2.245 mg/kg bodyweight) by oral gavage, cumulative excretion of radioactivity in feces after 48 hr amounted to about 93% of the dose, and no radioactivity was detectable in tissues. Similarly, none of the rats showed any clinical effects and no gross lesions were seen at necropsy. This study shows that ISMM residues are not significantly bioavailable and exhibit no subchronic toxicity.