Identification of structural fingerprints for in vivo toxicity by using Monte Carlo based QSTR modeling of nitroaromatics.

Monte Carlo based method by using either SMILES based or combination of SMILES and Graph-based descriptors is an important strategy to build the QSAR/QSTR model for prediction of different biological endpoints. In this study, Monte Carlo based QSTR approach was applied to the dataset of 90 nitroaromatic compounds related to their in vivo toxicity, represented by 50% lethal dose concentration for rats (LD50). Both classification and regression-based QSTR models were developed to get an idea about different fingerprints for promoters and hinderers of nitroaromatics toxicity. The best classification model was obtained by using SMILES and graph-based (GAO) descriptor with 1ECK connectivity (sensitivity = 0.7143, specificity = 1.0000, accuracy = 0.8889, and MCC = 0.7774). The best regression model calculated by using SMILES and hydrogen-suppressed graph descriptors with 0ECk connectivity (R2 = 0.7386, Q2 = 0.6315, S = 0.467, and MAE = 0.340). Finally, a consensus QSTR model was generated to predict efficiently the toxicity of new compounds. The study highlighted that the comparative QSTR models by using the Monte Carlo method can also be generated and will be a useful tool for structural fingerprint analysis in case of nitroaromatics for preliminary evaluation of its toxicity to mammals.

A simple approach for assessment of toxicity of nitroaromatic compounds without using complex descriptors and computer codes.

A simple approach is introduced to assess the toxicity of nitroaromatic compounds in terms of an oral LD50 dose (50% lethal dose) for rats. Most of the presented Quantitative Structure-Activity Relationship (QSAR) models for prediction of in vivo toxicity of nitroaromatics are calculated by quantum computing descriptors which are more difficult to interpret and apply, while the new model requires only the molecular structure of a desirable nitroaromatic compound. The novel model is based on the constitutional descriptors, such as the number of oxygen, sulphur, phosphorous and molecular fragments. Experimental data of 90 nitroaromatics are used to derive and test the new model as the logarithm of LD50 values, i.e. -log (LD50). Although it is based on only simple structural parameters, the reliability of the new model is also higher than the complex QSAR model because the values of the root-mean-square deviation (RMSD) of -log (LD50) for the new and the outputs of the latest QSAR method are 0.342 and 0.377, respectively.

Preparation and Measuring of Brain GABAA R-coupled Cl- /HCO3- - Activity for Integral Assessment of Aquatic Toxicity.

The wide use of aromatic hydrocarbons in various industries is having a negative effect on the environment and human health. Therefore, a key focus of current toxicology is the development and use of protein reporters with high sensitivity to various aromatic hydrocarbons (including phenolics and drugs). One molecular target for a wide range of pharmacology drugs and aromatic hydrocarbons (including phenol) is the neuronal GABAA R-coupled Cl- /HCO3- -ATPase. In this study, we present a protocol for isolation of the membrane-bound Cl- /HCO3- -ATPase from neuronal cells of animal brain. We then describe an uncomplicated in vitro method for measuring this ATPase activity for assessment of toxicity after interaction of this protein with an aquatic sample. This assay offers new avenues for using the Cl- /HCO3- -ATPase as a biomarker of water toxicity. This biotest is efficient, requires very little of the enzyme, and retains its sensitivity at low levels of various compounds. © 2019 by John Wiley & Sons, Inc.

Four selected high molecular weight heterocyclic aromatic hydrocarbons: Ecotoxicological hazard assessment, environmental relevance and regulatory needs under REACH.

Little is known about the ecotoxicity of heterocyclic aromatic hydrocarbons (NSO-HETs) to aquatic organisms. In the environment, NSO-HETs have been shown to occur in a strong association with their unsubstituted carbocyclic analogues, the polycyclic aromatic hydrocarbons (PAH), for which much more information is available. The present study addressed this issue by investigating the toxicity of four selected NSO-HETs in green algae (Desmodesmus subspicatus), daphnids (Daphnia magna) and fish embryos (Danio rerio). The four high molecular weight NSO-HETs dibenz[a,j]acridine (DBA), 7H-dibenzo[c,g]carbazole (DBC), benzo[b]naphtho[2,1-d]thiophene (BNT) and benzo[b]naphtho[1,2-d]furan (BNF) were selected, based on the results of a previous research project, indicating a lack of toxicity data and a high potential for persistence and bioaccumulation. The solubilities of the NSO-HETs in the test media were determined and turned out to be comparatively low (2.7-317 µg/L) increasing in the following order: DBA < BNT « DBC « BNF. Exposure concentrations during the toxicity tests were quantified with GC-MS and decreased strongly possibly due to sorption or metabolising during the test periods (48-96 h). Therefore, the estimated effect concentrations were related to the mean measured concentrations, as endpoints related to nominal concentrations would have underestimated the toxicity many times over. Within the range of the substance solubilities, BNF affected all test organisms with fish embryos being the most sensitive (fish: EC50 6.7 µg/L, algae: EC10 17.8 µg/L, daphnids: EC50 55.8 µg/L). DBC affected daphnids (EC50 2.5 µg/L,) and algae (EC10 3.1 µg/L), but not fish embryos. The lowest toxicity endpoint was observed for BNT affecting only algae (NOEC 0.556 µg/L) and neither daphnids nor fish embryos. DBA did not show any effects on the tested organisms in the range of the water solubility. However, we would expect effects in long-term toxicity studies to fish and aquatic invertebrates for all substances at lower concentrations, which needs further investigation. All four NSO-HETs were identified in mussels (Mytilus edulis) from the German coasts, in green kale (Brassica oleracea var. acephala) and in freshwater harbor sediment in concentrations between 0.07 and 2 µg/kg, highlighting their relevance as environmental contaminants. There is a need to regulate the four NSO-HETs within the REACH regulation due to their intrinsic properties and their environmental relevance. However, acquisition of additional experimental data appears to be pivotal for a regulation under REACH.

Toxicological Assessment of Mineral Hydrocarbons in Foods: State of Present Discussions.

The evaluation of mineral oils by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) provided high acceptable daily intakes for classes largely falling into the mass range strongly accumulated by humans. Because they are roughly 2 orders of magnitude above the present exposure, they authorize strongly increased exposure. An approach based on accumulation seems more adequate. Increased organ weights might be more critical than granulomas. Aromatic hydrocarbons with 1-2 aromatic rings should be distinguished from those with at least 3 aromatic rings. If mineral oil saturated hydrocarbon limits were low, no limit might be needed for the 1-2 ring aromatics. It should be considered to phase out substantial use of mineral oils in food application.

[Features of diurnal profile of blood pressure in workers having serum aromatic hydrocarbons level].

Features of diurnal profile of blood pressure in workers having serum level of benzol and ethylbenzene are high systolic and diastolic arterial blood pressure during the day, index of systolic arterial pressure time and index diastolic arterial pressure time was also high. These features should be considered in anti-hypertensives prescription.

Air pollution and general practitioner access and utilization: a population based study in Sarnia, 'Chemical Valley,' Ontario.

BACKGROUND: Health impacts of poor environmental quality have been identified in studies around the world and in Canada. While many of the studies have identified associations between air pollution and mortality or morbidity, few have focused on the role of health care as a potential moderator of impacts. This study assessed the determinants of health care access and utilization in the context of ambient air pollution in Sarnia, Ontario, Canada. METHODS: Residents of Sarnia participated in a Community Health Study administered by phone, while several ambient air pollutants including nitrogen dioxide (NO2), sulphur dioxide (SO2) and the volatile organic compounds benzene, toluene, ethylbenzene, mp- and o-xylene (BTEX) were monitored across the city. Land Use Regression models were used to estimate individual exposures to the measured pollutants and logistic regression models were utilized to assess the relative influence of environmental, socioeconomic and health related covariates on general practitioner access and utilization outcomes. RESULTS: The results show that general practitioner use increased with levels of exposure to nitrogen dioxide (NO2- Odds Ratio [OR]: 1.16, p < 0.05) and sulphur dioxide (SO2- OR: 1.61, p < 0.05). Low household income was a stronger predictor of having no family doctor in areas exposed to high concentrations of NO2 and SO2. Respondents without regular care living in high pollution areas were also more likely to report travelling or waiting for care in excess of 20 minutes (OR: 3.28, p < 0.05) than their low exposure counterparts (OR: 1.11, p > 0.05). CONCLUSIONS: This study provides evidence for inequitable health care access and utilization in Sarnia, with particular relevance to its situation as a sentinel high exposure environment. Levels of exposure to pollution appears to influence utilization of health care services, but poor access to primary health care services additionally burden certain groups in Sarnia, Ontario, Canada.

Developing tools for risk assessment in protected species: Relative potencies inferred from competitive binding of halogenated aromatic hydrocarbons to aryl hydrocarbon receptors from beluga (Delphinapterus leucas) and mouse.

Persistent organic pollutants such as halogenated aromatic hydrocarbons (HAHs) biomagnify in food webs and accumulate to high concentrations in top predators like odontocete cetaceans (toothed whales). The most toxic HAHs are the 2,3,7,8-substituted halogenated dibenzo-p-dioxins and furans, and non-ortho-substituted polychlorinated biphenyls (PCBs), which exert their effects via the aryl hydrocarbon receptor (AHR). Understanding the impact of HAHs in wildlife is limited by the lack of taxon-specific information about the relative potencies of toxicologically important congeners. To assess whether Toxic Equivalency Factors (TEFs) determined in rodents are predictive of HAH relative potencies in a cetacean, we used beluga and mouse AHRs expressed in vitro from cloned cDNAs to measure the relative AHR-binding affinities of ten HAHs from five different structural classes. The rank order of mean IC(50)s for competitive binding to beluga AHR was: TCDD

Validation of the target lipid model for toxicity assessment of residual petroleum constituents: monocyclic and polycyclic aromatic hydrocarbons.

A method is presented for developing scientifically defensible, numeric guidelines for residual petroleum-related constituents, specifically monocyclic aromatic hydrocarbons (MAHs) and polycyclic aromatic hydrocarbons (PAHs), in the water column. The guidelines are equivalent to a HC5 (i.e., hazard concentration to 5% of the tested species, or the concentration that protects 95% of the tested species). The model of toxicity used in this evaluation is the target lipid model (TLM) that was developed for assessing the toxicity of type I narcotic chemicals. An acute to chronic ratio is used for chronic expression and sublethal effects. The TLM is evaluated by comparing predicted and observed toxicity of these petroleum components. The methodology is capable of predicting both the acute and chronic toxicity of MAHs and PAHs in single exposures and in mixtures. For acute exposures, the TLM was able to predict the toxicity to within a factor of three to five. The use of toxic units was an effective metric for expressing the toxicity of mixtures. Within the uncertainty bounds, the TLM correctly predicted where sublethal effects of edemas, hemorrhaging, and other abnormalities were observed to occur in early life-stage exposure to PAHs. The computed HC5s were lower than no-observed-effect concentrations based on growth, reproduction, and mortality endpoints and sublethal effects. The methodology presented can be used by the oil spill community to compare residual concentrations of PAHs against defensible, numeric guidelines to assess potential ecological impacts.

Zebrafish whole-adult-organism chemogenomics for large-scale predictive and discovery chemical biology.

The ability to perform large-scale, expression-based chemogenomics on whole adult organisms, as in invertebrate models (worm and fly), is highly desirable for a vertebrate model but its feasibility and potential has not been demonstrated. We performed expression-based chemogenomics on the whole adult organism of a vertebrate model, the zebrafish, and demonstrated its potential for large-scale predictive and discovery chemical biology. Focusing on two classes of compounds with wide implications to human health, polycyclic (halogenated) aromatic hydrocarbons [P(H)AHs] and estrogenic compounds (ECs), we generated robust prediction models that can discriminate compounds of the same class from those of different classes in two large independent experiments. The robust expression signatures led to the identification of biomarkers for potent aryl hydrocarbon receptor (AHR) and estrogen receptor (ER) agonists, respectively, and were validated in multiple targeted tissues. Knowledge-based data mining of human homologs of zebrafish genes revealed highly conserved chemical-induced biological responses/effects, health risks, and novel biological insights associated with AHR and ER that could be inferred to humans. Thus, our study presents an effective, high-throughput strategy of capturing molecular snapshots of chemical-induced biological states of a whole adult vertebrate that provides information on biomarkers of effects, deregulated signaling pathways, and possible affected biological functions, perturbed physiological systems, and increased health risks. These findings place zebrafish in a strategic position to bridge the wide gap between cell-based and rodent models in chemogenomics research and applications, especially in preclinical drug discovery and toxicology.

Assessment of genotoxicity of methyl-tert-butyl ether, benzene, toluene, ethylbenzene, and xylene to human lymphocytes using comet assay.

Methyl-tert-butyl ether (MTBE) is a gasoline oxygenate and antiknock additive substituting for lead alkyls currently in use worldwide. Benzene, toluene, ethylbenzene, and xylene (BTEX) are volatile monoaromatic hydrocarbons which are commonly found together in crude petroleum and petroleum products such as gasoline. The aim of this study is to evaluate the genotoxic effects of these tested chemicals in human lymphocytes. Using the alkaline comet assay, we showed that all of the tested chemicals induce DNA damage in isolated human lymphocytes. This effect could follow from the induction of DNA strands breaks. The neutral version of the test revealed that MTBE, benzene, and xylenes induce DNA double-strand breaks at 200 microM. Apart from MTBE, the spin traps, 5,5-dimethyl-pyrroline-N-oxide (DMPO) and N-tert-butyl-alpha-phenylnitrone (PBN) can decrease the level of DNA damage in BTEX at 200 microM. This indicated that DNA damage could result from the participation of free radicals in DNA-damaging effect, which was further supported by the fact that post-treatment of formamidopyrimidine-DNA glycosylase (Fpg), enzyme recognizing oxidized DNA purines, gave rise to a significant increase in the extent of DNA damage in cells treated with benzene, and xylene at 200 microM. The results obtained suggested that MTBE and BTEX could induce a variety type of DNA damage such as single-strand breaks (SSBs), double-strand breaks (DSBs), and oxidative base modification.

Health risk assessment for polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and dibenzofurans, and polychlorinated naphthalenes in seafood from Guangzhou and Zhoushan, China.

This study determined the concentrations of organochlorine contaminants in common seafood in two Chinese coastal cities (Guangzhou and Zhoushan), and assessed the health risk due to the daily consumption of contaminated seafood. Twenty-six pooled samples, belonging to five food categories (fish, bivalves, shrimp, crab, and cephalopods), were purchased from local markets in Guangzhou and Zhoushan in 2003 and 2004. These samples were analyzed for total polychlorinated biphenyls (PCBs), non- and mono-ortho-PCBs (coplanar PCBs), polychlorinated naphthalenes (PCNs), and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/DFs). The concentrations of total PCBs and coplanar PCBs in fish samples were higher in Guangzhou than in Zhoushan, while the levels of PCNs and PCDDs/DFs were comparable between the two cities. The total daily intake values of dioxin-like compounds were 1.05 and 0.86pg WHO-TEQ/kg body weight in Guangzhou and Zhoushan, respectively. Hazard ratios of non-cancer risk in the two cities were all less than unity.

Toxicity assessment and microbial degradation of azo dyes.

Toxic effluents containing azo dyes are discharged from various industries and they adversely affect water resources, soil fertility, aquatic organisms and ecosystem integrity. They pose toxicity (lethal effect, genotoxicity, mutagenicity and carcinogenicity) to aquatic organisms (fish, algae, bacteria, etc.) as well as animals. They are not readily degradable under natural conditions and are typically not removed from waste water by conventional waste water treatment systems. Benzidine based dyes have long been recognized as a human urinary bladder carcinogen and tumorigenic in a variety of laboratory animals. Several microorganisms have been found to decolourize, transform and even to completely mineralize azo dyes. A mixed culture of two Pseudomonas strains efficiently degraded mixture of 3-chlorobenzoate (3-CBA) and phenol/cresols. Azoreductases of different microorganisms are useful for the development of biodegradation systems as they catalyze reductive cleavage of azo groups (-N=N-) under mild conditions. In this review, toxic impacts of dyeing factory effluents on plants, fishes, and environment, and plausible bioremediation strategies for removal of azo dyes have been discussed.

A review of hazardous chemical toxicity studies utilizing genetically-modified animals--their applications for risk assessment.

Studies on the mechanisms of chemical toxicity carried out using knockout mice lacking genes of enzymes for drug metabolism or nuclear receptor proteins were reviewed, and the studies were compared with the respective conventional mechanistic studies. While the toxicity of many hazardous chemicals was observed only in wild-type or knockout mice, which clearly showed that their toxicity was involved in the enzyme or receptor, some chemicals exhibited the same degree of toxicity in two genotypes, i.e., in both the wild strain and knockout mice, demonstrating that the enzymes or receptors are not involved in their toxicity. The use of genetically-modified animals presents not only the advantage of simultaneous evaluation of toxicity endpoints and mechanisms, but also suggests significant benefits over conventional methods using several chemicals to elucidate toxicity mechanisms. Elucidation of the mechanism of toxicity will provide useful information for risk assessment, and the use of genetically-modified animals for this purpose will lead to the advancement of this assessment.

Carcinogenicity of the aromatic amines: from structure-activity relationships to mechanisms of action and risk assessment.

Aromatic amines represent one of the most important classes of industrial and environmental chemicals: many of them have been reported to be powerful carcinogens and mutagens, and/or hemotoxicants. Their toxicity has been studied also with quantitative structure-activity relationship (QSAR) methods: these studies are potentially suitable for investigating mechanisms of action and for estimating the toxicity of compounds lacking experimental determinations. In this paper, we first summarized the QSAR models for the rodent carcinogenicity of the aromatic amines. The gradation of potency of the carcinogenic amines depended firstly on their hydrophobicity, and secondly on electronic (reactivity, propensity to be metabolically transformed) and steric properties. On the contrary, the difference between carcinogenic and non-carcinogenic aromatic amines depended mainly on electronic and steric properties. These QSARs can be used directly for estimating the carcinogenicity of aromatic amines. A two-step prediction is possible: (1) estimation of yes/no activity; (2) if the answer from step 1 is yes, then prediction of the degree of potency. The QSARs for rodent carcinogenicity were put in a wider context by comparing them with those for: (a) Salmonella mutagenicity; (b) general toxicity; (c) enzymatic reactions; (d) physical-chemical reactions. This comparative QSAR exercise generated a coherent global picture of the action mechanisms of the aromatic amines. The QSARs for carcinogenicity were similar to those for Salmonella mutagenicity, thus pointing to a similar mechanism of action. On the contrary, the general toxicity QSARs (both in vitro and in vivo systems) were mostly based on hydrophobicity, pointing to an aspecific mechanism of action much simpler than that for carcinogenicity and mutagenicity. The oxidation of the amines (first step in the main metabolic pathway leading to carcinogenic and mutagenic species) had identical QSARs in both enzymatic and physical-chemical systems, thus providing evidence for the link between simple chemical reactions and those in biological systems. The results show that it is possible to generate mechanistically and statistically sound QSAR models for rodent carcinogenicity, and indirectly that the rodent bioassay is a reliable source of good quality data.

Development of quantitative structure-activity relationships for the toxicity of aromatic compounds to Tetrahymena pyriformis: comparative assessment of the methodologies.

The purpose of this study was to develop quantitative structure-activity relationships (QSARs) for the toxicity of 268 aromatic compounds in the Tetrahymena pyriformis growth inhibition assay. The QSARs were developed using the response-surface (or two-parameter) approach, which was also modified using linear free-energy parameters to account for outliers. Subsequently, the data set was analyzed using partial least-squares (PLS). The results of the modeling using different methodologies were compared to the use of a Bayesian regularized neural network (BRANN) trained on the same data. Both response surface approaches, and PLS explained between 75 and 80% of the variance in the data; BRANN gave a higher statistical fit. In terms of the transparency of the approaches, the response surface clearly provides the simplest and easiest to use QSAR, it is readily interpreted in terms of mechanism of toxic action. PLS and BRANN are respectively less transparent. The use of atomistic and fragment-based indexes as descriptors in QSARs is assessed also, these are found not to be as useful as whole molecule parameters for the prediction of toxicity for molecules outside of the training set. The relative merits of the different approaches to the development of QSARs are described.

Bioassays for risk assessment of coal conversion products.

Traditional as well as biotechnological processing of coal leads to complex mixtures of products. Besides chemical and physical characterization, which provides the information for product application, there is a need for bioassays to monitor properties that are probably toxic, mutagenic or cancerogenic. Investigations carried out focused on the selection, adaptation and validation of bioassays for the sensitive estimation of toxic effects. Organisms like bacteria, Daphnia magna and Scenedesmus subspicatus, representing different complexities in the biosphere, were selected as test systems for ecotoxicological and mutagenicity studies. The results obtained indicate that bioassays are, in principle, suitable tools for characterization and evaluation of coal-derived substances and bioconversion products. Using coal products, coal-relevant model compounds and bioconversion products, data for risk assessment are presented.

Development validation and problems with the toxic equivalency factor approach for risk assessment of dioxins and related compounds.

Polychlorinated dibenzo-p-dioxins (PCDD), dibenzofurans (PCDF), and biphenyls (PCB) are industrial compounds or by-products that have been widely identified as environmental contaminants, and residues have been detected in fish, wildlife, and humans. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD; dioxin) is the most toxic member of this class of halogenated aromatic hydrocarbons (HAH); mechanistic studies indicate that the toxic and biochemical effects associated with exposure to TCDD are mediated via initial binding to the cytosolic aryl hydrocarbon (Ah) receptor protein present in target tissues and organs. Several other 2,3,7, 8-substituted PCDD and PCDF and non-ortho substituted PCB also bind to the Ah receptor and induce toxic responses similar to those for TCDD. Moreover, for these HAH there is a rank order correlation between their structure-Ah receptor binding and structure-toxicity relationships, and this supports a role for the Ah receptor in mediating these responses. Thus, the toxic equivalency factor (TEF) approach for HAH is based on the common mechanism of action for TCDD and related compounds in which a TEF value for a "dioxin-like" congener is defined as the potency of the individual (i) congener relative to TCDD ([EC50 [TCDD]/EC50 [test compound]). The toxic or dioxin equivalent (TEQ) for a mixture of HAH is defined by the following equation: TEQ = sigma [PCDDi] x TEFi + sigma [PCDFi] x TEFi. Industrial emissions and environmental and food residues contain complex mixtures of HAH (exodioxins) and the TEF/TEQ approach is used to regulate emissions and estimate the potential exposure and possible adverse health effects of exodioxins. The TEF approach for risk assessment of exodioxins makes a number of assumptions, including response additivity for individual compounds in a mixture of HAH. This review documents some of the following problems and limitations of the TEF approach: 1) environmental and food residues of HAH contain "non-dioxin-like" PCB that exhibit "antidioxin" activity for some responses; 2) the human diet contains endogenous Ah receptor ligands (endodioxins) such as polynuclear aromatic hydrocarbons (PAH), aromatic amines in cooked foods, indole-3-carbinol (I3C), and related hetero-PAH in cruciferous vegetables. Mass balance and mass potency estimates for human dietary intakes suggest that for some responses the effects of natural or endodioxins may be greater than those of exodioxins; and 3) I3C, a weak Ah receptor agonist, also exhibits Ah receptor antagonist activity, and interactions between I3C and endodioxins may inhibit or inactivate some toxic responses and decrease TEQ(Exodioxin).