Mode Of Toxic Action Research Articles

The mode of toxic action of ionic liquids: Narrowing down possibilities using high-throughput, in vitro cell-based bioassays

Growing concerns about the environmental impact of ionic liquids (ILs) have spurred research into their (eco)toxic effects, but studies on their mode of toxic action (MOA) still remain limited. However, understanding the MOA and identifying structural features responsible for enhanced toxicity is crucial for characterising the hazard and designing safer alternatives. Therefore, 45 ILs, with systematically varied chemical structures, were tested for cytotoxicity and two specific endpoints in reporter gene assays targeting the Nrf2-ARE mediated oxidative stress response (AREc32) and aryl hydrocarbon receptor activation (AhR-CALUX). While none of the ILs activated the reporter genes, cytotoxicity was high and markedly different between cell lines. Seven and 25 ILs proved more cytotoxic than predicted by baseline toxicity model in the AREc32 and the AhR-CALUX assays, respectively. The length of the side chain and headgroup structures of ILs altered the MOA of ILs. Cellular metabolism of the ILs, investigated by LC-MS/MS, showed side-chain oxidation of the long-chain quaternary ammonium compounds in AhR-CALUX cells and, to a lower extent, in AREc32 cells, however, this transformation could not explain the high cytotoxicity. Effect data for 72 ILs for ten endpoints retrieved from the Tox21 database identified the inhibition of aromatase activity and of mitochondrial membrane potential as potential MOAs. However, in vitro fluorimetric assays for these endpoints demonstrated that effects were activated in a non-specific manner, probably through cytotoxicity. Although many of the ILs tested induced cytotoxicity at concentrations lower than baseline toxicity, the specific MOAs responsible could not be identified. Alternatively, we suggest that the descriptors currently used may fail to define the affinity of ILs for cells. Testing of the affinity of ILs for a diverse range of biomolecules is needed to accurately describe their interactions with cells.

ToxProfiler: A novel human-based reporter assay for in vitro chemical safety assessment

In vitro chemical safety assessment often relies on simple and general cytotoxicity endpoint measurements and fails to adequately predict human toxicity. To improve the in vitro chemical safety assessment, it is important to understand the underlying mechanisms of toxicity. Here we introduce ToxProfiler, a novel human-based reporter assay that quantifies the chemical-induced stress responses at a single-cell level and reveals the toxicological mode-of-action (MoA) of novel drugs and chemicals. The assay accurately measures the activation of seven major cellular stress response pathways (oxidative stress, cell cycle stress, endoplasmic reticulum stress, ion stress, protein stress, autophagy and inflammation) that play a role in the adaptive responses prior to cellular toxicity. To assess the applicability of the assay in predicting the toxicity MoA of chemicals, we tested a set of 100 chemicals with well-known in vitro and in vivo toxicological profiles. Concentration response modeling and point-of-departure estimation for each reporter protein allowed for chemical potency ranking and revealed the primary toxicological MoA of chemicals. Furthermore, the assay could effectively group chemicals based on their shared toxicity signatures and link them to specific toxicological targets, e.g. mitochondrial toxicity and genotoxicity, and different human pathologies, including liver toxicity and cardiotoxicity. Overall, ToxProfiler is a quantitative in vitro reporter assay that can accurately provide insight into the toxicological MoA of compounds, thereby assisting in the future mechanism-based safety assessment of chemicals.

Evaluation of Short-Term Mussel Test for Estimating Toxicity.

Effect concentrations of ammonia, nickel, sodium chloride, and potassium chloride from short-term 7-day tests were compared to those from standard chronic 28-day toxicity tests with juvenile mussels (fatmucket, Lampsilis siliquoidea) to evaluate the sensitivities of the 7-day tests. The effect concentrations for nickel (59 µg Ni/L), chloride (316-519 mg Cl/L, a range from multiple tests), and potassium (15 mg K/L) obtained from the 7-day tests were within a range of effect concentrations for each corresponding chemical in the 28-day tests (41-91 µg Ni/L, 251->676 mg Cl/L, 15-23 mg K/L), whereas the 7-day ammonia effect concentration (0.40 mg/L total ammonia nitrogen; TAN) was up to 3.3-fold greater than the 28-day effect concentrations (0.12-0.36 mg TAN/L) but with overlapped 95% confidence limits. These results indicate that the 7-day tests produced similar estimates compared to the 28-day tests. Further studies are needed to evaluate the 7-day test sensitivity using additional chemicals with different modes of toxic action. Environ Toxicol Chem 2024;43:2020-2025. Published 2024. This article is a U.S. Government work and is in the public domain in the USA.

Plastic additive components of PM2.5 increase corrected QT interval: Screening for exposure markers based on airborne exposome.

The impact of industrial chemical components of ambient fine particles (e.g. PM2.5) on cardiovascular health has been poorly explored. Our study reports for the first time the associations between human exposure to complex plastic additive (PA) components of PM2.5 and prolongation of heart rate-corrected QT (QTC) interval by employing a screening-to-validation strategy based on a cohort of 373 participants (136 in the screening set and 237 in the validation set) recruited from 7 communities across China. The high-throughput airborne exposome framework revealed ubiquitous occurrences of 95 of 224 target PAs in PM2.5, totaling from 66.3 to 555 ng m-3 across the study locations. Joint effects were identified for 9 of the 13 groups of PAs with positive associations with QTC interval. Independent effect analysis also identified and validated tris(2-chloroisopropyl) phosphate, di-n-butyl/diisobutyl adipate, and 3,5-di-tert-butyl-4-hydroxybenzaldehyde as the key exposure markers for QTC interval prolongation and changes of selected cardiovascular biomarkers. Our findings highlight the important contributions of airborne industrial chemicals to the risks of cardiovascular diseases and underline the critical need for further research on the underlying mechanisms, toxic modes of action, and human exposure risks.

Using Multiple Metal Mixture Models to Predict Toxicity of Riverine Sediment Porewater to the Benthic Life Stage of Juvenile White Sturgeon (Acipenser transmontanus).

Five metal mixture dose-response models were used to predict the toxicity of porewater to young sturgeon at areas of interest in the Upper Columbia River (WA, USA/BC, Canada) and to evaluate these models as tools for risk assessments. Dose components of metal mixture models included exposure to free metal ion activities or metal accumulation by biotic ligands or humic acid, and links of dose to response used logistic equations, independent joint action equations, or additive toxicity functions. Laboratory bioassay studies of single metal exposures to juvenile sturgeon, porewater collected in situ in the fast-flowing Upper Columbia River, and metal mixture models were used to evaluate toxicity. The five metal mixture models were very similar in their predictions of adverse response of juvenile sturgeon and in identifying copper (Cu) as the metal responsible for the most toxic conditions. Although the modes of toxic action and the 20% effective concentration values were different among the dose models, predictions of adverse response were consistent among models because all doses were tied to the same biological responses. All models indicated that 56% ± 5% of 122 porewater samples were predicted to have <20% adverse response, 25% ± 5% of samples were predicted to have 20% to 80% adverse response, and 20% ± 4% were predicted to have >80% adverse response in juvenile sturgeon. The approach of combining bioassay toxicity data, compositions of field porewater, and metal mixture models to predict lack of growth and survival of aquatic organisms due to metal toxicity is an important tool that can be integrated with other information (e.g., survey studies of organism populations, life cycle and behavior characteristics, sediment geochemistry, and food sources) to assess risks to aquatic organisms in metal-enriched ecosystems. Environ Toxicol Chem 2024;43:62-73. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Interspecies scaling of toxicity reference values in human health versus ecological risk assessments: A critical review.

Risk assessments that focus on anthropogenic chemicals in environmental media-whether considering human health or ecological effects-often rely on toxicity data from experimentally studied species to estimate safe exposures for species that lack similar data. Current default extrapolation approaches used in both human health risk assessments and ecological risk assessments (ERAs) account for differences in body weight between the test organisms and the species of interest, but the two default approaches differ in important ways. Human health risk assessments currently employ a default based on body weight raised to the three-quarters power. Ecological risk assessments for wildlife (i.e., mammals and birds) are typically based directly on body weight, as measured in the test organism and receptor species. This review describes differences in the experimental data underlying these default practices and discusses the many factors that affect interspecies variability in chemical exposures. The interplay of these different factors can lead to substantial departures from default expectations. Alternative methodologies for conducting more accurate interspecies extrapolations in ERAs for wildlife are discussed, including tissue-based toxicity reference values, physiologically based toxicokinetic and/or toxicodynamic modeling, chemical read-across, and a system of categorical defaults based on route of exposure and toxic mode of action. Integr Environ Assess Manag 2024;20:749-764. © 2023 SETAC.

Transcriptional landscape of mitochondrial electron transport chain inhibition in renal cells

Analysis of the transcriptomic alterations upon chemical challenge, provides in depth mechanistic information on the compound’s toxic mode of action, by revealing specific pathway activation and other transcriptional modulations. Mapping changes in cellular behaviour to chemical insult, facilitates the characterisation of chemical hazard. In this study, we assessed the transcriptional landscape of mitochondrial impairment through the inhibition of the electron transport chain (ETC) in a human renal proximal tubular cell line (RPTEC/TERT1). We identified the unfolded protein response pathway (UPR), particularly the PERK/ATF4 branch as a common cellular response across ETC I, II and III inhibitions. This finding and the specific genes elaborated may aid the identification of mitochondrial liabilities of chemicals in both legacy data and prospective transcriptomic studies.Graphical abstract

Triphenyl phosphate proved more potent than its metabolite diphenyl phosphate in inducing hepatic insulin resistance through endoplasmic reticulum stress

Triphenyl phosphate (TPHP) is a widely used flame retardant and plasticizer and has been detected extensively in environmental media, wildlife and human bodies. Several epidemiological and animal studies have revealed that TPHP exposure is positively associated with glucose homeostasis disruption and diabetes. However, the effects of TPHP on hepatic glucose homeostasis and the underlying mechanisms remain unclear. The present work aimed to investigate the cytotoxicity and glucose metabolism disruption of TPHP and its metabolite diphenyl phosphate (DPHP) within hepatocytes. The cell viability assay undertaken on human normal liver (L02) cells showed that TPHP exhibited more potent hepatotoxicity than DPHP. RNA sequencing (RNA-seq) data showed that TPHP and DPHP presented different modes of toxic action. Insulin resistance is one of the predominant toxicities for TPHP, but not for DPHP. The insulin-stimulated glucose uptake and glycogen synthesis were impaired by TPHP, while DPHP exhibited no significant impairment on these factors. TPHP exposure induced endoplasmic reticulum (ER) stress, and the ER stress antagonist 4-PBA restored the impairment of insulin-stimulated glucose uptake and glycogen synthesis induced by TPHP. TPHP could also induce liver ER stress and insulin resistance in mice. Taken together, the results suggested that TPHP induces more potent insulin resistance through ER stress than its metabolite DPHP.

Differential contributions of the proteasome, autophagy, and chaperones to the clearance of arsenite-induced protein aggregates in yeast

The poisonous metalloid arsenite induces widespread misfolding and aggregation of nascent proteins invivo, and this mode of toxic action might underlie its suspected role in the pathology of certain protein misfolding diseases. Evolutionarily conserved protein quality-control systems protect cells against arsenite-mediated proteotoxicity, and herein, we systematically assessed the contribution of the ubiquitin-proteasome system, the autophagy-vacuole pathway, and chaperone-mediated disaggregation to the clearance of arsenite-induced protein aggregates in Saccharomyces cerevisiae. We show that the ubiquitin-proteasome system is the main pathway that clears aggregates formed during arsenite stress and that cells depend on this pathway for optimal growth. The autophagy-vacuole pathway and chaperone-mediated disaggregation both contribute to clearance, but their roles appear less prominent than the ubiquitin-proteasome system. Our invitro assays with purified components of the yeast disaggregating machinery demonstrated that chaperone binding to aggregates formed in the presence of arsenite is impaired. Hsp104 and Hsp70 chaperone activity was unaffected by arsenite, suggesting that this metalloid influences aggregate structure, making them less accessible for chaperone-mediated disaggregation. We further show that the defect in chaperone-mediated refolding of a model protein was abrogated in a cysteine-free version of the substrate, suggesting that arsenite directly modifies cysteines in non-native target proteins. In conclusion, our study sheds novel light on the differential contributions of protein quality-control systems to aggregate clearance and cell proliferation and extends our understanding of how these systems operate during arsenite stress.

Exploring specific biomarkers regarding neurobehavioral toxicity of lead dioxide nanoparticles in medaka fish in different water matrices

Nano-scale lead dioxide (nPbO2) is an industrial metal oxide nanoparticle that can be also formed as a corrosion by-product from chlorination of Pb-containing plumbing materials. nPbO2 governs release of toxic lead ion in drinking water and receiving organisms; however, its modes of toxic action regarding neurobehavioral toxicity remain unclear. This study evaluated the toxicity mechanism of nPbO2 (10 and 20 mg/L) versus its released Pb(II)aq (100 μg/L) in terms of aqueous chemistry, bioavailability and neurobehavioral toxicity to medaka fish in different water matrices. In very hard water (VHW), dissolved salts enhanced the aggregation and sedimentation of nPbO2, resulting in higher bioavailability and altered locomotion of treated fish than those fish exposed to nPbO2 in soft water with humic acid (SW + HA). Transcriptomic results identified six differentially expressed genes with greater altered expression with nPbO2 than the control or Pb(II)aq exposure. With VHW exposure, nPbO2 caused greater altered expression of genes involved in cell adhesion (nlgn1 and epd), cell cytoskeleton (α1-tubulin), and relevant apoptosis (c-fos, birc5.1-a and casp3), as compared with SW + HA or Pb(II)aq exposure. This study provides novel molecular mechanistic insights into the neurobehavioral nanotoxicity using nPbO2 and medaka fish as surrogates, suggesting nPbO2 promotes neurobehavioral dysfunction, leading to adverse outcomes from gene alteration to the organismal level. The identified biomarkers responded specifically to the nPbO2-induced neurotoxicity in different water matrices can be used for evaluating toxicity risks of small metal oxide particulates on human or aquatic life under environmentally relevant exposures.

Wastewater toxicity removal: Integrated chemical and effect-based monitoring of full-scale conventional activated sludge and membrane bioreactor plants

The literature is currently lacking effect-based monitoring studies targeted at evaluating the performance of full-scale membrane bioreactor plants. In this research, a monitoring campaign was performed at a full-scale wastewater treatment facility with two parallel lines (traditional activated sludge and membrane bioreactor). Beside the standard parameters (COD, nitrogen, phosphorus, and metals), 6 polynuclear aromatic hydrocarbons, 29 insecticides, 2 herbicides, and 3 endocrine disrupting compounds were measured. A multi-tiered battery of bioassays complemented the investigation, targeting different toxic modes of action and employing various biological systems (uni/multicellular, prokaryotes/eukaryotes, trophic level occupation). A traffic light scoring approach was proposed to quickly visualize the impact of treatment on overall toxicity that occurred after the exposure to raw and concentrated wastewater. Analysis of the effluents of the CAS and MBR lines show very good performance of the two systems for removal of organic micropollutants and metals. The most noticeable differences between CAS and MBR occurred in the concentration of suspended solids; chemical analyses did not show major differences. On the other hand, bioassays demonstrated better performance for the MBR. Both treatment lines complied with the Italian law's “ecotoxicity standard for effluent discharge in surface water”. Yet, residual biological activity was still detected, demonstrating the adequacy and sensitivity of the toxicological tools, which, by their inherent nature, allow the overall effects of complex mixtures to be taken into account.

Is Mixtures' Additivity Supported by Empirical Data? A Case Study of Developmental Toxicity of PFOS and 6:2 FTS in Wildtype Zebrafish Embryos.

Per- and polyfluoroalkyl substances (PFASs) are a major priority for many federal and state regulatory agencies charged with monitoring levels of emerging contaminants in environmental media and setting health-protective benchmarks to guide risk assessments. While screening levels and toxicity reference values have been developed for numerous individual PFAS compounds, there remain important data gaps regarding the mode of action for toxicity of PFAS mixtures. The present study aims to contribute whole-mixture toxicity data and advance the methods for evaluating mixtures of two key components of aqueous film-forming foams: perfluorooctanesulfonic acid (PFOS), and 6:2 fluorotelomer sulfonic acid (6:2 FTS). Wildtype (AB) zebrafish embryos were exposed to PFOS and 6:2 FTS, both as individual components and as binary mixtures, from 2 to 122 h post-fertilization. Five treatment levels were selected to encompass environmentally relevant exposure levels. Experimental endpoints consisted of mortality, hatching, and developmental endpoints, including swim bladder inflation, yolk sac area, and larval body length. Results from dose–response analysis indicate that the assumption of additivity using conventional points of departure (e.g., NOAEL, LOAEL) is not supported for critical effect endpoints with these PFAS mixtures, and that the interactions vary as a function of the dose range. Alternative methods for quantifying relative potency are proposed, and recommendations for additional investigations are provided to further advance assessments of the toxicity of PFAS mixtures to aquatic organisms.

Predicting joint toxicity of chemicals by incorporating a weighted descriptor into a mixture model: Cases for binary antibiotics and binary nanoparticles

A prediction method that integrated a mixture descriptor with an established mixture toxicology method was proposed for the joint toxicity of chemical pollutants. A weighted descriptor derived from the single descriptor of each component was employed to calculate a mixture descriptor, which was successfully embedded into the generalized concentration addition (GCA) model named the extended GCA (XGCA) model. To develop and validate the proposed approach, binary antibiotic mixtures (ciprofloxacin and oxytetracycline) and metal-oxide (copper oxide and zinc oxide) nanoparticle mixtures were selected to study their toxicity to freshwater green algae. The results showed that concentration-response curve (CRC) derived from the XGCA model was closer to the observed CRC than those from the GCA, Concentration Addition (CA), and Independent Action (IA) models. The difference between effect concentrations predicted by the XGCA model and observed did not exceed a factor of 1.6. The XGCA model was relatively more accurate at predicting joint toxicity (in terms of effect concentrations and effect errors) than the reference models, independent of component types and mixture ratios. The XGCA model predicts the joint toxicity through molecular structural or nanostructural characters, thus modes of toxic action are not preconditions for predicting the toxicity of the mixtures. This result demonstrates the practicability of using the XGCA method in toxicity assessments of mixture pollutants with unknown modes of action.

Aquatic toxicity and aquatic ecological risk assessment of wastewater-derived halogenated phenolic disinfection byproducts

Increasing number of wastewater-derived aliphatic and phenolic disinfection byproducts (DBPs) were discharged into aquatic environment with the discharge of disinfected wastewater. However, the currently available aquatic toxicity data and the aquatic ecological risk information of them are limited, especially for wastewater-derived phenolic DBPs. In this study, we investigated the acute toxicity of 7 phenolic DBPs that selected from the typical five groups of phenolic DBPs (2,4,6-trihalo-phenols, 2,6-dihalo-4-nitrophenols, 3,5-dihalo-4-hydroxybenzaldehydes, 3,5-dihalo-4-hydroxybenzoic acids and halo-salicylic acids) and 4 aliphatic DBPs to Gobiocypris rarus and also assessed their potential aquatic ecological risk. Experimental results indicated that the half lethal concentration (LC50) values of 2,4,6-trihalo-phenols and 2,6-dihalo-4-nitrophenols ranged from 1 to 10 mg/L; While that of 3,5-dihalo-4-hydroxybenzaldehydes was between 10 and 100 mg/L, and 3,5-dihalo-4-hydroxybenzoic acids and halo-salicylic acids was >100 mg/L. The toxicity mode of action (MOA) identification results from three methods suggested that no clear and consistent MOA were obtained for those 11 DBPs currently. The species-specific aquatic toxicity analysis results highlighted that no aquatic species would be considered as the most sensitive species for all 11 DBPs. However, crustacean and fish were more sensitive than that of algae for most of tested compounds. Lastly, the aquatic ecological risk assessment results of those 11 DBPs revealed that all 7 phenolic and 2 aliphatic DBPs (2-bromoacetamide and bromodichloromethane) had low aquatic ecological risk, while dichloroacetic acid and dibromoacetonitrile had high aquatic ecological risk. The low environmental concentration was the main reason why high toxic phenolic DBPs (2,4,6-trihalo-phenols and 2,6-dihalo-4-nitrophenols) exhibited low ecological risk. Their ecological risk may increase with the increases of corresponding environmental concentration. Thus, more efforts should be made to determine other potential harmful effects of those high toxic phenolic DBPs and to minimize their potential ecological risk by taking appropriate measures.

Classifying Modes of Toxic Action of Molecules with Electronic-structure Informatics. Application to Imbalanced Toxicity Data of Phenol Derivatives to Tetrahymena pyriformis

A set of electronic-structure informatics (ESI) descriptors was applied to classify modes of toxic action (MoA) of 245 phenol derivatives to the growth of Tetrahymena pyriformis. The extreme gradient boosting classification (XGBC) method was performed for classification where the synthetic minority oversampling technique (SMOTE) was employed to overcome the imbalance in the MoA classification data available in the literature. Performance evaluation of the machine learning procedure shows that it is able to classify the molecular set into the five MoA classes. The feature importance analysis suggests electronic factors determining characteristics for each MoA class.

Specific toxicity of azithromycin to the freshwater microalga Raphidocelis subcapitata

Pharmaceuticals are produced to inflict a specific physiological response in organisms. However, as only partially metabolized after administration, these types of compounds can also originate harmful side effects to non-target organisms. Additionally, there is still a lack of knowledge on the toxicological effects of legacy pharmaceuticals such as the antibiotic azithromycin. This macrolide occurs at high concentrations in the aquatic environment and can constitute a threat to aquatic organisms that are at the basis of the aquatic food chain, namely microalgae. This study established a high‐throughput methodology to study the toxicity of azithromycin to the freshwater microalga Raphidocelis subcapitata. Flow cytometry and pulse amplitude modulated (PAM) fluorometry were used as screening tools. General toxicity was shown by effects in growth rate, cell size, cell complexity, cell viability and cell cycle. More specific outcomes were indicated by the analysis of mitochondrial and cytoplasmatic membrane potentials, DNA content, formation of ROS and LPO, natural pigments content and photosystem II performance. The specific mode of action (MoA) of azithromycin to crucial components of microalgae cells was revealed. Azithromycin had a negative impact on the regulation of energy dissipation at the PSII centers, along with an insufficient protection by the regulatory mechanisms leading to photodamage. The blockage of photosynthetic electrons led to ROS formation and consequent oxidative damage, affecting membranes and DNA. Overall, the used methodology exhibited its high potential for detecting the toxic MoA of compounds in microalgae and should be considered for future risk assessment of pharmaceuticals.

Lethal, behavioral, growth and developmental toxicities of alkyl-PAHs and non-alkyl PAHs to early-life stage of brine shrimp, Artemia parthenogenetica

Alkyl-PAHs are the predominant form of PAHs in crude oils which are supposed to demonstrate different toxicities compared to non-alkyl PAHs. Little information is available about the toxicity of alkyl-PAHs on marine Artemia. This study addressed and compared the lethal, behavioral, growth and developmental toxicities of three alkyl-PAHs, namely 3-methyl phenanthrene (3-mPhe), retene (Ret) and 2-methyl anthracene (2-mAnt), to their non-alkyl forms, phenanthrene (Phe) and anthracene (Ant) using Artemia parthenogenetica (nauplii, <24 h) as test organism following a 48 h and a 7 d of exposure, respectively. Benzo-a-pyrene (Bap) was selected as a reference toxicant for the comparison with the above alkyl-PAHs and non-alkyl PAHs. Results showed that for all tested endpoints, A. parthenogenetica nauplii had the highest sensitivity to Bap while Ant had no significant effect on nauplii survival or development within given concentrations. Considering the aqueous freely dissolved PAH concentrations, the 48 h-LC50 (survival), 48 h-EC50 (immobility) and 7 d-LC10 (survival) of Bap were calculated as 0.321, 0.285 and 0.027 μg/L, respectively, which were twofold to fivefold lower than those of Phe, 3-mPhe, Ret, Ant and 2-mAnt. A higher acute toxicity of alkyl-PAHs (3-mPhe and 2-mAnt) than their non-alkyl forms (Phe and Ant) was observed. Not limited to Phe, the common non-polar narcotic mode of action was also observed for Bap, 3-mPhe, Ret and 2-mAnt, which was evident by the inhibited mobility of nauplii. The decreased body lengths were found for all PAH treatments compared to the solvent control, whereas instar retardations were only found in nauplii exposed to Bap, Phe and Ret. Our findings emphasized the sensitivity differences of A. parthenogenetica nauplii to selected alkyl PAHs and non-alkyl PAHs and confirmed the application of lethal, behavioral and growth indicators in the toxicity evaluation of selected PAHs other than Ant. However, the distinct toxicities of these PAHs suggested other toxic modes of action may play more important roles apart from narcotic mode of action and need to be elucidated in future studies. In addition, a strong correlation between the body length and the instar of A. parthenogenetica nauplii was observed for each PAH exposure, suggesting that body length can be representative for both growth and developmental indicators during biological monitoring of PAH pollution in marine environment.

Evidence for Specific Receptor-Mediated Toxicity of Pharmaceuticals in Aquatic Organisms Derived from Acute and Chronic Standard Endpoints.

The toxicity of 17 active pharmaceutical ingredients (APIs) was investigated using standardized acute and chronic tests with Daphnia magna and 2 algae species. Chronic toxicity was generally greater for Daphnia than for algae. Compilation of additional data resulted in 100 APIs for which the acute-to-chronic ratio (ACR) was determined for Daphnia. The frequency of high ACRs (~20% with ACRs > 100) indicates that specific receptor-mediated toxicity toward D. magna is rather common among APIs. The 11 APIs with ACRs > 1000 included lipid-modifying agents, immunosuppressants, antibiotics, antineoplastics, antiobesics, antivirals, and antihistamines. There was no consistent association between ACR and chronic toxicity, ionization status, or lipophilicity. High ACRs were not exclusively associated with the presence of orthologs of the pharmacological target in Daphnia. Statins, acetylcholinesterase inhibitors, and antihistamines are discussed in more detail regarding the link between targets and toxic mode of action. For acetylcholinesterase inhibitors, receptor-mediated toxicity was already apparent after acute exposure, whereas the high ACR and chronic toxicity of some antihistamines probably related to interaction with a secondary rather than the primary pharmacological target. Acute or modeled chronic toxicity estimates have often been used for prioritizing pharmaceuticals. This may be seriously misleading because chronic effects are currently not predictable for APIs with specific receptor-mediated toxicity. However, it is exactly these APIs that are the most relevant in terms of environmental risks. Environ Toxicol Chem 2022;41:601-613. © 2021 SETAC.

Essential concepts for interpreting the dose-response of low-level arsenic exposure in epidemiological studies

Scientifically robust selections of epidemiological studies and assessments of the dose-response of inorganic arsenic in the low-dose range must consider key issues specific to arsenic in order to reduce risk of bias. The abundance of toxicological, mechanistic, and epidemiological evidence on arsenic enables a nuanced assessment of risk of bias in epidemiological studies of low-level arsenic, as opposed to a generic evaluation based only on standard principles. Important concepts in this context include 1) arsenic metabolism and mode of action for toxicity and carcinogenicity; 2) effects of confounding factors such as diet, health status including nutritional deficiencies, use of tobacco and other substances, and body composition; 3) strengths and limitations of various metrics for assessing relevant exposures consistent with the mode of action; and 4) the potential for bias in the positive direction for the observed dose-response relationship as exposure increases in the low-dose range. As an example, evaluation of a recent dose-response modeling using eight epidemiological studies of inorganic arsenic and bladder cancer demonstrated that the pooled risk estimate was markedly affected by the single study that was ranked as having a high risk of bias, based on the above factors. The other seven studies were also affected by these factors to varying, albeit lesser, degrees that can influence the apparent dose-response in the low-dose range (i.e., drinking water concentration of 65 µg/L or dose of approximately ≤1 µg/kg-day). These issues are relevant considerations for assessing health risks of oral exposures to inorganic arsenic in the U.S. population, and setting evidence-based regulatory limits to protect human health.

Acute cerebrovascular effects in juvenile coho salmon exposed to roadway runoff

Urban stormwater is responsible for recurrent mortality events of coho salmon (Oncorhynchus kisutch) documented in the US Pacific Northwest. Currently, the toxic mode of action is unknown; however, mortality is often associated with large increases in hematocrit (Hct). We found that hemoconcentration and red cell swelling may offer a partial explanation for increased Hct, based on increased total hemoglobin and reduced mean cell hemoglobin concentration of the blood in runoff-exposed juvenile coho salmon, relative to controls. No significant changes were seen in plasma total thiols or antioxidant power, discounting osmoregulatory dysfunction and plasma dehydration as a driver of hemoconcentration. To test for plasma leakage, we injected Evans blue dye complexed with bovine serum albumin (EBD–BSA) intracardially and circulated for 5 min, followed by a 15 min washout with saline. Coho salmon exposed to roadway runoff showed significant accumulation of EBD–BSA in brain and olfactory rosette regions compared with controls, demonstrating plasma leakage from the cerebrovasculature. Our results suggest blood–brain barrier disruption underlies symptom development in coho salmon exposed to roadway runoff.