Potential Adverse Outcome Pathway Research Articles

Cyclosporin A toxicity on endothelial cells differentiated from induced pluripotent stem cells: Assembling an adverse outcome pathway

Cyclosporin A (CSA) is a potent immunosuppressive agent in pharmacologic studies. However, there is evidence for side effects, specifically regarding vascular dysfunction. Its mode of action inducing endothelial cell toxicity is partially unclear, and a connection with an adverse outcome pathway (AOP) is not established yet. Therefore, we designed this study to get deeper insights into the mechanistic toxicology of CSA on angiogenesis. Stem cells, especially induced pluripotent stem cells (iPSCs) with the ability of differentiation to all organs of the body, are considered a promising in vitro model to reduce animal experimentation. In this study, we differentiated iPSCs to endothelial cells (ECs) as one cell type that in other studies would allow to generate multi-cell type organoids from single donors. Flow cytometry and immunostaining confirmed our scalable differentiation protocol. Then dose and time course experiments assessing CSA cytotoxicity on iPS derived endothelial cells were performed. Transcriptomic data suggested CSA dependent induction of reactive oxygen species (ROS), mitochondrial dysfunction, and impaired angiogenesis via ROS induction which was confirmed by in vitro experiments. In order to put these data into a potential adverse outcome pathway (AOP) context, we performed a literature review for CSA-mediated endothelial cell toxicity and combined our experimental data with the publicly available knowledge. Such an AOP will help to design in vitro test batteries and to model events observed in human toxicity studies, as well in predictive toxicology.

Potential Adverse Outcome Pathways of Chlorinated Organophosphate Flame Retardants.

Chlorinated organophosphate flame retardants (Cl-OPFRs) are frequently detected chemicals in the environment and biological samples, yet there is a lack of systematic evaluation regarding the adverse effects and toxicological mechanisms of Cl-OPFRs. This study utilizes the adverse outcome pathway (AOP) framework to assess the health implications and mechanisms of Cl-OPFRs, identifying multi-system toxicity, with a particular emphasis on reproductive issues and the possible toxic mechanisms. These results enhance knowledge of the health hazards linked to Cl-OPFRs, supporting the creation of focused risk evaluations and suitable regulatory actions.

Baseline Toxicity Model to Identify the Specific and Nonspecific Effects of Per- and Polyfluoroalkyl Substances in Cell-Based Bioassays.

High-throughput screening is a strategy to identify potential adverse outcome pathways (AOP) for thousands of per- and polyfluoroalkyl substances (PFAS) if the specific effects can be distinguished from nonspecific effects. We hypothesize that baseline toxicity may serve as a reference to determine the specificity of the cell responses. Baseline toxicity is the minimum (cyto)toxicity caused by the accumulation of chemicals in cell membranes, which disturbs their structure and function. A mass balance model linking the critical membrane concentration for baseline toxicity to nominal (i.e., dosed) concentrations of PFAS in cell-based bioassays yielded separate baseline toxicity prediction models for anionic and neutral PFAS, which were based on liposome-water distribution ratios as the sole model descriptors. The specificity of cell responses to 30 PFAS on six target effects (activation of peroxisome proliferator-activated receptor (PPAR) gamma, aryl hydrocarbon receptor, oxidative stress response, and neurotoxicity in own experiments, and literature data for activation of several PPARs and the estrogen receptor) were assessed by comparing effective concentrations to predicted baseline toxic concentrations. HFPO-DA, HFPO-DA-AS, and PFMOAA showed high specificity on PPARs, which provides information on key events in AOPs relevant to PFAS. However, PFAS were of low specificity in the other experimentally evaluated assays and others from the literature. Even if PFAS are not highly specific for certain defined targets but disturb many toxicity pathways with low potency, such effects are toxicologically relevant, especially for hydrophobic PFAS and because PFAS are highly persistent and cause chronic effects. This implicates a heightened need for the risk assessment of PFAS mixtures because nonspecific effects behave concentration-additive in mixtures.

Associating prenatal phthalate exposure with childhood autistic traits: Investigating potential adverse outcome pathways and the modifying effects of maternal vitamin D

The association between prenatal phthalate mixture exposure and the risk of autism spectrum disorder (ASD) in children, as well as the potential mechanism and impact of maternal vitamin D, remains unclear. We analyzed data from 3209 mother–child pairs. The associations between prenatal phthalate exposure and autistic traits in children aged 1.5, 3, 5, and 6 years were explored. Furthermore, the modifying effects of maternal vitamin D and the adverse outcome pathway, which elucidates the contribution of phthalates to ASD, were estimated. Exposure to a phthalate mixture was associated with an increased risk of ASD in children aged 1.5–6 years. For mothers with 25(OH)D deficiency, an exposure‒response relationship was observed between phthalate mixtures in early to mid-pregnancy and autistic traits in children aged 3 years. However, this association was not observed for mothers with sufficient prenatal 25(OH)D levels. The potential mechanism of action of di(2-ethylhexyl) phthalate (DEHP) exposure may involve affecting GRIN2B, inhibiting NMDAR in the postsynaptic membrane, disrupting synaptic function, and impairing learning and memory, ultimately leading to ASD development. Importantly, maternal vitamin D supplementation was demonstrated to mitigate the risk of ASD associated with phthalate exposure. Reducing phthalate exposure during pregnancy may be associated with a decreased risk of autistic traits in children. Furthermore, adequate vitamin D supplementation could potentially mitigate the impact of phthalates on these traits. Additionally, the proposed biological mechanism provides insight into how phthalate exposure may contribute to the development of ASD.

Polystyrene micro- and nanoplastics induce gastric toxicity through ROS mediated oxidative stress and P62/Keap1/Nrf2 pathway

Microplastics (MPs) exist widely in the environment and can enter the human body indirectly through the food chain or directly through inhalation or ingestion. The primary organ that MPs contaminated food or water enters the human body through the digestive tract is the stomach. However, at present, the effects of MPs on the stomach and the related mechanism remain unclear. In this study, our results indicated that 50 nm and 250 nm polystyrene MPs (PS-MPs) at environmental related dose significantly decreased stomach organ coefficient, inhibited gastric juice secretion and mucus secretion, disrupted gastric barrier function and suppressed antioxidant ability in mice. In vitro experiments showed that PS-MPs inhibited cell viability, increased ROS generation, and induced apoptosis through mitochondria-dependent pathway. Simultaneously, PS-MPs also decreased mitochondrial membrane potential, ATP level, disrupted mitochondrial kinetic homeostasis, and activated P62 / Nrf2 / Keap1 pathway. Furthermore, blocking ROS (NAC) partially alleviated ROS and apoptosis caused by PS-MPs. Based on above findings, the potential adverse outcome pathway (AOP) of PS-MPs-caused gastric toxicity was proposed which provides a new insight into the risk assessment of MP related gastric damage. Our study unveils the gastric injury induced by PS MPs is dependent on ROS - mediated P62 / Nrf2 / Keap1 signaling pathway, and provides scientific basis for further exploration the mechanism of gastric toxicity of PS MPs.

A critical review of microplastics toxicity and potential adverse outcome pathway in human gastrointestinal tract following oral exposure

Microplastics (MPs) are typically produced via environmental degradation of larger plastics, where they enter the human food chain. MPs are complex materials containing chemical and physical characteristics that can potentially affect their hazard and exposure. These physical properties can be altered by environmental exposure potentially altering any risk assessment conducted on the primary material. We conducted a literature review using an Adverse Outcome Pathway (AOP)-based approach from Molecular Initiating Event (MIE) to cell effect event to identify multiple knowledge gaps that affect MPs hazard assessment. There is some convergence of key biological events but could relate to most lying along well-established biological effector pathways such as apoptosis which can respond to many MIEs. In contrast, MIEs of chemicals will be via protein interaction. As MPs may occur in the lumen of the alimentary canal for example to the mucus, therefore, not requiring translocation of MPs across the epithelial membrane. At the other end of the AOP, currently it is not possible to identify a single adverse outcome at the organ level. This work did establish a clear need to understand both external and internal exposure (resulting from translocation) and develop hazard data at both levels to inform on risk assessments.

Investigation on the potential adverse outcome pathway of the sensitive endpoint for nephrotoxicity induced by gardenia yellow based on an integrated strategy using bioinformatics analysis and in vitro testing validation.

To explore the potential the adverse outcome pathway of Gardenia Yellow (GY)-induced sensitive endpoint for nephrotoxicity, an integrated strategy was applied in the present study. Using bioinformatic analysis, based on the constructed Protein-protein interaction networks, Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis on the core target network were performed to illustrate the potential gene targets and signal pathways. Then, the most enriched pathway was validated with Cell counting kit-8 assays and Western blot analysis in embryonic kidney epithelial 293 cell models. According to the findings, GY may interact with 321 targets related to the endpoint. The five targets on the top ranking in the PPI network were STAT3, SRC, HRAS, AKT1, EP300. Among them, PI3K/Akt was the most enriched pathway. In vitro testing showed that GY exerted a proliferative effect on the cell variability in a dose-dependent manner. GY at concentration of 1000 μg/ml and stimulation for 30 min can significantly enhance the expression of phosphorylated Akt. Thus, after the quantitative weight of evidence evaluation, Akt phosphorylation induced PI3K/Akt activation was speculated as a molecular initiating event leading to a proliferative and inflammatory response in renal tubular epithelial cells.

Potential adverse outcome pathway (AOP) of emamectin benzoate mediated cardiovascular toxicity in zebrafish larvae (Danio rerio)

Emamectin benzoate (EMB) is an efficient insecticide which widely used as an anthelmintic drug additive in aquaculture fish. However, its extensive use has resulted in widespread pollution in the aquatic environment. Previous studies have identified the potential developmental and neurotoxic effects of EMB, however, systematic studies pertaining to the cardiovascular toxic effects of EMB on fish are scarce. In this study, zebrafish embryos were exposed to EMB at concentrations of 0, 0.1, 0.25, 0.5, 1, 2, 4, and 8 mg/L for 3 days, aiming to investigate the cardiovascular toxic effects of EMB via examining morphology, cardiac function, and vascular development phenotypes. It revealed that EMB exposure led to marked deteriorated effects, including adverse effects on mortality, hatching rate, and general morphological traits, such as malformation, heart rate, body length, and eye area, in zebrafish embryos/larvae. Furthermore, EMB exposure resulted in abnormal cardiac function and vascular development, triggering neutrophil migration and aggregation toward the pericardial and dorsal vascular regions, and finalized apoptosis in the zebrafish heart region, these phenomena were further deciperred by the transcriptome analysis that the Toll-like receptor pathway, P53 pathway, and apoptotic pathway were significantly affected by EMB exposure. Moreover, the molecular docking and aspirin anti-inflammatory rescue assays indicated that TLR2 and TLR4 might be the potential targets of EMB. Taken together, our study provides preliminary evidence that EMB may induce apoptosis by affecting inflammatory signaling pathways and eventually lead to abnormal cardiovascular development in zebrafish. This study provides a simple toxicological AOP framework for safe pesticide use and management strategies.

Identification of neurotoxic compounds in cyanobacteria exudate mixtures

Release of toxic cyanobacterial secondary metabolites threatens biosecurity, foodwebs and public health. Microcystis aeruginosa (Ma), the dominant species in global freshwater cyanobacterial blooms, produces exudates (MaE) that cause adverse outcomes including nerve damage. Previously, we identified > 300 chemicals in MaE. It is critical to investigate neurotoxicity mechanisms of active substances among this suite of Ma compounds. Here, we screened 103 neurotoxicity assays from the ToxCast database to reveal targets of action of MaE using machine learning. We then built a potential Adverse Outcome Pathway (AOP) to identify neurotoxicity mechanisms of MaE as well as key targets. Finally, we selected potential neurotoxins matched with those targets using molecular docking. We found 38 targets that were inhibited and eight targets that were activated, collectively mainly related to neurotransmission (i.e. cholinergic, dopaminergic and serotonergic neurotransmitter systems). The potential AOP of MaE neurotoxicity could be caused by blocking calcium voltage-gated channel (CACNA1A), because of antagonizing neurotransmitter receptors, or because of inhibiting solute carrier transporters. We identified nine neurotoxic MaE compounds with high affinity to those targets, including LysoPC(16:0), 2-acetyl-1-alkyl-sn-glycero-3-phosphocholine, egonol glucoside, polyoxyethylene (600) monoricinoleate, and phytosphingosine. Our study enhances understanding of neurotoxicity mechanisms and identifies neurotoxins in cyanobacterial bloom exudates, which may help identify priority compounds for cyanobacteria management.

Combined effect of polystyrene microplastics and bisphenol A on the human embryonic stem cells-derived liver organoids: The hepatotoxicity and lipid accumulation

Human are exposed to microplastics (MP) via inhalation or ingestion daily and inevitably. The liver is an important target organ of MP. Bisphenol A (BPA) is one of commonly used plasticizers. It is added in plastics, but also generally detected in the biological samples of human beings. However, the combined toxic effect of MP and BPA on human liver is unclear. In this study, a novel 3D in vitro model, the liver organoid (LO) derived from human-pluripotent stem cells, has been utilized to explore the 1 μm polystyrene (PS)-induced hepatotoxicity with BPA individually and jointly. Conclusively, all the changes in the cytotoxicity, cellular and molecular makers regarding the energy supplement, hepatic injury, oxidative stress, inflammatory response, disruption in the lipid accumulation, as well as epigenetics regulation induced by BPA or PS on the LOs individually were comparable to previous study. The BPA levels in the culture medium were declined by the added PS. The combined adverse effect of PS and BPA on the LOs was identified to be synergistic upon deteriorated hepatotoxicity and interfered the gene panels related to multiple processes of lipid metabolism, together with the proteins of HNF4A, CD36, ACC1, CPT1A, CYP2E1, ERα and ERβ. Specifically, PS didn't change the ERα or ERβ individually, but when the LOs were co-exposed to PS and BPA, the ERα further elevated significantly and synergistically. Our findings highlight the metabolic-related health risk due to co-exposure to MP and BPA, even at low-doses equivalent to human internal exposure level. Based on these findings, the potential adverse outcome pathway related to PS and BPA singly and jointly were proposed, predicting two possible outcomes to be hepatic steatosis. Moreover, the ERα and HNF4A were proposed to be potential candidate markers to investigate the “vector-like effect” of PS in the present of BPA.

Potential adverse outcome pathways with hazard identification of organophosphate esters

Data-driven analysis and pathway-based approaches contribute to reasonable arrangements of limited resources and laboratory tests for continuously emerging commercial chemicals, which provides opportunities to save time and effort for toxicity research. With the widespread usage of organophosphate esters (OPEs) on a global scale, the concentrations generally reached up to micromolar range in environmental media and even in organisms. However, potential adverse effects and toxicity pathways of OPEs have not been systematically assessed. Therefore, it is necessary to review the current situation, formulate the future research priorities, and characterize toxicity mechanisms via data-driven analysis. Results showed that the early toxicity studies focused on neurotoxicity, cytotoxicity, and metabolic disorders. Then the main focus shifted to the mechanisms of cardiotoxicity, endocrine disruption, hepatocytes, reproductive and developmental toxicity to vulnerable sub-populations, such as infants and embryos, affected by OPEs. In addition, several novel OPEs have been emerging, such as bis(2-ethylhexyl)-phenyl phosphate (HDEHP) and oxidation derivatives (OPAsO) generated from organophosphite antioxidants (OPAs), leading to multiple potential ecological and human health risks (neurotoxicity, hepatotoxicity, developmental toxicity, etc.). Notably, in-depth statistical analysis was promising in encapsulating toxicological information to develop adverse outcome pathways (AOPs) frameworks. Subsequently, network-centric analysis and quantitative weight-of-evidence (QWOE) approaches were utilized to construct and evaluate the putative AOPs frameworks of OPEs, showing the moderate confidences of the developed AOPs. In addition, frameworks demonstrated that several events, such as nuclear receptor activation, reactive oxygen species (ROS) production, oxidative stress, and DNA damage, were involved in multiple different adverse outcome (AO), and these AOs had certain degree of connectivity. This study brought new insights into facilitating the complement of AOP efficiently, as well as establishing toxicity pathways framework to inform risk assessment of emerging OPEs.

Characterization of Potential Adverse Outcome Pathways Related to Metabolic Outcomes and Exposure to Per- and Polyfluoroalkyl Substances Using Artificial Intelligence

Human exposure to per- and polyfluoroalkyl substances (PFAS) has been associated with numerous adverse health effects, depending on various factors such as the conditions of exposure (dose/concentration, duration, route of exposure, etc.) and characteristics associated with the exposed target (e.g., age, sex, ethnicity, health status, and genetic predisposition). The biological mechanisms by which PFAS might affect systems are largely unknown. To support the risk assessment process, AOP-helpFinder, a new artificial intelligence tool, was used to rapidly and systematically explore all available published information in the PubMed database. The aim was to identify existing associations between PFAS and metabolic health outcomes that may be relevant to support building adverse outcome pathways (AOPs). The collected information was manually organized to investigate linkages between PFAS exposures and metabolic health outcomes, including dyslipidemia, hypertension, insulin resistance, and obesity. Links between PFAS exposure and events from the existing metabolic-related AOPs were also retrieved. In conclusion, by analyzing dispersed information from the literature, we could identify some associations between PFAS exposure and components of existing AOPs. Additionally, we identified some linkages between PFAS exposure and metabolic outcomes for which only sparse information is available or which are not yet present in the AOP-wiki database that could be addressed in future research.

Polystyrene microplastics induce hepatotoxicity and disrupt lipid metabolism in the liver organoids

Microplastic particles (MP) has been detected in the environment widespread. Human beings are inevitably exposed to MP via multiple routines. However, the hazard identifications, as direct evidence of exposure and health risk, have not been fully characterized in human beings. Many studies suggest the liver is a potential target organ, but currently no study regarding the MP on human liver has been reported. In this study, we used a novel in vitro 3D model, the liver organoids (LOs) generated from human pluripotent stem cells, as an alternative model to the human liver, to explore the adverse biological effect of 1 μm polystyrene-MP (PS-MP) microbeads applying a non-static exposure approach. When the LOs were exposed to 0.25, 2.5 and 25 μg/mL PS-MP (the lowest one was relevant to the environmental concentrations, calculated to be 102 ± 7 items/mL). The potential mechanisms of PS-MP induced hepatotoxicity and lipotoxicity, in aspects of cytotoxicity, levels of key molecular markers, ATP production, alteration in lipid metabolism, ROS generation, oxidative stress and inflammation response, were determined. Specifically, it has been firstly observed that PS-MP could increase the expression of hepatic HNF4A and CYP2E1. Based on these findings, the potential adverse outcome pathways (AOPs) relevant to PS-MP were proposed, and the potential risks of PS-MP on liver steatosis, fibrosis and cancer were implicated. The combined application of novel LOs model and AOPs framework provides a new insight into the risk assessment of MP. Further studies are anticipated to validate the hepatotoxic molecular mechanism of PS-MP based on HNF4A or CYP2E1, and to investigate the MP-induced physical damage and its relationship to hepatic adverse effect for human beings. CapsuleMicroplastics cause hepatotoxicity and disrupt lipid metabolism in the human pluripotent stem cells-derived liver organoids, providing evidence for human implication.

Development of AOP relevant to microplastics based on toxicity mechanisms of chemical additives using ToxCast™ and deep learning models combined approach

Various additives are used in plastic products to improve the properties and the durability of the plastics. Their possible elution from the plastics when plastics are fragmented into micro- and nano-size in the environment is suspected to one of the major contributors to environmental and human toxicity of microplastics. In this context, to better understand the hazardous effect of microplastics, the toxicity of chemical additives was investigated. Fifty most common chemicals presented in plastics were selected as target additives. Their toxicity was systematically identified using apical and molecular toxicity databases, such as ChemIDplus and ToxCast™. Among the vast ToxCast assays, those having intended gene targets were selected for identification of the mechanism of toxicity of plastic additives. Deep learning artificial neural network models were further developed based on the ToxCast assays for the chemicals not tested in the ToxCast program. Using both the ToxCast database and deep learning models, active chemicals on each ToxCast assays were identified. Through correlation analysis between molecular targets from ToxCast and mammalian toxicity results from ChemIDplus, we identified the fifteen most relevant mechanisms of toxicity for the understanding mechanism of toxicity of plastic additives. They are neurotoxicity, inflammation, lipid metabolism, and cancer pathways. Based on these, along with, previously conducted systemic review on the mechanism of toxicity of microplastics, here we have proposed potential adverse outcome pathways (AOPs) relevant to microplastics pollution. This study also suggests in vivo and in vitro toxicity database and deep learning model combined approach is appropriate to provide insight into the toxicity mechanism of the broad range of environmental chemicals, such as plastic additives.

Early Life Exposures to Perfluoroalkyl Substances in Relation to Adipokine Hormone Levels at Birth and During Childhood.

Birth cohort studies have linked exposure to perfluoroalkyl substances (PFASs) with child anthropometry. Metabolic hormone dysregulation needs to be considered as a potential adverse outcome pathway. We examined the associations between PFAS exposures and concentrations of adipokine hormones from birth to adolescence. We studied 80 mother-child pairs from a Faroese cohort born in 1997 to 2000. Five PFASs were measured in maternal pregnancy serum and in child serum at ages 5, 7, and 13 years. Leptin, adiponectin, and resistin were analyzed in cord serum and child serum at the same ages. We fitted multivariable-adjusted generalized estimating equations to assess the associations of PFASs at each age with repeated adipokine concentrations at concurrent and subsequent ages. We observed tendencies of inverse associations between PFASs and adipokine hormones specific to particular ages and sex. Significant associations with all adipokines were observed for maternal and child 5-year serum PFAS concentrations, whereas associations for PFASs measured at ages 7 to 13 years were mostly null. The inverse associations with leptin and adiponectin were seen mainly in females, whereas the inverse PFAS associations with resistin levels were seen mainly in males. Estimates for significant associations (P value <0.05) suggested mean decreases in hormone levels (range) by 38% to 89% for leptin, 16% to 70% for adiponectin, and 33% to 62% for resistin for each twofold increase in serum PFAS concentration. These findings suggest adipokine hormone dysregulation in early life as a potential pathway underlying PFAS-related health outcomes and underscore the need to further account for susceptibility windows and sex-dimorphic effects in future investigations.

Biopesticide spinosad induces transcriptional alterations in genes associated with energy production in honey bees (Apis mellifera) at sublethal concentrations.

Bees experience substantial colony losses, which are often associated with pesticides. Besides synthetic insecticides biological compounds such as spinosad are used in agriculture and organic farming against insect pests. However, potential adverse effect at sublethal concentrations to pollinators are poorly known. Here we aim to determine potential adverse outcome pathways of spinosad and to identify molecular effects by investigating transcriptional alterations in the brain of honey bees. We experimentally exposed bees to three sublethal concentrations of 0.05, 0.5 and 5 ng spinosad/bee, and assessed transcriptional alterations of target genes. Additionally, we evaluated whether spinosad-induced transcriptional alterations were influenced by the time of the year. In April, alterations were most pronounced after 24 h exposure, while in June alterations occurred mostly after 48 h. In July, expressional alterations were often lower but the pattern was more similar to that in June than that in April. Down-regulation of genes encoding acetylcholine receptors, enzymes involved in oxidative phosphorylation (cox5a, ndufb7 and cox17), cytochrome P450 dependent monooxygenases (cyp9q1, cyp9q2 and cyp9q3) and insulin-like peptide-1 were among the most significant transcriptional alterations. This suggests adverse effects of spinosad to energy production and metabolism and thus negative consequences on foraging. Together, our study indicates that spinosad causes adverse effects at environmentally realistic concentrations, which may pose a risk to bee populations.

Next-Generation Complex Metal Oxide Nanomaterials Negatively Impact Growth and Development in the Benthic Invertebrate Chironomus riparius upon Settling.

Most studies of nanomaterial environmental impacts have focused on relatively simple first-generation nanomaterials, including metals or metal oxides (e.g., Ag, ZnO) for which dissolution largely accounts for toxicity. Few studies have considered nanomaterials with more complex compositions, such as complex metal oxides, which represent an emerging class of next-generation nanomaterials used in commercial products at large scales. Importantly, many nanomaterials are not colloidally stable in aqueous environments and will aggregate and settle, yet most studies use pelagic rather than benthic-dwelling organisms. Here we show thatexposure of the model benthic species Chironomus riparius to lithium cobalt oxide (Li xCo1- xO2, LCO) and lithium nickel manganese cobalt oxide (Li xNi yMn zCo1- y- zO2, NMC) at 10 and 100 mg·L-1 caused 30-60% declines in larval growth and a delay of 7-25 d in adult emergence. A correlated 41-48% decline in larval hemoglobin concentration and related gene expression changes suggest a potential adverse outcome pathway. Metal ions released from nanoparticles do not cause equivalent impacts, indicating a nanospecific effect. Nanomaterials settled within 2 days and indicate higher cumulative exposures to sediment organisms than those in the water column, making this a potentially realistic environmental exposure. Differences in toxicity between NMC and LCO indicate compositional tuning may reduce material impact.

Inhibition of growth in juvenile manila clam Ruditapes philippinarum: Potential adverse outcome pathway of TBBPA

Tetrabromobisphenol A (TBBPA) is ubiquitous and its contents showing an increasing trend in the coastal environment. In order to investigate the effects of TBBPA on marine bivalves, juvenile manila clams Ruditapes phillipinarum were exposed to TBBPA for 28 days. The results showed that shell growth rate of juvenile clams after exposure to 62.5–1000 μg L−1 TBBPA for 28 d were significantly inhibited (p < 0.05). Then in order to link the changes in filtration rate, mRNA expression of insulin-like growth factor homologue (IGF) and tissue thyroid hormone (TH) contents to growth, juvenile clams were exposed to 62.5 and 500 μg L−1 TBBPA for 14 days. The transcriptional levels of neuroendocrine signals (NPF and insulin homologue) associated with filter feeding regulation, and genes of TH synthesis-related enzymes were also examined. The results showed that filtration rates was significantly reduced to 44.1% and 14% of controls after 14 d of exposure. In parallel, exposure to TBBPA significantly increased the expression levels of insulin which may elicit the filter feeding inhibition. TBBPA exposure caused alterations in tissue content of THs and mRNA expression of TH synthesis-related enzymes. However, the data showed increased T3 content, T3/T4 ratio and mRNA expression of IGF. These data demonstrated that the most important key event of TBBPA could be linked to growth impairment in juveniles was the reduction of filtration rate. These results provide a robust framework towards revealing the underlying mechanism of the growth inhibition caused by TBBPA on bivalves and understanding the adverse outcome pathway across taxonomic phyla.

Investigation of binding and activity of perfluoroalkyl substances to the human peroxisome proliferator-activated receptor β/δ.

Previously, perfluoroalkyl substances (PFASs) have been found to be associated with many adverse effects mediated by the peroxisome proliferator-activated receptor α (PPARα) and PPARγ. Here, we found another subtype of the peroxisome proliferator-activated receptors (PPARs); the PPARβ/δ mediated pathway might also be a potential adverse outcome pathway for PFASs. We investigated the direct binding and transcriptional activity of PFASs toward human PPARβ/δ, and further revealed the structure-binding and structure-activity relationship between PFASs and PPARβ/δ. The receptor binding experiment showed that their binding potency was dependent on the carbon chain length and the terminal functional group. For twelve perfluoroalkyl carboxylic acids (PFCAs), an inverted U-shaped relationship existed between the PPARβ/δ binding potency and the carbon chain length, with perfluorododecanoc acid (C12) showing the highest binding potency. The three perfluoroalkane sulfonic acids (PFSAs) exhibited a stronger binding potency than their PFCA counterparts. The two fluorotelomer alcohols (FTOHs) showed no binding potency. In receptor transcriptional activity assays, they enhanced the PPARβ/δ transcriptional activity. Their transcriptional activity was also related to the carbon chain length and the terminal functional group. Molecular docking analysis showed the PFASs fitted into the ligand binding pocket of PPARβ/δ with a binding geometry similar to a fatty acid.

A Framework for Systematic Evaluation and Quantitative Integration of Mechanistic Data in Assessments of Potential Human Carcinogens.

Recently, the key characteristics of carcinogens (KCC) have been proposed as an organizational approach for the evaluation of mechanistic data related to carcinogenicity. Our objective was to develop a framework to systematically and quantitatively integrate KCC data using elements that are important to risk assessment. Methods for developing the framework included: defining objectives, identifying and accommodating key considerations for components, input, and output of the framework, and operational development via iterative testing by a multidisciplinary team. The proposed framework involves 3 steps: (1) a structured, yet flexible, appraisal of individual studies and endpoints, (2) a structured and transparent evaluation of the body of evidence for each key characteristic, and (3) an evaluation of all of the KCC-relevant data relative to tumors and/or cancer types. In step 1, data are assessed and scored for reliability, strength, and activity. In step 2, a mathematical algorithm is used to integrate (and weight) the quality, relevance, and activity for each of the KCCs. These scores facilitate subsequent evaluations related to the overall body of evidence in step 3 in which KCCs can be linked, assessing potential adverse outcome pathways or networks, and finally, considered in the context of observed carcinogenic responses in animals and/or humans. The output is an overall conclusion regarding KCC activity as it relates to carcinogenic responses. The proposed framework provides a flexible solution to quantitatively integrate KCC data in a systematic and transparent manner that provides weighting of data most well-suited for the assessment of potential human carcinogenicity.