ALTEX. 2026 Apr 9. doi: 10.14573/altex.2512011. Online ahead of print.
ABSTRACT
Bisphenol A is a high production volume chemical used extensively in the manufacture of polycarbonate plastics, epoxy resins, and thermal printer paper with a high potential for occupational and post-production dermal exposure. Bisphenol A-containing plastics were commonly used in food packaging resulting in significant public exposure through leaching into foodstuff. The public is also at risk of dermal exposure due to environmental contamination. Due to public health concerns regarding the potential for endocrine disrupting effects, efforts have been applied to replace bisphenol A with safer alternatives. Bisphenol A has been shown to cause skin sensitization in humans; however, there is a paucity of information available on the sensitizing potential of structural analogues which are increasingly being employed as substitutes. We utilized new approach methodologies (NAMs) addressing key events 1-3 of the adverse outcome pathway for skin sensitization to address the potential of bisphenol A substitutes to induce dermal sensitization. Defined approaches (DA) were applied to further classify and categorize potency according to OECD TG 497. The NAMs and DAs confirmed that bisphenol A was a skin sensitizer in potency category UN GHS 1B. Bisphenol B, AP, and E were also classified as UN GHS 1B sensitizers, and bisphenol AF as UN GHS 1A/1B depending on DA, while 2,4-bisphenol S and F were borderline sensitizers, and bisphenol S was classified as a non-sensitizer. These data provide evidence of skin sensitization hazard for the bisphenol structural analogues tested, except for bisphenol S, suggesting that they present risks for dermal allergy.
PMID:41964411 | DOI:10.14573/altex.2512011
Environ Sci Technol. 2026 Apr 8. doi: 10.1021/acs.est.6c00444. Online ahead of print.
ABSTRACT
Chemical persistence has long been recognized as a critical determinant of ecosystem and human exposure, exemplified by legacy pollutants such as DDT, PCBs, and, more recently, PFAS. Despite decades of regulation and research, robust experimental half-life data are available for only a fraction of chemicals in use, hampering their persistence assessment. Current testing frameworks, while refined, lack the efficacy to address these large data gaps, underscoring the need for innovative approaches. We argue that new approach methodologies for persistence assessment (P-NAMs)─including high-throughput (HT) experimental systems and advanced in silico models─are needed. HT-testing can bridge the gap between biodegradability screening tests and resource-intensive simulation studies. Simultaneously, HT-testing can generate large, consistent data sets needed to improve the mechanistic understanding of biotransformation and train more accurate predictive models. Integration of transformation product analysis and FAIR (findable, accessible, interoperable, and reusable) data repositories will further enhance mechanistic understanding and model reliability. We call for coordinated efforts across academia, industry, and regulatory bodies to establish standardized reporting, expand accessible data sets, and validate predictive tools. By advancing P-NAMs, the scientific community can ensure that persistence assessment evolves from a regulatory bottleneck into a driver of innovation, safeguarding human and ecosystem health and promoting safe chemical design.
PMID:41950227 | DOI:10.1021/acs.est.6c00444
J Cheminform. 2026 Mar 26. doi: 10.1186/s13321-026-01184-8. Online ahead of print.
ABSTRACT
Machine Learning (ML) and Artificial Intelligence (AI) approaches have potential to make better-informed decisions in chemical hazard identification while reducing animal testing. Their application in the context of New Approach Methodologies (NAMs) for Hazard Identification in Chemicals Risk Assessment (CRA) is challenging due to the limited knowledge, lack of experience, and uncertainty related to the use of these approaches. Therefore, to facilitate ML and AI approaches' potential acceptance for regulatory use, better standardization, guidelines for transparent reporting, validation, and frameworks are needed to understand their accessibility, verifiability, and usefulness criteria for predictions. An extensive literature review on the availability of ML and AI based NAMs for chemical hazard identification was conducted, focusing primarily on human health endpoints: specific target organ toxicity (STOT), genotoxicity and carcinogenicity, endocrine disruption, skin sensitization, developmental and reproductive toxicity (DART), and repeated dose or chronic toxicity. Nearly 2300 scientific articles were reviewed, and 274 publications with ML-QSAR models revealed that 60.9% of the models described in the scientific literature turned out to be non-usable, 21.9% were potentially usable, and 17.2% were directly usable, i.e., had available software solutions. By endpoint, the skin sensitization is best covered with the ML-QSAR models, followed by endocrine disruption, genotoxicity, and carcinogenicity models. The most derived ML-QSAR models are tree-based models such as random forests, and analogues, followed by artificial neural networks and support vector machine models, with other models being used to a lesser extent. The literature analysis led to a framework that helps model users to identify potentially suitable models for use in a regulatory context. In addition, the framework could help model developers better understand the expectations of model users in a regulatory context and use the framework as a reference when publishing their models, ensuring greater transparency, alignment with regulatory needs, and facilitating future acceptance.
PMID:41888966 | DOI:10.1186/s13321-026-01184-8
Ann Ist Super Sanita. 2026 Jan-Mar;62(1):40-52. doi: 10.4415/ANN_26_01_07.
ABSTRACT
INTRODUCTION: In recent years, the scientific community, together with regulatory authorities, institutions and stakeholders, joined in the common objective of changing the regulatory toxicology paradigm, moving from classical toxicological animal testing towards an exposure-driven risk assessment, based on mechanistic information derived from alternative methods, globally known as new approach methodologies (NAMs). The transition to the next generation risk assessment (NGRA) requires the collective effort of all involved actors. One approach addressing the needs of the NGRA is the adverse outcome pathway (AOP) concept. This construct describes a sequence of temporally and causally linked events at distinct levels of biological organization leading to an adverse health effect. The development, validation and implementation of AOPs at the regulatory level have become key objectives of several authorities.
METHODS: In this paper, the AOPs and AOP networks as applied by the European Food Safety Agency are analyzed, alongside the proposed batteries of NAMs.
RESULTS AND DISCUSSION: Two case studies, (1) Parkinson's disease and pesticide exposure, and (2) exposure to substances with endocrine-disrupting properties and the development of uterine adenocarcinoma, are described. The analysis of the entire AOP construct for each case and the assembly of the specific NAMs allowed us to identify the strengths and limitations of the AOP strategy, as well as of each proposed test. Their integration and status are assessed within the current regulatory framework.
PMID:41867154 | DOI:10.4415/ANN_26_01_07
Online 2026-02-25 to 2026-02-25
Food Chem Toxicol. 2026 Mar 1;212:116041. doi: 10.1016/j.fct.2026.116041. Online ahead of print.
ABSTRACT
Efficient strategies for the human health risk assessment of micro- and nanoplastics (MNPs) are urgently needed to address the complexity and diversity of these materials. Here, we propose a risk assessment framework for MNPs following the oral route of exposure. The framework is based on a flexible and modular approach, drawing on modern concepts in risk assessment, such as Integrated Approaches to Testing and Assessment (IATAs) or Adverse Outcome Pathways (AOPs), and substantially relies on New Approach Methodologies (NAMs). The framework is structured into three different main steps: i) basic physicochemical characterization of testing materials, ii) intestinal crossing, iii) hazard assessment testing. Each step of the framework is supported by scientifically sound methods, enabling a mechanistic and hypothesis-driven risk assessment of MNPs.
PMID:41775312 | DOI:10.1016/j.fct.2026.116041
Early last year, Republicans from the House Judiciary Committee were invited to a private dinner at the Justice Department. Representative Thomas Massie of Kentucky remembers asking his then girlfriend, now wife, what he should ask Attorney General Pam Bondi. She suggested he bring up Jeffrey Epstein. At first, he waved off the suggestion. But as he sat at the long formal table, set with formal china and surrounded by top Justice officials, he changed his mind.
“‘I saw you release the ‘Phase 1’ of the Epstein files,” Massie remembers asking Bondi at the April 28 dinner. “When do you think we might get ‘Phase 2’?’”
Bondi looked him in the eyes, Massie said, and explained that all that was left to be released was material she considered “child pornography” and that there was nothing more there. He didn’t follow up then, but her answer didn’t sit right.
“I suspected there was stuff that needed to come out,” he told me.
After the Justice Department announced in July that it would not share any more records, Representative Ro Khanna of California held a morning strategy meeting with his staff and asked for ideas for new bills. Sarah Drory, a young communications staffer who had never before spoken up in this meeting, had a suggestion: “What about a bill to release the Epstein files?”
The room was silent. Drory saw her more senior colleagues exchange confused or dismissive looks. One declared Epstein “a social-media thing.” On her walk home late that night, she called her boyfriend, second-guessing her suggestion. “Is that crazy?” she remembers asking him—and herself.
Three days later, Khanna introduced an amendment in the Rules Committee requiring the Department of Justice to release the Epstein files to gauge support and was surprised when a Republican congressman from South Carolina voted for it. After that vote, Massie called Khanna, and they discussed a long-shot idea late into the night: Could they convince other Republicans to do the same?
Khanna’s staff and consultants tried to talk him out of pursuing such legislation, explaining that it could make him look like a conspiracy theorist, instead of his desired image as a “serious economics guy.” But Khanna explained to his staff that the topic of Epstein kept coming up, especially when he appeared on podcasts or visited the more conservative parts of his district. He recalled a young man delivering a long rant at a recent town hall about how he didn’t trust the government because it was “protecting pedophiles” by not releasing the Epstein files or holding more people accountable. In that moment, Khanna told me, he realized that standing up for Epstein’s victims—and against the wealthy or powerful—was a way to build trust.
“You have to convince people that the government actually works,” Khanna said.
I have been reporting on Epstein—his criminal investigations, his network of powerful friends, and his hundreds, possibly thousands, of young victims—for almost a decade. I’ve chronicled the many false starts and dashed hopes for accountability following Epstein’s second arrest in July 2019, his death while in custody, and the efforts to bring others in his network to justice. I have spent years trying to figure out what, if anything, Donald Trump knew about Epstein’s abuse of young women, and whether the president was at all involved. On this question and many others, I was repeatedly told: “The story is over.”
But over the past several months, something extraordinary has happened: An unlikely duo of lawmakers—a progressive from Silicon Valley and a conservative from Kentucky—partnered with a group of Epstein survivors and a few female Republican lawmakers to successfully push for legislation mandating the release of the so-called Epstein files. It was the most significant act of defiance by Republicans in Congress against the president in his second term, and it transformed what had been treated as a fringe conspiracy theory into a populist rallying cry.
The release of the files over the past two months has led to new criminal investigations outside of the United States. In Britain, Andrew Mountbatten-Windsor, the former prince, and Peter Mandelson, the former U.K. ambassador to the United States, were arrested but haven’t been charged; in Norway, Thorbjørn Jagland, the former prime minister, was charged with “gross corruption.” More than a dozen other prominent people have stepped down from lucrative jobs or lost honorary titles or positions. This is the story, drawn from interviews with more than 40 people over the past eight months, of the unlikely coalition that came together to force some measur
Front Toxicol. 2026 Feb 3;8:1736963. doi: 10.3389/ftox.2026.1736963. eCollection 2026.
ABSTRACT
Endocrine disruptors (EDs) are implicated in adverse developmental and reproductive outcomes, yet their identification remains a major challenge in chemical safety assessment. Current testing strategies rely heavily on animal models, which are constrained by ethical concerns, interspecies differences, and limited mechanistic resolution but justified by the complexity of the endocrine system and its physiology. Capturing the complex biology of intact organisms and incorporating toxicokinetic properties in alternative test methods is challenging. To address this, the European Partnership for the Assessment of Risks from Chemicals (PARC) is advancing the development and regulatory integration of new approach methodologies (NAMs). This project specifically contributes by developing and validating human-relevant NAMs to identify key aspects of endocrine disruption relevant to developmental and reproductive toxicity (DART). Key innovative activities include predictive modeling, refinement of zebrafish and amphibian embryo assays, and establishment of advanced in vitro systems for assessing toxicity in the oocyte, testis, placenta, and brain. By combining mechanistic insights with multi-modality and high throughput testing strategies, this work aims to improve the predictive power and regulatory utility of NAMs for ED identification within the One Health paradigm.
PMID:41710520 | PMC:PMC12910832 | DOI:10.3389/ftox.2026.1736963
The EFSA Stakeholder Workshop on Genotoxicity Guidance Revision was held on 3‐4 November 2025 in Brussels, Belgium. The event aimed to inform stakeholders about the ongoing revision of EFSA's genotoxicity guidance framework and gather input on key scientific aspects, including the testing strategy, integration of new approach methodologies (NAMs), quantitative genotoxicity assessment and genotoxicity assessment of mixtures. The Workshop featured presentations from EFSA, sister agencies (ECHA, EMA), the European Commission (EC), OECD, as well as stakeholders from industry and academia.
bioRxiv [Preprint]. 2026 Jan 22:2026.01.19.700343. doi: 10.64898/2026.01.19.700343.
ABSTRACT
Development is a tightly regulated process that establishes body axes and orchestrates the spatial organization of tissues and organs. Although developmental programs contain inherent redundancies, they remain highly sensitive to environmental cues. Among environmental contaminants, per- and polyfluoroalkyl substances (PFAS), chemicals that resist degradation and bioaccumulate in the body, are of particular concern. These "forever chemicals" are widespread in our household products, including non-stick and waterproof materials, and drinking water remains a major source of exposure. PFAS accumulate in specific tissues and have been associated with developmental delays, childhood leukemia, and other adverse health outcomes, yet the cellular and molecular mechanisms by which they disrupt early development remain largely unknown. To address this, we employ zebrafish embryos as a New Approach Methodology (NAM) to investigate how perfluorooctanoic acid (PFOA), a prevalent environmental PFAS, alters early embryogenesis. Embryos were exposed to physiologically relevant low and high doses of PFOA and analyzed at 24 hours post-fertilization (hpf), a key stage of organogenesis. We also included a parental exposure group, in which adults were treated with PFOA and their offspring were collected to assess whether the effects of exposure were transmitted to the next generation. Developmental processes are inherently plastic, and we wanted to understand the extent to which PFOA impacts normal cellular processes as well as the redundancy in the system (different developmental signaling pathways) which ensures that an embryo develops properly. Towards this, we performed single-nucleus RNA sequencing at 24 hpf, and it revealed that neuronal and muscle tissue clusters are particularly sensitive to PFOA exposure. These molecular perturbations correspond with anxiety-like behavioral phenotypes we observed in the exposed larvae, linking early developmental disruptions to organism-level outcomes. Overall, our findings provide mechanistic insight into the way in which PFAS exposure alters development, disrupting gene expression patterns and chromatin organization in developing tissues, revealing how early molecular perturbations can give rise to long-term behavioral consequences.
PMID:41648621 | PMC:PMC12871824 | DOI:10.64898/2026.01.19.700343
Front Toxicol. 2025 Dec 17;7:1690491. doi: 10.3389/ftox.2025.1690491. eCollection 2025.
ABSTRACT
Read-across has matured from an expert-driven extrapolation based largely on structural analogy into a rigorously documented, mechanistically informed cornerstone of next-generation risk assessment. Three pivotal frameworks are compared that now shape its regulatory use: the European Food Safety Authority's (EFSA) 2025 guidance for food and feed safety, the European Chemicals Agency's (ECHA) Read-Across Assessment Framework (RAAF) for industrial chemicals under REACH, and the community-driven Good Read-Across Practice (GRAP) principles. Using five analytical lenses-conceptual structure, scientific rigor, implementation tools, regulatory acceptance, and practical impact-we identified areas of complementarity and divergence. EFSA provides a seven-step, uncertainty-anchored workflow that actively embeds new approach methodologies (NAMs) and adverse outcome pathway reasoning, offering applicants a transparent "how-to" template. RAAF, in contrast, operates as an evaluator's rubric: six scenario types and associated assessment elements delineate what evidence must be delivered, thereby standardizing regulatory scrutiny but leaving dossier construction to the registrant. GRAP supplies the conceptual glue, emphasizing mechanistic plausibility, exhaustive analogue selection, explicit uncertainty characterization, and the strategic use of NAMs; its influence is evident in both EFSA's and ECHA's evolving expectations. (Terminology note: the acronym "NAM" was popularized at an ECHA workshop in 2016; earlier documents such as RAAF and initial GRAP papers therefore may not use the term explicitly). Regulatory experience under REACH demonstrates that dossier quality and acceptance rates rise markedly when RAAF criteria are met, while EFSA's new guidance is poised to catalyze similar gains in food and feed assessments. Globally, the convergence of these frameworks-reinforced by OECD initiatives and NAM-enhanced case studies-signals an emerging international consensus on what constitutes defensible read-across. In conclusion, harmonizing EFSA's procedural roadmap with RAAF's evaluative rigor and GRAP's best-practice ethos can mainstream reliable, animal-saving read-across across regulatory domains, paving the way for fully mechanistic, AI-enabled chemical safety assessment.
PMID:41479674 | PMC:PMC12753468 | DOI:10.3389/ftox.2025.1690491
Open Res Eur. 2025 Dec 9;4:194. doi: 10.12688/openreseurope.18439.2. eCollection 2024.
ABSTRACT
Metabolism disrupting chemicals (MDCs) elicit negative effects on metabolically active organs such as the liver and the pancreas, altering normal metabolic processes. Chemicals that are known, or suspected MDCs include compounds found in everyday consumer products and food, making low-dose, continuous exposure inevitable for humans. Through the discovery of chemically induced metabolic disruption, a concern has surfaced whether and how MDCs impact human health and the development of metabolic diseases. This has accelerated research around the topic, and it has been found that exposure to MDCs is linked to increased incidence of metabolic diseases including obesity and liver steatosis. Effective regulatory action is hindered by the lack of accurate methods to identify MDCs. The NEMESIS project addresses this regulatory gap by investigating the mechanisms through which MDCs cause metabolic disruption. The project aims at identifying novel biomarkers of exposure and link exposure to disease outcomes. As chemical toxicity testing is rapidly moving towards new approach methodologies (NAMs), NEMESIS promotes non-animal methodologies by employing state-of-the-art in vitro methods, epidemiological data, systems biology approaches, and seeks to replace mammalian in vivo experiments with alternative models. By understanding mechanisms of MDC-induced metabolic health effects, and through the development of reliable effect biomarkers and testing strategies, the NEMESIS project aims to facilitate more effective regulatory measures to improve and protect the health and well-being of EU citizens. The project is particularly focused on maximizing its impact through effective dissemination and communication efforts, to ensure that the project's message and results reach a broad audience and are tailored to different population groups. These actions will improve the risk assessment of MDCs and ensure that the EU citizens are informed and protected from the harmful effects of MDCs and can adapt their consumer patterns and behaviors to prevent exposure.
PMID:41477515 | PMC:PMC12749563 | DOI:10.12688/openreseurope.18439.2
Toxics. 2025 Dec 3;13(12):1048. doi: 10.3390/toxics13121048.
ABSTRACT
The rapid growth in manufacturing and use of electrical and electronic equipment has led to unprecedented volumes of poorly managed e-waste, posing serious ecological risks. Although data on individual chemical substances in e-waste are available, evidence of ecotoxicity from actual e-waste materials remains scattered. This review consolidates organism-level ecotoxicity data on real e-waste samples (mixed fractions, fragments, leachates) and samples collected near e-waste facilities (soil, sediments, dust, water) across aquatic and terrestrial environments. It critically examines how methodological approaches influence reported outcomes and outlines research priorities. In aquatic environments, toxic responses vary with increased amounts of toxicants (dissolved metals, particles from dismantling operations) that mobilise to surface waters, while hydrophobic organic compounds cause sublethal behavioural and genotoxic effects. The few studies on terrestrial environments show impaired invertebrate growth and reproduction, along with changes in soil and "plastisphere" microbiota. However, tested concentrations, material complexity, and incomplete reporting of exposure chemistry, among other factors, limit the environmental relevance and comparability of the data. Uniformised procedures, combined with thorough chemical characterisation, environmentally realistic conditions, and cross-system bioassays (including different exposure routes and cumulative assessments), may provide mechanistic insights into e-waste toxicity, supporting evidence-based risk management strategies while contributing towards the development and validation of robust new approach methodologies (NAMs).
PMID:41441269 | PMC:PMC12737757 | DOI:10.3390/toxics13121048
Endocrinology. 2025 Dec 20:bqaf186. doi: 10.1210/endocr/bqaf186. Online ahead of print.
ABSTRACT
Endocrine mediators are essential for pregnancy maintenance, and their functional withdrawal is associated with normal term and preterm birth (PTB). Therefore, the disruption to endocrine functions or agents that can disrupt endocrine functions are naturally suspected as contributors to PTB. One of the well-studied endocrine-disrupting compounds is Polybrominated diphenyl ether(s) (PBDE). PBDE is a flame-retardant compound that is contained in several products and is a ubiquitous environmental contaminant. PBDE exists in several different congeners, many harmless compounds, but a few PBDE congeners are linked as endocrine disruptors contributing to adverse pregnancy outcomes like PTB. However, data ambiguity suggests that current platforms are insufficient to conclude PBDE's mechanisms of action as an endocrine disruptor at the feto-maternal interface (placenta/fetal membranes). The development of microfluidic-based new approach methods (NAMs) is being introduced to study PBDE and other environmental pollutants. Organs-on-a-chip (OOC) are an emerging class of NAMs that can replicate human organ-level functions in vitro. OOCs are microfluidic systems comprising multiple cell types from an organ that mimics the environment of a physiological organ. These devices are interconnected through microchannels to maintain intercellular interactions. OOC-based testing and development have accelerated globally as regulatory agencies now emphasize the need for reliable, humanized alternatives to traditional animal models. Multiple reproduction-associated OOCs are being developed, and their utility has been tested in assessing mechanisms of action and toxicological parameters of environmental pollutants. This review provides an overview of feto-maternal interface OOCs and uses PBDE as an example to demonstrate how OOCs can study endocrine-disrupting compounds.
PMID:41420538 | DOI:10.1210/endocr/bqaf186
The project Computational characterisation of ecological hazard and risk of environmental mixtures focused on two key priorities in advancing a next‐generation risk assessment (NGRA) workflow: (i) transitioning to assessing mixtures and (ii) utilising mechanism‐based hazard assessment based on new approach methodology (NAM) data.
New approach methodologies (NAMs) are gaining increasing attention within the scientific community as strategies to reduce the use of animal testing. Their integration into regulatory frameworks for chemical hazard and risk assessment in food and feed would directly support the goals of the European Food Safety Authority (EFSA).
Recently, I’m ashamed to admit, I received an email that initially made me feel warm, human, even grateful: a rejection for a job I’d applied to. But my thankful feelings quickly curdled into self-loathing—the nausea one gets when looking back over pathetic, paragraphs-long texts to an ex, whose monosyllabic responses suggested they’d clearly moved on. The rejection was a form letter, not even a late-round, personalized “we gave you serious consideration but ultimately decided to hire a VP’s nephew” message. I was so accustomed to being treated with indifference, I realized, that the barest acknowledgment of my existence felt like a win.
Putting aside the question of whether the job market itself is in good or bad shape right now (it’s bad), the code of what behavior is and isn’t acceptable seems to have broken down. Ghosting has become more rampant not just by employers but also by job seekers. In 2024, candidates reviewing employers on the website Glassdoor used the term ghosting nearly three times as much as they did in 2020. And a 2023 Indeed survey of job seekers found that 62 percent of respondents planned to ghost a prospective employer in a future job search, compared with only 37 percent in 2019. The disappearing act is not just in the early rounds, either. Employers routinely ask applicants to do multiple interviews and time-consuming test work, and are never heard from again; a survey this year from Greenhouse, a recruiting-software company, found that nearly two out of every three candidates in the United States had been ghosted after an interview. Meanwhile, some applicants who make it through the onerous hiring process and accept jobs never show up for their first day. One California recruiter told me that some of the candidates who ditched had even signed offers for positions that paid six-figure salaries. Today, many people on both sides of the hiring equation—whether because of convenience, self-protection, or resentment—have abandoned even the pretense of courtesy, resulting in a job market that’s as rude as it is dysfunctional.
[Read: The great ghosting paradox]
It might sound glib to blame a simple loss of manners for employment woes. The way cultural norms tell people to comport themselves can seem silly—like empty formality (Smile even if you don’t mean it) or even hypocrisy, because this behavior often hides the way people actually feel or what they actually want to say. But try going a single day telling a boss or a landlord, an employee or a co-worker, what you really think, and why everybody obscures their true opinions all the time would quickly become apparent.
Manners, it turns out, have long been smoothing over a lot of rough edges in the hiring process, hiding the uglier realities that can emerge from the unequal relationship between employers and the employed. In a society where many workers have little power, the fig leaf of societal niceties is often the only difference between a functioning system built on trust and a polarized one roiled by humiliation, disgust, and retribution.
Before a code of conduct was something that HR had you read and initial when you started a job, it was the main deterrent against open class warfare. Throughout the 18th and 19th centuries, in Europe and America, manners essentially boiled down to “Good fences make good neighbors.” People avoided class conflict mainly by avoiding each other; associating outside your class was strongly discouraged, according to Informalization, a history of manners by the sociologist Cas Wouters. When “class mixing” was necessary, people maintained psychological distance by acting with extreme formality, often making elaborate shows of haughty superiority or craven deference. When people clashed, norms dictated that the privileged classes were afforded the prerogative of violence; retribution from lower classes tended to be penalized. An 1859 guide to courtesy, The Habits of Good Society, for instance, advised that if a gentleman were confronted with a “dishonest cabman,” then “one well-dealt blow settles the whole matter.” The author goes on to specify that violence is only for pu
Comput Struct Biotechnol J. 2025 Nov 16;29:305-327. doi: 10.1016/j.csbj.2025.11.032. eCollection 2025.
ABSTRACT
Traditional in vivo methodologies have long formed the foundation of chemical and material safety assessment, yet they are increasingly inadequate to meet modern regulatory, ethical, and sustainability demands. These conventional approaches are resource-intensive, ethically questionable, and often fail to accurately predict human or environmental toxicity, particularly for emerging pollutants such as PFAS, (nano-) pesticides, and 2D materials. In response, the EU has launched initiatives like the Chemical Strategy for Sustainability and the Zero Pollution Action Plan under the European Green Deal to promote innovation in safer, and more sustainable chemicals. Central to this transformation is the Safe and Sustainable by Design (SSbD) framework, developed by the European Commission's Joint Research Center, which provides structured methodologies and metrics to integrate safety and sustainability into material innovation from the earliest stages of design. Building on this vision, the CHIASMA project aims to advance Next generation Safety Assessment (NGSA) by developing innovative New Approach Methodologies (NAMs) that combine experimental, computational, and Life Cycle Assessment (LCA) tools. Focusing on key biological systems and exposure routes, CHIASMA integrates Artificial Intelligence (AI), Machine Learning (ML), and Knowledge Graph (KG) technologies to enhance data interoperability and predictive accuracy. By embedding FAIR data principles and aligning with Good Laboratory Practice (GLP) standards, CHIASMA promotes transparency and regulatory acceptance. Fully aligned with SSbD principles, CHIASMA establishes a digital, interoperable infrastructure for predictive safety evaluation, that leverage on state-of-art experimental New Approach Methodologies (NAMs) bridging critical data gaps and supporting the transition towards sustainable, science-driven, and ethically responsible chemical and material innovation in Europe and beyond.
PMID:41362652 | PMC:PMC12682023 | DOI:10.1016/j.csbj.2025.11.032
Front Toxicol. 2025 Nov 19;7:1665163. doi: 10.3389/ftox.2025.1665163. eCollection 2025.
ABSTRACT
INTRODUCTION: Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants with potential immunotoxic effects. Most toxicological studies have focused on long-chain PFAS such as perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA). However, short-chain and ultra-short-chain alternatives, including trifluoroacetic acid (TFA), are increasingly used despite limited toxicological data.
METHODS: This study evaluated and compared the immunotoxic effects of PFAS with varying chain lengths-long-, short-, ultra-short-chain compounds, and fluoropolymer representatives (polytetrafluoroethylene, PTFE)-using human-relevant new approach methodologies (NAMs). Two complementary in vitro models were employed. Peripheral blood mononuclear cells (PBMCs) from healthy donors to assess antibody (IgG and IgM) production. THP-1-derived dendritic cells (DCs) to evaluate maturation marker expression (CD83, CD86, HLA-DR). Environmentally and occupationally relevant PFAS concentrations were tested.
RESULTS: PFOS, PFOA, and perfluorononanoic acid (PFNA) suppressed antibody production and impaired DC maturation in a concentration-dependent manner, consistent with previous in vivo and epidemiological data. Short-chain PFAS (PFHxS, PFBS, PFHxA, PFBA) showed modest to intermediate immunomodulatory activity, with subtle immunosuppressive trends in female donors. Notably, TFA reduced antibody production at levels comparable to PFOS, indicating that chain length alone is not a reliable predictor of immunotoxic potential. PTFE exhibited no suppressive effects; instead, increased antibody release was observed in female donors, suggesting possible sex-dependent immunostimulation.
DISCUSSION: Findings support a nuanced, compound-specific approach to PFAS risk assessment rather than a simple long- vs. short-chain distinction. In vitro NAMs provided mechanistic, human-relevant insights and reinforce their integration into regulatory frameworks.
PMID:41347043 | PMC:PMC12672224 | DOI:10.3389/ftox.2025.1665163
Toxicol Sci. 2025 Dec 4:kfaf166. doi: 10.1093/toxsci/kfaf166. Online ahead of print.
ABSTRACT
The EPA continues to evaluate strategies to implement new approach methods (NAMs) for screening chemicals that disrupt the thyroid endocrine system. Validation of NAMs is a critical milestone toward establishing confidence in data sources that could be used in a regulatory decision-making context. The objective of this study was to conduct an interlaboratory validation of the human thyroid microtissue assay to evaluate its relevance and reliability. In coordination with the U.S. validation authority, NICEATM, and collaboration with industry partners (LifeNet Health, Bayer Crop Science, Corteva Agrisciences), the study aims were to 1) define the study design and establish standard operating procedures, 2) conduct test method transfer, training, and within-laboratory model performance evaluation, 3) perform interlaboratory reference chemical testing and assay performance evaluation. Progress was independently monitored by a validation management team comprised of an international group of experts in thyroid physiology, in vitro test methods, and regulatory toxicology. Results indicated the thyroid microtissue model could be reliably transferred to new laboratories with reproducible effects on thyroid hormone synthesis. Interlaboratory testing of four blinded reference chemicals (three true positive, one true negative) across three independent human donors revealed consistent bioactivity across the reference set and performance metrics (dynamic range, precision, screening quality) that met acceptance criteria and shed insight into areas for improvement. In the context of a NAM-based testing strategy, a validated human thyroid microtissue assay enables direct measurement of thyroid hormone synthesis perturbations, reducing reliance on animal testing and addressing a critical mode-of-action that is of regulatory concern.
PMID:41344865 | DOI:10.1093/toxsci/kfaf166
Environ Sci Technol. 2025 Nov 21. doi: 10.1021/acs.est.5c11122. Online ahead of print.
ABSTRACT
The global restriction of legacy per- and polyfluoroalkyl substances (PFASs), such as perfluorooctanoic acid (PFOA), has led to increased production and environmental occurrence of emerging alternatives. This study applied a next generation risk assessment (NGRA) framework to evaluate and compare the local environmental risks of eight PFOA alternatives using multiple lines of evidence from new approach methodologies (NAMs). Targeted chemical analysis was conducted to quantify environmental concentrations in wastewater and surface waters near an industrial source. Hazard characterization integrated in silico modeling, in vitro assays using zebrafish liver cells, in vivo zebrafish embryo toxicity tests, and transcriptomic profiling. Multiple points of departure (PODs), including conventional effect concentrations, benchmark concentrations (BMC), transcriptomic POD (tPOD), and species sensitivity distributions (SSD), were derived. The results showed that HFPO-TA and HFPO-TeA were cytotoxic, while HFPO-DA, HFPO-TA, and HFPO-TeA caused significant embryo toxicity. HFPO-TeA emerged as the most potent alternative, with transcriptomic signatures implicating disruption of metabolic and cardiovascular functions. Risk quotients (RQ) for HFPO-DA exceeded concern thresholds across end points, while those for HFPO-TA and HFPO-TeA declined with appropriate assessment factor adjustments. These findings highlight potential local environmental risks from PFOA alternatives and demonstrate the utility of NAM-based NGRA frameworks.
PMID:41270243 | DOI:10.1021/acs.est.5c11122
Toxicol Sci. 2025 Nov 19:kfaf162. doi: 10.1093/toxsci/kfaf162. Online ahead of print.
ABSTRACT
Zebrafish (danio rerio) are an ideal system for understanding developmental toxicity as they display similar toxicity outcomes to other vertebrates. Further, many molecules have been tested for developmental toxicity in zebrafish providing an opportunity for machine learning model development. We curated 1345 small molecules from ToxCast, flame retardant compounds, per- and polyfluoroalkyl substances (PFAS), and industrial chemicals published by the Superfund Research Program (SRP). Following curation, we trained machine learning models on the zebrafish toxicity endpoints ANY_ = any effect including mortality, ANY_BUT_MORT = any effect excluding mortality, MORT = mortality ie did the embryo die, EDEM = did an edema form, CRAN = Craniofacial malformation. We demonstrated that these models were better than random when compared to shuffled data. We also fine-tuned the molecular SMILES encoder MolBART to predict on all zebrafish toxicity endpoints and found it generally matched the performance of classical machine learning models for ANY_BUT_MORT, CRAN, and EDEM endpoints. We present new toxicity data for Proteolysis Targeting Chimeras (PROTACs) in Zebrafish and machine learning models for these data by fingerprinting different parts of the molecule individually, yielding predictive performance (AUROC 0.6-0.7). If we are to reduce animal testing with new approach methodologies (NAMs) like these Zebrafish toxicity models they need to be able adapt to new molecular classes like PROTACs.
PMID:41259055 | DOI:10.1093/toxsci/kfaf162
International Journal of Toxicology, Ahead of Print.
The U.S. Food and Drug Administration (FDA), Center for Drug Evaluation and Research (CDER), and Office of New Drugs (OND) has continuously encouraged the submission of nonclinical tests utilizing new approach methodologies (NAMs) and sponsor engagement ...
International Journal of Toxicology, Volume 45, Issue 2, Page 136-156, March/April 2026.
The U.S. Food and Drug Administration (FDA), Center for Drug Evaluation and Research (CDER), and Office of New Drugs (OND) has continuously encouraged the submission of nonclinical tests utilizing new approach methodologies (NAMs) and sponsor engagement ...
Front Toxicol. 2025 Oct 8;7:1657708. doi: 10.3389/ftox.2025.1657708. eCollection 2025.
ABSTRACT
Scientific confidence frameworks (SCFs) are alternatives to traditional validation for new approach methodologies (NAMs). The SCFs adapted by the Interagency Coordinating Committee for the Validation of Alternative Methods (ICCVAM) and the American Chemistry Council (ACC) both address inference performance-the ability of NAMs to predict or inform the biological effect of interest. Inference performance is a distinct evaluation procedure in ACC's SCF but is blended into several steps of ICCVAM's SCF. Here, we first reproduce the previously derived human relevant potency threshold (HRPT) for the estrogen receptor alpha (ERα) agonism of Borgert et al. (2018) using guideline and guideline-like studies; we found that a HRPT of 1 to 10-1 positively and consistently predicted clinical endometrial and endocervical effects. We next mapped inference performance to ICCVAM's SCF and found that it can be used as an effective initial screen prior to performing more detailed characterizations in their SCF. We first conclude that a HRPT for ERα agonism of 10-2 to 10-4 is a health-protective NAM based on an established mode of action that could potentially be used in early screening, much like the threshold of toxicological concern. We then conclude that inference performance is a core requirement for SCFs.
PMID:41132625 | PMC:PMC12540326 | DOI:10.3389/ftox.2025.1657708
Mar Pollut Bull. 2025 Oct 17;222(Pt 3):118790. doi: 10.1016/j.marpolbul.2025.118790. Online ahead of print.
ABSTRACT
Coastal marine environments receive large influxes of anthropogenic contaminants from land-based sources such as metal(loid) s and emerging persistent organic pollutants (POPs). These contaminants can accumulate in marine sediments, seagrass and herbivorous marine megafauna such as dugongs, manatees and green turtles. Using the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) approach, we evaluated 131 publications on the bioaccumulation and effects of metal(loid)s and organic contaminants in seagrass, dugongs, manatees and green turtles. The global distribution of these publications was mapped, with concentration and effects data extracted to calculate chemical risk quotients (RQ). Metal(loid)s were the most studied contaminant class, while research on other chemicals including per - and polyfluoroalkyl substances (PFAS) and pharmaceuticals and personal care products (PPCP) were under-represented. The link between the concentration of a contaminant measured in tissue and its biological effect was often lacking. New Approach Methods (NAMs), which include in vitro assays and omics approaches, were highlighted as valuable but not widely used. The reported bioaccumulated concentrations of contaminants in the literature varied greatly, with little data on the cumulative risk of mixtures. The calculation of RQ suggest a risk to many wild populations to the adverse effects of some contaminants. This review emphasises the need for standardised methods and data sharing, which will foster a more unified and global approach to better assess the risk contaminants pose to seagrass and the megafauna they support.
PMID:41109174 | DOI:10.1016/j.marpolbul.2025.118790
Toxicol Sci. 2025 Sep 29:kfaf136. doi: 10.1093/toxsci/kfaf136. Online ahead of print.
ABSTRACT
Environmental exposure to industrial chemicals, endocrine disruptors, and pharmaceuticals has been increasingly linked to the global decline in male reproductive health. To address the urgent need for efficient and mechanistically informed toxicity screening, we developed a high-throughput, high-content screening (HTS/HCA) platform using a 3D in vitro mini-testis model. This system was used to evaluate 87 structurally diverse compounds from the National Toxicology Program (NTP) chemical library. The model incorporates murine-derived spermatogonia, Sertoli, and Leydig cells embedded in an extracellular matrix, providing a physiologically relevant environment for mechanistic toxicology. Each compound was tested across ten phenotypic endpoints, including nuclear morphology, cytoskeletal integrity (F-actin), DNA damage (γH2AX), and cell viability by using high-content imaging. Quantitative Points of Departures (PODs) were calculated and integrated into a High-content Assay Index (HCAI). Toxicological Priority Index (ToxPi) scores, derived from the PODs, enabled compound ranking and clustering. Compared to existing in vivo reproductive toxicity data, the 3D model demonstrated 91.5% sensitivity, 93.8% specificity, and 93.6% concordance (n = 64 compounds). Notably, 22 compounds lacking reproductive toxicity data were identified as potentially reproductive toxicants. Mechanistic analyses revealed that nuclear morphology, F-actin intensity, and γH2AX were the most sensitive indicators of reproductive toxicity. Cluster and category-level analysis showed that flame retardants and pesticides ranked highest in toxicity. The integration of multi-parametric data via ToxPi facilitated high-resolution chemical prioritization. Given current ethical and technical challenges in sourcing human testicular tissue or differentiating stem cells into testicular cell types, murine cells provide a reproducible and practical alternative for complex multicellular testis modeling. Our results demonstrate that the HCA-integrated 3D mini-testis model offers a robust, scalable, and mechanistically insightful platform for male reproductive toxicity screening, supporting its adoption as New Approach Methodologies (NAMs) aligned with regulatory and ethical testing goals.
PMID:41017718 | DOI:10.1093/toxsci/kfaf136
Front Endocrinol (Lausanne). 2025 Sep 12;16:1654965. doi: 10.3389/fendo.2025.1654965. eCollection 2025.
ABSTRACT
INTRODUCTION: Intrauterine exposure to endocrine disrupting chemicals (EDCs), particularly anti-androgens, has been implicated in hypospadias by disrupting fetal masculinization of the genital tubercle (GT). Other pathways, including estrogen signaling, may also contribute but remain poorly characterized, especially in rats - a key model in chemical toxicity testing. Estrogen signaling has also been linked to hypospadias in mice, raising questions about androgen-estrogen interactions in guiding GT differentiation.
METHODS: We induced hypospadias in male rat offspring via intrauterine exposure to the antiandrogenic drug flutamide and characterized androgen and estrogen receptor expression.
RESULTS: We observed key structural and transcriptional changes in the developing penis, including altered estrogen receptor a (ERa, Esr1) expression. Notably, beyond this established androgen-estrogen relationship in hormone-sensitive tissues, anti-androgenic exposure also induced spatial changes in Esr1 expression in specific regions of the GT.
DISCUSSION: Future toxicological testing using new approach methodologies (NAMs) should consider androgen-estrogen balance and crosstalk in reproductive tissues as a mechanism of action.
PMID:41019341 | PMC:PMC12464887 | DOI:10.3389/fendo.2025.1654965
Mol Reprod Dev. 2025 Sep;92(9):e70058. doi: 10.1002/mrd.70058.
ABSTRACT
Visfatin, a multifunctional adipokine, plays a crucial role in ovarian physiology. This study examines its effects on the level of selected angiogenesis and apoptosis factors in the porcine corpus luteum (CL). Using in vitro cultures of luteal cells, we analyzed the effects of visfatin on key angiogenic factors (VEGF family, PDGF, bFGF2, ANG, and iNOS) and their receptors. Additionally, we assessed luteal cell viability and apoptosis by measuring caspase activity, DNA fragmentation, and transcript levels of apoptotic factors. We noted that visfatin mostly decreases the expression of angiogenic factors and their receptors during the luteal phase except for VEGFC, VEGFR3 on Days 2-3 of the estrous cycle, and VEGFB, VEGFD, VEGFR1, VEGFR2, FGFR2 on Days 14-16 of the estrous cycle, which were upregulated. Moreover, visfatin inhibits apoptosis by downregulating caspase-3, -8, -9, BAX, and caspase 3/7 activity while increasing BCL-2 and luteal cells viability. Treatment with FK866, nicotinamide phosphoribosyltransferase (NAMPT) inhibitor, largely disrupted observed effects, highlighting the NAMPT-dependent function of visfatin. Moreover, visfatin suppressed VEGF-A levels via insulin receptor and MAPK, while insulin receptor, MAPK, and AKT pathways mediated the inhibition of caspase 3/7 activity. These findings suggest that visfatin regulates apoptosis and angiogenesis in the porcine CL.
PMID:41015905 | DOI:10.1002/mrd.70058
International Journal of Toxicology, Volume 44, Issue 6, Page 499-506, November/December 2025.
Technological advances and the desire to reduce dependence on animal models have brought human-relevant models to the forefront of drug development. This paradigm shift is leveraging the advances inin vitrosystems and new approach methodologies (NAMs), ...
International Journal of Toxicology, Ahead of Print.
Technological advances and the desire to reduce dependence on animal models have brought human-relevant models to the forefront of drug development. This paradigm shift is leveraging the advances inin vitrosystems and new approach methodologies (NAMs), ...
Enzyme Microb Technol. 2025 Jul 24;191:110724. doi: 10.1016/j.enzmictec.2025.110724. Online ahead of print.
ABSTRACT
Poly and per fluorinated substances (PFAS) are emerging contaminants of concern that are thought to be involved in causing numerous adverse health effects, such as immunosuppression, increased chance of cancer development, and altered levels of hepatic enzyme levels in humans. However, PFAS are considered highly persistent and resistant to biodegradation given the fact that the C-F bond can have a bond dissociation energy of up to 544 kJ/mol. Though many studies have reported PFAS biodefluorination by bacterial isolates and microbial communities, little is known regarding the molecular foundations for biodefluorination. In this study, a novel defluorinase was identified, that is responsible for the biodefluorination of 6:2 fluorotelomer carboxylic acid (6:2 FTCA) in R.jostii RHA1 using the combination of transposome-based insertional mutagenesis and heterologous expression. From a library of 417 R.jostii RHA1. mutants, 3 individual mutants lost their ability for defluorination when they were exposed to 6:2 FTCA (mutant # 15, 32 and 38 - Table S2). The disruption of the genetic locus in all 3 non-defluorinating mutants was identified coding for a putative MhPC superfamily protein. The MhPC superfamily of proteins is known to harbor other proteins such as fluoroacetate dehalogenase (UniProt - Q6NAM1) that are capable of -C-F bond cleavage. This identified gene was cloned into the heterologous expression host M. smegmatis MC2-155. After induction, the M. smegmatis MC2-155 transformant exhibited the ability to defluorinate 6:2 FTCA at a rate of 13 µmol/h (Vmax = 80.9 µmol/min and Km = 5.04 mM in Michaelis-Menten models). In contrast, defluorination was not observed in either abiotic or biotic controls. Further characterization of the novel defluorinase indicated that it could moderately defluorinate the unsaturated PFAS compound 6:2 FTCUA (4.9 µmol/h fluoride) and minimally defluorinate 5:2 sFTOH (1.3 µmol/h fluoride). The novel defluorinase indicated a maximal specific activity of 12.9 ± 1.9 µmol F/hr/g protein, against its primary PFAS substrate, 6:2 FTCA. However, it showed no activity with 5:3 FTCA or sulfonated PFAS compounds such as 6:2 FTS and 8:2 FTS. The wild-type Rhodococcus could defluorinate 6:2 FTCA at a rate of 2.2 µmol/h. The discovery of this MhPC class novel defluorinase in WT R.jostii RHA1. has substantial value since it is responsible for the critical step that initiates defluorination of PFAS compounds such as 6:2 FTCA.
PMID:40714482 | DOI:10.1016/j.enzmictec.2025.110724
J Toxicol Environ Health B Crit Rev. 2025 Jul 6:1-69. doi: 10.1080/10937404.2025.2521615. Online ahead of print.
ABSTRACT
Per- and polyfluoroalkyl substances (PFAS) comprise a large class of human-made chemicals that are in widespread use and present concerns for persistence, bioaccumulation and toxicity. Whilst a handful of PFAS have been characterized for their hazard profiles, the vast majority of PFAS have not been extensively studied. A comprehensive evaluation to characterize the hazard profiles of the thousands of available PFAS would require extensive resources in terms of cost, number of animals and time. An alternative and more efficient approach is to develop a structural chemical categorization approach to prioritize which PFAS or categories of PFAS should be subject to additional study. To that end, the U.S. Environmental Protection Agency (EPA), in collaboration with the National Institute of Environmental Health Sciences (NIEHS) Division of Translational Toxicology (DTT), initiated a research project in 2018 to screen approximately 150 PFAS through a battery of alternative model organisms, in vitro cell and biochemical assays, and in vitro toxico kinetic (TK) assays in order to inform chemical category and read-across approaches. The aim of this review summarizes the experimental testing undertaken, how data were processed, what insights were derived from a category perspective and how these might potentially inform subsequent tiered testing.
PMID:40619890 | DOI:10.1080/10937404.2025.2521615
Regul Toxicol Pharmacol. 2025 Nov;162:105884. doi: 10.1016/j.yrtph.2025.105884. Epub 2025 Jun 6.
ABSTRACT
Read-across is a widely used adaptation to address information requirements under REACH. However, registrants submitting for the 2010 and 2013 deadline have often failed to satisfy regulatory requirements from ECHA's point of view. Due to their complex composition and unique physicochemical properties, surfactants are posing major challenges in this respect. With the aim to improve future submissions and prevent unnecessary animal testing, read-across-related discussions of 72 ECHA Final Decisions on Compliance Checks and Testing Proposal Evaluations of 24 major surfactant groups were analysed in-depth, and causes for acceptance or rejection were identified. Key drivers of regulatory acceptance/rejection were presence or absence of composition information, considerations on structural similarity as well as availability and nature of bridging studies. Several elements were identified that may be easily improved in future REACH dossiers without the need for additional animal testing. Other cases revealed uncertainty of expectations by ECHA, highlighting the need for improved communication during the dossier preparation. Notably, no example for acceptance of read-across based on non-animal New Approach Methodologies (NAMs) was identified in this analysis. Owing to the benefits that non-animal NAMs may present as supporting information, all stakeholders are encouraged to contribute to an increase of regulatory acceptance.
PMID:40484111 | DOI:10.1016/j.yrtph.2025.105884
The American Chemistry Council’s Long-Range Research Initiative (LRI) is proud to announce its participation in the NIH Common Fund’s Complement Animal Research In Experimentation (Complement-ARIE) public-private partnership (PPP) to accelerate the scientific development and evaluation of New Approach Methodologies (NAMs).
Sci Total Environ. 2025 Jul 1;984:179690. doi: 10.1016/j.scitotenv.2025.179690. Epub 2025 May 24.
ABSTRACT
Aquatic ecosystems provide essential services, yet they face increasing pressures from anthropogenic activities, including land-use change, eutrophication, browning, and contaminant pollution. While the ecological effects of these stressors are documented, the impacts of complex contaminant mixtures, particularly those from wastewater treatment plant (WWTP) effluents, remain poorly understood. Mixtures effects are typically assessed using traditional species-by-species toxicological approaches, which, though the gold standard, are time-intensive, require test animals, and have limited extrapolability. New Approach Methodologies (NAMs), such as environmental DNA (eDNA), offer a non-invasive alternative, enabling broader assessments of taxa responses across trophic levels. Here, we apply an eDNA approach to assess community-wide responses to effluent discharge in the St. Lawrence River, one of North America's most diverse freshwater ecosystems. We sampled water and aquatic communities along the effluent plume of the Montréal WWTP, analyzing taxa-specific responses across trophic levels using high-throughput sequencing. We evaluated the influence of water physico-chemistry and per- and polyfluoroalkyl substances (PFAS) on aquatic beta diversity and network structure. To validate our eDNA results, we compared fish-specific detections with traditional fishing surveys. Our findings highlight how wastewater-derived contaminants influence biodiversity patterns and species interactions, with taxonomic responses varying across trophic levels. Network analyses revealed shifts in ecological stability, with changes in species connectivity and modularity influenced by effluent exposure. This study demonstrates the value of eDNA for characterizing biodiversity responses to anthropogenic stressors and provides insights into the broader implications of point-source pollution for freshwater ecosystem resilience.
PMID:40414060 | DOI:10.1016/j.scitotenv.2025.179690
Neurotoxicology. 2025 Jul;109:11-26. doi: 10.1016/j.neuro.2025.05.006. Epub 2025 May 19.
ABSTRACT
With the societal interest to decrease experimental animal testing for regulatory purposes, the need for reliable new approach methods (NAMs) has become evident. To ensure the continued safe use of chemicals, NAMs should perform ideally at a comparable or better level of sensitivity and specificity as the in vivo modalities that they aim to replace, especially if they are to be used for hazard assessment. The use of relevant reference substances, selected with transparent criteria, forms a cornerstone of developing and validating (in silico and in vitro) NAMs. Claims on sensitivity and specificity should be based on results generated with reference chemicals that were previously scrutinised by independent expert panels on whether the substance has a hazardous property. CLP (Regulation (EC) No 1272/2008 on the classification, labelling and packaging of substances and mixtures) forms a key pillar in EU chemicals management. The evaluation of all available information by the Committee for Risk Assessment (RAC) and their comparison to CLP classification criteria creates the opportunity to objectively compile lists of positive reference substances. We collated a reference list of neurotoxic substances to aid in the development of neurotoxicity NAMs. We screened CLP Annex VI and reflected on existing reference lists and mode of action data. The identified substances included industrial chemicals and active substances in plant protection products and biocidal products. This list of neurotoxicants is not an exhaustive consensus list, which ideally would be the result of combining this list with those generated by other authorities or expert groups.
PMID:40398565 | DOI:10.1016/j.neuro.2025.05.006
Deadline: 2025-06-03
Approximate launch date: 05/06/2025
Deadline to register interest: 03/06/2025
In 2023 EFSA launched a project to promote the use of New Approach Methods (NAMs) into the risk assessment of pesticide residues metabolites (NAMs4PestRes) through the use of the OECD QSAR toolbox, a free available in silico tool managed by ECHA, to enhance connectivity of IUCLID and MetaPath and to implement embedded in silico models. The result will be an automated and standardized workflow for the assessment of mutagenicity (bacterial gene mutation) of pesticide residue metabolites to be implemented in a...
The 2nd Commission Conference on the Roadmap towards Phasing out Animal Testing for Chemical Safety Assessments, held on 25 October 2024, brought together key stakeholders from regulatory bodies, industry, research organisations, and NGOs to discuss the future of chemical safety assessments done using non-animal methods (NAMs) in the EU. This second conference aimed to collect input from various stakeholders on how they address the transition from animal-based testing to innovative, reliable non-animal alternatives and to report progress with the development of the roadmap, which will set out actions and milestones to drive this transformation across chemical safety assessments.
Toxicology. 2025 Aug;515:154167. doi: 10.1016/j.tox.2025.154167. Epub 2025 Apr 27.
ABSTRACT
Chronic liver diseases, including metabolic dysfunction-associated steatosic liver disease (MASLD) and hepatocellular carcinoma (HCC), are on the rise, potentially due to daily exposure to complex mixtures of chemical compounds forming part of the exposome. Understanding the mechanisms involved in hepatotoxicity of these mixtures is essential to identify common molecular targets that may highlight potential interactions at the molecular level. We illustrated this issue with two families of environmental contaminants, per- and polyfluoroalkyl substances (PFAS) and heterocyclic aromatic amines (HAAs), both of which could be involved in the progression of chronic liver diseases, and whose toxicity may be potentiated by interactions at the level of xenobiotic metabolism. In the study of exposome effects on chronic liver disease, New Approach Methodologies (NAMs) including omics analyses and data from various in vitro, in vivo and in silico approaches, are crucial for improving predictivity of toxicological studies in humans while reducing animal experimentation. Additionally, the development of complex in vitro human liver cell models, such as organoids, is essential to avoid interspecies differences that minimize the risk for humans. All together, these approaches will contribute to construct Adverse Outcome Pathways (AOPs) and could be applied not only to PFAS mixtures but also to other chemical families, providing valuable insights into mixture hepatotoxicity prediction in the study of the exposome. A better understanding of toxicological mechanisms will clarify the role of environmental contaminant mixtures in the development of MASLD and HCC, supporting risk assessment for better treatment, monitoring and prevention strategies.
PMID:40300710 | DOI:10.1016/j.tox.2025.154167
This editorial provides an update on research & innovation (R&I) needs that can support EFSA's regulatory science in the coming years. The paper presents research needs for EFSA's work in a number of domains: omics technologies; gut microbiome; new approach methodologies (NAMs); allergenicity risk assessment; aggregate exposure assessment and environmental risk assessment (ERA). In briefly describing R&I needs, the document also addresses emerging challenges and opportunities.
Comput Toxicol. 2024 Jun;30:1-15. doi: 10.1016/j.comtox.2024.100304.
ABSTRACT
Read-across is a well-established data-gap filling technique used within analogue or category approaches. Acceptance remains an issue, mainly due to the difficulties of addressing residual uncertainties associated with a read-across prediction and because assessments are expert-driven. Frameworks to develop, assess and document read-across may help reduce variability in read-across results. Data-driven read-across approaches such as Generalised Read-Across (GenRA) include quantification of uncertainties and performance. GenRA also affords opportunities on how New Approach Method (NAM) data can be systematically incorporated to support the read-across hypothesis. Herein, a systematic investigation of differences in expert-driven read-across with data-driven approaches was pursued in terms of establishing scientific confidence in the use of read-across. A dataset of expert-driven read-across assessments that made use of registration data as disseminated in the public International Uniform Chemical Information Database (IUCLID) (version 6) of Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) Study Results were compiled. A dataset of ~5000 read-across cases pertaining to repeated dose and developmental toxicity was extracted and mapped to content within EPA's Distributed Structure Searchable Toxicity database (DSSTox) to retrieve chemical name and structural identification information. Content could be mapped to ~3600 cases which when filtered for unique cases with curated quantitative structure-activity relationship-ready SMILES resulted in 389 target-source analogue pairs. The similarity between target and the source analogues on the basis of different contexts - from structural similarity using chemical fingerprints to metabolic similarity using predicted metabolic information was evaluated. An attempt was also made to quantify the relative contribution each similarity context played relative to the target-source analogue pairs by deriving a model which predicted known analogue pairs. Finally, point of departure values (PODs) were predicted using the GenRA approach underpinned by data extracted from the EPA's Toxicity Values Database (ToxValDB). The GenRA predicted PODs were compared with those reported within the REACH dossiers themselves. This study offers generalisable insights on how read-across is already applied for regulatory submissions and expectations on the levels of similarity necessary to make decisions.
PMID:38993812 | PMC:PMC11235147 | DOI:10.1016/j.comtox.2024.100304
Front Toxicol. 2024 Apr 26;6:1359507. doi: 10.3389/ftox.2024.1359507. eCollection 2024.
ABSTRACT
In the European regulatory context, rodent in vivo studies are the predominant source of neurotoxicity information. Although they form a cornerstone of neurotoxicological assessments, they are costly and the topic of ethical debate. While the public expects chemicals and products to be safe for the developing and mature nervous systems, considerable numbers of chemicals in commerce have not, or only to a limited extent, been assessed for their potential to cause neurotoxicity. As such, there is a societal push toward the replacement of animal models with in vitro or alternative methods. New approach methods (NAMs) can contribute to the regulatory knowledge base, increase chemical safety, and modernize chemical hazard and risk assessment. Provided they reach an acceptable level of regulatory relevance and reliability, NAMs may be considered as replacements for specific in vivo studies. The European Partnership for the Assessment of Risks from Chemicals (PARC) addresses challenges to the development and implementation of NAMs in chemical risk assessment. In collaboration with regulatory agencies, Project 5.2.1e (Neurotoxicity) aims to develop and evaluate NAMs for developmental neurotoxicity (DNT) and adult neurotoxicity (ANT) and to understand the applicability domain of specific NAMs for the detection of endocrine disruption and epigenetic perturbation. To speed up assay time and reduce costs, we identify early indicators of later-onset effects. Ultimately, we will assemble second-generation developmental neurotoxicity and first-generation adult neurotoxicity test batteries, both of which aim to provide regulatory hazard and risk assessors and industry stakeholders with robust, speedy, lower-cost, and informative next-generation hazard and risk assessment tools.
PMID:38742231 | PMC:PMC11089904 | DOI:10.3389/ftox.2024.1359507
Sci Total Environ. 2024 Feb 20;912:168738. doi: 10.1016/j.scitotenv.2023.168738. Epub 2023 Nov 27.
ABSTRACT
Per- and polyfluoroalkyl substances (PFAS) are ubiquitously distributed in the aquatic environment. They include persistent, mobile, bioaccumulative, and toxic chemicals and it is therefore critical to increase our understanding on their adsorption, distribution, metabolism, excretion (ADME). The current study focused on uptake of seven emerging PFAS in zebrafish (Danio rerio) and their potential maternal transfer. In addition, we aimed at increasing our understanding on mixture effects on ADME by developing a physiologically based kinetic (PBK) model capable of handling co-exposure scenarios of any number of chemicals. All studied chemicals were taken up in the fish to varying degrees, whereas only perfluorononanoate (PFNA) and perfluorooctanoate (PFOA) were quantified in all analysed tissues. Perfluorooctane sulfonamide (FOSA) was measured at concerningly high concentrations in the brain (Cmax over 15 μg/g) but also in the liver and ovaries. All studied PFAS were maternally transferred to the eggs, with FOSA and 6:2 perfluorooctane sulfonate (6,2 FTSA) showing significant (p < 0.02) signs of elimination from the embryos during the first 6 days of development, while perfluorobutane sulfonate (PFBS), PFNA, and perfluorohexane sulfonate (PFHxS) were not eliminated in embryos during this time-frame. The mixture PBK model resulted in >85 % of predictions within a 10-fold error and 60 % of predictions within a 3-fold error. At studied levels of PFAS exposure, competitive binding was not a critical factor for PFAS kinetics. Gill surface pH influenced uptake for some carboxylates but not the sulfonates. The developed PBK model provides an important tool in understanding kinetics under complex mixture scenarios and this use of New Approach Methodologies (NAMs) is critical in future risk assessment of chemicals and early warning systems.
PMID:38030006 | DOI:10.1016/j.scitotenv.2023.168738
Arch Toxicol. 2023 Dec;97(12):3075-3083. doi: 10.1007/s00204-023-03601-5. Epub 2023 Sep 27.
ABSTRACT
In Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) the criterion for deciding the studies that must be performed is the annual tonnage of the chemical manufactured or imported into the EU. The annual tonnage may be considered as a surrogate for levels of human exposure but this does not take into account the physico-chemical properties and use patterns that determine exposure. Chemicals are classified using data from REACH under areas of health concern covering effects on the skin and eye; sensitisation; acute, repeated and prolonged systemic exposure; effects on genetic material; carcinogenicity; and reproduction and development. We analysed the mandated study lists under REACH for each annual tonnage band in terms of the information they provide on each of the areas of health concern. Using the European Chemicals Agency (ECHA) REACH Registration data base of over 20,000 registered substances, we found that only 19% of registered substances have datasets on all areas of health concern. Information limited to acute exposure, sensitisation and genotoxicity was found for 62%. The analysis highlighted the shortfall of information mandated for substances in the lower tonnage bands. Deploying New Approach Methodologies (NAMs) at this lower tonnage band to assess health concerns which are currently not covered by REACH, such as repeat and extended exposure and carcinogenicity, would provide additional information and would be a way for registrants and regulators to gain experience in the use of NAMs. There are currently projects in Europe aiming to develop NAM-based assessment frameworks and they could find their first use in assessing low tonnage chemicals once confidence has been gained by their evaluation with data rich chemicals.
PMID:37755502 | PMC:PMC10567824 | DOI:10.1007/s00204-023-03601-5
Regul Toxicol Pharmacol. 2022 Dec;136:105278. doi: 10.1016/j.yrtph.2022.105278. Epub 2022 Oct 21.
ABSTRACT
The Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) regulation was created to protect human health and the environment through the better and earlier identification of harmful intrinsic properties of chemical substances on the European market. One of its central aims was the promotion of alternatives to animal testing, yet it has instead become a long tick-box list of in vivo experiments questionable relevance to human health outcomes despite a global trend towards new approach methods (NAMs) in chemical safety assessment. The Chemicals Strategy for Sustainability (CSS), proposed by the European Commission in 2020, is a golden opportunity to revise REACH in a significant and impactful way, yet proposals presented so far have significant negative animal welfare consequences. There is still time to correct the course of the ongoing REACH revision - proposals made herein offer a path towards the promising future intended by the CSS. These proposals are anchored in three vectors of action, varying in level of complexity - from changes that ECHA can implement to improve existing processes, through technical changes aimed at minimizing animal testing and increasing NAM acceptance, to deeper structural changes to establish non-animal testing strategies as the basis for risk assessment.
PMID:36280152 | DOI:10.1016/j.yrtph.2022.105278
Regul Toxicol Pharmacol. 2022 Nov;135:105261. doi: 10.1016/j.yrtph.2022.105261. Epub 2022 Sep 11.
ABSTRACT
New Approach Methodologies (NAMs) are considered to include any in vitro, in silico or chemistry-based method, as well as the strategies to implement them, that may provide information that could inform chemical safety assessment. Current chemical legislation in the European Union is limited in its acceptance of the widespread use of NAMs. The European Partnership for Alternative Approaches to Animal Testing (EPAA) therefore convened a 'Deep Dive Workshop' to explore the use of NAMs in chemical safety assessment, the aim of which was to support regulatory decisions, whilst intending to protect human health. The workshop recognised that NAMs are currently used in many industrial sectors, with some considered as fit for regulatory purpose. Moreover, the workshop identified key discussion points that can be addressed to increase the use and regulatory acceptance of NAMs. These are based on the changes needed in frameworks for regulatory requirements and the essential needs in education, training and greater stakeholder engagement as well the gaps in the scientific basis of NAMs.
PMID:36103951 | DOI:10.1016/j.yrtph.2022.105261