Chemosphere

Alternaria mycotoxins represent an emerging concern due to their frequent occurrence in food and feed. However, available toxicological data remain limited. Under the current EU regulatory framework, and in line with the EFSA/ECHA/JRC guidance for the identification of endocrine disruptors (EDs), assessment of endocrine activity relies on standardized assays performed according to OECD Test Guidelines (TGs) for the estrogen-, androgen- and steroidogenesis- (EAS) modalities. Within the framework of the European Partnership for the Assessment of Risks from Chemicals (PARC), standardized in vitro methods of regulatory relevance were performed for six chemically characterized Alternaria toxins, aiming to address current regulatory gaps on EAS-mediated activity. Alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), altertoxin-I (ATX-I), tentoxin (TEN) and altenuene (ALT) were assessed over a broad concentration range, from 0.001 up to 60 M, depending on cytotoxicity and solubility profile of each compound. Our findings indicate estrogenic activity for AOH (PC50: 3.9 - 4.6 {micro}M) and AME (PC50: 5.2 - 8.5 {micro}M) in the estrogen receptor transactivation assay (OECD TG 455), as well as an anti-estrogenic activity for ATX-I (IC30: 0.27 - 0.37 {micro}M). Minimal positive responses were observed at high concentrations for AOH (from the dose of 3 {micro}M) and for AME (from the dose of 10 {micro}M) in the agonistic part of the androgen receptor transactivation assay (OECD TG 458), which may also reflect glucocorticoid receptor activation. No effects on estradiol or testosterone production were observed for any of the tested Alternaria compounds in the steroidogenesis assay (OECD TG 456).

Alternaria toxins are naturally occurring food contaminants with limited and often inconsistent genotoxicity and mutagenicity data. Within the European Partnership for the Assessment of Risks from Chemicals (PARC), an OECD-aligned in vitro testing strategy was applied to fill existing data gaps and to characterize the genotoxic potential of major Alternaria toxins using high-purity test materials. Mutagenicity was assessed using bacterial reverse mutation test (OECD TG 471) and SOS/umu assay, while chromosomal damage was assessed using the in vitro micronucleus (MN) assay (OECD TG 487) in TK6 and HepG2 cells, complemented by fluorescence in situ hybridization (FISH) and {gamma}H2AX assay in HepaRG cells. Alternariol (AOH), alternariol monomethyl ether (AME), and altertoxin-I (ATX-I) showed clear mutagenicity in bacteria, whereas altenuene (ALT), tenuazonic acid (TeA), and tentoxin (TEN) were negative under the tested conditions. In mammalian cells, AOH, AME, and ATX-I induced MN formation in TK6 cells at concentrations [≥]5.5 {micro}M, [≥]2.5 {micro}M, and [≥]0.21 {micro}M, respectively, with FISH analysis supporting a clastogenic mode of action. In HepG2 cells, all tested toxins induced chromosomal damage, with effect threshold ranging from [≥]6.25 {micro}M (AOH) to [≥]50 {micro}M (TeA). {gamma}H2AX induction confirmed DNA damage for AOH and ATX-I, and at higher concentrations for TeA (1000 {micro}M). Overall, the data indicate clear in vitro genotoxic potential for AOH, AME, and ATX-I and provide evidence of chromosomal damage for ALT, TEN, and TeA, thereby reducing critical data gaps for hazard assessment.

Depleted uranium (DU) is an environmental contaminant with a 30 g/L (ppb; parts per billion) EPA maximum contaminant level (MCL) for drinking water. The mining of uranium and use of DU in modern weapons underly human exposure that disproportionally impacts military and tribal communities in the United States. Uranium's radiotoxic characteristics are understood, but its chemical hazards much less so. In zebrafish (Danio rerio) and human cell cultures we test the hypothesis that exposure to DU negatively impacts cellular function and development through disruption of mitochondrial metabolism. Using a novel shrapnel model with TEM/SEM+EDS, we showed uranium microparticles caused proximity-dependent mitochondrial disruption. In waterborne exposure paradigms, larval movement was reduced and hatching delayed as a result of reduced movement and not enzyme deficiencies in response to 18 ppb DU, below the MCL. Increased DNA damage accumulation was detected in exposed larva and cells. DNA-damage quantitative PCR of DU-exposed larvae showed increased damage in the ahr1 locus (nuclear gene) and decreased mitochondrial DNA (mtDNA) copy number, but mtDNA damage levels varied across experiments. Mitochondrial function was assessed using a resazurin-based assay in the presence and absence of antioxidants and showed diminished cytoplasmic reductive capacity. DU exposure alone did not enrich antioxidant gene expression, contrasting with arsenic exposure, a known ROS-inducer and Nrf2-activator. Sulforaphane (SFN), a potent Nrf2-activator, did not blunt the effects of DU exposure, despite activation of antioxidant response element (ARE) genes (gstp and gss), but did blunt the effects of arsenic exposure. The most enriched transcript in DU-exposed larvae coded for slingshot protein phosphatase (ssh), further exploration revealed ssh1b as the zebrafish-specific ortholog activated in response to DU, and inhibition using an identified SSH1 inhibitor, Sennoside A, partially rescued the metabolic and hatching defects observed. Our data points to a cytotoxic mechanism in which DU disrupts mitochondrial function through ssh1b enrichment that impairs normal mitophagy, leading to decreased cellular reductive potential independent of either ROS production or ARE-activation. Our results suggest that health impacts from DU exposure may be directly linked to impaired mitochondrial functions.

High concentrations of steroidal hormone compounds are a growing source of concern for environmental pollution in aquatic ecosystems. In this study, we examine the effects of two estrogenic compounds (estriol and 17-ethinylestradiol) on fitness traits in the aquatic microcrustacean, Daphnia magna, a key bioindicator species for toxicology studies. The impacts were compared of two forms representing a natural and synthetic estrogenic compound. Growth and reproduction traits were assayed by exposing Daphnia to each estrogen type at four concentrations reflecting potential environmental exposure conditions up to acute toxicity levels (ranging from 0.1 - 50 {micro}g/L). Assaying the effects at a variety of concentrations is important given that it is known that hormone exposures can often result in non-monotonic responses. Both forms of estrogen impact a subset of the traits assessed, in some cases leading to beneficial changes and others causing harm. Estriol, the naturally-occurring estrogen, and EE2, the synthetic version, at high doses shift fitness traits in opposite directions such as adult growth rate as do at low doses for fecundity. In conclusion, our results support the need to assay a wide array of traits using multiple forms of steroidal hormones at a range of doses in order to assess non-monotonic patterns and their impact on an organismal fitness. In particular, assays that extend beyond the conventional measurements of lethality during acute exposure windows will be essential for understanding the impact of increased levels of hormone pollution on aquatic organisms and ecosystem health.

Background The placenta serves a critical role in nutrient uptake and waste elimination for the developing fetus. The placenta is also responsible for the uptake and/or exchange of xenobiotics, including medications, between the maternal and fetal bloodstreams. An estimated 40-80% of women take medications or drugs during pregnancy for a variety of conditions. Very little is understood about fetal drug and nutrient exposure during pregnancy and how it may change over the course of fetal development. Objective This study aimed to characterize the abundance of transport proteins in placental tissue, which are important in modulating fetal nutrient and drug exposure, over the duration of pregnancy. Mass spectrometry-based global proteomic analysis revealed trends in the expression of thousands of proteins throughout gestation. Focusing on the membrane-associated proteome enabled an increased emphasis on the solute carrier and ATP-binding cassette families of transporter proteins that are critical for nutrient and xenobiotic transport across the maternal-fetal barrier. Study Design Using data-independent acquisition proteomics, relative abundance of proteins in placental tissue samples was profiled across all three trimesters of pregnancy (Trimester 1 = 16, Trimester 2 = 9, and Term = 9). Membrane fractions were generated to enrich membrane-associated proteins for proteomic analysis. Placental samples were grouped into randomized batches for membrane fraction generation and mass spectrometry analysis. Proteomic search results from each batch were imported into the R programming environment from Skyline, concatenated, and normalized as one data set for downstream analysis. Results A total of 6,331 proteins were detected across all samples with 4,210 proteins identified in every sample. Pathway analysis revealed that as gestational age increases, membrane-associated proteins involved in more complex metabolic pathways increase in relative abundance while those involved in extracellular remodeling events and simple organic ion transport tended to decrease. A total of 139 solute carrier and ATP-binding cassette transport proteins were identified in all samples, and 80 were identified in every sample. In general, membrane-associated proteins, including solute carrier and ATP-binding cassette transport proteins, were significantly enriched in placental tissue collected during early gestation compared to term placental tissue. Conclusion This study presents a comprehensive profiling of membrane-associated proteomic changes during gestation and identifies significant gestational age associated abundance changes at the protein level in several transport protein families. The application of data-independent acquisition global proteomic techniques enabled in-depth analysis of thousands of proteomic changes across pregnancy in a single experiment. These data provide critical information to support future studies into the understanding of fetal exposure to xenobiotics and nutrients circulating in the maternal bloodstream.

Nanoplastics (NPs) are emerging environmental contaminants able to cross biological barriers, disrupt cellular and organelle homeostasis, and alter the brain microenvironment. This study investigated whether NPs affect microglia to neuron communication, a key mechanism underlying neuronal resilience, via the nuclear factor erythroid 2 like 2 (NFE2L2) pathway. Using an in vitro model, we evaluated the effects of polystyrene nanoplastics on microglial metabolic fitness and microglia-mediated neuronal stress responses. Increasing NP concentrations induced a dose dependent biphasic effect. Low to intermediate concentrations increased intracellular adenosine triphosphate (ATP) levels in microglia and enhanced microglia-mediated activation of neuronal NFE2L2. In contrast, high NP concentration impaired microglial metabolism, reduced ATP availability, and decreased microglia to neuron communication. These findings indicate that NPs alter microglial energetic status and modulate neuroprotective signalling, potentially contributing to impaired neuron to microglia interactions and increased susceptibility to neurotoxicity.

Background: Lead exposure is estimated to cause approximately 3.5 million premature deaths a year, yet the key ongoing sources of lead exposure are unclear. Methods: We estimated the contribution of dietary lead intake to global blood lead levels (BLLs) for 7-year-old children and 22-year-old adults by applying the All-Ages Lead Model (AALM) to calculate blood lead levels (BLLs) based on 25 total diet studies (TDS) that quantify dietary lead intake across 46 countries. Results: For children, the population-weighted average dietary lead intake in low- and middle-income countries (LMICs) (32.0 g/day) was found to be more than three times higher than in high-income countries (HICs) (9.3 g/day), and more than 10 times higher than the FDA reference level for children (2.2 g/day). The average impact on BLLs for children is estimated to be near 29 g/L in LMICs and near 12 g/L in HICs. Averaged across the TDS data, vegetables (27%) and cereals (24%) were found to contribute the most to dietary lead. Conclusions: While there are limitations associated with biokinetic modelling and the TDS data from LMICs, these results suggest that the contribution of dietary lead intake to global lead exposure is in the region of 40 to 50%, suggesting, in turn, that dietary lead intake is likely a major global driver of lead poisoning. Lead absorbed from the environment into food crops is expected to be the key driver of dietary lead. Current regulatory levels for maximum lead concentrations in foods (0.05-0.3 mg/kg) are out-of-date and may imply a dietary lead intake of 200 g/day, far higher than the FDA reference level (2.2 g/day). Collecting representative TDS data in high lead burden countries should be a priority. Further research is also recommended on upstream lead sources and pathways of lead uptake in plants, driving global food contamination.

Evidence of negative health and environmental effects of glyphosate-based herbicides (GBHs) has led to marketing of glyphosate-free formulations. A frequent glyphosate replacement is pelargonic acid, which is rapidly degraded, leading to claims of greater safety and less environmentally damaging than GBHs. However, toxicity of commercial pelargonic acid formulations containing several co-formulants have not been determined. Using Roundup NL, a representative pelargonic acid-based herbicide, we undertook tissue culture cell assays measuring viability, plasma membrane integrity, DNA damage, and activation of stress-response pathways. In human hepatoma HepG2 cells, Roundup NL was more cytotoxic than pelargonic acid, and more toxic than the GBH Roundup ProBio and glyphosate as shown by reduced viability underpinned by plasma membrane damage. Pelargonic acid and Roundup NL did not induce oxidative stress. However, comet assays revealed that pelargonic acid but not Roundup NL caused a modest but significant increase in DNA damage at sub-cytotoxic concentrations. The murine embryonic stem cell-based ToxTracker system confirmed Roundup NL as not directly genotoxic but triggered oxidative stress and protein damage (ER stress, impaired proteostasis) indicating cell and assay dependency of oxidative stress pathway activation. Our results suggest that exposure to pelargonic acid-based herbicides constitutes a health hazard and that co-formulants present in Roundup NL contribute substantially to its overall toxicity.

Tenuazonic acid (TeA) is an emerging Alternaria mycotoxin frequently detected in food and feed commodities, raising concerns about its toxicological relevance. Chronic oral exposure to TeA has been reported to induce dysplastic alterations in the esophageal mucosa of mice, while human biomonitoring data indicate an association between TeA exposure and esophageal cancer, although a causal relationship has not yet been established. At a mechanistic level, the effects of TeA in esophageal cells remain poorly characterized. Therefore, this study investigated the impact of TeA on cytotoxicity, oxidative stress, DNA damage, mitochondrial homeostasis, cell-cycle distribution and transcriptomic stress responses in human esophageal KYSE-510 cells. TeA induced a concentration-dependent reduction in metabolic activity and total protein content after 24 h exposure to 0.1-100 M. Significant cytotoxicity was measured starting from 20 M. At sub-cytotoxic concentrations, TeA triggered rapid ROS formation within 5-30 min exposure and induced formamidopyrimidine-DNA glycosylase (FPG) sensitive DNA damage after 1 h exposure (5-7.5 M), indicating oxidative DNA lesions. In addition, TeA altered mitochondrial morphology after 4 h exposure at 7.5 M, manifested by shrinkage of the mitochondrial network area and perinuclear redistribution, while mitochondrial respiration showed only a non-significant tendency towards reduced respiratory capacity. RNA sequencing after 6 h exposure to 10 M TeA revealed oxidative stress-associated transcriptional changes, impaired antioxidant and stress-adaptive responses, and p53-associated stress signaling. Furthermore, TeA induced significant G2/M phase accumulation after 24 h exposure to 1-10 M.

Background: Current chemical mixture approaches are largely data-driven without considering shared biological mechanisms among mixture components, highlighting the need for biology-informed approaches. Objectives: We constructed an integrated measure of a chemical mixture's oxidative stress potential and assessed its association with mortality in the US population. Methods: The sample comprised 4,574 adults ([≥] 20 years) from National Health and Nutrition Examination Survey (NHANES) 2005-2010. To obtain robust estimates, we performed 1,000 repeated random 50:50 splits into training and testing sets. In each training set, we used survey-weighted quantile g-computation to model serum gamma-glutamyl transferase (GGT), an oxidative stress biomarker, as a function of a 30-chemical mixture (blood metals, urinary polycyclic aromatic hydrocarbons (PAHs), pesticides, phenols/parabens, and phthalates), adjusting for sociodemographic, behavioral, and dietary factors. We then applied the fitted model from each training set to the corresponding testing set to derive the environmental risk score for oxidative stress (ERSOS), defined by predicted GGT values. Associations of ERSOS with all-cause, cardiovascular, and cancer mortality over 11 years of follow-up were estimated in the testing sets using survey-weighted Cox proportional hazards models and summarized across the 1,000 repeated splits. Results: Chemicals with the largest positive weights in quantile g-computation included mono-(2-ethyl-5-hydroxyhexyl) phthalate, mono-2-ethyl-5-carboxypentyl phthalate, 2-hydroxyfluorene, methyl paraben, and benzophenone-3; chemicals with the largest negative weights included mono-(2-ethyl-5-oxohexyl) phthalate and PAH metabolites (1-hydroxynaphthalene, 3-hydroxyphenanthrene, and 3-hydroxyfluorene). The median correlation between observed and predicted GGT in the testing sets was 0.43 (2.5th, 97.5th percentiles: 0.40-0.48). A one standard deviation increase in ERSOS was associated with a median hazard ratio of 1.60 (2.5th, 97.5th percentiles: 1.01-2.57) for cardiovascular mortality. No associations were found for all-cause mortality or cancer mortality. Discussion: The proposed survey-weighted quantile g-computation approach may help estimate biology-informed chemical mixture effects in complex survey data, supporting the potential utility for population-generalizable environmental mixture research.

Human hair is a common contaminant in GMP-controlled manufacturing environments, and its identification is important for contamination source investigation and corrective action. Because human hair can originate from multiple body sites, it is often necessary to determine not only the species of origin but also the anatomical source of the hair. Conventional forensic approaches distinguish scalp hair from body hair by microscopic examination of cuticle patterns, medullary structure, cross-sectional morphology, and pigment distribution. However, these methods depend on examiner expertise, are difficult to apply to damaged specimens, and provide limited quantitative information. In this study, we developed a proteomics-based approach for distinguishing scalp hair from pubic hair using identical sample preparation and analytical workflows. Comparative proteomic analysis identified keratin-associated proteins KAP 4-3 and KAP 9-6 as enriched in scalp hair, whereas cuticular keratins Ha7 and Ha8 were strongly enriched in pubic hair. Amino acid composition analysis further revealed that scalp hair-enriched proteins were highly cysteine-rich, consistent with sulfur-rich cross-linking matrix proteins, whereas pubic hair-enriched proteins exhibited characteristics of structural keratin filaments. These results demonstrate that proteomic signatures can provide a quantitative and objective means of determining the anatomical origin of human hair and may contribute to contamination source tracing in GMP manufacturing and forensic investigations.

Tenuazonic acid (TeA) is one of the most frequently detected Alternaria mycotoxins in contaminated food. Despite its frequent occurrence, its immunomodulatory effects remain insufficiently characterized. Therefore, the present study investigated the impact of TeA on inflammatory signaling and cytokine regulation in monocytes and intestinal epithelial cell (IEC) models. NF-{kappa}B activity was assessed using a reporter gene assay in THP1-Lucia monocytes, while cytokine mRNA expression and protein secretion were quantified in Caco-2 and HCEC-1CT cells by qRT-PCR and ELISA, respectively. In THP-1 monocytes, TeA significantly suppressed lipopolysaccharide (LPS)-induced NF-{kappa}B activation in a concentration-dependent manner starting at 25 M, while cytotoxicity occurred only at concentrations [≥]100 M. In HCEC-1CT and differentiated Caco-2 cells, TeA increased IL-6, IL-8, and TNF- mRNA levels at non-cytotoxic concentrations ([≥]10 M). In Caco-2 cells, these transcriptional changes were accompanied by increased cytokine secretion, whereas HCEC-1CT cells showed only partial effects on the protein level after short-term exposure. Following prolonged incubation, TNF- secretion was increased and IL-6 and IL-8 secretion were slightly reduced. IL-10 remained unaffected under all conditions. Overall, TeA exerted cell type-dependent immunomodulatory effects characterized by immunoinhibitory activity in monocytes and pro-inflammatory responses in IECs, highlighting the complex immunotoxic potential of this Alternaria mycotoxin.

Background: Men with prostate cancer (PCa) may be especially vulnerable to per- and polyfluoroalkyl substances (PFAS) exposure due to their endocrine-disrupting and cardiometabolic impacts and cardiotoxicity and immune suppression of treatments. Objective: A pilot study was launched to measure serum and tap water PFAS concentrations in PCa survivors. Methods: Men with PCa were recruited from Rutgers Cancer Institute between February 2025 and March 2026, with ongoing enrollment and follow-up. Eligible men were aged [≥]40 years and either on active surveillance or within 3-12 months of initial definitive treatment. Participants provided blood and residential tap water samples, which were analyzed using mass spectrometry (serum) and modified EPA method 537 (water). Geometric means were used to summarize PFAS concentrations by race and assess serum-tap water correlations. Results: Of 235 eligible patients, 124 (60%) enrolled. Median age was 64 years; 63% were non-Hispanic White, 43% had a Gleason score [≤]6. Roughly half of participants provided serum and/or tap water samples. In serum, six PFAS analytes had >80% detection; of these analytes, median concentrations ranged from 0.13 ng/mL (IQR: 0.07-0.20) for PFHpS to 2.55 ng/mL (IQR:1.54-3.82) for nPFOS. Among 74 tap water samples, 9 PFAS analytes had >60% detection; of these, median concentrations of PFNA (0.56 ng/L; IQR: 0.33-0.75), PFOA (3.75 ng/L; IQR: 1.21-5.27), and PFOS (2.29 ng/L; IQR: 0.46-2.89), were below New Jersey Maximum Contaminant Levels. Non-White participants had significantly higher levels of multiple PFAS analytes in both serum and tap water. Serum-tap water correlations were modest (r=0.22-0.41). Significance: The pilot study has demonstrated both the feasibility and importance of studying PFAS exposure pathways as well as potential impacts of PFAS exposure in diverse populations. Keywords: Prostatic Neoplasms, Per- and Polyfluoroalkyl Substances (PFAS), Biomonitoring, Environmental Exposure, Cohort Studies, Pilot study Impact Statement: This study provides some of the first estimates of PFAS exposure among prostate cancer patients in serum and tap water, showing moderate correlations between tap water and serum concentrations of specific PFAS analytes. These findings can support larger studies to identify environmental exposure sources and evaluate the role of PFAS in prostate cancer progression and outcomes.

Bisphenol S (BPS), a widely used substitute for bisphenol A, is increasingly detected in aquatic environments; however, its neurodevelopmental effects remain insufficiently understood. This study investigated whether developmental exposure to an environmentally relevant concentration of BPS disrupts social behaviour and underlying neurobiological pathways in zebrafish (Danio rerio). At 21 days post-fertilization, BPS-exposed larvae exhibited a significant reduction in social preference, indicating impaired conspecific interactions. Neurochemical analysis revealed a marked increase in serotonin (5-HT) levels, whereas lipid peroxidation (MDA) remained unchanged, suggesting the absence of overt oxidative damage. Gene expression profiling demonstrated a dysregulated antioxidant response, suppression of apoptotic signaling, and pronounced upregulation of serotonergic receptors and transporters. To resolve system-level mechanisms, protein-protein interaction (PPI) network analysis identified BDNF and CREB1 as dominant regulatory hubs, with the serotonergic synapse pathway as the most significantly enriched term. Molecular docking further demonstrated direct binding of BPS to multiple serotonergic targets, including HTR1A and TPH2, supporting receptor-level interference. Expanded network and pathway analyses revealed coordinated enrichment of monoamine GPCR, oxidative stress, and inflammatory pathways. These findings demonstrate that BPS induces serotonergic dysregulation and network-level reprogramming rather than significant oxidative damage, leading to behavioural impairment. This study provides a multi-scale mechanistic framework linking molecular perturbations to neurobehavioural outcomes, identifying serotonergic signaling and BDNF-CREB1 pathways as central targets of BPS neurotoxicity.

Copper (Cu) is an essential metal involved in neurobiological processes including energy metabolism and neurotransmission, yet dysregulated Cu levels may adversely affect brain health and olfactory performance. Although olfactory dysfunction has primarily been studied in older adults and neurodegenerative disease, adolescence is a critical period of brain maturation during which the olfactory system may be particularly vulnerable. This cross-sectional study examined associations between Cu exposure, olfactory bulb (OB) volume, and olfactory performance in 200 adolescents and young adults (64% female; ages 13 - 25) from the Public Health Impact of Metals Exposure cohort. Cu concentrations in blood, urine, hair, and saliva were measured using inductively coupled plasma mass spectrometry. T2-weighted magnetic resonance imaging scans estimated left, right, and total OB volumes using a three-stage deep learning pipeline. Olfactory performance was assessed using the Sniffin Sticks test. Weighted quantile sum regression evaluated associations between a Cu mixture index and OB outcomes, while standard linear regression models assessed individual Cu biomarkers. Models were adjusted for age and sex. A higher Cu index was associated with reduced left (Beta= -0.72, 95% CI [-1.42, -0.02]), right (Beta = -0.79, 95% CI [-1.43, -0.15]), and total OB volume (Beta= -1.55, 95% CI [-2.85, -0.25]), as well as lower odor threshold scores (Beta = -0.23, 95% CI [-0.42, -0.03]). Individual biomarkers were not independently associated with outcomes. These findings suggest that Cu exposure may adversely affect olfactory neurodevelopment during adolescence and highlight the importance of studying environmental exposures relevant to long-term neurological health.

Interest in microbiota-host co-metabolism and the effects of its derived co-metabolites on biological processes is increasing rapidly. In addition to their demonstrated associations with mammalian metabolic health and cognition, microbiota-host co-metabolites (MHCMs) represent lifelong contributors to the endogenous exposome. We have previously shown the MHCM trimethylamine N-oxide (TMAO) to exert beneficial effects on murine blood-brain barrier integrity and cognition. Here we investigated whether these positive neural effects of TMAO extended to humans, analysing how TMAO exposure associates with neurodevelopmental outcomes in children and whether an in vitro human neuronal-astrocyte co-culture could contribute to further investigation of the underlying mechanism(s) and neuronal processes related to these associations. In a cohort study of childhood mental health (N=1,203), TMAO was associated with fewer internalising problems, while its precursor microbial metabolite trimethylamine was associated with more behavioural problems in both the cross-sectional and an independent longitudinal study from 1 to 15 years of age (N=630-820). Given prior associations between TMAO exposure and exposure to the environmental pollutants mercury and arsenic, we investigated how the effects of TMAO interacted with these known neurotoxicants. TMAO had a protective effect, modifying the relationship between arsenic exposure and poorer neurodevelopmental outcomes. Furthermore, TMAO activated synaptogenesis-related gene expression and was functionally protective against the negative effects of mercury in our in vitro model. Together, our findings emphasise the importance of interdisciplinary approaches to evaluate associations and potential pathways of MHCMs (endogenous) and environmental (exogenous) metabolites on neurodevelopment in exposome studies.

Per- and polyfluoroalkyl substances (PFAS) are persistent toxicants with immunological, metabolic and epithelial effects, but their relevance to inflammatory skin disease remains unclear. We developed a computational toxicology framework to test whether perfluoroalkyl sulfonate programs, especially perfluorooctanesulfonic acid (PFOS) and perfluorohexanesulfonic acid (PFHxS), converge with psoriasis-associated keratinocyte inflammation. Exposure transcriptomes were derived from GSE236956, in which human embryonic stem cell-derived epithelial-lineage models were exposed to 10 M PFAS for 8-16 days. Six PFAS were prioritized using descriptors, Tanimoto similarity, toxicology evidence, adverse outcome pathway (AOP)-like key events, exposure differentially expressed gene burden and read-across support. PFAS signatures were integrated with psoriasis bulk transcriptomes, single-cell RNA sequencing, keratinocyte-state mapping, regulator and communication inference, spatial transcriptomics and token-aware Geneformer-compatible virtual perturbation. PFOS ranked highest in integrated prioritization, followed by PFHxS and perfluorooctanoic acid. PFHxS produced a smaller but directionally informative signature within a PFOS-dominant perfluoroalkyl sulfonate footprint. The shared PFOS and PFHxS program converged with psoriasis through inflammatory keratinocyte, epidermal-stress, cytoskeletal and lipid-related modules. Single-cell and spatial analyses localized the program to activated keratinocytes and inflammatory epidermal niches, with strong spatial co-localization with inflammatory keratinocyte and epidermal stress scores. Virtual perturbation prioritized S100A9, S100A8, KRT16, IL36G, CCL20, CXCL8, FABP5, KRT17, FOS, JUN and NFKBIZ as candidate effectors. These findings support an exposure-informed, experimentally testable hypothesis linking persistent perfluoroalkyl sulfonate programs to keratinocyte inflammatory niches in psoriasis.

Metabolites from Plantago lanceolata (plantain) biomass have been linked with biological nitrification inhibition (BNI) in soil. After grazing, leaf metabolite chemistry is altered via digestion, and a suite of secondary metabolites are then delivered onto soil via dung and urine. The purpose of this study was to establish if urine from sheep grazed on plantain had BNI activity when added to pasture soil, and to identify the metabolite profile(s) that most likely contribute to the BNI effects observed. Groups of sheep (n=5) were grazed on one of nine different plantain cultivars in autumn and spring with analysis of leaf material, urine, soil incubation and BNI bioassay data used to identify potential metabolite candidates implicated with BNI. The urinary nitrogen and metabolite composition of sheep fed plantain varied significantly between cultivars and season. After 28 days of incubation, all soil microcosms treated with plantain-derived urine had up to 35% less nitrate than comparative ryegrass urine controls in both seasons, except one in autumn. The key phytochemistry associated with lower soil nitrate concentrations was phenylethanoid and iridoid glycosides resulting in a higher output of glucuronidated, methylated and sulfated secondary metabolites in the urine. Among 19 secondary metabolites identified in the urine, hydroxytyrosol-related metabolites as well as catechol glucuronide, 2-methoxyphenyl sulfate and guaiacol-{beta}-D-glucuronide appear to be the most likely target compounds with respect to the BNI effects observed. Variation in metabolites from different plantain cultivars affected the ratio of metabolite derivatives in urine, which ultimately affected soil nitrification rates. Cultivar phytochemistry is therefore an important consideration with respect to BNI under urine patches. HighlightsO_LISheep grazing different plantain cultivars had different urine compositions C_LIO_LIUrines elicited biological nitrification inhibition (BNI) in soil and in vitro C_LIO_LIDifferent BNI response was related to differential expression of urine metabolites C_LIO_LIKey urine metabolites associated with BNI are derived from glycosidic compounds C_LI

While exposure to volatile organic compounds such as ethylene dichloride and vinyl chloride monomer is a well-established cause of liver disease, particularly hepatic hemangiosarcoma, characterizing real-world exposure profiles in communities surrounding industrial centers remains challenging. Calvert City, Kentucky (population ~2,500), provides a unique setting characterized by both active industrial emissions and legacy sources of air toxics. To address these complexities, this method paper describes the framework for the Biomonitoring and Environmental Assessment for Community Outreach and Neighborhood Safety (BEACON) study. By utilizing a novel, multi-dimensional exposure assessment strategy, BEACON aims to characterize air toxic exposures and provide actionable data for community health and safety. For the BEACON study, we will leverage Kentucky Department of Air Quality measures of air toxics, analyze urine samples in a small cohort of community volunteers, analyze community urine via wastewater in an adjacent community, geocode citizen odor reporting, assess blood markers in wildlife, survey small and large animal veterinarians in the area for anomalies in morbidity and mortality, and work with the regional health system to enhance vigilance for health issues associated with toxicants present in the area. In addition, blood samples will be collected at three time points and biobanked for future analyses. Efforts will be made to link this study to additional large-scale long-term cohorts where possible. Throughout the project, community engagement will play a critical role by raising awareness, fostering collaboration, and ensuring that the voices of affected residents are heard.

Abstract Background. Chronic low-level exposure to lead, cadmium, mercury, and arsenic remains a determinant of premature mortality in the U.S. general population, but previous hazard-ratio analyses do not characterize how exposure shifts the lower tail of the survival distribution, where premature mortality is concentrated. Objectives. We estimated the association of whole-blood lead, whole-blood total mercury, urinary cadmium, and the sum of urinary inorganic and methylated arsenic species with the 10th, 25th, and 50th conditional quantiles of follow-up time to all-cause mortality among U.S. adults aged 40 years and older. Methods. NHANES Continuous 1999 to 2018 was linked to the National Death Index through December 31, 2019 (n = 29,652). Censored quantile regression was fit per metal on the log2 scale at quantiles {tau}{0.10, 0.25, 0.50}. A restricted-cubic-spline (RCS) censored-quantile-regression was fit for blood lead and urinary cadmium to investigate the threshold effect. Results. Over a median follow-up of 9.1 years, 7,215 deaths were ascertained. A doubling of urinary cadmium was associated with -1.57 years of follow-up (95% CI: -2.08, -1.07) at the 10th conditional quantile, -1.50 (-2.04, -0.96) at the 25th, and -1.49 (-1.93, -1.04) at the median (Benjamini Hochberg q < 0.001 throughout). A doubling of whole-blood lead was associated with -0.70 years (95% CI: -0.99, -0.40) at the 10th conditional quantile, -0.62 (-0.92,-0.31) at the 25th, and -0.61 years (-0.89, -0.34) at the median; the absolute loss was largest at {tau} = 0.10 for both metals. Urinary arsenic-metabolite sum was not associated with conditional follow-up at the estimable quantiles. Despite adjustment for dark and fatty-fish intake or DHA/EPA, whole-blood total mercury was associated with longer follow-up (i.e., negatively associated with mortality risk), possibly due to residual confounding by broader dietary or socioeconomic factors, rather than a true protective effect. The cadmium association was additionally robust to the mutual adjustment of lead. Discussion. Low-to-moderate urinary cadmium and whole-blood lead were associated with fewer years of follow-up survival at the lower-tail and median conditional quantiles of survival, with the largest absolute losses at the lower tail of the conditional survival distribution, where premature mortality is concentrated. These findings support continued reductions in U.S. cadmium exposure and lead with particular benefit for adults most vulnerable to premature death.