Chemosphere

The Fusarium mycotoxin deoxynivalenol (DON) is one of the most frequently occurring food contaminants. Nearly all individuals are exposed to DON, due to it widespread presence in grains and grain-based products. Chronic DON poisoning is associated with growth retardation, immunotoxicity as well as impaired reproduction and fetal development. At the molecular level, DON alters intracellular signaling by activating mitogen-activated protein kinases (MAPKs) that modulate cell growth, differentiation, and apoptosis. Of note, these MAPKs are also critical mediators of gonadotrophin-releasing hormone (GnRH)-induced synthesis and secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by pituitary gonadotrope cells. So far, no research has explored the potential endocrine-disrupting effects of DON on pituitary gonadotropins production. Herein, we show the first evidence that DON affects LH production by the immortalized gonadotrope-like cell line L{beta}T2 in a concentration-dependent manner. Taken together, our experiments demonstrated that low concentrations of DON affect GnRH-induced signaling through a mechanism that, at least in part, involves apoptosis and inhibition of GnRH-induced phosphorylation of ERK-MAPK. Consequently, DON also affects the GnRH-induced expression of Cga and Lhb, two critical genes for LH synthesis and secretion by gonadotrope cells in mammals. This research broadens our knowledge of the toxicity of DON and brings a new depth to the potential neuroendocrine implications for reproduction. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/607800v2_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@78d077org.highwire.dtl.DTLVardef@1be5bdaorg.highwire.dtl.DTLVardef@804d49org.highwire.dtl.DTLVardef@1520136_HPS_FORMAT_FIGEXP M_FIG C_FIG

Per- and polyfluoroalkyl substances (PFAS) are manmade chemicals that are persistent in the environment and have been linked to various physiological and neurobehavioral outcomes, including anxiety disorders. Trifluoroacetic acid (TFA), a short chain PFAS and the most common PFAS degradation product, is increasingly detected in water, soil, and human blood, raising significant concerns about its developmental toxicity. However, the impact of early-life TFA exposure on neurodevelopment and behavior remain insufficiently characterized. In this study, we employed Zebrafish (Danio rerio) embryos as a New Approach Methodology (NAM), to evaluate the development, behavior, and protein expression changes in response to early-life TFA exposure. Embryos were exposed to environmentally relevant low and high concentrations of TFA beginning at one-cell stage. Early developmental physiology was assessed by measuring viability, tail twitch response, hatching rates, and chorion diameters during embryogenesis. Anxiety-like behaviors were evaluated at 5- and 6-days post-fertilization using validated behavioral assays such as the Light-Dark Test and Startle Response. Each test evaluates distinct anxiety-related behaviors by measuring locomotor activity, thigmotaxis (wall preference), and stimulus reactivity, with anxious zebrafish larvae showing increased movement in light and greater wall preference. Then to identify molecular pathways underlying observed developmental phenotypes with TFA exposure, proteomic analyses were performed on embryos at 24- and 48-hours post-fertilization. Our results indicate that TFA exposure altered developmental physiology, evidenced by reduced chorion diameters, and lead to increased anxiety-like behaviors with larvae exhibiting thigmotaxis. These phenotypic changes were accompanied by detectable alterations in the embryonic proteome. Collectively, our findings provide insight into how short-chain PFAS exposure during critical windows of development may contribute to neurobehavioral dysfunction, highlighting potential risks relevant to inform public health policies and environmental regulations.

Rising concerns about Bisphenol A (BPA) toxicity have prompted the search for safer alternatives. However, concerns persist regarding the safety of replacements like bisphenol TMC (BPTMC), a rapidly emerging BPA substitute. Utilizing the in vivo model organism Caenorhabditis elegans (C. elegans), whose shared genes mirror human biology, we aim to unveil the potential toxicity of BPTMC on a live animal. C. elegans exposed to 1 mM BPTMC exhibited developmental delays, reduced reproduction, and diminished longevity. Furthermore, an investigation into mortality at various animal stages, oxidative stress, and thermal stress revealed additional compromised toxicity. Notably, exposure to BPTMC resulted in mitochondrial abnormalities, including reduced oxygen consumption, lowered mitochondrial membrane potential, and decreased ATP levels. Additionally, BPTMC increased ROS levels but decreased mitochondrial population. Transcriptome analysis revealed that BPTMC induces alterations in the expression of genes associated with mitochondrial biogenesis. Our findings raise crucial concerns about BPTMC as a safe BPA alternative. The observed widespread toxicity across key life stages suggests a need for further investigation into the potential toxicity of BPTMC on human health and environmental consequences.

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. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=61 SRC="FIGDIR/small/700343v1_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@1fec69aorg.highwire.dtl.DTLVardef@1a0b3beorg.highwire.dtl.DTLVardef@17883e2org.highwire.dtl.DTLVardef@11a7571_HPS_FORMAT_FIGEXP M_FIG C_FIG

There is increasing evidence that endocrine disrupting chemicals (EDCs) are contributing to the rise in metabolic disorders and obesity. Humans are constantly exposed to numerous EDCs, thus human exposure entails complex EDC mixtures. In this study, we examined the effects of an EDC mixture, mixture G, composed of four phthalate esters, triclosan, and three poly- och perfluorinated alkyl substances. Mixture G had previously been defined based on its association with lower birth weight in a pregnancy cohort, where low birth weight is an early risk factor for metabolic morbidities later in life. Here, we studied its effects on adipogenesis and uncovered their underlying transcriptional changes. Human mesenchymal stem cells (hMSCs) were exposed to mixture G in concentrations and mixing ratios that reflect those measured in human serum. Mixture G induced adipogenesis in hMSCs, as evidenced by a dose-dependent increase in lipid droplet accumulation after 14-21 days. Notably, significant adipogenic effects were observed at concentrations comparable to those detected in humans. RNA-sequencing upon exposure for 48 h revealed dose-dependent transcriptional changes in over 1000 genes. Mixture G-induced differentially expressed genes (DEGs) showed significant overlap with genes involved in osteogenesis, with glucocorticoid-regulated genes, and with genes associated with birth weight alterations and diabetes type II. These results indicate that exposure to an environmentally relevant EDC mixture induces adipogenesis and leads to transcriptional alterations that might change the balance between adipogenic and osteogenic differentiation as well as the functionality of MSCs, possibly via interference with glucocorticoid signalling. Thus our findings underscore the role of EDCs as metabolic disruptors and shed light on the molecular mechanisms underlying their potential contribution to the development of metabolic disorders. HighlightsO_LIAn endocrine disruptor mixture linked to lower birth weight increases adipogenesis C_LIO_LIThe mixture induced transcriptomic changes at low doses C_LIO_LIAffected genes are associated with birth weight and diabetes type II C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/638050v1_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@12428bforg.highwire.dtl.DTLVardef@ef28b3org.highwire.dtl.DTLVardef@159024corg.highwire.dtl.DTLVardef@a605b7_HPS_FORMAT_FIGEXP M_FIG C_FIG

Bats are potentially exposed to pesticides via foraging in croplands. Common pesticides like organophosphates are neurotoxic for vertebrates and even low doses can impair essential processes such as locomotion and cognition. These sublethal effects are usually studied using molecular biomarkers with limited ecological relevance. Behavioral studies, in contrast, represent a more informative yet sensitive approach. Spatial navigation, for example, is an ecologically relevant behavior that is modulated by cellular pathways potentially targeted by neurotoxicants. We evaluated whether bats ability to memorize and navigate novel spaces was negatively affected by environmental relevant doses of chlorpyrifos, a common organophosphate insecticide. We also tested how the behavioral response correlated with molecular biomarkers. We orally dosed captive big brown bats (Eptesicus fuscus) with chlorpyrifos and studied exploratory behavior in two testing arenas. We evaluated similarity of stereotype flight trajectories in a flight tent, and associative memory in a Y-maze. We quantified brain cholinesterase (ChE) activity as a cellular biomarker and employed non-targeted proteomics as molecular biomarkers. Bats exposed to chlorpyrifos were less explorative and made more incorrect choices in the Y-maze, but the consistency of their flight trajectories was unaffected. Exposed bats had 30% lower ChE activity, showed down-regulation of proteins involved in memory (VP37D), learning and sound perception (NOX3). Other important nervous system processes such as synaptic function, plasticity, oxidative stress, and apoptosis were enriched in chlorpyrifos-exposed bats. These results support the sensitivity of behavior as a biomarker of toxicity and the importance of considering other levels of organization to help explain the mechanisms underlying altered behavior due to human activities.

Humans are constantly exposed to many environmental pollutants, some of which have been largely acknowledged as key factors in the development of metabolic disorders such as diabetes and obesity. These chemicals have been classified as endocrine-disrupting chemicals (EDCs) and, more recently, since they can interfere with metabolic functions, they have been renamed as metabolism-disrupting chemicals (MDCs). MDCs are present in many consumer products, including food packaging, personal care products, plastic bottles and containers, and detergents. The scientific literature has ever-increasingly focused on insulin-releasing pancreatic {beta}-cells as one of the main targets for MDCs. Evidence highlights that these substances may disrupt glucose homeostasis, altering pancreatic {beta}-cell physiology. However, their potential impact on glucagon-secreting pancreatic -cells remains poorly known despite the essential role that this cellular type plays in controlling glucose metabolism. In the present study, we have selected seven paradigmatic EDCs representing major toxic classes, including bisphenols, phthalates, perfluorinated compounds, metals, and pesticides. By using an in vitro cell-based model, the pancreatic -cell line TC1-9, we have explored the effects of these compounds on pancreatic -cell viability, gene expression, and secretion. Our results indicated that most of the selected chemicals studied caused functional alterations in pancreatic -cells. Moreover, we revealed, for the first time, their direct effects on key molecular aspects of pancreatic -cell biology.

Chronic exposure to pesticide mixtures through diet is common, yet their combined metabolic effects and interactions with dietary factors remain unclear. We identified four pesticides prevalent in human exposure (imazalil, thiabendazole, boscalid, lambda-cyhalothrin) and assessed their combined impacts on hepatic metabolism and metabolic homeostasis using human liver cells and male mice fed standard chow or western diets. We found that the pesticide mixture induced metabolic perturbations in human hepatocytes. In addition, the pesticide mixture altered hepatic gene expression in chow-fed mice and exacerbated western diet-induced glucose intolerance, fasting hyperglycemia, and insulin resistance without affecting body weight or liver steatosis. These findings reveal that dietary context influences the metabolic consequences of pesticide mixtures, highlighting the need to consider nutritional status when evaluating environmental contaminant risks. Our results suggest that pesticide mixtures at reference doses may contribute to metabolic dysregulation, particularly under obesogenic dietary conditions. Highlights- Four common pesticides in mixture disrupt metabolism in liver cells - Dietary exposure to this pesticide mixture alters hepatic gene expression in mice - The pesticide mixture exacerbates WD-induced disruptions in glucose homeostasis - Pesticides and diet interact in producing the metabolic effects of a pesticide mixture

Organophosphorus pesticides (OPs) are a chemically diverse class of commonly used insecticides. Epidemiological studies suggest that low dose chronic prenatal and infant exposures can lead to life-long neurological damage and behavioral disorders. While inhibition of acetylcholinesterase (AChE) is the shared mechanism of acute OP neurotoxicity, OP-induced developmental neurotoxicity (DNT) can occur independently and/or in the absence of significant AChE inhibition, implying that OPs affect alternative targets. Moreover, different OPs can cause different adverse outcomes, suggesting that different OPs act through different mechanisms. These findings emphasize the importance of comparative studies of OP toxicity. Freshwater planarians are an invertebrate system that uniquely allows for automated, rapid and inexpensive testing of adult and developing organisms in parallel to differentiate neurotoxicity from DNT. Effects found only in regenerating planarians would be indicative of DNT, whereas shared effects may represent neurotoxicity. We leverage this unique feature of planarians to investigate potential differential effects of OPs on the adult and developing brain by performing a comparative screen to test 7 OPs (acephate, chlorpyrifos, dichlorvos, diazinon, malathion, parathion and profenofos) across 10 concentrations in quarter-log steps. Neurotoxicity was evaluated using a wide range of quantitative morphological and behavioral readouts. AChE activity was measured using an Ellman assay. The toxicological profiles of the 7 OPs differed across the OPs and between adult and regenerating planarians. Toxicological profiles were not correlated with levels of AChE inhibition. Twenty-two "mechanistic control compounds" known to target pathways suggested in the literature to be affected by OPs (cholinergic neurotransmission, serotonin neurotransmission, endocannabinoid system, cytoskeleton, adenyl cyclase and oxidative stress) and 2 negative controls were also screened. When compared with the mechanistic control compounds, the phenotypic profiles of the different OPs separated into distinct clusters. The phenotypic profiles of adult vs regenerating planarians exposed to the OPs clustered differently, suggesting some developmental-specific mechanisms. These results further support findings in other systems that OPs cause different adverse outcomes in the (developing) brain and build the foundation for future comparative studies focused on delineating the mechanisms of OP neurotoxicity in planarians.

PurposeTo investigate follicular fluid (FF) phthalate levels in adolescents undergoing fertility preservation compared to oocyte donors and explore its association with ovarian reserve and cumulus cell gene expression. Methods20 Adolescents (16.7 {+/-} 0.6 years old) and 24 oocyte donors (26.2 {+/-} 0.4 years old) undergoing fertility preservation were included in the study. Patient demographics, ovarian stimulation and oocyte retrieval outcomes were analyzed for each group. FF levels of 9 phthalate metabolites were assessed individually and as molar sums representative of common compounds (all phthalates: Phthalates; DEHP: DEHP), exposure sources (plastics: Plastic; personal care products: PCP), and modes of action (anti-androgenic: AA) and compared between the two groups. ResultsFollicular fluid Plastic and PCP levels were significantly higher in adolescents compared to oocyte donors (p<0.05). Follicular fluid DEHP, Plastic, PCP, AA, and Phthalates levels were positively associated with antral follicle count (AFC) (p<0.05) in oocyte donors when adjusted for age, BMI, and race/ethnicity. RNA-seq analysis revealed 248 differentially expressed genes (DEGs) in cumulus cells of adolescents within the top quartile (n=4) of FF Phthalates levels compared to the adolescents within the bottom half (n=9). Genes enriched in pathways involved in cell motility and development were significantly downregulated. ConclusionAdolescents undergoing fertility preservation cycles demonstrate higher levels of phthalate metabolites in their follicular fluid compared to oocyte donors. Phthalate metabolite levels in FF are associated with higher AFC levels in oocyte donors. Higher phthalate levels in FF are associated with alterations in the cumulus cells transcriptome in adolescents. Capsule SummaryPhthalates are detected in the follicular fluid of adolescents and oocytes donors, and the levels are increased in the follicular fluid of adolescents. Higher total phthalate levels in follicular fluid are associated with altered cumulus cells transcriptome in adolescents.

Environmental exposure to polycyclic aromatic hydrocarbons (PAHs) causes metabolic dysfunction, but reliable biomarkers are still needed to assess human health effects. This study used 21-day matured human HepG2 spheroids, a metabolically competent three-dimensional (3D) liver model, to assess metabolic responses to graded, non-cytotoxic concentrations of benzo[a]pyrene (BaP) and benzo[b]fluoranthene (BBF) after 24- and 96-h exposure. Untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics, combined with multivariate and network analyses, identified compound- and time-specific metabolic signatures. At 24 hours, no metabolites showed significant changes. In contrast, at 96 hours, both PAHs consistently altered seven robust metabolites linked to polyamine metabolism, membrane dynamics, mitochondrial energy, and DNA-repair pathways. Network analysis showed BBF caused broader and more connected changes than BaP, indicating distinct toxicodynamics. These findings underscore the importance of extended exposure in revealing metabolic disruption and support a set of candidate biomarkers for future low-dose studies and improved risk assessment of airborne toxicants.

The exposome encompasses all lifetime environmental exposures affecting health. Its complexity and high data dimensionality make it challenging to link specific exposure combinations to adverse health outcomes. Establishing relevant exposome criteria is key to addressing current knowledge gaps. This study evaluated contaminant levels in Wisconsin groundwater and their effects on host health. We focused on three co-occurring chemicals that were detected at concentrations exceeding groundwater standards, nitrate, atrazine and imidacloprid, and the emerging contaminant, microplastics. In this study, broilers were exposed to a low dose chemical mixture (35,000 ppb nitrate + 1.7 ppb atrazine + 0.58 ppb imidacloprid) and high dose chemical mixture (100,000 ppb nitrate + 3,000 ppb atrazine + 3,000 ppb imidacloprid) or polyethylene microplastics (PE MPs) for 49 days. We observed that both ternary mixtures and PE fiber MPs significantly altered the cecal microbiomes as determined by the enrichment of genera, Fournierella, Ruminococcus and an unclassified genus in the family Coriobacteriaceae. In addition, +PE fiber presence dysregulated metabolic pathways associated with bile acid biosynthesis and lipid metabolism. Similarly, perturbations to cecal microbial activity for both ternary chemical mixtures were confirmed via modulation of six metabolites including methylisopelletierine which had a higher total ion intensity than the control group. Interestingly, there were no detectable pathological effects to either the +PE fiber or ternary mixture treatment groups. Overall, the data presented here demonstrates that low doses of environmental contaminants are sufficient to dysregulate cecal taxonomic composition and microbial activity without inducing detectable pathological effects. ImportanceWe found that exposure to mixtures of environmental toxins caused gut dysbiosis observed by changes to the chicken cecal microbiome and metabolome. This highlights the importance of conducting such studies with environmentally relevant mixtures of contaminants at detected concentrations to understand the actual risks associated with exposures like drinking contaminated groundwater over a long period of time. Our findings demonstrate that gut microbial metabolites, now known to be key regulators and signaling molecules in a wide range of host health issues, are the source of the negative health outcomes; superseding cell death or pathological damage that are caused by acute exposures. These changes have implications for predicting negative long-term chronic health outcomes.

The past several decades have seen alarming declines in the reproductive health of humans, animals and plants. While humans have introduced numerous pollutants that can impair reproductive systems (such as well-documented endocrine disruptors), the potential for microplastics (MPs) to be contributing to the widespread declines in fertility is particularly noteworthy. Over the same timespan that declines in fertility began to be documented, there has been a correlated shift towards a "throw-away society" that is characterised by the excessive consumption of single-use plastic products and a concomitant accumulation of MPs pollution. Studies are showing that MPs can impair fertility, but data have been limited to rodents that were force-fed hundreds of thousands of times more plastics than they would be exposed in the environment. As a first step to link in vitro health effects with in vivo environmental exposure, we quantified microplastics in the follicular fluid of women and domestic cows. We found that the concentrations of polystyrene microplastics that naturally occurred in follicular fluid were sufficient to compromise the maturation of bovine oocytes in vitro. Collectively, these findings demonstrate that microplastics may also be contributing to the widespread declines in fertility that have been occurring over recent Anthropocene decades.

Polychlorinated biphenyls (PCBs) are persistent environmental toxicants that bioaccumulate in the food chain and readily cross the placenta, raising concerns for developmental toxicity. While PCB exposure has been associated with metabolic and neurodevelopmental disorders, its cell type-specific effects on liver development remain poorly understood. This study aimed to investigate how maternal exposure to an environmentally relevant Fox River PCB mixture affects liver development in female offspring at single-cell resolution. We hypothesized that early-life PCB exposure disrupts hepatic metabolic and immune function in a cell type-specific manner. Using single-cell RNA sequencing (scRNA-seq) on liver tissue from postnatal day 28 female mice perinatally exposed to PCBs, we identified major hepatic and immune cell populations and assessed cell-specific transcriptional responses. PCB exposure significantly altered the proportions of endothelial cells and Kupffer cells and reduced neutrophil abundance. Transcriptomic analysis revealed that PCBs dysregulated key pathways in hepatocytes and non-parenchymal cells, including ER stress responses, drug metabolism, and glucose/insulin signaling. Notably, hepatocytes exhibited upregulation of phase-I drug-metabolizing enzymes and uptake transporters, but downregulation of phase-II enzymes and efflux transporters. Kupffer cells and endothelial cells had altered immune and metabolic gene expression, and intercellular communication analysis predicted disrupted fibronectin, collagen, and chemokine signaling due to PCB exposure. RT-qPCR validation confirmed increased hepatic ER stress marker expression. Together these findings demonstrate that perinatal PCB exposure induces persistent, cell type-specific transcriptomic reprogramming in the liver, impairing metabolic and immune functions. This study highlights the utility of single-cell transcriptomics for revealing toxicant effects with cellular precision during critical windows of development.

Exposure of cells or tissues to chemical compounds can be analyzed through transcriptomic signatures, which can be used to classify chemical agents. This information can also enrich Adverse Outcome Pathways (AOP). Transcriptional signatures have generally been obtained using "bulk" analysis, by which the global gene expression pattern of an entire tissue is determined. Although this approach has been useful in toxicology, some information is lost, especially when tissues containing multiple cell types are considered. With the advent of single-cell transcriptomics (scRNA-seq), it is now possible to obtain higher resolution, cell type-specific responses in complex tissues. The aim of the present study was to evaluate the added value of scRNA-seq in analysis of the acute response of human bronchial epithelial cells grown at the air/liquid interface (ALI) to a known toxic compound, CdCl2, with well described transcriptional signatures of exposure. Fully differentiated mucocilliary epithelia obtained from three independent donors were exposed to 10 {micro}M CdCl2 and scRNA-seq analysis was performed on a total of 18255 cells to obtain cell type-specific signatures. Our results show that the contribution of each cell type to the overall transcriptomic bulk response varies. For example, the classical heavy metal detoxification response was only detected in multiciliated and secreting cells, while absent in basal cells. The data demonstrate that scRNA-seq provides high-resolution transcriptional signatures with unexpected features. This added information is likely to have implications for the refinement of AOPs and could serve as a basis for a new generation of tests in predictive toxicology.

Phthalate monoesters have been identified as endocrine disruptors in a variety of models, yet understanding of their exact mechanisms of action and molecular targets in cells remains incomplete. Here, we set to determine whether epidemiologically relevant mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) can affect biological processes by altering cell plasma membrane fluidity or formation of cell-cell contacts. As a model system, we chose endometrial stromal cell lines, one of which was previously used in a transcriptomic study with MEHHP or MEHHP-containing mixtures. A short-term exposure (1 h) of membrane preparations to endocrine disruptors was sufficient to induce changes in membrane fluidity/rigidity, whereas different mixtures showed different effects at various depths of the bilayer. A longer exposure (96 h) affected the ability of cells to form spheroids and highlighted issues with membrane integrity in loosely assembled spheroids. Finally, in spheroids assembled from T-HESC cells, MEHHP interfered with the formation of tight junctions as indicated by the immunostaining of zonula occludens 1 protein. Overall, this study emphasized the need to consider plasma membrane, membrane-bound organelles, and secretory vesicles as possible biological targets of endocrine disruptors and offered an explanation for a multitude of endocrine disruptor roles documented earlier.

Endocrine disrupting compounds (EDC) are ubiquitous in the human environment, displaying a highly relevant research topic. The impact of EDC on the differentiation of primitive cells, e.g. in hematopoiesis, is of particular interest. We found profound inhibitory effects of di-2-ethylhexyl phthalate (DEHP) on erythropoiesis and dendropoiesis, mediated via reactive oxygen species (ROS) generation. Neutrophil differentiation, however, was not affected by DEHP. ROS leads to a shift from glycolysis to the pentose phosphate pathway and diminishes ATP generation from glycolysis, ultimately resulting in apoptosis in both cell types. In neutrophils, ATP generation is held constant by active fatty acid oxidation (FAO), rendering these cells highly resistant against ROS. This relationship also holds true in HUVEC and HepG2 cells, also in combination with other organic peroxides. We, therefore, uncover a key mechanism for ROS quenching which further explains the distinct ROS quenching ability of different tissues.

Exposure to plastic-derived estrogen-mimicking endocrine-disrupting bisphenols can have a long-lasting effect on bone health. However, gestational exposure to below tolerable daily intake (TDI) of bisphenol A (BPA) and its substitute, bisphenol S (BPS), on offsprings bone mineralization is unclear. This study examined the effects of in-utero bisphenol exposure on the growth and bone density of the offspring rats. Pregnant Wistar rats were exposed to BPA and BPS (0.0, 0.4 g/kg bw) via oral gavage from gestational day 4 to 21. The bone density, IGF-1, osteocalcin, and calcium levels were measured by DEXA, ELISA and AAS, respectively. The bisphenols action on canonical BMP signaling was examined in osteoblast SaOS-2 cells. Maternal exposure to bisphenols (BPA and BPS) increased the body weight, bone mineral content, and density in the offspring aged 30 and 90 days (p<0.05). Plasma IGF-1, calcium, osteocalcin, and alkaline phosphatase activities were altered in BPA-exposed offspring (p<0.05). The bisphenols exposure to SaOS-2 cells decreased its viability in a dose-dependent manner and promoted the cell cycle progression of the S/G2-M phase (p<0.05). The expression of BMP1, BMP4, and intracellular signalling mediators SMAD1, SMAD5, and RUNX2 mRNAs was altered upon bisphenol exposure in these cells (p<0.05). The bone mineralization index and expression of extracellular matrix proteins such as ALPL, COL1A1, DMP1, and FN1 were downregulated (p<0.05). Bisphenol co-incubation with noggin decreased TGF-{beta}1 expression, indicating its involvement in bone mineralization. Overall, exposure to bisphenols (BPA and BPS) during gestation altered growth and bone mineralization in the offspring by modulating canonical BMP/ TGF-{beta}1 signalling mediators. HighlightsO_LIGestational exposure to low doses of bisphenol increases whole-body BMC and BMD in the offspring. C_LIO_LIIn-utero BPA exposure increased plasma IGF-1 and gla-type osteocalcin, a marker of osteoblast activity in the offspring. C_LIO_LIBisphenol exposure modulates Smad-dependent BMP signaling in the SaOS-2 cells. C_LI

Premature depletion of ovarian reserve is a cause of female infertility. Plasticizer Bisphenol AF (BPAF) residues have been found in human reproductive-related samples. However, little is known about its influence on the ovarian reserve. Here, we demonstrate that BPAF exposure causes excessive activation of primordial follicles through Hippo signaling in young females, resulting in rapid exhaustion of the ovarian reserve and, ultimately, premature ovarian insufficiency. We found that oral ingestion of BPAF disrupts normal estrous cyclicity and induces constant estrus in a dose-dependent manner. BPAF can upregulate the expression of YAP transcriptional coactivator. Intriguingly, only the high dose of BPAF inhibits Hippo signaling by eliciting a substantial decrease in phospho-YAP levels. Through such regulatory effects, BPAF causes YAP translocation into the nucleus and triggers the overactivation of primordial follicles. Collectively, this study proposes a novel toxicological mechanism explaining the negative impact of BPAF on the ovarian health of young females.

There is a need to develop identification tests for Metabolism Disrupting Chemicals (MDCs) with diabetogenic activity. Here we used the human EndoC-{beta}H1 {beta}-cell line, the rat {beta}-cell line INS-1E and dispersed mouse islet cells to assess the effects of endocrine disruptors on cell viability and glucose-stimulated insulin secretion (GSIS). We tested six chemicals at concentrations within human exposure (from 0.1 pM to 1 M). Bisphenol-A (BPA) and tributyltin (TBT) were used as controls while four other chemicals, namely perfluorooctanoic acid (PFOA), triphenylphosphate (TPP), triclosan (TCS) and dichlorodiphenyldichloroethylene (DDE), were used as "unknowns". Regarding cell viability, BPA and TBT increased cell death as previously observed. Their mode of action involved the activation of estrogen receptors and PPAR{gamma}, respectively. ROS production was a consistent key event in BPA- and TBT-treated cells. None of the other MDCs tested modified viability or ROS production. Concerning GSIS, TBT increased insulin secretion while BPA produced no effects. PFOA decreased GSIS, suggesting that this chemical could be a "new" diabetogenic agent. Our results indicate that the EndoC-{beta}H1 cell line is a suitable human {beta}-cell model for testing diabetogenic MDCs. Optimization of the test methods proposed here could be incorporated into tier protocols for the identification of MDCs.