Journal of Xenobiotics doi: 10.3390/jox16030083

Authors: Hassan Omary Kumenya Irene Nalumansi Christopher Angiro Ivan Kiganda Timothy Omara Emmanuel Ntambi

In equatorial Africa, rapid urbanization has increased city populations and particulate matter emissions. Street dust is a visual indicator that can be used to track urban pollution. In the present study, the total concentration and speciation of 10 potentially toxic elements (PTEs; As, Cd, Cu, Cr, Ni, Mn, Fe, Pb, Co, and Zn) in dust (n = 36) sampled from three streets of Mbarara City, Uganda, were determined using Energy Dispersive X-ray Fluorescence and Inductively Coupled Plasma-Optical Emission Spectrometry. The concentration of PTEs (0.27–36,401.50 mg/kg) geostatistically indicated moderate to extremely high enrichment of Cd, Cu, and Co in street dust. According to principal component and hierarchical cluster analyses, As, Pb, Cu, Zn, and Cd originated mainly from anthropogenic inputs, Fe and Mn came from geogenic sources, while Cr, Ni, and Co were from both natural and anthropogenic contributions. The mobility of the PTEs followed a general trend, Zn > Co > Cd > Ni > Cr, with Zn and Co being more environmentally mobile. Human health risk assessments indicated that discernible non-carcinogenic health risks may result from ingestion of dust by both children and adults. Children could also experience cancer health effects through the same exposure pathway.

Journal of Xenobiotics doi: 10.3390/jox16030082

Authors: Elena Vakonaki Eleftheria Hatzidaki Stella Baliou Maria Marmara Athanasios Alegakis Eleftheria Mylonaki Zoi Volonaki Fanourios Makrygiannakis Aristides Tsatsakis Manolis N. Tzatzarakis

Background: Exposure of pregnant women to stress and endocrine-disrupting chemicals (EDCs) during pregnancy can have a substantial impact on mother and infant health. We investigated the concentrations of EDCs, such as parabens (PBs) and triclosan (TCS), as well as stress hormones (cortisone and cortisol), across pregnancy trimesters and examined their associations with maternal average telomere length (TL). Methods: Hair samples from 49 postpartum women were analyzed using liquid chromatography–mass spectrometry (LC-MS) to quantify EDCs and stress hormone concentrations. Results: The mean methyl paraben concentrations in the hair of postpartum women were prevalent across all pregnancy trimesters, while butyl paraben was detected at the lowest levels. The mean concentration of PBs followed the order methyl > propyl > ethyl > benzyl > butyl paraben across pregnancy trimesters. We found that ethyl paraben and triclosan were each positively and significantly associated with cortisol levels in postpartum women’s hair. Consistent with this, the mean cortisone concentration gradually increased from the first to the third pregnancy trimester, whereas cortisol reached the highest mean concentration at the second trimester. A significant positive association between cortisol and cortisone levels was observed. Further analyses revealed that mothers’ average TL was positively associated with ethylparaben and triclosan levels and inversely associated with benzylparaben levels. Last but not least, we found that cortisol/cortisone levels were positively associated with postpartum women’s TL in a statistically significant manner. Conclusions: In the present study, prenatal exposure to stress hormones and EDCs appears to exert a statistically significant impact on maternal TL dynamics.

Journal of Xenobiotics doi: 10.3390/jox16030081

Authors: Edward Heinen James V. Cizdziel Boluwatife S. Olubusoye Ruojia Li Matthew T. Moore Lindsey M. Witthaus

Microplastic (MP) pollution in agricultural settings is an emerging field of study, with interest focusing on potential resource contamination of soil and water due to the use of plastic materials in farming practices. The Mississippi Delta, a highly agricultural region, is prone to both natural and intentional flooding, potentially exacerbating this issue. This exploratory study investigated MP (>30 µm) concentrations, sizes, and polymer compositions in floodwater, irrigation water, and surface runoff from soybean fields across two counties in the Mississippi Delta using micro-Fourier transform infrared spectroscopy (µ-FTIR). Mean ± SE concentrations (MPs/L) were 72 ± 66 in floodwater (n = 18), 169 ± 121 in source (irrigation pond) water (n = 4), and 30 ± 37 in runoff (n = 3) in Sunflower County, MS. In Coahoma County, MS, mean ± SE runoff concentration was 88 ± 76 MPs/L (n = 24). Mean concentrations were elevated as compared to other MP studies in agricultural environments. The most common polymers present were polyethylene terephthalate (PET), polyethylene (PE), polyethylene-ethyl vinyl acetate (PE/EVA), and thermoplastic elastomers (TPEs), which are commonly used in the manufacturing of agricultural materials. MPs from the smallest size fraction measured (30–100 µm) were the most common in all floodwater samples, ranging from 75.5–91% abundance. Using Attenuated Total Reflectance (ATR)-FTIR, larger plastic litter was identified as mostly PE and PET, which is consistent with polymer distributions in floodwater samples. Overall, MPs were prevalent in both floodwater and runoff, with relatively consistent concentrations and polymer compositions across samples. However, further research is needed to fully elucidate their fate and potential impacts on agricultural systems.

Journal of Xenobiotics doi: 10.3390/jox16030080

Authors: Paweł Szpot Olga Wachełko Kaja Tusiewicz Marcin Zawadzki

Aim: The aim of this study was to develop and validate a reliable HS-GC-FID/FID method for the determination of ethanol and other low-molecular-weight volatile compounds in biological fluids for forensic applications. Method: The method is based on headspace gas chromatography with dual-column and dual flame ionization detection (HS-GC-FID/FID), using Zebron ZB-BAC1 and ZB-BAC2 columns. The procedure was validated in terms of linearity, limits of detection and quantification, precision, and accuracy, as well as carryover. Results: The method demonstrated linearity over the concentration range of 0.05–5.0‰, with R2 values of 0.997–0.999. The limit of quantification (LOQ) was 0.05‰, and the limit of detection (LOD) was 0.025‰. Precision and accuracy were both below 5%. Retention times (min) on the two columns were as follows: methanol (1.70/1.81), ethanol (2.06/2.29), acetone (2.60/2.59), isopropanol (2.45/2.73), n-propanol (3.24/3.98), and n-butanol (6.35/8.45). The developed method was successfully evaluated through international proficiency testing and is routinely applied in our laboratory for forensic casework. Conclusions: The developed HS-GC-FID/FID method provides accurate and reliable determination of ethanol and other relevant volatile compounds in biological fluids (i.a., blood) and meets the requirements for forensic toxicology. Additionally, the literature review conducted in this study highlights that globally unified principles for forensic alcohol analysis are still lacking, and that certain inappropriate methodological approaches remain in use. The present paper also provides recommendations defining essential methodological requirements to ensure the evidential reliability of ethanol analysis in forensic toxicology.

Journal of Xenobiotics doi: 10.3390/jox16030079

Authors: Yanpeng Gao Weijie Lu Yibo Zhang Mingze Geng Xianglong Luo Yuemeng Ji Yang-Guang Gu

Halogenated polycyclic aromatic hydrocarbons (HPAHs) are persistent contaminants with elevated toxicity, yet dietary exposure data remain limited. Here, we systematically assessed dietary HPAHs using 87 duplicate diet samples collected from populations living in and around a coking industrial area by applying the duplicate diet method, a gold-standard approach that provides precise individual-level exposure information. Thirty-one HPAHs were detected, including seven previously unreported congeners, with mean concentrations of 62.51, 33.68, and 16.52 ng/g in the coking plant, nearby residential, and control areas, respectively. Lipid-rich foods, particularly meats, exhibited the highest HPAH burdens, and thermal processing approximately doubled concentrations in meals collected from the coking plant area. Dietary cancer risk was evaluated using a toxicity equivalency-based incremental lifetime cancer risk (ILCR) framework. Although several HPAHs occurred at low concentrations, congeners with high relative effect potency contributed disproportionately to cumulative cancer risk. Population-level risk distributions revealed that 25.8% of dietary samples exceeded the benchmark ILCR threshold of 10−4 in the coking plant area. In silico toxicity predictions further indicated potential organ-specific toxicological relevance for the blood, liver, kidney, and cardiovascular systems, supporting the health relevance of dietary HPAH exposure. In general, these results suggest that industrial influence, food composition, and cooking practices jointly contribute to dietary HPAH exposure and toxicity-weighted cancer risk. Our findings highlight the importance of incorporating halogenated congeners into routine monitoring programs and health risk assessments in industrialized regions.

Journal of Xenobiotics doi: 10.3390/jox16030078

Authors: Claudia Cipollone Riccardo Mastrantonio Paola Mozzoni Giada Mastrangeli Massimo Corradi Stefano Renzetti Veronica Saponara Maria Nicastro Delia Cavallo Raffaele Maiello Marco Gentile Diana Poli Mario Muselli Alessia Romantini Giorgia Di Gennaro Gloria Cenci Carmela Protano Matteo Vitali Giuseppe De Palma Cinzia Lucia Ursini Leila Fabiani

Firefighters are potentially exposed to multiple harmful substances, and their activities are classified as carcinogenic to humans. This case study assessed early genotoxic damage (fpg-comet and BMCyt assays) and epigenetic alterations (seven circulating miRNAs) in 35 firefighters compared to 45 non-exposed workers. Occupational exposure to fire smoke was self-reported via questionnaire. Firefighters showed higher median genotoxic DNA damage with respect to the non-exposed group (%DNA tail Buff 19.4 vs. 16.8; %DNA tail Enz 22.2 vs. 19.3; Tail moment 5.5 vs. 4.5; % of apoptotic cells 1.13 vs. 0.97). miRNAs related to air pollution, oxidative stress, tumor suppression, and immune responses, like mir-16, mir-15a, mir-29a, mir-125b, and mir-142, showed significant downregulation (p < 0.001) in the exposed group. Mean percentages of early apoptosis biomarkers and composite DNA damage indices among FF also differed significantly from the other participants (‰Condensed chromatin 0.46 vs. 0.06; ‰Tot anomalies 5.15 vs. 3.82). Multiple correlations emerged, particularly between miRNAs and comet assay parameters, and between comet assay and BMCyt indicators. The implemented integrated approach provides information about the existence of a relationship between genotoxic and epigenetic effects in firefighters, also influenced by time since exposure. Future studies with bigger sample sizes are required.

Journal of Xenobiotics doi: 10.3390/jox16030077

Authors: Dimitra Florou Thalia Vlachou Amvrosios Orfanidis Vasilios Sakkas Vassiliki A. Boumba

Toxicological testing for drugs of abuse (DOAs) is an essential tool for healthcare practitioners and law enforcement agencies. Oral fluid (OF) is an alternative biological fluid for detecting recent DOA intake and is widely employed in forensic investigations. In the current study, a relatively novel and “green” fabric phase sorptive extraction (FPSE) procedure for sample preparation was coupled to liquid chromatography–tandem mass spectrometry (LC–MS/MS) to provide simplicity, cost-effectiveness, rapidity, low solvent consumption, and high analytical performance for the quantitative determination of ten commonly encountered DOAs and metabolites: amphetamine, benzoylecgonine, cocaine, codeine, ecgonine methyl ester, methadone, methamphetamine, 3,4-methylenedioxyamphetamine, 6-monoacetylmorphine, and morphine. The FPSE procedure was optimized by testing different filters, pH, extraction time, and solvents. The validated method demonstrated excellent linearity for all analytes, selectivity, acceptable precision, and high sensitivity (ranges for limits of detection (LODs) and quantification (LOQs) were 0.01–2 ng/mL and 0.03–6 ng/mL, respectively). Autosampler and short-term freeze stability exceeded 95% and 90% for all analytes, respectively. Overall, the combination of FPSE with LC–MS/MS provided a sensitive, selective, and environmentally friendly innovative analytical approach for the determination of DOA in OF and is suitable for both screening and confirmatory forensic and clinical applications.

Journal of Xenobiotics doi: 10.3390/jox16030076

Authors: Apurva Lad Jason Kindle Prajwal Hegde Gabriel G. Kleer Andrew L. Kleinhenz Johnna A. Birbeck Judy Westrick Nicholas J. Peraino Terry D. Hinds Neeraja Purandare Andrew M. Fribley Steven T. Haller David J. Kennedy

Microcystin-LR (MC-LR) is a potent hepatotoxin that has been shown to cause liver damage even at doses lower than the established Low Observable Adverse Effect Level (LOAEL) of 200 μg/kg in animal models. We have previously observed that low-dose exposure to MC-LR in animals with diet-induced Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) and subsequent treatment with antioxidants like N-acetylcysteine (NAC) and the Na+/K+ ATPase-Src kinase inhibitor pNaKtide significantly alleviated hepatic infiltration of immune cells, downregulated markers of inflammation and hepatotoxicity, increased the breakdown of the toxin molecule, and restored phase I and II drug metabolism pathways, including the glutathione pathway. Because the liver is composed of heterogeneous cell types, this study aimed to determine the specific role of hepatocytes in the uptake and metabolism of MC-LR, especially in the setting of MASLD. To address this, we used two well-established hepatocyte cell lines—AML-12 murine hepatocytes and human Hep3B hepatocytes. Preliminary dose comparison studies with AML-12 cells showed that MC-LR at 10 μM concentration showed a significant upregulation in the genetic expression of the markers of hepatotoxicity—OSMR (p ≤ 0.01) and SerpinE (p ≤ 0.0001)—in comparison to Vehicle. Treatment with pNaKtide (1 µM) and/or NAC (10 mM) in the presence of MC-LR significantly reduced the expression of both OSMR (p ≤ 0.0001) and SerpinE (p ≤ 0.01 and p ≤ 0.0001, respectively). To model steatotic hepatocytes characteristic of the MASLD phenotype, Hep3B hepatocytes were first treated with 500 µM of oleic acid (OA) before exposing them to the toxin in the presence and absence of antioxidants. MC-LR exposure, induced markers of inflammation and hepatotoxicity to be elevated significantly in the presence of OA as compared to MC-LR exposure alone. This elevation of the genetic markers of inflammation and hepatotoxicity was significantly attenuated on treatment with pNaKtide (1 µM) and NAC (10 mM). Quantification of human SERPINE1 (PAI1) and 8-OHdG, a stable marker of oxidative stress, in the spent media of Hep3B cells corroborated the trends observed in the genetic markers of hepatotoxicity. These observations support the central role that hepatocytes play in the uptake and metabolism of MC-LR, which is complicated by the presence of MASLD-like conditions and can help in the development of future therapeutic strategies.

Journal of Xenobiotics doi: 10.3390/jox16030075

Authors: Tamara Lazarević-Pašti Tamara Terzić Andreja Leskovac Sandra Petrović Vedran Milanković Nevena Radivojević Jugoslav Krstić Katarina Kokanov Stanković Ana Jocic Snežana Brković Igor Pašti

Organophosphate pesticides are widely used agricultural chemicals that pose significant environmental and health risks due to their neurotoxicity, which is associated with inhibition of acetylcholinesterase. In this study, carbon materials derived from rabbit litter-based precursors were investigated as sustainable adsorbents for the removal of organophosphate pesticides from aqueous systems. The prepared materials exhibited a broad range of textural properties, with specific surface areas ranging from 10 to 487 m2 g−1, depending on the precursor composition. Adsorption experiments demonstrated measurable removal of chlorpyrifos, malathion, and dimethoate, with maximum adsorption capacities reaching 71.8 mg g−1 for malathion, although adsorption performance varied among materials, indicating a combined influence of pore accessibility and surface chemical heterogeneity. Evaluation of acetylcholinesterase inhibition before and after adsorption showed a consistent decrease in enzyme inhibition across all systems, with values reduced from 40% to as low as 20% for chlorpyrifos, from 35% to as low as 11% for malathion, and from 20% to as low as 10% for dimethoate, indicating a reduction in the neurotoxic potential of the treated solutions. In addition, the genotoxicity of the carbon materials varied with their structural and compositional characteristics, underscoring the importance of considering both adsorption performance and biological interactions. These findings demonstrate that waste-derived carbon materials can contribute to the removal of organophosphate contaminants while simultaneously reducing their associated neurotoxic effects.

Journal of Xenobiotics doi: 10.3390/jox16030074

Authors: Louisa Bechohra Chahrazed Makhlouf Hassina Khaldoun Samira Aouichat Amina Settar Dalila Tarzaali Nacera Lemlikchi Amina Bouhallel Yasmine Oularbi Schahinez Terkmane Nacima Djennane

The widespread use of modern insecticide formulations underscores the need for mechanistic evaluation of their potential renal toxicity. This study investigated the nephrotoxic effects of Ampligo® 150 ZC, a binary formulation of lambda-cyhalothrin and chlorantraniliprole, in female rabbits under subacute exposure conditions, with particular emphasis on apoptosis-related and epithelial integrity biomarkers, and evaluated the protective effects of thyme essential oil (TEO) and vitamin C. Rabbits were allocated into four groups: control, AP, AP + TEO, and AP + TEO + vitamin C. Ampligo (AP) exposure resulted in significant renal dysfunction, as evidenced by elevated biochemical biomarkers and marked histopathological lesions. At the molecular level, AP induced p53 upregulation alongside Bcl-2 and Cyclin D1 downregulation, suggesting apoptosis induction and cell cycle dysregulation. Moreover, reduced E-cadherin and β-catenin expressions indicated disruption of epithelial junction integrity and impaired renal structural homeostasis. Notably, co-administration of TEO and vitamin C markedly attenuated these alterations, improving biochemical, histopathological, and immunohistochemical parameters. Overall, these findings suggest that AP-driven nephrotoxicity may involve apoptotic and epithelial pathways under subacute exposure conditions, whereas antioxidant co-treatment may mitigate kidney injury, supporting the potential of natural antioxidants as adjuncts against pesticide-induced renal injury.

Journal of Xenobiotics doi: 10.3390/jox16030073

Authors: George Jîtcă László-István Bába Ingrid Evelin Mehelean Ana Natalia Maier Ioana-Irina Popoviciu Tudor-Nicolae Cotruş Erzsébet Májai

According to the most recent data published by the World Health Organization (WHO), it is estimated that approximately 332 million persons worldwide suffer from depression. The relationship between depression and alcohol consumption is complex and bidirectional. This study aimed to investigate the effects of cannabidiol (CBD) on behavior and malondialdehyde (MDA) imbalance in female Wistar rats exposed to chronic stress and alcohol. Sixteen intact cycle female 5-month-old Wistar rats were randomly assigned to two groups: the Control group (n = 8), and the CBD group (n = 8), which received CBD at a dose of 10 mg/kg. Following chronic stress induction, during the three-week treatment period, the animals were exposed to alcohol on three separate occasions. CBD-treated females showed increased freezing time in the Open Field test with no clear anxiolytic effect. In the Y maze and Morris Water Maze, they exhibited improved memory-related performance. Brain MDA levels were reduced, while plasma MDA was unchanged. Cortisol tended to be higher in the CBD group. CBD administration showed potential cognitive and central antioxidant effects, but no clear anxiolytic effect.

Journal of Xenobiotics doi: 10.3390/jox16030072

Authors: Scott A. Helgeson Hossny Alaws Mohamed I. Ibrahim Augustine S. Lee Danielle H. W. Vlecken Hassan Z. Baig

Microplastics may pose health risks, particularly for chronic lung diseases. Clarifying the link between circulating microplastics and pulmonary disease is vital for shaping research and interventions. The objective of this study was to evaluate whether microplastics in peripheral blood are associated with COPD or IPF vs. no lung disease. In this pilot prospective case-control study, participants were grouped as control (n = 10), COPD (n = 9), or IPF (n = 10). Relevant comorbidities and exposures were obtained from records and questionnaires. All underwent standardized blood collection (PlasticTox©). Samples were analyzed for total microplastic concentration, stratified by size (<10 µm, 10–30 µm, 30–70 µm). The primary outcome was to show a difference in total microplastic burden between lung disease and controls. Secondary measures were to determine size-specific concentrations and associations with demographic variables and smoking history. Among 29 participants (median age 70 (IQR 64–80); 14 women (48.3%)), COPD/IPF groups had significantly higher total microplastic concentrations vs. controls (median 26.0 vs. 3.5 particles/100 µL; p < 0.01). Particle burdens <10 µm and 10–30 µm were particularly elevated (both p < 0.01). After adjusting for smoking, only the <10 µm fraction remained independently associated with lung disease (adjusted odds ratio 1.94 (95% CI, 1.23–7.04)). In this pilot exploratory study, individuals with COPD or IPF showed greater circulating microplastic levels than controls. These findings should be interpreted as hypothesis-generating, and larger analytically validated studies are needed to clarify directionality, causal mechanisms, contamination control, and the clinical relevance of circulating microplastic burden.

Journal of Xenobiotics doi: 10.3390/jox16030071

Authors: Olivia Torres-Bugarín Alejandro Sánchez-González María Luisa Ramos-Ibarra Idalia Yazmín Castañeda-Yslas Nina Bogdanchikova Alexey Pestryakov María Evarista Arellano-García

The intensive use of agricultural inputs and the increasing incorporation of nano-materials into crop management practices raise concerns about their ecotoxicological interactions in plant systems. This study evaluated phytotoxicity, cytotoxicity, and genotoxicity in Allium cepa L. under experimental nano-agrochemical exposure scenarios combining two conventional nitrogen fertilizers—ammonium sulfate (AS) and urea—with silver nanoparticles (AgNPs). Biological responses were assessed across fertilizer concentrations (0.03–0.5 g/L), applied individually, simultaneously, and sequentially, to identify modulatory effects of AgNPs on plant proliferative activity and genomic stability. Results showed the relative stability of morphophysiological indicators associated with root growth, whereas cytogenetic biomarkers exhibited selective alterations under specific conditions. Significant increases in genetic damage markers were detected at intermediate ammonium sulfate concentrations, suggesting sublethal phytotoxicity windows not reflected by macroscopic growth parameters. In addition, modulation of the mitotic index and absence of generalized genotoxic effects in most combined or sequential treatments indicate that AgNPs primarily acted as modulators of proliferative responses rather than direct cytotoxic agents. Overall, these findings highlight the dynamic and non-linear nature of nano-agrochemical interactions in plant systems and underscore the importance of multibiomarker approaches for the early detection of genomic instability. The results provide experimental evidence relevant to the environmental risk assessment of nano-enabled fertilization strategies under realistic mixed-exposure scenarios. This study contributes to advancing the ecotoxicological understanding of emerging agricultural technologies and supports the need for further mechanistic research and field-based evaluations to guide the safe and sustainable use of nanomaterials in crop production.

Journal of Xenobiotics doi: 10.3390/jox16020070

Authors: Syed Raju Ali Yasir Anwar Hani Mohammed Ali

Fluorene poses ecological and health hazards that originate from biomass combustion and petroleum. However, some microorganisms can counter fluorene through complex enzymatic degradation pathways. This research aimed to explore the catalytic efficiency of enzymes on metabolites and their toxicity levels throughout the fluorene biodegradation pathway. Several web servers and software were used to characterize them and analyse molecular dockings between ligands and proteins. Fluorene and its metabolites have mild toxicities to the brain, lung, neurons, and kidneys, and consequent endpoints cause mutations, cancer, and ecotoxicity at different levels. The catalytic enzymes are well-folded, single-chained, medium-sized proteins that are acidic, thermostable, and with few exceptions, hydrophilic, cytoplasmic, non-allergenic, and nonvirulent, possessing multiple active sites. The ERRAT, PROCHECK, and VERIFY 3D tools successfully validated the SWISS-modelled 3D structures of proteins. Molecular docking results showed moderate binding affinities between proteins and ligands, ranging from −9.4 to −6.1 kcal/mol, indicating potential activities of the enzymes. This computational study supports the conventional fluorene degradation pathway and may provide a new avenue for further research.

Journal of Xenobiotics doi: 10.3390/jox16020069

Authors: Ningbo Wang Yingying Zhong

Fluopyram (FO), a widely used succinate dehydrogenase inhibitor (SDHI) fungicide, poses a potential risk to aquatic ecosystems due to its mitochondrial toxicity in non-target organisms. This study investigated its toxic effects on zebrafish (Danio rerio). Embryos (n = 30 per concentration) were exposed to FO (0, 0.375, 0.75, 1.5 mg/L) for 96 h, resulting in concentration-dependent developmental toxicity, including increased malformations, reduced heart rate, and inhibited swimming behavior. Adult zebrafish were chronically exposed to lower concentrations (0, 0.01, 0.1, 1.0 mg/L; n = 20 per concentration per replicate) for 28 days. Biochemical analyses across both life stages revealed that FO significantly inhibited succinate dehydrogenase (SDH) activity and mitochondrial complex II, reduced ATP levels, and induced oxidative stress. Integrated transcriptomic and metabolomic analyses showed that FO profoundly perturbed specific metabolic pathways, primarily glutathione metabolism, cytochrome P450-mediated detoxification, and core nutrient metabolism pathways involving carbohydrates, lipids, and amino acids. In adults, chronic exposure induced significant hepatotoxicity, evidenced by histopathological damage, altered liver enzyme activities (GPT/GOT), and activation of autophagy and PPAR/FoxO signaling pathways. Our results demonstrate that FO induces multifaceted toxicity in zebrafish, from developmental defects to hepatic metabolic dysfunction, primarily driven by mitochondrial impairment and oxidative stress. This study provides crucial mechanistic hazard data and insights for the ecological risk assessment of SDHI fungicides.

Journal of Xenobiotics doi: 10.3390/jox16020068

Authors: Albert Stuart Reece Gary Kenneth Hulse

Atrial septal defect (ASD) has become increasingly common in the USA and now affects 1 in 11.3 children in some places, but space–time analysis has not been applied to this emerging trend. ASD rate (ASDR) data were obtained from the National Birth Defects Prevention Network 2003–2020. Substance (cigarettes, alcohol, cannabis, analgesics, cocaine) use data were obtained from the National Survey of Drug Use and Health. Income data were obtained from the US Census. Analysis was limited to the Non-Hispanic White population by technical factors. Time-sequential univariate and bivariate maps were prepared for both covariates and outcomes and their combinations. Spatial regression of the ASDR was performed using the R package splm. A total of 7.6% of data was interpolated by linear regression. A total of 110,107 ASD cases were identified amongst 17,751,437 live births in 27 US states across 10 reporting periods. Time series maps showed that ASDR showed concordant patterns with indices of cannabis use rather than other substances. This was confirmed by multivariate spatial regression where cannabis and cannabinoids alone were found to significantly relate to ASDR, with p = 0.00002 for cannabidiol. Cannabis legal status similarly tracked with ASDR. Compared to states where cannabis was not legal, ASDR was more prevalent in cannabis-legal states (OR = 2.73 (2.66, 2.80); E-Value 4.90 (lower C.I. 4.76)). Twenty-seven of 34 (79.4%) E-values were >9 (high range) and 34/34 were > 1.25 (causal threshold). Data show that cannabis, including cannabis legalization, is driving the US ASD epidemic. While most high-ASDR states have high rates of cannabis use, Midwestern states where cannabis is farmed, such as Kentucky, Tennessee and Missouri, do not, suggesting other routes of exposure, potentially implicating environmental contamination. ASD is a bellwether marker for cannabinoid teratogenicity, indicating that communities should carefully control cannabinoid exposure and limit transgenerational cannabinoid genotoxicity more generally.

Journal of Xenobiotics doi: 10.3390/jox16020067

Authors: Ravikumar Jagani Jasmin Chovatiya Hiraj Patel Sandipkumar Teraiya Divya Pulivarthi Syam S. Andra

A comprehensive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the simultaneous quantification of 50 pesticide biomarkers across nine current-use chemical classes in human urine. These classes include organophosphorus insecticides (which encompass dialkyl phosphates and specific metabolites), pyrethroid insecticides, fungicides, neonicotinoid insecticides, herbicides, insect repellents, organochlorine pesticide metabolites, and plant growth regulators. The method employs solid-phase extraction (SPE) for sample preparation, requiring only 0.2 mL of urine. Chromatographic separation was optimized using a Hypersil Gold AQ column, achieving a total run time of 18 min. Mass spectrometric detection utilized polarity switching in electrospray ionization mode with multiple reaction monitoring. Method validation demonstrated satisfactory linearity (R2 > 0.99), high sensitivity with limits of detection ranging from 0.01 to 0.88 ng/mL, and extraction efficiencies between 85% and 113%. Precision and accuracy were within acceptable ranges, with relative standard deviations generally below 15%. The method’s robustness was confirmed through participation in external quality assessment schemes. Application to real samples revealed significant inter-individual variability in pesticide biomarker concentrations, with total measured biomarker levels ranging from 89 to 1242 ng/mL across the 10 individuals analyzed. This method offers comprehensive coverage of current-use pesticide chemical classes, including 30 biomarkers from the U.S. National Health and Nutrition Examination Survey (NHANES) biomonitoring program, and demonstrates improved sensitivity and broader analyte coverage compared to existing methods. The developed assay provides a valuable tool for large-scale biomonitoring studies and environmental health research.

Journal of Xenobiotics doi: 10.3390/jox16020066

Authors: Carlos Sanchez-Martin Mariagrazia D’Agostino Stefano Miglietta Veronica Cocetta Luna Laera Isabella Giacomini Martina Lanza Marica Mennini Maria Maddalena Storelli Ettore Cicinelli Monica Montopoli Alessandra Castegna

Background: The extensive use of insecticides in modern agriculture has raised concerns about potential chronic effects on human health beyond acute toxicity. Limited evidence exists regarding their impact on immune regulation and intestinal barrier integrity, two key components of host-environment interactions. Methods: Human in-vitro models were used to investigate the immunomodulatory and intestinal effects of several commonly used agricultural insecticides. Primary human macrophages derived from peripheral blood mononuclear cells were exposed to insecticides to assess cell viability and polarization status. Intestinal barrier function was evaluated using Caco-2 cell monolayers by measuring oxidative stress, epithelial integrity, paracellular permeability, and tight junction organization. Results: The tested insecticides induced a pro-inflammatory macrophage phenotype, characterized by increased expression of M1 markers and reduced M2 markers, without affecting cell viability. In Caco-2 cells, insecticide exposure compromised epithelial barrier integrity and disrupted tight junction organization. In this context, neither Spinetoram nor Spirotetramat induced notable oxidative stress under pro-oxidant conditions. However, Spirotetramat caused a significant increase in paracellular permeability. Conclusions: These findings indicate that commonly used insecticides can modulate immune responses and impair intestinal barrier function, suggesting potential mechanisms by which chronic low-level exposure may contribute to immune dysregulation and epithelial dysfunction in humans.

Journal of Xenobiotics doi: 10.3390/jox16020065

Authors: Phuong D. Tran Kyoungtae Kim

Perfluorooctane sulfonate (PFOS) is a persistent environmental pollutant associated with potential hepatoxic effects and other health risks. Despite its widespread distribution, the mechanisms underlying its toxicities remain to be fully understood. To investigate PFOS toxicology, our study utilized HepG2 and THLE-2 human hepatic cell models to replicate conditions reflecting PFOS accumulation in the liver. Cell viability, cell stress, and cell death assays were conducted to assess the toxicological influence of the chemical on both cell lines. Total RNA extraction was performed, followed by cDNA sequencing, and rt-qPCR. The XTT viability assay revealed a dose-dependent decrease in the number of viable cells when incubated with increasing concentrations of PFOS. The inhibitory concentration (IC50) values were approximately 100 micromolar, which led to morphological changes, elevated reactive oxygen species (ROS), and induced early apoptosis in liver cells after 6 h. Based on the transcriptomic analysis for HepG2 cells, mitochondrial genes involved in oxidative phosphorylation were downregulated, including COX, ND, and the ATP synthase family. Additionally, significant alterations of transcripts implicated in cell adhesion molecules (CAMs) were observed. In conclusion, PFOS inhibited cell growth, induced oxidative stress, and elevated apoptotic levels via transcriptomic alteration, including gene transcripts required for mitochondrial activity and cell adhesion.

Journal of Xenobiotics doi: 10.3390/jox16020064

Authors: Eirini Baira Evangelia N. Tzanetou Electra Manea-Karga Kyriaki Machera Konstantinos M. Kasiotis

Despite the undisputable ecosystem importance of honeybees, human activities have a substantial impact on their health. Since foraging is directly linked to a wide range of crops and bee-attracting flowers, plant protection products are at the forefront of chemical scrutiny, along with contamination of pollen, nectar, beehive components and water by other xenobiotics. In this study, a non-targeted Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS) screening was applied to 25 honeybee samples collected after reported death incidents in Greece. This approach led to the tentative annotation of over 50 compounds across various chemical classes, including pesticides, PFAS candidates not included in the EFSA “PFAS-4”, pharmaceuticals, antibiotics, industrial chemicals, and natural product constituents. In parallel, targeted pesticide residue analysis using liquid and gas chromatography coupled to tandem mass spectrometry (LC-MS/MS and GC-MS/MS) was performed, covering more than 250 active substances and providing direct quantitative results, revealing 11 active substances in concentrations ranging from <limit of quantification (LOQ) to 0.95 mg/kg, overlapping substantially with the HRMS detection. Overall, this study does not allow concrete causal attribution of mortality to specific chemicals; however, it documents complex co-occurrence patterns (pesticides together with other xenobiotics and plant bioactives), not excluding sublethal and mixture-toxicity effects. Quantified pesticide concentrations were below acute LD50-based thresholds, yet selected samples combined neonicotinoid/pyrethroid/fungicide signatures and other contaminants, supporting the need for mixture-toxicity frameworks and effect-based follow-ups.

Journal of Xenobiotics doi: 10.3390/jox16020063

Authors: Jelena Radovanović Sanja Milutinović-Smiljanić Biljana Antonijević Katarina Baralić Marijana Ćurčić Đurđica Marić Zoran Mandinić

The dose–response relationship for fluoride (F−) exposure remains largely unexplored. Hence, the current study assessed the hepatotoxic and nephrotoxic effects of subacute exposure (28 days) to increasing F− concentrations in Wistar rats via the benchmark dose (BMD5) method. Thirty male rats were assigned to six groups (n = 5): a control group (tap water) along with five groups that received F− via drinking water at increasing concentrations (10, 25, 50, 100, and 150 mg/L). F− toxicity was determined via water intake, weight gain, histological analyses, redox status, and essential element levels. PROASTweb 70.1 software was utilized to investigate the external and internal F− dose–response relationships. Specified major cytoarchitecture damage and superoxide anion (O2·−), total oxidative status (TOS), superoxide dismutase (SOD) activity, total thiol groups (SH), and advanced oxidation protein product (AOPP) level alterations were detected in both sets of tissues. Moreover, F− caused an imbalance in copper (Cu), zinc (Zn), iron (Fe), and manganese (Mn). The most sensitive parameters were O2·− (0.06 mg F−/kg) in the liver and AOPP (6.5 × 10−6 mg F−/L) in the kidneys. These findings contribute to the limited risk assessment of fluorides and highlight the dose-dependent relationship between redox status parameters and bioelements in the liver and kidneys.

Journal of Xenobiotics doi: 10.3390/jox16020062

Authors: Kleyson Willames da Silva Emilly Cruz da Silva Giulian César da Silva Sá Joane de Almeida Alves Darlisson de Alexandria Santos Alexandre Orsato Karoline Leite Dante Santos da Silva Adriano Richard Santos da Silva Zanderluce Gomes Luis Flavia Karoliny Araujo dos Santos José Augusto Pires Bitencourt Cristina Maria Quintella Pamela Dias Rodrigues Doumit Camilios-Neto Paul R. Race James E. M. Stach Sidnei Cerqueira dos Santos

The pervasive environmental dispersal of glyphosate has established this herbicide as a dominant anthropogenic xenobiotic, necessitating advanced bioremediation strategies to restore soil integrity. This study assessed the bioremediation efficacy of biosurfactants produced by Serratia ureilytica BM01-BS in glyphosate-contaminated soils, establishing their adsorption dynamics and ecotoxicological safety. The strain S. ureilytica BM01-BS gave a biosurfactant yield of 3.7 g·L−1 with promising surface properties, utilizing babassu (Attalea speciosa) waste as the sole nutrient source. Whole-Genome Sequencing and Biosynthetic Gene Cluster mining identified a Nonribosomal Peptide Synthetase cluster homologous to rhizomide-type lipopeptides responsible for biosurfactant production. Bioremediation assays in glyphosate-contaminated soils demonstrated a removal efficiency exceeding 95% in approximately 60 min, outperforming the synthetic surfactant SDS (20–30% efficiency). Kinetic and isothermal modeling suggest that the bioremediation process is governed by chemisorption, adhering to a pseudo-second-order model (R2 = 0.998) with a maximum adsorption capacity of 845 µg·kg−1. Fourier-Transform Infrared spectroscopy confirmed that the biosurfactant effectively removes glyphosate and restores the soil’s mineral integrity, as evidenced by the complete disappearance of glyphosate-associated phosphonic and carboxylic bands. Ecotoxicological assessments verified the environmental safety of the bioremediation process. These findings position the BM01-BS biosurfactant as a sustainable, biodiversity-based adjuvant for enhancing ecological resilience in glyphosate-impacted landscapes.

Journal of Xenobiotics doi: 10.3390/jox16020061

Authors: Ioana-Cezara Caba Raluca Stefănescu Alexandra-Andreea Botnaru Ionela Daniela Morariu Liliana Vereștiuc Florina-Daniela Cojocaru Bogdan Caba Oana Cioancă Alexandra Jităreanu Luminița Agoroaei

Glyphosate is the most widely used pesticide globally, raising concerns about its environmental persistence and biological impacts. Therefore, monitoring pesticide use is essential for assessing agricultural practices and the risks to human health associated with chemical use. This research examined glyphosate contamination in water (40 samples) and soil (28 samples) from northeastern Romania, an important agricultural region. Glyphosate concentrations in environmental water and soil samples were quantified using a spectrophotometric method based on ninhydrin derivatization, with good linearity over the concentration range 1–30 µg/mL (R2 = 0.9981). Glyphosate was detected at concentrations above the LOQ in one water sample. Also, the study proposes a UHPLC-MS/MS method for the confirmation of glyphosate presence in the analyzed sample. Additionally, this study contributes to the characterization of the toxicity profiles of glyphosate and a commercial glyphosate-based formulation (Roundup®) in primary human gingival fibroblast (hGF) cell lines. The commercial product Roundup, containing glyphosate, exhibited cytotoxicity similar to that of the active compound at low and intermediate doses; a significant cytotoxic effect was observed at a maximum concentration of 1 mM, with prolonged exposure. These findings demonstrate minimal cytotoxicity under the examined conditions and underscore the need for dose- and time-dependent assessments to evaluate the biological impact of herbicide formulations.

Journal of Xenobiotics doi: 10.3390/jox16020060

Authors: Jingyun Ding Stefanie Krais Zequn Li Rita Triebskorn Heinz-R. Köhler

Although chronic contamination by silver ions (Ag+) can persist in aquatic systems over long periods of time and can therefore have an impact on population developments, regulatory testing commonly relies on single-generation endpoints. Here, we used Daphnia magna to quantify long-term effects of pg/L to ng/L concentrations of Ag+ across generations and to test whether recovery depends on exposure history. Using 21 d life-cycle assays over up to seven consecutive generations, we quantified survival, key life-history traits, and population fitness (intrinsic rate of natural increase, r). In our study, low environmental concentrations of Ag+ caused minimal mortality, but sublethal effects persisted or multiplied over generations. Notably, continuous exposure led to significant reductions in body length and r at 50 pg/L (nominal LOEC) by the fourth generation exposed, representing population-relevant effects of Ag+ at very low concentrations which should be given consideration in the assessment of both water quality and the chemical itself. Recovery was concentration-dependent: low-concentration-exposed lineages recovered within a few generations, whereas 15 ng/L exposure resulted in persistent deficits even through the recovery period of three generations. Exposure-history patterns indicated that long-term outcomes were dominated by the cumulative number of exposed generations. These findings highlight the limitations of acute and single-generation assays and emphasize the importance of considering information on the effects of chemicals, including Ag+, across multiple generations in risk assessments. They also highlight the need to include expectations regarding recovery after the removal of pollutants in these assessments.

Journal of Xenobiotics doi: 10.3390/jox16020059

Authors: Maria-Nefeli Georgaki Despoina Ioannou Elpis Chochliourou Kanellos Skourtsidis Theodora Papamitsou Dimosthenis Sarigiannis

Background: Brain-derived neurotrophic factor (BDNF) is central to synaptic plasticity and neurodevelopment. Toxic metal exposure is linked to oxidative stress and neuroinflammation, yet its effects on BDNF signaling remain unclear. Objectives: To systematically synthesize evidence from human and experimental studies on the association between environmental or occupational metal exposure and BDNF alterations, and to highlight research gaps with an emphasis on hexavalent chromium (Cr(VI)). Methods: PubMed, Scopus, and ScienceDirect were searched following PRISMA guidelines. Eligible studies included human observational research and animal models reporting quantitative associations between metal exposure (biomarkers/environmental measures) and BDNF outcomes (protein or gene expression). Data were extracted on exposure assessment, BDNF measurement, and neurobehavioral outcomes. Study quality was assessed using NOS (human studies) and SciRAP (experimental studies). Results: Nineteen studies were included. Across metals such as Pb, Hg, Cd, As, Mn, and mixtures, exposure was associated with altered BDNF levels in blood or brain tissue, often alongside oxidative stress markers, inflammatory changes, and cognitive or behavioral impairment in animal models. Most human studies reported decreased circulating BDNF with higher exposure, while experimental evidence suggested context-dependent regulation across exposure windows and brain regions. Conclusions: The available evidence supports a biologically plausible link between metal exposure and BDNF dysregulation. No eligible studies evaluated BDNF in relation to Cr(VI), indicating a major research gap. Future studies should integrate neurotrophic biomarkers with exposome-oriented designs to clarify chromium-related neurotoxicity and support Adverse Outcome Pathway (AOP)-informed frameworks.

Journal of Xenobiotics doi: 10.3390/jox16020058

Authors: Hai-Tao Shen Jian-Long Han Xiao-Min Xu Xiao-Dong Pan

This study presents a comprehensive investigation of OPE residues, distribution patterns, and dietary exposure risks in crustaceans from southeast China. OPEs were detected in over 90% of samples, with mean total concentrations (ΣOPEs) of 5.80 μg/kg wet weight (ww) in freshwater shrimp, 6.52 μg/kg ww in marine prawn, and 1.25 μg/kg ww in marine crab. Tributyl phosphate (TiBP), triethyl phosphate (TEP), and tris(2-chloroethyl) phosphate (TCEP) emerged as the dominant congeners, accounting for 68.1% of ΣOPEs, which indicates inputs from industrial emissions, plastic waste leaching, and aquaculture equipment. Spatial analysis revealed striking regional differences: coastal industrial cities (Zhoushan, Taizhou) exhibited ΣOPE levels up to 12-fold higher than inland mountainous areas (Quzhou, Lishui), while no significant temporal variations were observed. Human health risk evaluation, based on estimated daily intake (EDI) and target hazard quotient (THQ), demonstrated negligible non-carcinogenic risks for the general population (HI < 1), though children and frequent seafood consumers have slightly elevated exposure. These findings indicate the value of crustaceans as bioindicators for OPE contamination and require long-term monitoring of emerging OPEs and their synergistic effects with co-occurring pollutants.

Journal of Xenobiotics doi: 10.3390/jox16020057

Authors: Claudia A. Rocha Luís M. Félix Dércia Santos Sandra M. Monteiro Carlos Venâncio

Wildlife acts as a sentinel of environmental pollution, providing critical insights into potential risks to human health within the One Health framework. However, knowledge on the occurrence of legacy and emerging contaminants in wildlife, as well as their potential trophic transfer to humans, remains limited. Thus, monitoring contaminants in terrestrial wildlife, particularly in game species, is especially relevant, as game meat represents an important source of high-quality protein that must be safeguarded. This review summarizes current evidence on chemical contaminant levels in terrestrial wildlife from a “One Health” perspective. Despite the growing relevance of this approach, few studies have explicitly applied this term, and even fewer have focused on game meat, resulting in an incomplete picture of contamination. Although reported contaminants—metals, metalloids, pesticides, microplastics, and mycotoxins—originate from overlapping natural and anthropogenic sources, such as ammunition, agriculture, and industrial activities, a strong dependence on local environmental conditions continues to hamper cross-regional comparisons and the establishment of representative exposure levels. Overall, this review highlights the need for systematic monitoring of contaminants in terrestrial wildlife, with emphasis on emerging pollutants that are currently underrepresented in literature, to improve risk assessment, protect food safety, and better understand the impacts of environmental contamination on animal and human health.

Journal of Xenobiotics doi: 10.3390/jox16020056

Authors: Maya Radeva-Ilieva Stanila Stoeva-Grigorova Ivanesa Yarabanova Ivelina Panayotova Georgi Bonchev Nadezhda Hvarchanova Mario Milkov Simeon Marinov Petko Marinov Snezha Zlateva

Caffeine is widely consumed and generally considered safe at customary doses. How-ever, high-dose preparations available online pose a risk of severe and potentially fatal intoxication. Although uncommon, lethal caffeine poisoning is associated with profound cardiovascular and neurological toxicity. A rare case of intentional acute caffeine intoxication with fatal outcome is presented. A 25-year-old woman ingested an estimated 60 tablets containing 200 mg of caffeine each, purchased online. She was admitted to hospital shortly after ingestion of the caffeine tablets with palpitations, agitation, dizziness, and repeated vomiting. On examination, she presented with arterial hypotension (90/60 mmHg) and marked sinus tachycardia (150 beats/min), accompanied by psychomotor agitation. Her blood caffeine concentration measured by means of high-performance liquid chromatography (HPLC) was 177 µg/mL. The patient’s condition rapidly deteriorated, with the development of convulsive syndrome progressing to coma, extreme ventricular tachycardia, exotoxic shock, and toxic cardiomyopathy. Despite intensive care management, including mechanical ventilation and advanced cardiopulmonary resuscitation, the patient died several hours after admission. In conclusion, this case underscores the life-threatening potential of acute high-dose caffeine ingestion and highlights the risk associated with unrestricted access to concentrated caffeine products. Early recognition and aggressive management are crucial, yet may be insufficient in cases of massive overdose.

Journal of Xenobiotics doi: 10.3390/jox16020055

Authors: Jéssica Veiga-Matos Daniel J. V. A. dos Santos Andreia Palmeira Emília Sousa Ana I. Morales Marta Prieto Fernando Remião Renata Silva

P-glycoprotein (P-gp), an efflux transporter highly expressed in renal tubules, plays a crucial role in the detoxification and protection of barrier/excretory tissues from harmful xenobiotics. Xanthones and thioxanthones (TXs) are known for their antimicrobial and antitumor activities and for their ability to modulate membrane transporters such as P-gp. Previous studies have reported that (thio)xanthonic derivatives enhance P-gp expression and/or activity in intestinal cells, reducing the intracellular accumulation of toxic substrates; however, their capacity to modulate P-gp in renal cells remains poorly explored. This study aimed to predict, in silico, TXs’ binding sites within P-gp and to evaluate, in vitro, in human kidney (HK)-2 cells, the effects of selected TXs (TX1–5) on P-gp activity and expression, and protection against cisplatin-induced cytotoxicity. Computational studies identified preferential TX1–5 binding to the drug-binding pocket, particularly the rhodamine 123 (R) or modulator (M) sites, and to nucleotide-binding domain 1. In vitro, rhodamine 123 accumulation assays revealed increased P-gp transport activity after 120 min or 24 h exposure to TX1–5, except TX4. TX2 elicited the strongest effect (141% increase, p < 0.0001), upregulated P-gp expression (24 h, p < 0.0001), and significantly protected HK-2 cells from cisplatin-induced cytotoxicity (increased IC50, p < 0.0001). Altogether, these findings position thioxanthones as promising scaffolds for the development of P-gp-targeted strategies to mitigate drug-induced nephrotoxicity.

Journal of Xenobiotics doi: 10.3390/jox16020054

Authors: Hannah M. Starnes Scott M. Belcher

Per- and polyfluoroalkyl substances (PFAS) constitute a large and chemically diverse class of synthetic compounds characterized by one or more fully fluorinated methyl or methylene groups. Many PFAS are toxic, environmentally persistent, bioaccumulative, and highly mobile, resulting in widespread contamination and biological exposure. Across taxa PFAS exhibit affinity for proteins and preferentially accumulate in protein-rich, highly perfused tissues. Protein binding critically influences PFAS distribution, bioaccumulation, toxicity, and elimination. A variety of different approaches for determining binding affinity have existed for decades; however, depending on experimental conditions, calculated affinities can vary over multiple orders of magnitude which limits comparison of protein–PFAS binding affinities across studies and across PFAS chemical space. Addressing this limitation requires robust and standardized experimental platforms capable of rapidly generating quantitative binding data. Among the most important targets is serum albumin—the principal transport protein in vertebrate blood—which plays a central role in governing PFAS toxicokinetics. This review summarizes current methodologies for measuring protein–PFAS binding affinities, evaluates the strengths and limitations of each approach, synthesizes the existing literature on serum albumin–PFAS interactions, and highlights differential scanning fluorimetry as a rapid, reproducible, and sensitive technique for in vitro assessment of relative protein–PFAS binding.

Journal of Xenobiotics doi: 10.3390/jox16020053

Authors: Olawale Razaq Ajuwon Damilola Rebecca Oladejo Akinwunmi Oluwaseun Adeoye John Adeolu Falode Basiru Olaitan Ajiboye Foluso Oluwagbemiga Osunsanmi Babatunji Emmanuel Oyinloye

Diabetes mellitus (DM) is a major disorder contributing to human mortality and morbidity globally. The use of medicinal plants in the management of diabetes is gaining global popularity due to their accessibility and cost-effectiveness. In this study, we evaluated the ameliorative potential of Vernonia amygdalina leaves crude extract (CE), free phenol (FP), and bound phenol (BP) fractions (50 mg/kg body weight) in a rat model of streptozotocin (STZ)-induced type 1 diabetes (T1DM). The effects of these treatments for 28 days on glucose, insulin, glycated hemoglobin, hepatic injury indices, and lipid profile were assessed in the serum. Furthermore, redox biomarkers (liver) and inflammatory mediators (serum and liver) were analyzed. Our results indicated that CE, FP, and BP fractions of Vernonia amygdalina inhibited the deleterious effects of T1DM by attenuating hyperglycaemia, insulin deficiency, hepatic injury, and dyslipidemia. Also, CE, FP, and BP fractions differentially improved antioxidant enzymes activity and reduced oxidative and inflammatory markers production. Specifically, CE showed superior effects compared with FP, BP, and metformin across multiple biomarkers, including glycated hemoglobin, α-amylase, α-glucosidase, hepatic glycogen, total cholesterol, LDL-cholesterol, protein carbonyl, SOD, IL-1β, and IL-10. The antidiabetic effects produced by CE, FP, and BP fractions of Vernonia amygdalina may be ascribed to the presence of different bioactive phytochemicals as revealed by HPLC analysis. Overall, our data would suggest a potential therapeutic role for Vernonia amygdalina leaves extracts in addressing hepatic complications due to T1DM.

Journal of Xenobiotics doi: 10.3390/jox16020052

Authors: Chantalle Moulton Anna Baroni Ennio Tasciotti

Micro- and nanoplastics (MNPs) are pervasive environmental contaminants with growing relevance for human health across the lifespan. Older adults may be especially vulnerable to their effects due to cumulative lifetime exposure, age-related physiological changes, and a higher burden of chronic disease. Adopting a One Health perspective, this review synthesizes current evidence on the sources, exposure pathways, and biological effects of MNPs, integrating findings from environmental, animal, and human studies with a specific focus on aging populations. Experimental studies consistently show that MNP exposure triggers oxidative stress, inflammation, mitochondrial dysfunction, and cellular senescence, mechanisms central to biological aging. These processes are linked to dysfunction of the cardiovascular, nervous, gastrointestinal, and immune systems, suggesting that MNPs may contribute to the development or progression of age-related diseases. Within the One Health framework, MNPs also act as carriers of chemical additives and environmental pollutants, potentially amplifying health risks through combined and cumulative exposures along food chains and ecosystems. Despite increasing mechanistic evidence, direct epidemiological data in older adults remain limited. This review highlights key knowledge gaps and emphasizes the need for integrative, longitudinal research to clarify the role of MNPs in aging and to inform public health and environmental policy.

Journal of Xenobiotics doi: 10.3390/jox16020051

Authors: Divya Mishra

Glyphosate, one of the most widely used herbicides worldwide, has raised significant concerns regarding its potential involvement in hepatotoxicity and molecular changes associated with liver cancer biology. These concerns highlight the need to better understand its underlying molecular mechanisms in hepatoma cells. Emerging evidence suggests that glyphosate exposure may increase the risk of liver cancer and chronic liver disease. However, the precise molecular alterations and promising biomarkers associated with glyphosate-induced hepatic toxicity and disease remain largely unexplored. In this study, an RNA-Seq-based in silico systems biology approach was employed to elucidate glyphosate-induced differential transcriptional profiling in hepatoma cells. This analysis revealed significant transcriptional profiling characterized by the upregulated hub genes ATF3, JUNB, ALDOA, FOSB, PFKFB3, G6PD, ENO2, HK2, FOS and PGK1. These genes were primarily associated with glucose metabolism, TNF-α/NF-κB signaling, epithelial–mesenchymal transition (EMT) and cellular stress responses. Conversely, several key genes were significantly downregulated, including PIK3R1, FYN, CEBPA, MLXIPL, PPARA, CD36, PCK2, PNPLA3, NR1H4 and MGLL, which were involved in lipid metabolism, immune regulation and non-alcoholic fatty liver disease (NAFLD) pathways. Notably, all hub genes demonstrated strong diagnostic performance, highlighting their potential as sensitive biomarkers of glyphosate exposure. Collectively, this study provides comprehensive insights into gene expression changes associated with glyphosate exposure in hepatoma cells, linking them to hepatic metabolic dysregulation and immune modulation and suggesting a panel of hub genes with potential diagnostic and therapeutic significance.

Journal of Xenobiotics doi: 10.3390/jox16020050

Authors: Sosana Bdir İrfan Çapan Mohammed Hawash Süleyman Servi Mohammad Qneibi

Background: Fast excitatory transmission in the central nervous system is carried out by AMPA-type glutamate receptors. Neuronal hyperexcitability and epilepsy have been associated with the dysregulation of AMPA receptor function. Modulation of the gating kinetics of AMPA receptor function has been proposed to be a desirable target for therapy, especially when the modulation is transmembrane AMPA receptor regulatory protein (TARP)-dependent and AMPA receptor subunit composition-dependent. Methods: Eight dibenzobarrelene-based heterocycles were characterized for their effects on the human embryonic kidney cells expressing homomeric GluA1 and heteromeric GluA1/2 AMPA receptors, either alone or co-expressed with the TARPγ8 auxiliary subunit, using whole-cell patch-clamp electrophysiological recordings, and the current amplitude and kinetics of desensitization and deactivation were measured after rapid glutamate application. Results: Each chemical evaluated suppressed glutamate-induced currents via AMPA receptors and augmented both desensitization and deactivation, indicating a negative allosteric modulatory effect. The co-expression of TARPγ8 diminished, but did not eradicate, the inhibition and acceleration induced by the compounds. The observations indicate that the chemicals diminish agonist-bound open states and facilitate transitions to non-conducting states while maintaining effectiveness. Conclusions: The present study describes a specific kinetic mechanism by which dibenzobarrelene derivatives impair the function of the AMPA receptor and its dependence on auxiliary proteins. The present study provides a mechanistic understanding of AMPA receptor gating modulation and establishes a pharmacological framework for future investigations in more physiologically relevant systems.

Journal of Xenobiotics doi: 10.3390/jox16020049

Authors: Emoke Dalma Kovacs Nguyen Khoi Nghia Melinda Haydee Kovacs

Background and Objectives: Polyethylene (PE) mulching enhances crop productivity through microclimate optimization but introduces synthetic polymer-derived compounds into agricultural soils. Despite widespread use, biochemical and microbial impacts of PE mulch emissions remain poorly understood. This study investigated the impact of PE mulch emissions on soil metabolomes and microbial communities during field pea (Pisum sativum L.) cultivation. Methods: A 75-day field experiment compared PE-mulched and non-mulched soils across five temporal sampling points (T0–T4). Headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry was used to identify PE-derived organic compounds in mulched soils. Microbial community structure was assessed through the phospholipids derived fatty acids (PLFA) approach, whereas mass spectrometric untargeted metabolomics was used to characterize the soil biochemical profiles. Results: Analysis identified 18 PE-derived organic compounds (n-alkanes, phthalates, and additives) in the mulched soils. PE mulching significantly increased bacterial abundance (anaerobic bacteria, actinomycetes, and aerobic bacteria) but suppressed all functional fungal guilds, particularly saprotrophic fungi (30% reduction) and arbuscular mycorrhizal symbionts. PE-derived organic compounds were associated primarily with the first RDA axis (RDA1), which alone explained 44.6% of the metabolome variance. These compounds presented strong positive correlations with organic nitrogen compounds and lipids and negative correlations with benzenoids and nucleotides. Pathway analysis revealed perturbations in energy metabolism, lipid metabolism, and xenobiotic degradation pathways. Conclusions: PE mulch emissions differentially shift soil microbial communities and metabolic networks, with bacterial proliferation contrasting with fungal suppression. These findings highlight the complex trade-offs between agronomic benefits and soil biological impacts, emphasizing the need for sustainable mulching alternatives.

Journal of Xenobiotics doi: 10.3390/jox16020048

Authors: Denis Oberiukhtin Anton Chernitskiy Desheng Hu Alexey Sarapultsev

Cyclophosphamide (CP) is extensively used in oncology and as an immunosuppressant, but dose-limiting renal injury remains a major constraint. We systematically reviewed in vivo rodent models of CP nephrotoxicity (2010–2025) and meta-analysed core outcomes while separating the model effect (CP vs. control) from the treatment effect (intervention + CP vs. CP-only). Fifty-four studies met eligibility criteria, and random-effects syntheses were feasible for serum creatinine, serum urea, and renal oxidative stress markers. CP produced a marked functional deterioration, increasing serum creatinine by 1.059 mg/dL (95% CI 0.517–1.601; k = 9) and serum urea by 39.852 mg/dL (95% CI 6.557–73.148; k = 9). Across intervention studies, protective effects were most consistently expressed in oxidative endpoints (MDA/TBARS reduction and glutathione preservation), whereas functional recovery estimates were more variable and frequently limited by incomplete reporting and between-study heterogeneity. Overall, the evidence base supports CP as a robust preclinical model of combined functional and redox-mediated renal injury and indicates that multiple mechanistic classes of interventions can partially mitigate injury, but current reporting and design heterogeneity preclude reliable ranking of candidate agents. The protocol was registered on OSF.

Journal of Xenobiotics doi: 10.3390/jox16020047

Authors: Mohammed Hawash Benay Mahmutoğlu Murad Abualhasan Deniz Cansen Kahraman Sultan Nacak Baytas

The indole scaffold is common in natural products and bioactive compounds, including anti-cancer and anti-inflammatory medicines. In this work, a series of indole-acrylamide derivatives was synthesized, and their antiproliferative and anti-inflammatory effects were evaluated on COX enzymes and against a panel of cancer cell lines. All the final compounds were characterized via HRMS and (1H & 13C)-NMR. Anticancer and anti-inflammatory activities were evaluated using standard biomedical techniques by SRB, MTS, and COX kit assays. Additionally, the molecular docking analysis was conducted using the AutoDock Vina tool. The results demonstrated that the produced compounds displayed significant inhibitory effects on the COX-2 enzyme, with IC50 values of 128 nM to 1.04 µM. 6a demonstrated significant COX-2 selectivity with an IC50 of 128 nM and an SI of 352, highlighting its preference for COX-2 over COX-1. 6c exhibited potent COX-2 inhibition with an IC50 of 0.215 µM and an SI of 10.6. The assessed compounds exhibited substantial cytotoxic effects on cancer cells, especially against liver cancer cell lines (Huh7, HepG2, Mahlavu, and SNU475), and breast cancer (MCF-7). 6d compound was the most COX-1 selective inhibitor, which observed potent activity against hepatocellular carcinoma, with IC50 values as low as 3.5 µM, and was highly effective against MCF-7. Additionally, COX-2 selective inhibitors, 6a and 6b, exhibited strong antiproliferative effects against both breast cancer (MCF-7) and melanoma (B16F1), with IC50 values ranging from 4.75 to 15.4 µM. Furthermore, the molecular docking of 6a demonstrated a strong affinity for the COX-2 enzyme, with energy scores (S) of −8.392 kcal/mol, comparable to celecoxib’s score of −10.96 kcal/mol. The findings suggest a possible correlation between COX-2 inhibition and anticancer efficacy, especially for compounds 6a and 6c, which demonstrate excellent COX-2 selectivity and notable antiproliferative effects, positioning them as prospective candidates for further advancement in cancer treatment.

Journal of Xenobiotics doi: 10.3390/jox16020046

Authors: Gabriel Pérez-Lucas Simón Navarro

Farmers rely on pesticides to keep their crops safe from pests, diseases, and weeds. However, if pesticides are not used properly, they can have serious consequences for human and environmental health. Many pesticides are not easily biodegradable and persist in the environment for a long time. Their residues, including toxic metabolites, pose risks to non-target organisms, contaminate surface- and groundwater sources, and may affect future crops. Among other soil remediation actions, it is important to highlight the impact of agricultural waste and agro-industrial byproducts on the behavior of pesticides as a strategy to eliminate or at least minimize soil pollution by their residues. Waste from various food industries and agriculture poses a severe threat to the ecosystem and is difficult to manage properly. Agriculture and food production waste accounts for over 30% of total global agricultural output. Therefore, managing agri-food waste from different sources is crucial to promoting sustainable development with minimal environmental impact. Key components of waste management interventions in the agricultural circular and bioeconomy include incorporating crop residues and food waste into soils. For these reasons, we present an updated review of the impact of agricultural waste and agro-industrial byproducts on the behavior of pesticides in soil. The goal of this review is to promote the sustainable use of these wastes within the context of a circular economy.

Journal of Xenobiotics doi: 10.3390/jox16020045

Authors: Carlos J. A. Campos Louis A. Tremblay Olivier Champeau Gregory Goblick

Fluorescent dyes are commonly used as tracers in hydrological investigations to quantify transport pathways, residence times, mixing behavior, and connectivity in surface water, groundwater, and coastal systems. Despite their long history of application, the ecological implications of deliberate dye releases are not well understood. This review synthesizes current knowledge on the physico-chemical characteristics, environmental behavior, and ecotoxicological effects of major dye classes, with emphasis on rhodamines, fluorescein derivatives, and sulfonated xanthene dyes commonly used in water tracing studies. Toxicity data for algae, cyanobacteria, invertebrates, and fish show large inter-specific variability. Some dyes, particularly rhodamine B and eosin Y, show acute or sub-lethal effects at concentrations detected during poorly controlled applications. By contrast, dyes with high polarity and extensive sulfonation (e.g., rhodamine WT, sulforhodamine B, pyranine, and fluorescein) show consistently low toxicity and minimal bioaccumulation potential. Environmental fate processes, including photolysis, sorption, and transformation into potentially more reactive products, influence exposure dynamics, especially in clear, shallow, or slow-moving systems. This review also evaluates regulatory frameworks and operational guidance for safe use, identifies gaps in toxicological and fate data, and proposes recommendations for minimizing environmental impact through dye selection, mass optimization, injection design, and monitoring. The findings support the continued use of fluorescent dyes but highlight the need for more systematic assessment of transformation products, chronic and sub-lethal responses, and cumulative exposure in sensitive environments.

Journal of Xenobiotics doi: 10.3390/jox16020044

Authors: Thanakorn Sawangchart Sutee Chutipaijit Bunyarit Meksiriporn Worapat Narueban Worrathon Tilokkarn Pattareewan Imsuwan Thanawat Sutjaritvorakul

Fungal transformation is increasingly recognized as an important process influencing metal solubilization and immobilization in soil environments. In this study, a fungal strain (PTW4) isolated from mining-contaminated soil was molecularly identified as Aspergillus aculeatus. The strain was evaluated for its ability to solubilize and transform several heavy metal oxides, including ZnO, Pb3O4, Cu2O, and MoO3. PTW4 produced consistent halo formation across all tested oxides, accompanied by progressive acidification of the culture medium, suggesting organic acid-mediated solubilization. Characterization of extracellular precipitates by SEM-EDS and XRD indicated mineral phases consistent with oxalate-associated biominerals, including zinc oxalate dihydrate (ZnC2O4·2H2O), lead oxalate (PbC2O4), and copper oxalate hydrate (CuC2O4·xH2O). These minerals represent low-solubility phases that may reduce metal mobility in the surrounding environment. In contrast, molybdenum did not precipitate under the experimental conditions, suggesting metal-specific constraints in fungal biomineralization processes. Although organic acid production was not directly quantified, identification of oxalate mineral phases supports an oxalate-associated mineralization mechanism. Overall, the results provide evidence for heavy metal solubilization and selective extracellular precipitation consistent with oxalate biomineral formation by A. aculeatus PTW4, highlighting its potential relevance to fungal-mediated bioremediation and selective bioleaching processes.

Journal of Xenobiotics doi: 10.3390/jox16020043

Authors: Albert Stuart Reece Gary Kenneth Hulse

Atrial septal defect (ASD) affects 1:11.3 children in some US states; however, the antecedents of these trends are yet to be identified. A total of 1882 ASD rates (ASDRs) for 2003–2020 were sourced from the National Birth Defects Prevention Network reports. A total of 406,893 ASDs are reported. Substance (cigarettes, binge alcohol, cannabis, cannabinoids, analgesics, cocaine) exposure data were taken from the National Survey of Drug Use and Health. Income and ethnicity data were derived from the US Census. Adjustment was performed by mixed effects, survey and generalized additive regression. Causal analysis was by inverse probability weighting and E-values. Data were analyzed in RStudio. The highest ASDR of 884/10,000 live births was amongst Non-Hispanic Asians and Pacific Islanders in Nevada in 2016–2020. The 2005–2018 median ASDR rose >12-fold in Nevada and New Mexico, >6-fold in New York, and 4.2-fold nationally 1989–2020; it doubled in NY from 2012–2016 to 2016–2020. The average state ASDR rose supra-exponentially (p = 0.0075) and was associated with higher cannabis use states (p = Zero, Cohen’s D = 1.24), apparently driven by cannabis legalization (p = Zero). Estimated exposures to Δ9THC, cannabidiol and cannabigerol were implicated (from p = 2.67 × 10–68). Cannabis-legal states were compared with others (mean ASDR (C.I.) 178.15 (131.68, 224.62) vs. 74.28 (70.60, 77.96), p = Zero; O.R. 1.82 (1.81, 1.84), E-values 3.04 (lower C.I. 3.02), Cohen’s D 1.29 (0.96, 1.62)). Overall, 29/39 (74.4%) E-value estimates were >4; 39/39 (100%) were >1.25. Cannabis, cannabinoids and cannabis legalization are strong candidates for driving the US ASDR supra-exponentially. Estimates of many cannabinoids, including cannabidiol, Δ9THC, and cannabigerol, are implicated. The results are consistent with other large epidemiological studies. The importance of the results is magnified by the increasing legalization and penetration of cannabinoids into the US population. Since therapeutic abortion is not practiced for ASD, it may be used as a bellwether index of heritable transgenerational cannabinoid genotoxicity and epigenotoxicity associated with cannabinoid exposure.

Journal of Xenobiotics doi: 10.3390/jox16020042

Authors: Eleonora Conterosito Mariachiara Lo Scalzo Marysol Ferretti Andrea Rosmino Simona Stradella Mauro Mottin Erika Mottin Valentina Gianotti

In the context of plastic recycling, legislation is evolving and varies across regions, but it remains largely nonspecific. In the European context, producers of post-industrial and post-consumer recycled plastics must guarantee the same wholesomeness as virgin materials. However, they cannot maintain such strict control over incoming materials, because, since the secondary raw materials derived from separate waste collection, they are subjected to high variability in composition and heterogeneity over time. In this frame, a rapid, and easy-to-apply GC-MS method was developed. It employs a liquid–liquid extraction with acetone, followed by quantitative analysis with gas chromatography coupled to mass spectrometry (GC-MS). A combination of total ion chromatograms (TICs) and extracted ion chromatograms (EICs) was used. Adequate sensitivity was demonstrated in the selected concentration ranges for most of the analytes, with limits of quantification (LOQs) lower than the legislative limit, when existing. The results showed that the method is sufficiently accurate with recoveries ever higher than 68.3% and relative standard deviations (RSDr) smaller than 4.2%. This method allows, for the first time, the simultaneous quantification of 40 molecules at levels of a few ng/g. It ensures the possibility of obtaining real-time data for the production control system about the safety of the input materials, allowing immediate corrective action in the event of anomalies. This method is focused on PE and PP recycled plastics and is to be considered a screening method that allows for highlighting batches of incoming materials that are too contaminated to control the output material. This method was successfully tested analyzing some batches of plastics both in input and post-recycling.

Journal of Xenobiotics doi: 10.3390/jox16020041

Authors: Iria Osa-Subtil Teolincacihuatl Romero-Rosales María José Dios-Duarte

Background/objective: The widespread use of pesticides in modern agriculture has raised increasing concern about their potential adverse effects on human health. Exposure to these compounds has been linked to multiple negative health outcomes. This systematic review aims to evaluate and synthesise the available scientific evidence on the effects of pesticide exposure on human health during agricultural production—with particular emphasis on alterations in gut microbiota and intestinal membrane permeability—by integrating results from experimental and observational studies conducted on animals and humans. Methods: This systematic review was conducted in accordance with PRISMA guidelines. A systematic literature search was carried out using the main databases Medline/PubMed, Embase and Web of Science, introducing the search algorithm “pesticides” AND “gut microbiota”, from which a total of seven systematic reviews that met our inclusion criteria were found and subsequently analysed. The quality assessment was based on the principles of evidence-based medicine. This systematic review was registered in the OSF. Results: The findings indicate that prenatal exposure to pesticides is linked to adverse outcomes in foetal development. Additionally, pesticide exposure affects metabolic, immune, and nervous system function due to alterations in gut microbiota composition and membrane permeability. Evidence from animal model studies complements human data by providing insight into the underlying biological mechanisms, such as oxidative stress, liver dysfunction, alterations in hormonal signalling and activation of the inflammatory response. Conclusions: Public health strategies should prioritise reducing pesticide exposure, strengthening environmental protection and supporting further research on gut microbiota modulation and intestinal membrane permeability. Such measures may contribute to the prevention and mitigation of pesticide-related health disorders. Limitations: Human data are insufficient to establish clear causal relationships. Moreover, substantial variability among pesticide types and the difficulty of distinguishing the effects of complex mixtures from those of individual compounds complicate interpretation of the findings.

Journal of Xenobiotics doi: 10.3390/jox16020040

Authors: Veronika Svitková Ivana Horáková Viliam Kolivoška Eva Vaněčková Olívia Dakošová Eva Melníková Dušan Žabka Zuzana Imreová Alexandra Tulipánová Alexandra Paulína Drdanová Marek Haššo Peter Nemeček Michal Hatala Tomáš Mackuľak Miroslav Gál

The rapid, field-ready detection of methamphetamine (MET) directly in sewage under flow remains a bottleneck for public health and law enforcement surveillance. We engineered a low-cost, 3D-printed flow-through electrochemical cell that houses a commercial screen-printed carbon electrode and operates in both non-flow and flow regimes. The platform was validated using the [Ru(NH3)6]3+/2+ couple, confirming negligible kinetic hindrance and suitability for voltammetric sensing under convective transport. Using square wave voltammetry and chronoamperometry, MET was quantified in filtered wastewater, with limits of detection of 15.9 µg L−1 in non-flow and 211.2 µg L−1 in flow conditions. Specificity tests yielded well-separated faradaic responses for the pre precursor α-phenylacetoacetonitrile (APAAN) and for MET, while amphetamine produced only a weak signal, enabling side-by-side discrimination in a single run. To our knowledge, this is the first demonstration of direct electrochemical sensing of MET in flowing wastewater using a 3D-printed flow-through platform. The simple, disposable design provides an actionable foundation for portable, near-real-time sewer surveillance and motivates antifouling/auto-cleaning strategies for long-term deployment.

Journal of Xenobiotics doi: 10.3390/jox16020039

Authors: Francisco Alejandro Lagunas-Rangel

Environmental pollutants are persistent chemicals that pose substantial risks to human health, contributing to global mortality and economic burden. In real-world situations, exposure rarely occurs to single compounds; instead, people are chronically exposed to complex mixtures at low concentrations. However, most regulatory frameworks still rely on single-substance risk assessments, potentially underestimating the hazards associated with combined exposures. This study investigated the cytotoxic interactions of binary mixtures of five environmentally relevant pollutants: bisphenol A (BPA), bisphenol A diglycidyl ether (BADGE), dibutyl phthalate (DBP), di(2-ethylhexyl) phthalate (DEHP), and perfluorooctanoic acid (PFOA), using the human lymphoblast cell line NALM-6. Cells were exposed for 72 h to each compound individually and to all possible binary combinations, reflecting concentrations reported in human plasma or serum. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and interactions were analyzed using the Bliss model of independence and two-way analysis of variance (ANOVA). Intracellular reactive oxygen species were measured using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe to explore the involvement of oxidative stress. Synergistic interactions were observed under specific conditions, although not all statistically identified interactions corresponded to biologically significant effects. The BPA-DBP combination produced the highest cytotoxicity when both pollutants were present at 100 nM (31%), consistent with a strong synergistic effect. A similar pattern was observed for BADGE-BPA. ROS production was partially associated with cytotoxicity in these selected mixtures. Overall, these findings highlight the importance of distinguishing statistical synergy from toxicological relevance.

Journal of Xenobiotics doi: 10.3390/jox16010038

Authors: Antonio Peña-Fernández Manuel Higueras Roberto Valiente Borox M. Carmen Lobo-Bedmar

Rare earth elements (REEs) and technology-related trace elements are increasingly used in modern products and processes, but biomonitoring data in healthy children and adolescents remain scarce; scalp hair provides a practical, integrative matrix for assessing multi-element patterns over time. Scalp hair collected in April–May 2001 from children (6–9 years; n = 120) and adolescents (13–16 years; n = 97) living in Alcalá de Henares (Spain) was retrieved from archival storage and analysed in 2025 using a single QA/QC-controlled ICP–MS workflow. Seven REEs (Ce, La, Pr, Nd, Gd, Er, and Y) and nine technology-related trace elements (Bi, Sb, Th, U, Pd, Pt, Rh, Ir, and Rb) were quantified after rigorous decontamination; left-censored data were treated using Kaplan–Meier, regression on order statistics, and maximum-likelihood approaches, and population reference values were derived as percentile-based upper limits (P95, 95% CI). In children, REEs were frequently detected and showed strong within-suite covariation, with medians in the low ng g−1 range (e.g., Ce ≈ 0.011 µg g−1; La ≈ 0.007 µg g−1), whereas in adolescents, most REEs were near reporting limits. Sb and U were ubiquitous in both age groups, while platinum-group elements were largely undetected. Shale-normalised REE patterns were subparallel across normalisers, La/Ce anomalies were centred below unity, and weak soil–hair correlations suggested multiple microenvironmental exposure pathways. These data provide a robust pre-diffusion baseline for REE metals in European youth, offering a benchmark for future urban exposome assessments.

Journal of Xenobiotics doi: 10.3390/jox16010037

Authors: Raluca Maria Stirbescu Cristiana Radulescu Raluca Maria Bucur (Popa) Andreea Laura Banica Ioan Alin Bucurica Ioana Daniela Dulama

This research aimed to quantify and investigate the morphology of microplastics in skincare and treatment creams related to their chemical composition and the potential risks to human health associated with exposure to microplastics by dermal contact. A total of 21 skincare and treatment cream samples, indicating the target audience (men, women, and children) for each product, and potential diseases were analyzed in terms of the hidden risk of microplastics. To determine the exact number of microplastics to which adults and children are exposed over the course of a year, in-depth research was conducted on the cosmetic care and treatment products used by over 354 respondents from Romania. This study used a free, self-reported questionnaire method, which took into account consumer habits and preferences, as well as any potential medical conditions that could affect exposure. Optical microscopy and micro-FTIR revealed a total of 109 microplastics, with different sizes, colors, and shapes (i.e., fragments and fibers) and various chemical compositions, including mixtures of polymeric and natural structures, as well as 100% synthetic materials, e.g., polyethylene and polyester. The potential health risk of exposure to microplastics in certain cosmetic formulations for adults was assessed by calculating various risk indices, such as the polymer risk index (H), pollution load index (PLI), dermal plastic absorption (DPA), chronic daily dermal exposure (CDDE), risk to human health from dermal absorption (RHHDA), and estimated annual dermal absorption (EADA). These indices were calculated based on the medical conditions and application areas indicated on the labels of the analyzed creams (i.e., skincare and treatment), for both adult and children’s categories, using the fingertip unit (FTU) method for estimating the cream amount. The plastic toxicity of the analyzed samples was assessed using the H and PLI indices. The risk of microplastics to human health from dermal exposure was assessed using the DPA, CDDE, RHHDA, and EADA indices, which showed concerning results regarding the presence of these particles in cosmetic formulations.

Journal of Xenobiotics doi: 10.3390/jox16010036

Authors: Kazuma Naito Ayame Tomii Katsunori Ishii Hidehisa Shimizu

The escalating consumption of red meat is a potent environmental risk factor for inflammatory bowel disease (IBD), which is characterized by compromised expression of the xenobiotic transporter P-glycoprotein (MDR1/ABCB1). While gut microbiota metabolize dietary tryptophan into diverse indole derivatives that function as aryl hydrocarbon receptor (AhR) ligands, their differential regulation of MDR1 remains an unresolved AhR paradox. Here, we investigated the mechanisms by which two distinct metabolites, indole-3-acetic acid (IAA) and skatole, regulate MDR1 expression in human colonic epithelial Caco-2 cells. We observed that IAA selectively enhances MDR1 protein stability via an AhR-dependent pathway without inducing de novo transcription, suggesting a mechanism we term enhanced proteostasis mediated by the AhR-Hsp90 complex. Conversely, skatole, a toxic dysbiotic metabolite linked to red meat intake, triggered a time-dependent depletion of MDR1 and potently abrogated the protective efficacy of IAA. Our findings are consistent with a model in which skatole acts as a putative structural disruptor, potentially destabilizing the chaperone complex essential for MDR1 integrity. This destruction is facilitated by a key bacterial enzyme, indoleacetate decarboxylase (IAD), which is a pH-dependent metabolic switch in the gut. The modern Western diet, characterized by high protein and low fiber content, elevates colonic pH, thereby activating IAD to convert protective IAA into toxic skatole. These findings provide a molecular framework for the red meat–microbiome–barrier failure axis and highlight the restoration of the IAA/skatole balance through dietary intervention as a promising therapeutic strategy.

Journal of Xenobiotics doi: 10.3390/jox16010035

Authors: Teresa D’Amore

Pesticides and biocides remain indispensable chemicals for agriculture, food safety, public health, and industrial applications, as they safeguard crop yields, control disease vectors, and maintain high hygiene standards [...]

Journal of Xenobiotics doi: 10.3390/jox16010034

Authors: Jakub Kufel Miłosz Korbaś Julita Janiec Zofia Pankowska Marta Młynek Aleksandra Gaweł Adam Mitręga

Environmental pollution with micro- and nanoplastics (MNPs) is an escalating global health concern. Despite growing evidence of MNPs’ presence in the human body, their impact on cerebrovascular diseases remains poorly understood. This study aimed to systematically assess the presence of MNPs in the vascular system and their association with the risk and progression of stroke. A systematic review was conducted in accordance with PRISMA 2020 guidelines and registered in PROSPERO (CRD420251272759). PubMed, Scopus, Web of Science, and Embase databases were searched for original research articles published in the last 10 years. Five studies were included (2 human observational, 3 animal in vivo), comprising 287 patients and rodent models. Methodological quality was assessed using ROBINS-E and SYRCLE’s RoB tools. The analysis confirmed the presence of MNPs, particularly polyethylene and polyvinyl chloride, in key human pathological structures, including carotid atherosclerotic plaques and stroke thrombi. Notably, the presence of MNPs in plaques was associated with a 4.5-fold increase in the risk of major cardiovascular events and death. Animal model studies provided a biological rationale for these observations, demonstrating that MNP exposure may lead to microembolization in cerebral circulation, blood–brain barrier disruption, and exacerbated ischemic injury. Importantly, MNP burden may reflect cumulative environmental exposure and vascular disease severity rather than a direct causal factor in stroke pathogenesis. Nevertheless, MNPs may still represent a novel, modifiable risk factor for stroke through their association with adverse vascular outcomes. Available evidence confirms their accumulation in the cardiovascular system and suggests an association with adverse clinical outcomes. Due to the limited number of studies, further standardized research on larger populations is required to establish whether a causal relationship exists.

Journal of Xenobiotics doi: 10.3390/jox16010033

Authors: Kouthulgama Veekshith Reddy Majid Shafi Abhinav Madari Sharath Chandra Goud Shayaib Ahmad Kamil Akeel Bashir Masood Saleem Mir Mir Nadeem Hassan Mudasir Ali Rather Zahoor Ahmad Wani Showkeen Muzamil Bashir Atif Khurshid Wani Showkat Ahmad Shah

Monosodium glutamate (MSG), a widely used flavor enhancer, has been implicated in oxidative stress-mediated systemic and reproductive toxicity, particularly affecting the male gonadal system. This study evaluated the ameliorative potential of eugenol, a phenolic compound with potent antioxidant properties, against MSG-induced reproductive toxicity in male Wistar rats. Thirty rats were randomly divided into five groups: group 1 (control), group 2 (MSG 2.5 g/kg), group 3 (eugenol 200 mg/kg), group 4 (MSG + eugenol 100 mg/kg), and group 5 (MSG + eugenol 200 mg/kg). Treatments were administered orally for 28 days. Hematological, biochemical, hormonal, antioxidant, gross, and histopathological assessments were conducted after sacrifice on Day 29. MSG exposure significantly reduced testicular weight, testosterone levels, TEC, Hb, PCV, serum proteins, and testicular GSH and SOD, while markedly elevating TLC, AST, ALT, BUN, creatinine, and TBARS. Severe testicular degeneration, vascular congestion, germ-cell loss, and disrupted seminiferous tubules were observed histologically. Co-administration of eugenol resulted in significant and dose-dependent amelioration of MSG-induced alterations, restoring hematological and biochemical parameters, improving antioxidant status, and elevating testosterone levels. Gross pathology and histopathology demonstrated progressive structural recovery, with the higher eugenol dose showing near-normal testicular architecture and active spermatogenesis. Eugenol alone produced no adverse effects and remained comparable to the control group across all parameters. The findings indicate that eugenol confers strong protective effects against MSG-induced reproductive toxicity, primarily through its antioxidant and cytoprotective actions. Eugenol may serve as a promising natural therapeutic agent for mitigating chemically induced male reproductive impairments.

Journal of Xenobiotics doi: 10.3390/jox16010032

Authors: Jhon Fredy Narváez Valderrama Juan José García Londoño Daniel Gil Ramírez Clara S. Arias-Monsalve Jorge L. Gallego

Polycyclic aromatic hydrocarbons (PAHs) can enter water bodies and bioaccumulate in fish, leading to biomagnification; therefore, their monitoring is necessary. Passive sampling is easy to handle and shows potential for this purpose. However, studies in vivo are scarce, and kinetic parameters governing analyte partitioning between tissue and samplers remain poorly characterized. In this study, the silicone rubber membranes (SRMs) were exposed to fish fillet from common carp (Cyprinus carpio) to determine bioaccumulation parameters based on dissipation modelling using performance reference compounds (PRCs). The SRM was implanted in vivo in fish, and the dissipated PRCs were measured and applied to a mono-compartmental model. The results in fish fillet showed a pseudo-first kinetic order, and the plateau was attained at a time > 30 h. However, the equilibrium may not be ensured because of the low lipid fraction (fl) in fish (4.5%), which could lead to a local saturation of the tissue in contact with the SRM. The ratio between elimination and uptake constants (Ke/Ku) showed faster PAHs–SRM sorption than PAHs-fish tissue sorption (200 times); thus, fish with low fl will lead to faster SRM sorption. By contrast, in fish with higher fl, the long-term exposures will be necessary. The percentage of released deuterated PAHs from SRM during in vivo fish exposure was 1.6 times higher than that observed in the fish fillet, indicating an active clearance process. Therefore, during implantation, the rate of clearance and the fl should be considered to ensure detectable levels for applying the integrative equation based on dissipation modelling.

Journal of Xenobiotics doi: 10.3390/jox16010031

Authors: Graziana Assalve Paola Lunetti Vincenzo Zara Alessandra Ferramosca

Background: Human exposure to endocrine-disrupting chemicals (EDCs) is increasingly linked to male reproductive dysfunction, but underlying mechanisms remain unclear. This study aimed to evaluate how selected pharmacological (dihydroxyflutamide, 2OH-FTA; bicalutamide, BIC) and agrochemical (lindane, βHCH; permethrin, PERM; mancozeb, MNZ; tributyltin oxide, TBTO) EDCs affect mitochondrial function in human spermatozoa with parameters within World Health Organization (WHO) reference ranges. Methods: Human sperm cells were exposed ex vivo to 0.1–1000 nM of each compound. Mitochondrial respiration was measured using polarography, assessing oxygen consumption in active (V3) and resting (V4) states, and the respiratory control ratio (RCR) was calculated as an index of mitochondrial coupling. Results: Both 2OH-FTA and BIC reduced RCR in a concentration-dependent manner, mainly due to increases in V4, with BIC showing the strongest effect. βHCH produced a similar pattern, elevating V4 and decreasing RCR. In contrast, PERM, MNZ, and TBTO caused near-complete collapse of both V3 and V4 even at sub-nanomolar concentrations, indicating severe, concentration-independent mitochondrial toxicity. Conclusions: Sperm mitochondria are highly sensitive to EDCs, and distinct compounds exert different bioenergetic effects. Mitochondrial respiration assays provide a useful tool for ex vivo toxicological screening and risk assessment.

Journal of Xenobiotics doi: 10.3390/jox16010030

Authors: Anastasios Potiris Panagiotis Antsaklis Panagiotis Christopoulos Nikolaos Kathopoulis Efthalia Moustakli Ismini Anagnostaki Eirini Drakaki Nefeli Arkouli Aikaterini-Lydia Vogiatzoglou Athanasios Zikopoulos Sofoklis Stavros Charalampos Theofanakis

A significant and persistent issue in assisted reproduction is recurrent implantation failure (RIF), which is often observed even after the transfer of embryos of high morphological and/or genetic quality. Accumulating data suggest that exposure to chemicals with endocrine-disrupting effects (EDCs) may be associated with adverse implantation outcomes. Many environmentally widespread substances have the potential to interfere with the regulation of the endocrine system, affecting critical mechanisms involved in implantation, such as endometrial receptivity, steroid hormone receptor signaling, immune tolerance at the maternal–fetal interface, and the epigenetic regulation of genes that are essential for successful implantation. Experimental studies have shown that exposure to EDCs can alter gene expression in the endometrium, inflammatory pathways, and the dynamics of early embryonic development, while clinical and epidemiological data have associated increased levels of EDCs in the body with lower implantation rates in assisted reproductive technology (ART) cycles. This narrative review examines the implications of these findings in reproductive medicine, summarizes recent experimental and clinical data, and highlights the molecular mechanisms linking exposure to endocrine disruptors with recurrent implantation failure. Recognizing environmental chemical exposure as a potentially modifiable risk factor may offer new perspectives for the prevention of RIF and the development of more personalized therapeutic strategies.

Journal of Xenobiotics doi: 10.3390/jox16010029

Authors: Carlos Alberto-Silva Felipe Assumpção da Cunha e Silva Brenda Rufino da Silva Leticia Ribeiro de Barros Adolfo Luis Almeida Maleski Maricilia Silva Costa

Oxidative and nitrosative stress are central mechanisms in the pathogenesis of neurodegenerative diseases, where excessive production of reactive oxygen and nitrogen species (ROS/RNS) leads to mitochondrial dysfunction, membrane damage, and neuronal death. In this study, we established and compared short-term (2 h) and long-term (20 h) exposure paradigms to sodium nitroprusside (SNP), used as a xenobiotic nitric oxide donor, in two neuronal cell lines (mHippoE-18 and PC12) and zebrafish larvae, aiming to provide a preclinical framework for neurodegenerative drug discovery. In vitro, SNP exposure caused concentration-dependent reductions in viability and alterations in oxidative balance, with mHippoE-18 cells exhibiting higher susceptibility than PC12 cells. In the short-term exposure paradigm, cytotoxicity was primarily associated with membrane disruption at higher concentrations, whereas oxidative stress contributed more strongly at intermediate doses. In the long-term exposure, mHippoE-18 cells showed strong integrated correlations between ROS, LDH release, and viability loss, highlighting their increased vulnerability to nitrosative stress. In zebrafish, SNP exposure impaired metabolic activity and swimming behavior in both paradigms. Long-term exposure led to consistent dose-dependent increases in ROS, accompanied by locomotor deficits tightly linked to energy metabolism. Overall, the higher sensitivity of mHippoE-18 cells compared with PC12 cells, together with the dose-dependent metabolic and behavioral impairments observed in zebrafish, indicates that cellular responses partially mirror in vivo outcomes. This integrative approach underscores the value of combining neuronal cell lines with zebrafish larvae to capture complementary aspects of SNP-induced neurotoxicity and to strengthen preclinical evaluation of candidate compounds with protective or therapeutic potential. These findings support the use of SNP as a xenobiotic model to probe nitrosative stress-driven neurotoxicity across cellular and organismal systems.

Journal of Xenobiotics doi: 10.3390/jox16010028

Authors: Aqsa Aufa Syauqi Sadana Saekhol Bakri Shinji Tokonami Eka Djatnika Nugraha Hasnawati Amqam Muflihatul Muniroh

Genetic polymorphisms can modulate susceptibility to mercury (Hg) toxicity by altering metabolic and detoxification pathways. This review evaluated the association between genetic variants, Hg exposure, and obstetric outcomes. A systematic search of Scopus, PubMed and ScienceDirect through May 2025 identified 12 eligible studies (n = 4995), conducted in accordance with PRISMA guidelines, with methodological quality assessed using the Newcastle–Ottawa Scale. Meta-analysis was selectively performed only for genetically and methodologically comparable studies. The most frequently examined genes were GSTP1, GCLC, GCLM, GPX1, MT1A, ALAD, and APOE. Meta-analysis of GSTP1 rs1695, showed no statistically significant association between the Val105 allele and hair mercury concentrations (MD = −0.08 µg/g; 95% CI: −0.18 to 0.02; p = 0.13), although the direction of effect suggested a potential protective trend. Polymorphisms in other glutathione-related genes (GCLC, GCLM, and GPX1) were consistently associated with increased risks of small-for-gestational-age infants, preeclampsia, and impaired neurodevelopmental outcomes in offspring. In contrast, the APOE ε4 allele appeared to be associated with reduced fetal mercury burden, whereas polymorphisms in ALAD and MT1A were linked to higher mercury levels and adverse pregnancy-related outcomes. By integrating epidemiological evidence with mechanistic insights within a gene–environment interaction framework, this review helps to address important gaps in the existing literature. These findings underscore the importance of incorporating genetic susceptibility into Hg risk assessment and precision-based prenatal interventions.

Journal of Xenobiotics doi: 10.3390/jox16010027

Authors: Francisco Esteves Helena Caria

The central mechanistic hypothesis underlying multifactorial hearing loss posits that genetic susceptibility and environmental exposures act synergistically to disrupt cochlear homeostasis through redox imbalance, mitochondrial dysfunction, and pro-inflammatory mechanisms. This gene–environment paradigm has significant translational implications: elucidating the molecular crosstalk between genetic variants and environmental factors may enable precision risk stratification and the development of targeted otoprotective strategies. The present review provides a comprehensive examination of the major determinants implicated in hearing loss. The manuscript is organized into six main sections that encompass the most relevant domains of current research. First, it offers (I) an overview of epidemiological patterns and the multifactorial nature of hearing impairment. This is followed by (II) a background synthesis of the complex genetic architecture underlying hearing loss. Next, the authors present (III) an outline of environmental determinants and exposure profiles associated with auditory dysfunction, highlighting prominent pollutant/xenobiotic classes (e.g., organic solvents and volatile aromatic hydrocarbons, heavy metals, pesticides, and especially organophosphates and persistent organochlorine compounds), followed by (IV) an analysis of oxidative stress, mitochondrial impairment, and inflammatory pathways involved in cochlear injury. Subsequently, (V) translational perspectives and integrated therapeutic approaches are discussed, with emphasis on epidemiological prevention and precision-based interventions. Finally, (VI) this review addresses current challenges and future directions in elucidating gene–environment interactions in hearing loss.

Journal of Xenobiotics doi: 10.3390/jox16010026

Authors: Quan Shi Jamie Dunn Juan-Carlos Carrillo Michael G. Penman Robert H. Powrie Corinne Haines Hua Shen Yuan Tian Sophie Jia Fabienne Hubert Peter J. Boogaard

Higher olefins are a class of alkenes widely used as intermediates in the production of essential consumer and industrial products. This radiolabel disposition and partial mass balance study investigated the distribution and excretion of four 14C-radiolabelled alpha higher olefins (i.e., 1-octene, 1-decene, 1-hexadecene, and 1-eicosene) in male Wistar rats following a single oral dose (100 mg/kg). Blood, liver, kidney, adipose tissue, urine, and faeces were collected and analysed for total 14C-derived radioactivity. Urinary elimination was rapid, with approximately 70% and 90% of total radioactivity recovered in urinary excreted within 24 and 48 h, respectively. Excretion patterns showed a clear chain-length-dependent trend: shorter-chain olefins (C8, C10) exhibited higher urinary excretion, indicating greater systemic absorption, while longer-chain olefins (C16, C20) were primarily eliminated via faeces, suggesting limited intestinal uptake. Tissue distribution was minimal in blood, liver, and kidney, but adipose tissue retention increased with chain length. Total recovery of administered radioactivity in the analysed matrices was low, ranging from 17% to 60%. Importantly, because exhaled 14CO2 and volatile parent compounds were not captured, the missing fraction cannot be quantified and the balance cannot be considered closed. All in all, the current study describes the partial disposition of higher olefins and highlights the influence of molecular size and lipophilicity on the biological fat, though further studies are required to fully characterise their metabolic profile and total elimination kinetics.

Journal of Xenobiotics doi: 10.3390/jox16010025

Authors: Mila Milutinovic Stuart Lutimba Mohammed A. Mansour

Cancer metastasis, the spread of tumour cells from the primary site to distant organs, is responsible for over 90% of cancer deaths, yet effective treatments remain elusive due to incomplete understanding of the molecular drivers involved. Podocalyxin (PODXL), a protein overexpressed in many aggressive cancers, links the cell membrane to the internal skeleton through its interaction with Ezrin, an actin cytoskeleton cross-linker. Despite its therapeutic relevance, the PODXL–Ezrin interface remains structurally uncharacterised and pharmacologically intractable. Here, we employed an integrated computational approach combining protein–protein docking, molecular dynamics (MD) simulations, and virtual screening to investigate the structural basis of the PODXL–Ezrin interaction. Using AlphaFold-predicted structures, we modelled PODXL and Ezrin complexes, revealing that PODXL’s cytoplasmic domain stabilises upon Ezrin binding, with Arg495 mediating temporally distinct electrostatic interactions essential for initial complex assembly. Particularly, we characterised the R495W missense mutation in PODXL’s Ezrin-binding domain, demonstrating that substitution of arginine with bulky, hydrophobic tryptophan may allosterically destabilise Ezrin’s dormant conformation. This mutation slightly increases the intramolecular distance between the F3 subdomain and C-terminal domain from 2.59 Å to 3.40 Å, thus leading to potential partial unmasking of the Thr567 phosphorylation site that could plausibly prime Ezrin for activation. Molecular dynamics simulations in the WT state with a total simulation time of 100 ns revealed enhanced structural rigidity and reduced radius of gyration fluctuations in the mutant complex, consistent with a potential “locked,” activation-prone state that amplifies oncogenic signalling. Through virtual screening, we identified NSC305787 as a selective destabiliser of the R495W mutant complex by disrupting key Trp495–pre-C-terminal loop Ezrin interactions and causing steric hindrance to PIP2 recruitment. Our findings identified mutation-dependent changes in drug binding that can guide the development and repurposing of compounds for targeting PODXL-related cancers and improve patient outcomes in PODXL-altered malignancies.

Journal of Xenobiotics doi: 10.3390/jox16010024

Authors: Alba Lara-Moreno Belen Rodriguez-Morillo Fernando Madrid Pedro M. Martin-Sanchez Jaime Villaverde Carmen Mejías Esteban Alonso Juan Luis Santos Esmeralda Morillo

Diclofenac (DCF) is a widely used non-steroidal anti-inflammatory drug whose presence in environmental matrices has led to its classification as an emerging contaminant. Developing effective and sustainable removal strategies is therefore essential. In this study, Pseudomonas aeruginosa CSWD.1, Pseudomonas sp. CSWD.2, and a microbial consortium (MC) were isolated from activated sludge through enrichment cultures with DCF and employed as laboratory models to investigate DCF biodegradation capacity under a biosafety-aware framework. Biodegradation assays supplemented with glucose showed limited removal (45.5%) by CSWD.1, whereas CSWD.2 and the MC achieved complete elimination (100%) of 10 mg L−1 DCF in 21 and 5 days, respectively. Three extracellular metabolites, 4’-hydroxy-diclofenac (4’-OH-DCF), 5-hydroxy-diclofenac (5-OH-DCF), and putative NO2-DCF, were detected, with concentrations varying during degradation. Persistence of 4’-OH-DCF and tentatively identified NO2-DCF after 28 days was potentially associated with increased toxicity relative to the abiotic control. Overall, the results suggest that evaluating metabolites and their toxicity is essential, requiring isolation of additional microorganisms able to degrade 4’-OH-DCF and NO2-DCF to combine with the microorganisms isolated in this study. Metabarcoding analysis of the microbial consortium after bioremediation revealed the dominant bacterial population of Burkholderia (88.9% relative abundance) and a predominant fungal genus Talaromyces (80.1%), indicating that both bacteria and fungi may be associated with DCF transformation. These results provide insights into microbial community dynamics and their potential application in designing effective consortia for DCF bioremediation.

Journal of Xenobiotics doi: 10.3390/jox16010023

Authors: Govindhasamay R. Varatharajan Martina Coletta Santosh Kumar Daizy Bharti Arnab Ghosh Shikha Singh Amit C. Kharkwal Francesco Dondero Antonietta La Terza

Heavy metals (HMs) and metal-oxide nanoparticles (NPs) frequently co-occur in freshwater systems, yet their combined effects on microbial predators remain poorly understood. Here, the freshwater ciliate Coleps hirtus was used to evaluate the cytotoxicity of single and binary mixtures of HMs (Cd, Cu, Zn) and NPs (ZnO, CuO, TiO2, SiO2), and to characterize associated antioxidant responses. Acute toxicity was assessed after 24 h by estimating LC20 and LC50 values, while mixture toxicity for Cd + Zn and Cd + ZnO was analyzed using the Toxic Unit approach and the MixTOX framework. Non-enzymatic (TPC, DPPH, HRSA) and enzymatic (CAT, GST, GPx, SOD) antioxidants were quantified as sublethal biomarkers at concentrations below lethal thresholds. HMs were markedly more toxic than NPs, with a toxicity ranking of Cu > Cd >> Zn, whereas NPs followed ZnO > CuO >> TiO2 >> SiO2. Cd + Zn mixtures showed predominantly antagonistic or non-interactive effects, while Cd + ZnO mixtures exhibited strong synergistic toxicity with a non-linear dependence on mixture composition, as supported by MixTox modeling. Exposure to HMs and NPs induced significant and often coordinated changes in antioxidant biomarkers, with binary mixtures eliciting stronger responses than single contaminants. Together, these findings indicate that mixture composition strongly influences both toxicity outcomes and oxidative stress responses in C. hirtus. The combination of clear, mixture-dependent toxicity patterns and robust oxidative stress responses makes C. hirtus a promising bioindicator for freshwater environments impacted by HMs and NPs.

Journal of Xenobiotics doi: 10.3390/jox16010022

Authors: Yi-Fu Xing Xuan-Yi Zhu Hong-Biao Dong Jian-Hua Huang Ya-Fei Duan Jia-Song Zhang

Nitrite is a common toxic substance in aquaculture, and microplastics are environmental pollutants capable of adsorbing small molecules/particles. Shrimp rely mainly on the hepatopancreas to accomplish detoxification metabolism. In this study, we investigated the individual and combined effects of nitrite and microplastics on the physiological function of the P. vannamei hepatopancreas. The results demonstrated that both nitrite and microplastics induced morphological damage, with the combined stress exacerbating tissue damage. Oxidative stress biochemical indicators were disrupted, and most enzyme activities and gene expression levels were upregulated to varying degrees in each experimental group. The expression levels of immune genes (cytC, CASP-3, Crus, ALF, and proPO), detoxification metabolism genes (CYP450, EH1, SULT, and UGT), and oxidative-stress-related genes (ROMO1, SOD, GPx, and Trx) exhibited different fluctuations. Nitrite and microplastic stress resulted in altered hepatopancreatic function, mainly involving amino acid biosynthesis and metabolism, ABC transporters, oxidative phosphorylation, and the mTOR pathway. We identified 17 metabolic biomarkers, including 6 lipids (Oleic acid, Prostaglandin G2, Linoleic acid, Palmitic acid, Docosahexaenoic acid, Docosapentaenoic acid), 6 amino acids (L-Leucine, Agmatine, L-Arginine, L-Tyrosine, Ornithine, N-Acetylornithine), and 5 carbohydrates (Glyceric acid, Citric acid, D-Mannose, Sorbitol, Fumaric acid). These findings suggest that nitrite and microplastic stresses cause hepatopancreatic tissue damage and induce oxidative stress, physiological and metabolic dysfunction in the shrimp P. vannamei, thereby impacting its normal physiological functions.

Journal of Xenobiotics doi: 10.3390/jox16010021

Authors: Roberto Bargagli Emilia Rota

The deposition of airborne microplastics (MPs) poses potential risks to human health and terrestrial ecosystems. Therefore, suitable mitigation efforts are needed, as is knowledge of their deposition patterns in inhabited and remote regions. Currently, there are no standardized protocols for monitoring airborne MPs, and implementing and managing automatic monitoring systems would be costly and feasible only in a few fixed locations. Over the past few decades, several species of cryptogams have proven to be reliable biomonitors of persistent atmospheric contaminants. Due to the lack of standardized methodologies, the results of preliminary biomonitoring surveys for MPs have been inconsistent and difficult to compare. However, they clearly indicate higher MP concentrations in epigeic mosses than in epiphytic lichens (collected at the same site or experimentally exposed in parallel in bags). This review discusses the morphophysiological features that favor the entrapment and retention of intercepted MPs in mosses, as well as the field and laboratory activities necessary to determine whether these organisms progressively accumulate airborne MPs as a function of the exposure time. Steps for future research needed to develop a cost-effective, reliable and easily applicable biomonitoring methodology are suggested. Evaluating the advantages of active moss biomonitoring over sampling atmospheric bulk deposition or exposing suitable commercial materials is recommended.

Journal of Xenobiotics doi: 10.3390/jox16010020

Authors: Saereh Mohammadpour Thainara Viana Rosa Freitas Eduarda Pereira Bruno Henriques

As demand for rare earth elements (REEs) rises and environmental concerns about the extraction of primary resources grow, biological methods for removing these elements have gained significant attention as eco-friendly alternatives. This study assessed the ability of the green macroalga Ulva lactuca to remove europium (Eu) from aqueous solutions, evaluated the cellular partition of this element and investigated the toxicological effects of Eu exposure on its biochemical performance. U. lactuca was exposed to variable concentrations of Eu (ranging from 0.5 to 50 mg/L), and the amount of Eu in both the solution and algal biomass was analyzed after 72 h. The results showed that U. lactuca successfully removed 85 to 95% of Eu at low exposure concentrations (0.5–5.0 mg/L), with removal efficiencies of 75% and 47% at 10 and 50 mg/L, respectively. Europium accumulated in algal biomass in a concentration-dependent manner, reaching up to 22 mg/g dry weight (DW) at 50 mg/L. The distribution of Eu between extracellular and intracellular fractions of U. lactuca demonstrated that at higher concentrations (5.0–50 mg/L), 93–97% of Eu remained bound to the extracellular fraction, whereas intracellular uptake accounted for approximately 20% at the lowest concentration (0.5 mg/L). Biochemical analyses showed significant modulation of antioxidant defenses. Superoxide dismutase activity increased at 10 and 50 mg/L, while catalase and glutathione peroxidase activities were enhanced at lower concentrations (0.5–1.0 mg/L) and inhibited at higher exposures. Lipid peroxidation levels remained similar to controls at most concentrations, with no evidence of severe membrane damage except at the highest Eu level. Overall, the results demonstrate that U. lactuca is an efficient and resilient biological system for Eu removal, combining high sorption capacity with controlled biochemical responses. These findings highlight its potential application in environmentally sustainable remediation strategies for REE-contaminated waters, while also providing insights into Eu toxicity and cellular partitioning mechanisms in marine macroalgae.

Journal of Xenobiotics doi: 10.3390/jox16010019

Authors: Vesela Yancheva Stela Stoyanova Elenka Georgieva Eleonora Kovacheva Bartosz Bojarski László Antal Ifeanyi Emmanuel Uzochukwu Krisztián Nyeste

The intensive use of pesticides in agriculture poses serious risks to aquatic ecosystems and non-target organisms, yet toxicological data remain limited. This study evaluated the acute effects of three widely used pesticides—pirimiphos-methyl (10 and 60 μg/L), propamocarb hydrochloride (40 and 80 μg/L), and 2,4-D (50 and 100 μg/L)—on the liver of common carp (Cyprinus carpio Linnaeus, 1758), a sentinel species in aquaculture, but also a species equally important in risk assessment and environmental monitoring. Fish were exposed for 96 h under controlled conditions, and histopathological, histochemical, and biochemical biomarkers were analyzed. All tested pesticides induced significant histopathological alterations, predominantly circulatory and degenerative changes, with severity increasing at higher concentrations. Propamocarb hydrochloride and 2,4-D caused more pronounced and partly irreversible hepatotoxicity compared to pirimiphos-methyl. The histochemical assessment revealed altered glycogen metabolism, while the biochemical assays showed inhibition of key liver enzymes, including ALAT, ASAT, ChE, and LDH, indicating disrupted metabolic processes. These findings highlight the vulnerability of aquatic organisms to pesticide exposure and support the use of fish liver biomarkers as effective tools in ecotoxicology research. The study also emphasizes the need for stricter regulation and environmental monitoring of pesticide contamination in aquatic ecosystems.

Journal of Xenobiotics doi: 10.3390/jox16010018

Authors: Andrea Binelli Alberto Cappelletti Cristina Cremonesi Camilla Della Torre Giada Caorsi Stefano Magni

This study presents the first evaluation of plastic particle contamination along a complete drinking water supply chain within the distribution system of Milan, Northern Italy. Fourteen grab water samples were collected from various points, including groundwater extraction, post-treatment stages, a public fountain, and ten household taps. Plastic particles were identified using µFTIR spectroscopy and characterized by polymer type, shape, size, and color. Overall, low concentrations of plastic particles were detected, ranging from 0.3 ± 0.5 particles/L in the accumulation tank to an average of 1.9 ± 1.4 particles/L in household tap water, with no significant increase observed along the supply chain. The entire data set was dominated by cellulose particles (76%), as plastics accounted for only 8%. Microplastics (1 µm–1 mm) were the most commonly detected (90%), while the remaining 10% were large microplastics (1–5 mm). Qualitatively, polyester fibers were the most prevalent particles identified. However, greater variability and higher concentrations were found in private residence samples, suggesting that internal plumbing systems may be a primary source of contamination. Estimated human exposure through this supply system, based on a current theoretical model, was minimal compared to global benchmarks. These findings highlight the necessity of integrating private distribution infrastructure into future regulatory frameworks to assist stakeholders in making informed decisions to mitigate potential plastic contamination.

Journal of Xenobiotics doi: 10.3390/jox16010017

Authors: Antonio Peña-Fernández Manuel Higueras Gevorg Tepanosyan M. Ángeles Peña Fernández M. C. Lobo

Nine technology-related metal(loid)s (Ag, Co, Fe, Mo, Pt, Rh, Sb, Se and Y) were monitored in 137 topsoil samples from urban parks, industrial areas and gardens in Alcalá de Henares (Spain) using ICP–MS. Selenium was not detected, while Mo, Sb and Rh showed a high proportion of values below the detection limit, indicating generally low contamination. In contrast, Fe, Co and Y were detected in all samples, with industrial soils showing about two-fold higher median Co and Fe than urban soils. Garden soils displayed marked silver enrichment (median 0.439 vs. 0.068 mg kg−1 in urban soils), with Ag pollution indices up to 71 and enrichment factors up to 69; around 17% of garden samples exceeded EF > 40, and more than one-quarter had EF > 10. Principal component analysis suggested a predominantly geogenic association for Co, Fe and Y and an anthropogenic component for Ag, Mo, Rh and Sb, while Pt was mainly linked to vehicular emissions. Under standard US EPA exposure scenarios applied to the 2001 topsoil concentrations, oral and inhalation hazard quotients for elements with available benchmarks remained <0.2 and inhalation cancer risks for Co were ≤2.5 × 10−7, indicating low estimated risk within the model assumptions. However, quantitative risk characterisation remains constrained by benchmark gaps for Pt and Rh and by limited consensus toxicity values for Y, which introduces uncertainty for these technology-related elements. These results should therefore be interpreted primarily as a baseline (2001) in surface soils for Alcalá de Henares rather than as a direct representation of current exposure conditions.

Journal of Xenobiotics doi: 10.3390/jox16010016

Authors: Chien-Yu Lin Hui-Ling Lee Ta-Chen Su

Background and hypothesis: Per- and polyfluoroalkyl substances (PFAS) are highly persistent synthetic chemicals that can accumulate in renal tissue and potentially disrupt kidney function. Most prospective studies on PFAS–renal associations have focused on middle-aged or older adults, leaving uncertainty about whether similar patterns exist in younger populations. Methods: We investigated decade-long trajectories of plasma concentrations of 11 PFAS and their longitudinal associations with estimated glomerular filtration rate (eGFR) among 529 Taiwanese adolescents and young adults (aged 12–30 years) enrolled in the prospective YOung TAiwanese Cohort (YOTA), with measurements obtained in 2006–2008 and 2017–2019. Results: Nearly all plasma PFAS declined significantly over the 10-year period. Despite these reductions, higher baseline levels and greater annualized increases (Δln-PFAS/Δt) in linear perfluorooctanoic acid (PFOA), linear and branched perfluorooctane sulfonic acid (PFOS), perfluorononanoic acid (PFNA), and perfluorodecanoic acid (PFDA) were consistently associated with larger eGFR gains over time (β = 0.33–0.40, q < 0.05). In complementary models using follow-up eGFR as the outcome, both baseline and cumulative PFAS changes (Δln-PFAS) remained positively associated with higher eGFR (β = 1.71–3.84, q < 0.05). Polynomial analyses further indicated mild non-linear exposure–response patterns for several PFAS, suggesting that renal effects may deviate from linearity across exposure ranges. The composite PFAS exposure index (mean of standardized ln-PFAS concentrations) was robustly associated with higher eGFR across sensitivity analyses excluding participants with chronic conditions. These associations were more pronounced among individuals with greater metabolic or physiological vulnerability. Conclusions: Higher PFAS exposure was associated with elevated eGFR in young adults, which may be consistent with early glomerular hyperfiltration or other renal hemodynamic alterations. These findings raise the hypothesis of early renal stress in early life and underscore the need for ongoing biomonitoring and longitudinal follow-up with additional kidney injury markers to clarify long-term renal consequences.

Journal of Xenobiotics doi: 10.3390/jox16010015

Authors: Edwina Brennan Priya Das Pearl Wasif Xianyu F. Wang Jochen F. Mueller Chang He Jean V. Varghese Alexandra E. Butler Stephen L. Atkin Naji Alamuddin

Background: Plasticisers with endocrine-disrupting potential are ubiquitous and associate with obesity and type-2 diabetes (T2D), with higher levels reported in the Middle East. However, no data exist on plasticiser exposure in Bahrain where T2D affects 15% of the national population. Methods: An observational exploratory study in T2D (n = 60) and controls (n = 96), analysed for 24 h urinary plasticiser levels (liquid chromatography tandem mass spectrometry (LC-MS/MS)). Correlation and generalised linear model (GLM) analyses were employed to examine associations. Results: T2D were older (p < 0.001), had higher body mass index (BMI) (p < 0.001), body weight (p < 0.001) and glycosylated haemoglobin A1c (HbA1c) (p < 0.001). Correlation analysis revealed differences in inter-plasticiser, and plasticiser and biomarker relationships, with loss or reversal in T2D compared to controls. Mono-n-butyl phthalate (MnBP) levels were higher in T2D (p = 0.04); however, regression analysis revealed significant association with age. The GLM analyses demonstrated marked differences in the levels of mono(3-carboxypropyl) phthalate (MCPP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), monoethyl phthalate (MEP) and bisphenol S (BPS), with lower levels in T2D versus controls (B = −3.41, p = 0.01; B = −5.28, p < 0.001; B = −8.94, p < 0.001; B = −6.09, p = 0.006, respectively); however, these contrasts appeared to be substantially confounded by BMI and/or age. Positive influence of age and negative influence of BMI when observed across the full dataset were generally reversed in T2D. Levels were complementary to those previously reported for the Middle East. Conclusions: The study indicates the phthalate levels in Bahrain are elevated though complementary to studies of phthalates in the Middle East; within those levels, the study indicates differential exposure–response relationships with plasticisers, influenced by age and BMI, in those with T2D compared to healthy controls.

Journal of Xenobiotics doi: 10.3390/jox16010014

Authors: Michele De Rosa Silvia Canepari Giovanna Tranfo Ottavia Giampaoli Adriano Patriarca Agnieszka Smolinska Federico Marini Lorenzo Massimi Fabio Sciubba Mariangela Spagnoli

Foundries represent complex exposure scenarios where metals, particulate matter, and combustion by-products coexist, posing potential cumulative biological effects. Urinary metabolic profiles from 64 foundry workers and 78 residents living in surrounding areas were investigated using multivariate statistical modeling. Differences in urinary metabolite patterns were observed between the two groups, including lower levels of several amino acids (e.g., valine, alanine, tyrosine, and tryptophan) and tricarboxylic acid intermediates (e.g., citrate and succinate), together with higher levels of selected branched-chain amino acid catabolites (e.g., 3-hydroxyisobutyrate and erythro-2,3-dihydroxybutyrate) in workers. Variations in gut microbiota-related metabolites, such as phenylacetylglycine and p-cresol sulphate, were also detected. Based on these metabolic patterns, potential molecular mechanisms related to energy metabolism, oxidative stress and host–microbiome interaction are discussed as interpretative hypotheses. The comparison between workers and residents was interpreted, taking into account differences in demographic and lifestyle characteristics between groups. Overall, the results indicate that occupational exposure in foundries is associated with measurable differences in urinary metabolic profiles, demonstrating that the applied NMR-based metabolomic strategy is capable of capturing early biological effects and supporting its potential as a non-invasive and holistic biomonitoring tool for evaluating the health impact of complex occupational exposures.

Journal of Xenobiotics doi: 10.3390/jox16010013

Authors: Can Wang Jingxian Xu Tingting Gao Xiaomei Hong Fakang Pan Fuwei Sun Kai Huang Dejian Wang Tianhu Chen Ping Zhang

The rising levels of Co(II) in aquatic environments present considerable risks, thereby necessitating the development of effective remediation strategies. This study introduces an innovative pre-hydration method for synthesizing carbonated calcined clay dolomite (CCCD) to efficiently remove Co(II) from contaminated water. This pre-hydration treatment successfully reduced the complete carbonation temperature of the material from 500 °C to 400 °C, significantly enhancing energy efficiency. The Co(II) removal performance was systematically investigated by varying key parameters such as contact time, initial Co(II) concentration, pH, and solid/liquid ratio. Optimal removal was achieved at 318 K with pH of 4 and a solid/liquid ratio of 0.5 g·L−1. Continuous flow column experiments confirmed the excellent long-term stability of CCCD, maintaining a consistent Co(II) removal efficiency of 99.0% and a stable effluent pH of 8.5 over one month. Isotherm and kinetic models were used to empirically describe concentration-dependent and time-dependent uptake behavior. The equilibrium data were best described by the Langmuir model, while kinetics followed a pseudo-second-order model. An apparent maximum removal capacity of 621.1 mg g−1 was obtained from Langmuir fitting of equilibrium uptake data. Mechanistic insights from Visual MINTEQ calculations and solid phase characterizations (XRD, XPS, and TEM) indicate that Co(II) removal is dominated by mineral water interface precipitation. The gradual hydration of periclase (MgO) forms Mg(OH)2, which creates localized alkaline microenvironments at particle surfaces and drives Co(OH)2 formation. Carbonate availability further favors CoCO3 formation and retention on CCCD. Importantly, this localized precipitation pathway maintains a stable, mildly alkaline effluent pH (around 8.5), reducing downstream pH adjustment demand and improving operational compatibility. Overall, CCCD combines high Co(II) immobilization efficiency, strong long-term stability, and an energy-efficient preparation route, supporting its potential for scalable remediation of Co(II) contaminated water.

Journal of Xenobiotics doi: 10.3390/jox16010012

Authors: Cinzia Lucia Ursini Giorgia Di Gennaro Giuliana Buresti Raffaele Maiello Anna Maria Fresegna Aureliano Ciervo Marco Gentile Virginia Di Basilio Sabrina Beltramini Daniela Gaggero Nicoletta Rigamonti Erica Maccari Giorgia Zorzetto Piera Maiolino Pasquale Di Filippo Maria Concetta Bilancio Paolo Baldo Valeria Martinello Andrea Di Mattia Chiara Esposito Patrizia Nardulli Mariarita Laforgia Maria Vittoria Visconti Matteo Vitali Emanuela Omodeo-Salè Delia Cavallo

Cancer cases have been estimated that will increase in the next years with consequent increase of antineoplastic (AD) drug treatments and workers handling these hazardous chemicals. We aimed to evaluate genotoxic/oxidative effects of AD exposure by fpg-comet assay on a large size sample of workers (214 exposed and 164 controls) involved in preparation; administration, including Hyperthermic intraperitoneal chemotherapy (HIPEC) and pressurized intraperitoneal aerosol chemotherapy (PIPAC); and disposal. With the final aim to identify suitable early biomarkers of genotoxic effect useful to health surveillance, we correlated fpg-comet assay (blood) and Buccal Micronucleus Cytome (BMCyt) assay data. Fpg-comet parameters resulted higher in the exposed group vs. controls, demonstrating direct and oxidative DNA damage in workers handling ADs. Fpg-comet direct DNA damage and genotoxic parameters of BMCyt assay demonstrated a weak statistically significant correlation. This cross-sectional study is one of the few available evaluating both direct and oxidative DNA damage due to ADs on a large sample size of workers and correlating fpg-comet and BMCyt assay results. It highlights the need to evaluate genotoxic effects by both the biomarkers and furnishes a contribution to their validation. Moreover, we demonstrate for the first time oxidative DNA damage on workers performing HIPEC and PIPAC administration.

Journal of Xenobiotics doi: 10.3390/jox16010011

Authors: Sergio R. Ribone Mario Alfredo Quevedo

Human carboxylesterases (CES) are enzymes that play a central role in the metabolism and biotransformation of diverse endogenous substances and xenobiotics. The two most relevant isoforms, CES1 and CES2, are crucial in clinical pharmacotherapy as they catalyze the hydrolysis of numerous approved drugs and prodrugs. Elucidating the structural basis of CES isoform substrate specificity is essential not only for understanding and anticipating the biological fate of administered drugs, but also for designing prodrugs with optimized site-specific bioactivation. Additionally, this knowledge is also important for the design of biomedically useful molecules such as subtype-targeted CES inhibitors and fluorescent probes. In this context, both experimental and computational methodologies have been used to explore the mechanistic and thermodynamic properties of CES-mediated catalysis. Experimental designs commonly employ recombinant CES or human tissue microsomes as enzyme sources, utilizing quantification methods such as spectrophotometry (UV and fluorescence) and mass spectrometry. Computational approaches fall into two categories: (1) modeling substrate: CES recognition and affinity (molecular docking, molecular dynamics simulation, and free-energy binding calculations), and (2) modeling substrate: CES reaction coordinates (hybrid QM/MM simulations). While experimental and theoretical approaches are highly synergistic in studying the catalytic properties of CES subtypes, they represent distinct technical and scientific fields. This review aims to provide an integrated discussion of the key concepts and the interplay between the most commonly used wet-lab and dry-lab strategies for investigating CES catalytic activity. We hope this report will serve as a concise resource for researchers exploring CES isoform specificity, enabling them to effectively utilize both experimental and computational methods.

Journal of Xenobiotics doi: 10.3390/jox16010010

Authors: Alla P. Toropova Andrey A. Toropov Sofia Mescieri Alessandra Roncaglioni Emilio Benfenati

Background: Pesticide toxicity to insects is an important adverse effect with a potentially large ecological impact when considering the effect on beneficial insects, as pollinators. The assessment of this endpoint is necessary to avoid applying ecologically dangerous pesticides. Aim of the study: Assessment of the availability of the Monte Carlo method for the development of a model for toxicity (pLD50) towards bees and other pollinators. In addition, the index of ideality of correlation is examined as a possibility to increase the statistical quality of quantitative structure–activity relationships (QSARs) for the toxicity of pesticides to pollinators. Main results and novelty: models with good performance on the toxic effect of pesticides towards different pollinators, wrapping acute and chronic effects, using the Monte Carlo method for QSAR analysis.

Journal of Xenobiotics doi: 10.3390/jox16010009

Authors: Monia Cecati Valentina Pozzi Veronica Pompei Valentina Schiavoni Stefania Fumarola Alice Romagnoli Giovanni Tossetta Angelo Montana Alessandro Polizzi Davide Sartini Roberto Campagna

Cisplatin, a platinum-based compound, is a cornerstone of modern chemotherapy and remains widely used against a variety of solid tumors, including testicular, ovarian, lung, bladder, and head and neck cancers. Its anticancer activity is primarily attributed to the formation of DNA crosslinks, which obstruct replication and repair, ultimately leading to apoptosis. However, the clinical value of cisplatin is constrained by two major challenges: its toxic profile and the development of resistance. Cisplatin toxicity arises from its interaction not only with tumor DNA but also with proteins and nucleic acids in healthy tissues, resulting in a range of adverse effects, including, but not limited to, nephrotoxicity, ototoxicity, neurotoxicity, and gastrointestinal injury. In pediatric patients, permanent hearing loss represents a particularly debilitating complication. On the other hand, tumor cells can evade cisplatin cytotoxicity through diverse mechanisms, including reduced intracellular drug accumulation, enhanced DNA repair, detoxification by thiol-containing molecules, and alterations in apoptotic signaling. These resistance pathways severely compromise treatment outcomes and often necessitate alternative or combination strategies. This review examines the chemical structure of cisplatin, the molecular mechanisms of cisplatin cytotoxicity and cisplatin-induced resistance, as well as the main applications in cancer management and the complications associated with its clinical use.

Journal of Xenobiotics doi: 10.3390/jox16010008

Authors: Komal Pandey Baljeet Singh Saharan Yogender Singh Pardeep Kumar Sadh Joginder Singh Duhan Dilfuza Jabborova

Agricultural biomass has potential as a renewable and versatile carbon feedstock for developing eco-friendly and biodegradable polymers capable of replacing conventional petrochemical plastics. To address the growing environmental concerns associated with plastic waste and carbon emissions, lignocellulosic residues, edible crop by-products, and algal biomass were utilized as sustainable raw materials. These biomasses provided carbohydrate-, lipid-, and lignin-rich fractions that were deconstructed through optimised physical, chemical, and enzymatic pretreatments to yield fermentable intermediates, such as reducing sugars, organic acids, and fatty acids. The intermediates were subsequently converted through tailored microbial fermentation processes into biopolymer precursors, primarily polyhydroxyalkanoates (PHAs) and lactate-based monomers. The resulting monomers underwent polymerization via polycondensation and ring-opening reactions to produce high-performance biodegradable plastics with tunable structural and mechanical properties. Additionally, the direct extraction and modification of naturally occurring polymers, such as starch, cellulose, and lignin, were explored to develop blended and functionalized bioplastic formulations. Comparative evaluation revealed that these biomass-derived polymers possess favourable physical strength, thermal stability, and biodegradability under composting conditions. Life-cycle evaluation further indicated a significant reduction in greenhouse gas emissions and improved carbon recycling compared to fossil-derived counterparts. The study demonstrates that integrating agricultural residues into bioplastic production not only enhances waste valorization and rural bioeconomy but also supports sustainable material innovation for packaging, farming, and consumer goods industries. These findings position agriculture-based biodegradable polymers as a critical component of circular bioeconomy strategies, contributing to reduced plastic pollution and improved environmental sustainability.

Journal of Xenobiotics doi: 10.3390/jox16010007

Authors: Patrícia Hockicková Alžbeta Kaiglová Marie Korabečná Soňa Kucharíková

Endocrine disruptors, including bisphenol A, S, AF, and F, have been demonstrated to exhibit endocrine-disrupting activity. This phenomenon has been associated with a variety of health problems, including (but not limited to) neurological and reproductive disorders. Given the potential hazards, it is essential to have effective tools to assess their toxicity. The nematode Caenorhabditis elegans has become a widely used model organism for studying bisphenols because of its genetic simplicity and the conservation of its fundamental biological processes. This review article summarizes current knowledge of bisphenol toxicity and the use of the model organism C. elegans as a high-throughput system for investigating the toxicological profiles of BPA and its emerging alternatives. Furthermore, we highlight the specific methodologies for assessing the toxic effects of bisphenols in C. elegans. While highlighting its advantages, we critically discuss its limitations, including the absence of specific metabolic organs, which constrain direct extrapolation to mammalian systems. Based on available evidence, we conclude that C. elegans serves as an essential bridge between in vitro assays and mammalian models, offering a powerful platform for the early hazard identification and mechanistic screening of bisphenol analogues.

Journal of Xenobiotics doi: 10.3390/jox16010006

Authors: Ekaterina Sergeevna Osipova Evgeny Aleksandrovich Gladkov Dmitry Viktorovich Tereshonok

Heracleum sosnowskyi Manden., or H. sosnowskyi, of the Apiaceae was first cultivated in the USSR in 1947 as a potential fodder plant. Due to the development of cold-resistant cultivars and the characteristics of H. sosnowskyi, it quickly became feral. As a result, H. sosnowskyi began to spread as an aggressive invasive species in the 1970s and 1980s. By the 90s it had become an ecological disaster. As well as forming monocultures and displacing native species, H. sosnowskyi contains furanocoumarins, photosensitizing compounds that increase skin sensitivity to ultraviolet rays and cause severe burns. In addition, furanocoumarins have cytotoxic, genotoxic, mutagenic and estrogenic effects. H. sosnowskyi also contains essential oils, which are particularly active during flowering and can irritate the mucous membranes of the eyes and respiratory tract, as well as cause allergic reactions in the form of bronchospasm in people with asthma and hypersensitivity. When released in high concentrations, these biologically active compounds have an allelopathic effect on native plant species, displacing them and reducing biodiversity. As H. sosnowskyi is not native; the biologically active compounds it secretes have a xenobiotic effect, causing serious damage to the ecosystems it occupies. However, in parallel with these negative properties, furanocoumarins have been found to be effective in the treatment of cancer and skin diseases. Furanocoumarins possess antimicrobial antioxidant osteo- and neuroprotective properties. Essential oils containing octyl acetate, carboxylic acid esters, and terpenes can be used in the pharmaceutical industry as antiseptic and anti-inflammatory agents. Additionally, essential oils can be used as biofumigants and natural herbicides. A comprehensive approach allows H. sosnowskyi to be viewed in two ways. On the one hand, it is an aggressive alien species that causes significant damage to ecosystems and poses a threat to human health. On the other hand, it is a potentially valuable natural resource whose biomass can be used within the principles of the circular economy. It is hoped that the use of H. sosnowskyi for economic interests can be a partial compensation for the problem of its aggressive invasion, which is of anthropogenic origin.

Journal of Xenobiotics doi: 10.3390/jox16010005

Authors: Raymond F. Palmer David Kattari

Background: Chemical Intolerance (CI), Autism Spectrum Disorder (ASD) and Attention Deficit Hyperactive Disorder (ADHD) are conditions with rising incidence rates not fully explained by greater awareness or changes in diagnostic practices. It is now generally accepted that the interaction between genetic and environmental exposures plays a role in all of these conditions. Prior studies show that these conditions co-occur. This study seeks to explore previous findings using an international sample. Methods: A five-country (N = 5000) stratified panel survey was used to assess self-reported CI in themselves, and ASD and ADHD in their children. A generalized linear model was used to estimate Odds Ratios. Age- and sex-adjusted logistic models used CI as a predictor of ASD and ADHD in separate models. Results: Compared to those classified as Low CI, High levels of CI were associated with greater Odds Ratios (OR) of reporting a child with ASD and ADHD in all countries except Japan. Italy, India, and the USA had over twice the OR of reporting a child with ASD. Mexico had over 1.9 times the OR. The results with ADHD are similar to the ASD results. Conclusions: The results of this study are consistent with two prior U.S. studies, showing an association between ASD and ADHD among women who have CI. However, cross-cultural comparisons, especially prevalence estimates for ASD and ADHD, cannot be interpreted as epidemiologic rates due to serious limitations of the survey methodology. No causal relationship should be inferred from this study.

Journal of Xenobiotics doi: 10.3390/jox16010004

Authors: Krzesimir Ciura

Modern pesticide risk assessment relies on data on bioaccumulation and acute aquatic toxicity, yet generating such data is labour-intensive and animal-demanding. This study evaluated whether phospholipid affinity of pesticides, quantified by the chromatographic hydrophobicity index CHIIAM obtained from high-throughput gradient biomimetic chromatography, can serve as a surrogate descriptor of these endpoints. Nineteen pesticides representing different chemical and functional classes were analyzed on IAM.PC.DD2 columns, and CHIIAM values were determined. Bioconcentration factors (BCF) in fish and acute toxicity data (96 h LC50 for fish, 48 h EC50 for Daphnia magna) were retrieved from the Pesticide Properties DataBase. CHIIAM ranged from −12.1 to 54.8 and correlated strongly with log10BCF (r = 0.84) and log10LC50 in fish (r = −0.84), and moderately with log10EC50 for Daphnia (r = 0.76). Highly lipophilic pesticides with high CHIIAM showed elevated BCF and low LC50/EC50 values, whereas polar compounds with low CHIIAM exhibited negligible bioconcentration and low acute toxicity. Deviations from these trends, for compounds with specific modes of action, highlighted the contribution of mechanisms beyond membrane toxicity. Overall, CHIIAM measured under high-throughput conditions retains prognostic value for ecotoxicological assessment and may serve as a rapid experimental descriptor to support preliminary screening.

Journal of Xenobiotics doi: 10.3390/jox16010001

Authors: Pratik Shriwas Abigail M. Noonchester Andre Revnew Thomas R. Lane Christopher M. Hadad Sean Ekins Craig A. McElroy

Of the Cytochrome P450 enzymes, the CYP2C9 variant is very important in the metabolism of several human drugs, acting as a natural bioscavenger. Previously, CYP2C9 was shown to convert the thion (P=S) to the oxon (P=O) form for some organophosphorus (OP) pesticides, such as dimethoate, diazinon, and parathion. In this study, we tested the ability of CYP2C9 to degrade other OP compounds. We investigated the metabolism of OP compounds by CYP2C9 using LC-MS/MS as well as time-dependent inhibition using the previously developed pFluor50 fluorogenic assay. We found that CYP2C9 metabolizes thions preferentially over oxons, and that many OP compounds inhibit CYP2C9 activity in a time-dependent manner. Additionally, we performed molecular docking based on the crystal structure (1OG5) of the CYP2C9 receptor. We observed a positive, though moderate, correlation between the calculated binding energy and the CYP2C9 metabolism of various OP compounds (R = 0.59). These in vitro data, combined with further analysis and additional OP derivatives, could potentially be used to develop artificial intelligence (AI)/machine learning (ML) models to predict the metabolism of specific OP compounds by CYP2C9. This type of approach could be particularly relevant for the prediction of the metabolism of current and emerging chemical warfare agents.

Journal of Xenobiotics doi: 10.3390/jox16010003

Authors: Chesmi Kumbalatara Lindsey Johnson Matthew MacArthur Meungguk Park Wasantha Jayawardene

Electronic Nicotine Delivery Systems (ENDSs) expose users to nicotine, volatile organic chemicals, and ultrafine particles that pose emerging toxicological concerns for youth. The prevalence of vaping among college students quadrupled between 2017 and 2019. The Vaping Initiation, Continuation, Termination, or Resumption in Youth (VICTORY) study explored a random sample of 543 undergraduate students at a Midwestern university, using an anonymous online survey, for factors associated with initiation and regular inhalation of vape-derived aerosols. Results showed that 50% of participants had ever used a vape, and 67% had used tobacco, vape, or marijuana. The mean age of first use of tobacco was 15.16 years, significantly younger than the mean ages for vaping (16.33) and marijuana (16.60). There were no significant gender differences in ENDS use, although more males reported tobacco as their first substance (18% difference). Notably, 40% reported non-alcoholic substance or alcohol use in the past 30 days. Decision-tree analysis revealed complex relationships between vaping aerosols, tobacco, alcohol use, marijuana use, and living arrangements. Logistic regression identified key predictors of regular vaping, including higher school year, lower household income, employment status, and younger age at first use. These findings highlight the need for tailored public health interventions and continued monitoring to address the growing trend of youth vaping.

Journal of Xenobiotics doi: 10.3390/jox16010002

Authors: Alice N. Mafe Dietrich Büsselberg

Although cancer biology has advanced considerably, the impact of environmental toxins on carcinogenesis remains underrecognized and scattered across disciplines. Evidence increasingly shows that chronic exposure to a broad range of toxins—including persistent organic pollutants, heavy metals, pesticides, phthalates, microplastics, and fine particulate matter (PM2.5), which significantly contributes to cancer initiation, progression, and treatment resistance. This review synthesizes mechanistic, molecular, and epidemiological findings from 2015 to 2025, identified through systematic searches of PubMed, Scopus, Web of Science, and MeSH. Key pathways include oxidative stress-mediated DNA damage, epigenetic reprogramming (DNA methylation, histone modifications, miRNA dysregulation), hormone receptor modulation, chronic inflammation, immune evasion, and tumor microenvironment remodeling. Case studies of benzene, arsenic, aflatoxins, pesticides, and microplastics detail exposure routes, molecular targets, and associated cancers, highlighting significant public health risks. Ongoing debates persist regarding safe exposure thresholds, latency periods, and the effects of mixed toxin exposures. The review also highlights recent innovations in environmental oncology, including AI-based predictive models, CRISPR screens for susceptibility genes, organoid/3D models, green chemistry interventions, and real-time exposure monitoring, which provide mechanistic insight and inform early detection and personalized prevention strategies. Additionally, regional data gaps, particularly in low- and middle-income countries, indicate the need for stronger interdisciplinary collaboration. By integrating molecular mechanisms, epidemiology, and technological advances, this review offers a comprehensive framework for understanding toxin-induced carcinogenesis and guiding future research, public health policy, and preventive strategies.

Journal of Xenobiotics doi: 10.3390/jox15060211

Authors: Shuqin Li Guohua Ni

The metallurgical industry generates substantial amounts of heavy metal-containing solid waste, posing significant environmental and health risks. This study systematically evaluates the environmental behavior and ecological risks of heavy metals in four typical metallurgical wastes: jarosite slag (SW1), electric arc furnace ash (SW2), chromium-containing sludge (SW3), and acid-base sludge (SW4). We demonstrate that particle size fundamentally governs heavy metal mobility, with fine-structured SW1 and SW2 (D50 = 4.76 µm and 1.34 µm) exhibiting enhanced metal mobility and bioavailability. In contrast, coarser SW3 and SW4 particles (D50 = 268.83 µm and 133.94 µm) retain heavy metals in more stable forms. Among all metals analyzed, cadmium (Cd) presents the most severe ecological threat, with acid-extractable fractions reaching 52% in SW2 and 45% in SW3—indicating high release potential under changing pH conditions. Risk assessment confirms high to very high ecological risks for Cd in both SW2 and SW3. Moreover, under acidic leaching conditions, SW1 and SW2 show significantly higher cumulative toxicity than SW3 and SW4. These findings highlight the critical role of waste-specific properties in controlling heavy metal fate and provide a scientific basis for targeted risk management and sustainable remediation strategies.

Journal of Xenobiotics doi: 10.3390/jox15060210

Authors: Susana I. L. Gomes Janeck J. Scott-Fordsmand Mónica J. B. Amorim

Silver nanomaterials (Ag NMs) are widely used, including in consumer products, and they inevitably enter the environment, with the soil compartment acting as a major sink. However, most available toxicity data focus on the reference Ag NM300K and rely on standard tests, even though long(er)-term exposure tests are recognized as particularly important for assessing the risks to soil invertebrates. Hence, the aim of the present study was to investigate the toxicity of commercial Ag NPs (Ag-Sigma, NPs < 150 nm) to the soil ecotoxicology model Folsomia candida (Collembola). Effects were assessed based on the standard OECD reproduction test (28 days) and beyond, with exposure prolonged for a second generation (56 days). Results showed that, based on the standard test (50% reproduction effect concentration—EC50 = 988 mg Ag/kg soil), the commercial Ag NPs were less toxic than the reference Ag NM300K and the ionic form AgNO3 (from literature). However, the toxicity dramatically increased (ca. 4 times) during the second-generation exposure (EC50(56d) = 234 mg Ag/kg soil), surpassing the toxicity of Ag NM300K. The decrease in adults’ size indicates that moulting might be affected. Overall, increased toxicity in prolonged exposure was not expected based on the available and standard test results, which highlights the importance of long(er)-term exposures to fully assess the risks of NMs to soil communities.

Journal of Xenobiotics doi: 10.3390/jox15060209

Authors: Volodymyr V. Tkach Tetiana V. Morozova Isabel Gaivão Ana Martins-Bessa Yana G. Ivanushko José Inácio Ferrão de Paiva Martins Ana Novo Barros

The use of sweeteners in e-cigarette liquids has become increasingly common, aiming to enhance the sensory appeal of vaping products. Compounds like aspartame, saccharin, and sucralose are added to provide a sweet taste without any calories, especially in flavored e-liquids popular among younger users. However, recent studies suggest that these additives may pose significant health risks when vaporized and inhaled. Sucralose, in particular, can break down into potentially harmful chlorinated by-products at high temperatures typical of vaping devices. Moreover, there is growing concern about the synergistic effects of sweeteners like sucralose, one sweetener with another and when combined with other e-liquid components. It has been observed that the presence of sucralose may amplify oxidative stress; genotoxicity, including mutations; and overall toxicity, along with environmental impact. This is not limited to nicotine- and smoke-related harm, as it may strengthen the toxic effect of the substances used in e-liquids that are not present in traditional cigarettes. The combined exposure to these heated compounds can intensify cytotoxicity, potentially increasing the risk of respiratory, cardiovascular, and neurological effects over time. While marketed as safer alternatives to tobacco, e-cigarettes containing sweeteners like sucralose may introduce new and poorly understood toxicological hazards that deserve urgent regulatory attention.

Journal of Xenobiotics doi: 10.3390/jox15060208

Authors: Constantina-Bianca Vulpe Adina-Daniela Iachimov-Datcu Andrijana Pujicic Bianca-Vanesa Agachi

Triazole fungicides are widely used in agriculture but may pose risks to human health through occupational, accidental, or environmental exposure. This systematic review aimed to evaluate the toxicity of ten European Union-approved triazole fungicides in rodent models and cell lines. A total of 70 studies were included, reporting quantitative in vivo oral, dermal, or inhalation toxicity in mammals or quantitative in vitro cytotoxicity in human or mammalian cell lines; the exclusion criteria comprised publications not in English or not accessible. Literature searches were conducted in Web of Science, Google Scholar, and the Pesticide Properties DataBase (PPDB), and risk of bias in included studies was assessed using ToxRTool. Due to heterogeneity in study designs, reporting formats, and endpoints, data were synthesized descriptively. Quantitative endpoints included LD50/LC50 values for in vivo studies and LOEC, IC50, LC50, and EC50 values for in vitro studies, while mechanistic endpoints highlighted apoptosis, oxidative stress, genotoxicity, and endoplasmic reticulum stress. Difenoconazole and tebuconazole were the most extensively studied compounds, whereas several triazoles had limited data. The limitations included heterogeneity of data and incomplete reporting, which restrict cross-study comparisons. Overall, the findings provide a comprehensive overview of potential human health hazards associated with EU-approved triazole fungicides and highlight critical knowledge gaps. The review was registered in Open Science Framework.

Journal of Xenobiotics doi: 10.3390/jox15060207

Authors: Minatullah Al-Ani Yassir Al-Ani Shahad Sabaawi Ibrahim Raghad Sabaawi Ibrahim Peter Kubatka Dietrich Büsselberg

Bisphenol A (BPA), a synthetic industrial compound widely found in plastics and other materials, has been linked to cancer development. As human exposure increases, BPA may pose potential carcinogenic concerns. Although BPA binds to estrogen receptors with much lower affinity than natural estrogens, its accumulation in human tissues can cause harmful effects. This review summarizes current evidence on BPA’s role in cancer initiation and progression, with a focus on its effects on cancer signaling pathways. These effects involve modulating pathways involved in cell growth, movement, invasion, survival, and adhesion. BPA acts as an estrogen ligand, binding to estrogen receptors and activating related pathways. The main route of exposure is through dietary intake of canned and plastic-packaged foods, with migration rates increasing at higher temperatures. To raise awareness of BPA’s harmful effects, industries have proposed “BPA-free” alternatives, some of which use derivatives like bisphenol S (BPS) and bisphenol F (BPF), which, unfortunately, may have even worse effects on human health. Given the ongoing challenges of eliminating BPA and similar harmful compounds, future research should focus on identifying safe substitutes, developing more effective removal technologies, and strengthening stringent regulations to mitigate public health risks.

Journal of Xenobiotics doi: 10.3390/jox15060206

Authors: Alevtina Y. Grishanova Maria L. Perepechaeva

Pharmacological compounds can disrupt glucose homeostasis, leading to impaired glucose tolerance, hyperglycemia, or newly diagnosed diabetes, as well as worsening glycemic control in patients with pre-existing diabetes. Traditional risk factors alone cannot explain the rapidly growing global incidence of diabetes. Therefore, prevention of insulin resistance could represent an effective strategy. Achieving this goal requires a deeper understanding of the mechanisms underlying the development of insulin resistance, with particular attention to the aryl hydrocarbon receptor (AhR). AhR, a transcription factor functioning as a xenobiotic sensor, plays a key role in various molecular pathways regulating normal homeostasis, organogenesis, and immune function. Activated by a range of exogenous and endogenous ligands, AhR is involved in the regulation of glucose and lipid metabolism as well as insulin sensitivity. However, current findings remain contradictory regarding whether AhR activation exerts beneficial or detrimental effects. This narrative review summarizes recent studies exploring the role of the AhR pathway in insulin secretion and glucose homeostasis across different tissues, and discusses molecular mechanisms involved in this process. Considering that several drugs act as AhR ligands, the review also compares how these ligands affect metabolic pathways of glucose and lipid metabolism and insulin sensitivity, producing either positive or negative effects.

Journal of Xenobiotics doi: 10.3390/jox15060205

Authors: Rita Sofia Vilela Francisco Pina-Martins Célia Ventura

Alternaria mycotoxins represent a significant and emerging concern in the field of food safety due to their widespread occurrence in diverse food and feed commodities, including cereals, tomatoes, oilseeds, and dried fruits. Among these, alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), and altertoxin-I (ATX-I) are the most frequently detected, often co-occurring at varying concentrations, thereby increasing the complexity of exposure and risk assessment. The gastrointestinal tract (GIT) is a crucial target of these toxins, as well as the liver, particularly considering its detoxifying role. Nevertheless, despite being a source of possible gastrointestinal and hepatic toxicity, there is still scarce data on the toxicokinetics of Alternaria toxins, on their mode of action, and respective toxic effects. To date, in vitro studies have shown that different Alternaria mycotoxins exhibit diverse toxicological effects, which may be dependent on their chemical structure. AOH and ATX-I have shown genotoxicity and cytotoxicity, mainly through interaction with the DNA and apoptosis, respectively. Tentoxin (TEN) has displayed hepatotoxic potential via impairment of detoxification pathways, and altenuene (ALT) has revealed lower toxicity. In vivo, AME and ATX-II revealed genotoxicity, while AOH and ATX-I showed context-dependent variability in their effects. Altogether, this review emphasizes that there is still a great lack of knowledge on these mycotoxins and an urgent need for more comprehensive toxicological and occurrence data to support proper risk assessment and, ultimately, regulatory decision-making.

Journal of Xenobiotics doi: 10.3390/jox15060204

Authors: Jiangang Wang Nannan Ma Gancong Mo Xian Qin Jin Zhang Xiangping Yao Jiahua Guo Zewei Sun

Triclosan (TCS) is a widely used antimicrobial agent found in personal care products and household cleaners. While valued since the 1960s for its ability to inhibit bacterial fatty acid synthesis, its environmental persistence, ecotoxicity, and bioaccumulative potential have raised significant global concern. The increased use of disinfectants during the COVID-19 pandemic has further exacerbated its prevalence as an aquatic pollutant. In the environment, TCS is distributed through water bodies and sediments, undergoing processes such as biodegradation and photochemical degradation. Its bioaccumulation poses a substantial threat to aquatic organisms, particularly fish. A growing body of research indicates that TCS acts as an endocrine disruptor and developmental toxicant, with documented adverse effects encompassing impaired embryonic and larval development, skeletal malformations, and induction of oxidative stress, mitochondrial dysfunction, DNA damage, and inflammatory responses. Furthermore, TCS exposure is linked to reproductive toxicity, including altered sex hormone levels and diminished reproductive capacity. This review consolidates current knowledge on the chemical properties, environmental fate, biodegradation pathways, and ecotoxicological impacts of TCS, with a specific emphasis on its multifaceted health risks to fish. The synthesis aims to provide a foundation for future research, inform environmental risk assessments, and support the development of evidence-based regulatory measures.

Journal of Xenobiotics doi: 10.3390/jox15060203

Authors: Francesca Rombolà Sara Bartoletti Sabrine Bilel Patrizia Hrelia Matteo Marti Monia Lenzi

In recent years, the expansion of the illicit market for Novel Psychoactive Substances (NPS) has resulted in the emergence of numerous synthetic recreational drugs specifically designed to evade legal control and analytical detection. Among these, nitazenes represent one of the most potent classes of new synthetic opioids, although information regarding their toxicological properties remains limited. The present study aimed to assess the genotoxic potential of four nitazenes: clonitazene, etonitazene, isotonitazene and metonitazene in human lymphoblastoid TK6 cells using a flow cytometric version of the In Vitro Mammalian Cell Micronucleus Test, following OECD Guideline No. 487. Cells were exposed to concentrations ranging from 12.5 to 100 μM, and cytotoxicity, cytostasis, and apoptosis were evaluated to identify appropriate doses for micronucleus frequency assessment. Vinblastine, a well-established mutagen, was included as positive control. Our findings demonstrated that clonitazene and isotonitazene exhibit mutagenic potential, suggesting an increased long-term risk of developing chronic degenerative diseases. Furthermore, the results revealed that structurally related molecules can induce markedly different cellular effects, underscoring the importance of compound-specific toxicological evaluations to achieve a comprehensive understanding of the risks associated with their illicit use—risks often presumed to involve only addiction or acute toxicity.

Journal of Xenobiotics doi: 10.3390/jox15060202

Authors: Aderonke Gbemi Adetunji Emmanuel Obeng-Gyasi

Exposure to environmental pollutants, including toxic metals, volatile organic compounds (VOCs), and per- and polyfluoroalkyl substances (PFAS), has been increasingly linked to impaired kidney function. However, the combined effects of these exposures, along with essential elements, on kidney health remain poorly understood. This study aimed to evaluate the independent and cumulative or mixture effects of toxic metals (cadmium, lead, and mercury), essential elements (iron, manganese, and selenium), PFAS (PFOA and PFOS), and VOCs (m-/p-xylene and o-xylene) on kidney function as measured by estimated glomerular filtration rate (eGFR). Using data from the National Health and Nutrition Examination Survey (NHANES), we applied multiple imputation to address missing data and implemented statistical techniques, including Bayesian Kernel Machine Regression (BKMR), quantile g-computation, and Weighted Quantile Sum Regression (WQSR) to assess complex exposure–response relationships, including non-linear, potential synergistic, and antagonistic effects. The results indicated that several exposures were correlated, particularly o-xylene with m-/p-xylene (r = 0.77), Cd with Pb (r = 0.46), and PFOS with PFOA (r = 0.61). eGFR was negatively associated with Pb, PFOS, PFOA, and Hg. In the BKMR analysis, overall posterior inclusion probabilities (PIPs) highlighted PFOS, Cd, Se, Mn, and Fe as the most influential exposures. Quantile g-computation highlighted Cd and Mn as major contributors, while WQSR modeling confirmed Mn as a key contributor. The findings underscore the importance of considering complex interactions in environmental exposure assessments. While essential elements may offer protective effects, toxic metals, PFAS, and VOCs remain critical contributors to kidney dysfunction. These insights highlight the need for integrative risk assessment approaches and public health strategies aimed at mitigating harmful exposures while promoting optimal nutrient balance.

Journal of Xenobiotics doi: 10.3390/jox15060200

Authors: Maura Calliera Ettore Capri Nicoleta Alina Suciu Marco Trevisan

This paper explores the evolving integration of the One Health framework into the European regulatory landscape for plant protection products, focusing on key scientific and procedural challenges. The analysis addresses three fundamental components of risk evaluation—regulatory complexity, hazard identification, and characterisation—and exposure assessment, while providing an up-to-date overview of emerging policies and challenges affecting the sustainable use of plant protection products in Europe. Addressing these issues requires interdisciplinary collaboration among toxicologists, epidemiologists, ecologists, regulatory authorities, industry stakeholders, and public health experts, working synergistically to tackle complex risks. It emphasises that transitioning to more sustainable and resilient agricultural systems in line with One Health principles requires critically reviewing existing policies. By integrating evolving scientific knowledge with communication and agricultural production needs across diverse European contexts, this approach offers valuable insights to inform future policy development and risk management innovation.

Journal of Xenobiotics doi: 10.3390/jox15060201

Authors: Yasser Essadki Antonio Casas-Rodríguez Antonio Cascajosa-Lira Leticia Diez-Quijada Alexandre Campos Vitor Vasconcelos Fatima El Khalloufi Brahim Oudra Ana M. Cameán Angeles Jos

Lichens are complex symbiotic systems known for synthesizing diverse secondary metabolites with documented antimicrobial, antioxidant, and antiproliferative activities. The present study focused on Pseudevernia furfuracea, a species widely distributed across Moroccan habitats. Two hydrodistillation-derived extracts (HE1 and HE2) were analyzed through ultra-high-Performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) to characterize their metabolite composition, and their effects were evaluated on Jurkat cells, a representative human cell line of the immune system. As the results of the characterization, the main compounds identified were Caprolactam, N,N-Diethylaniline, Erucamide, and 4-Isopropylaniline. Cytotoxicity assessment revealed that both HE1 and HE2 decreased the viability of Jurkat cells in a concentration-dependent manner. The mean effective concentrations (EC50) after 24 h of treatment were 53.79 ± 2.92 µg/mL for HE1 and 59.76 ± 2.01 µg/mL for HE2. Cell death mechanisms were further examined by flow cytometry, revealing that apoptosis predominated after 24 h of treatment, progressing mainly to late apoptotic stages after 48 h. In parallel, the expression levels of key cytokine genes, including IL-2, TNF-α, and IFN-γ, were quantified at the mRNA level to evaluate potential immunomodulatory effects. Up-regulation was observed in IL-2 after exposure to both extracts for 24 and 48 h, and in the case of IFN-γ after exposure to HE2 for 24 h; in contrast, HE1 and HE2 produced down-regulation in TNF-α at 24 h. These findings suggest that HE1 and HE2 have immunomodulatory activity in Jurkat cells. Further investigations are needed to elucidate the underlying mechanisms and to clarify how HE1 and HE2 influence immune responses in human systems.

Journal of Xenobiotics doi: 10.3390/jox15060199

Authors: Soisungwan Satarug David A. Vesey Tanaporn Khamphaya Donrawee Waeyeng Supabhorn Yimthiang

This study investigates the relationship between kidney function and exposure to low-level cadmium (Cd) and lead (Pb) in individuals with and without diabetes. Specifically, it tests the hypothesis that the nephrotoxicity of Cd and Pb reduces the tubular degradation of filtered proteins, namely β2-microglobulin (β2M). Data were obtained from a Thai cohort of 137 people, of which 65 were diagnosed with diabetes. Blood Cd, blood Pb, and urinary excretion of Cd (ECd) were used as exposure indicators, while urinary N-acetylglucosaminidase (ENAG) and fractional tubular degradation of β2M (FrTDβ2M) reflected kidney tubular cell injury and the function of tubular cells, respectively. Spearman’s rank correlation revealed that FrTDβ2M varied directly with the estimated glomerular filtration rate (eGFR; r = 0.434), and inversely with fasting plasma glucose (r = −0.215), ECd (r = −0.527), ENAG (r = −0.536), and Cd/Pb exposure (r = −0.249). In a multiple regression model analysis adjusting for potential confounders, the association between FrTDβ2M and eGFR in those with diabetes was particularly strong (β = 0.476) compared to controls (β = 0.360), whereas an inverse association of FrTDβ2M and ECd (β = −0.295) was found only in those with diabetes, along with a positive association of ENAG with ECd (R2 = 0.071). A mediation analysis has revealed that tubular injury (ENAG) mediated 26% of the FrTDβ2M decrease associated with Cd/Pb exposure. These findings suggested that tubular protein degradation pathways may be compromised under combined metabolic and environmental stressors, Cd, and Pb.

Journal of Xenobiotics doi: 10.3390/jox15060197

Authors: Jana Růžičková Helena Raclavská Marek Kucbel Pavel Kantor Barbora Švédová Karolina Slamová

Volatile organic compounds (VOCs) emitted from indoor surface coatings can significantly impact indoor air quality and health. This study compared emissions from water-based polyurethane (PUR) and acrylate–polyurethane (ACR–PUR) coatings, identifying 94 VOCs across 16 chemical classes. Time-resolved concentrations were analysed via Principal Component Analysis (PCA), which revealed distinct temporal emission patterns and chemically coherent clusters. Aromatic hydrocarbons, alcohols, esters, and isocyanates dominated the emission profiles, with ACR–PUR releasing markedly higher concentrations of symptom-relevant compounds. Acute exposure was linked to toluene, styrene, phenol, and methyl butyl ketone (MBK), which decreased sharply within 60 days, while compounds such as 1,3-dioxolane, isopropylbenzene, and ethenyl acetate exhibited persistent emissions, suggesting increased chronic risk. Although total VOC levels remained below the German UBA “excellent” threshold (<200 µg/m3), neurotoxic and carcinogenic compounds remained detectable. The combination of PCA-based temporal insights with toxicological profiling and emission transfer dynamics offers a refined framework for indoor air risk assessment. These results underscore the need to complement total VOC indices with symptom-oriented, time-resolved screening protocols to better evaluate SBS risk in indoor environments using water-based coatings.

Journal of Xenobiotics doi: 10.3390/jox15060198

Authors: François Gagné Chantale André

There is an urgent need to evaluate the toxicity of xenobiotics and environmental mixtures for preventing loss in water quality for the sustainability of aquatic ecosystems. A simple prebiotic chemical pathway based on malate formation from pyruvate (pyr) and glyoxalate (glyox) is proposed as a quick and cheap screening tool for toxicity assessment. The assay is based on the pyr and glyox (aldol) condensation reactions, leading to biologically relevant precursors such as oxaloacetate and malate. Incubation of pyr and glyox at 40–70 °C in the presence of reduced iron Fe(II) led to malate formation following the first 3 h of incubation. The addition of various xenobiotics/contaminants (silver, copper, zinc, cerium IV, samarium III, dibutylphthalate, 1,3-diphenylguanidine, carbon-walled nanotube, nanoFe2O3 and polystyrene nanoparticles) led to inhibitions in malate synthesis at various degrees. Based on the concentration inhibiting malate concentrations by 20% (IC20), the following potencies were observed: silver < copper ~ 1.3-diphenylguanidine ~ carbon-walled nanotube < zinc ~ samarium < dibutylphthalate ~ samarium < Ce(IV) < nFeO3 < polystyrene nanoplastics. The IC20 values were also significantly correlated with the reported trout acute lethality data, suggesting its potential as an alternative toxicity test. The pyr-glyox pathway was also tested on surface water extracts (C18), identifying the most contaminated sites from large cities and municipal wastewater effluents dispersion plume. The inhibition potencies of the selected test compounds revealed that not only pro-oxidants but also chemicals hindering enolate formation, nucleophilic attack of carbonyls and dehydration involved in aldol-condensation reactions were associated with toxicity. The pyr-glyox pathway is based on prebiotic chemical reactions during the emergence of life and represents a unique tool for identifying toxic compounds individually and in complex mixtures.

Journal of Xenobiotics doi: 10.3390/jox15060196

Authors: Doyinsola Suliat Mustapha Olga Rodríguez-Díaz Miren P. Cajaraville Amaia Orbea

Conventional plastics are widely utilised across industrial sectors and in consumer products. However, the growing use of plastics has led to plastic pollution, including the formation of nanoplastics (NPs), which are harmful to aquatic organisms. Bioplastics are emerging alternatives. They are renewable and/or biodegradable and are supposed to be more environmentally friendly. However, the toxicity and environmental fate of bioplastics are not yet fully understood. This study evaluated the toxicity and fate of commercially available plain or fluorescent polylactic acid (PLA) NPs (250 nm) on aquatic organisms. Confocal microscopy demonstrated the uptake of fluorescent PLA NPs by the test organisms, marine rotifers (Brachionus plicatilis), brine shrimps (Artemia salina) and zebrafish (Danio rerio) embryos. However, the results of the bioassays indicate that plain PLA NPs did not induce acute toxicity in either of the two zooplankton species and did not cause substantial mortality, malformations, or hatching delays in zebrafish embryos at the tested concentrations (up to 100 mg/L). However, brine shrimp showed a significant decrease in ingestion capability. The biochemical biomarkers, catalase activity induction, as an indicator of oxidative stress, and acetylcholinesterase inhibition, as a marker of neurotoxicity, showed no significant alterations compared to the control of both zooplankton species and that of zebrafish embryos. Overall, the findings suggest a pattern of no acute and low sublethal toxicity for the tested plain PLA NPs in the studied organisms. Nonetheless, further research is imperative to comprehensively assess the environmental fate of bioplastics found in various consumer products, as these may contain harmful chemical additives, as well as the effects of prolonged exposure and their impact on physiological parameters, ensuring informed decisions before their widespread commercialisation and presence in the environment.

Journal of Xenobiotics doi: 10.3390/jox15060195

Authors: Darija Klinčić Karla Jagić Nemčić Ivana Jakovljević Marija Jelena Lovrić Štefiček Marija Dvoršćak

Indoor exposure to polybrominated diphenyl ethers (PBDEs), particularly those bound to fine particulate matter (PM1, particles < 1 µm), may pose a health concern, especially in light of prolonged indoor occupancy and the capacity of ultrafine particles to reach the lower respiratory tract. This study investigates indoor exposure to PBDEs associated with PM1 in residential homes in Zagreb, Croatia, across warm and cold seasons. BDE-47 was consistently detected in all samples, while BDE-183 was consistently absent. Elevated concentrations and increased detection frequencies of BDE-99 and BDE-100 were observed during the colder season. Consequently, total PBDE (ΣPBDE) levels in the cold season were approximately 2.5 times higher than in the warm season. Although estimated daily inhalation intakes were below established oral reference doses, the potential for deep pulmonary deposition and systemic distribution underscores the need for further investigation. These findings represent the first reported data on indoor PM1-associated PBDEs in Europe, emphasizing the impact of seasonal dynamics on inhalation exposure due to variation on indoor contaminant levels.