Journal of Xenobiotics doi: 10.3390/jox16040125

Authors: Maria João Gouveia Nuno Vale

Drug repurposing represents a rational and resource-efficient strategy to expand the oncological armamentarium by leveraging the established pharmacology, clinical experience, and safety-monitoring frameworks of approved non-oncological agents. Clozapine (CZP), an atypical antipsychotic characterized by broad receptor pharmacology, complex biotransformation, and clinically relevant toxicological liabilities, has emerged as a candidate of interest following preclinical evidence of context-dependent anticancer activity across multiple tumor types. As such, CZP provides an informative case study at the interface between therapeutic drug action and xenobiotic behavior. This review provides a critical and integrated synthesis of the current evidence supporting the repurposing of CZP in oncology, with particular emphasis on the relationship between its molecular mechanisms, dose–exposure requirements, pharmacological complexity, and potential toxicity. Analysis of in vitro and in vivo studies across glioblastoma, non-small cell lung cancer, breast cancer, and melanoma brain metastasis models indicates that CZP can impair tumor cell proliferation and survival through a form of mechanistic plasticity. Rather than acting through a single conserved pathway, CZP appears to disrupt shared upstream processes related to pro-survival signaling, cellular stress tolerance, and metabolic homeostasis, while engaging tumor-specific downstream responses, including autophagic cell death, mitochondria-dependent apoptosis, oxidative stress, and coordinated modulation of survival and angiogenic pathways. Despite this mechanistic rationale, translation remains substantially constrained, most notably by the order of magnitude gap between anticancer-effective concentrations in vitro and clinically achievable plasma exposures, requiring careful distinction between potentially useful anticancer pharmacology and nonspecific xenobiotic-induced cellular stress and clinically unacceptable toxicity. Key limitations include the discrepancy between anticancer-effective concentrations observed in vitro and exposures achievable during standard psychiatric dosing, the limited understanding of how CZP metabolism and metabolite formation may influence efficacy and toxicity, the absence of integrated pharmacokinetic–pharmacodynamic and toxicokinetic modeling, and the lack of dedicated clinical trial evidence. To address these challenges, this review examines complementary translational strategies, including patient-derived organoids, co-culture systems, microphysiological platforms, pharmacokinetic and toxicological modeling, and computational digital twin frameworks. Together, these approaches may support a biologically informed and risk-aware evaluation of CZP, helping to identify responsive tumor contexts, anticipate exposure-related liabilities, and prioritize rational combination strategies. By integrating therapeutic potential with xenobiotic pharmacology and toxicology, this review positions CZP within the evolving landscape of precision oncology and evidence-driven drug repurposing.

Journal of Xenobiotics doi: 10.3390/jox16040124

Authors: Jhon Fredy Narváez-Valderrama Juan José García-Londoño Juan David González-Calderón Yileni Argoti-Ospina Gabriel Jaime Maya Jorge L. Gallego Ana Luisa Urrego Carlos Daniel Ramos-Contreras

Maternal exposure to polycyclic aromatic hydrocarbons (PAHs) during pregnancy has been associated with adverse obstetric and perinatal outcomes, including miscarriage, low birth weight, intrauterine growth restriction, and spontaneous abortion. Exposure occurs through multiple pathways, including dietary intake and inhalation, which ultimately determine the final body burden. PAHs may reach relevant levels in the blood, representing the initial step in their internal distribution to the placenta, umbilical cord, and breast milk, thereby compromising maternal–fetal health. In this exploratory study, maternal blood samples were collected from pregnant women residing in different regions of Antioquia, Colombia. Serum was isolated from whole blood, subsequently extracted using freezing-assisted liquid–liquid extraction, purified by solid-phase extraction, and analyzed by GC–MS. Method performance showed PAH recoveries between 60 and 120%, limits of detection (LOD) ranging from 0.5 to 3.3 ng·mL−1, and limits of quantification (LOQ) ranging from 1.4 to 9.9 ng·mL−1. Airborne PAH concentrations were measured using a photoelectric aerosol sensor, and higher levels were observed in municipalities intersected by major highways, indicating a strong vehicular contribution, with an average concentration of 72.6 ± 39.2 ng·m−3. Low and medium-molecular weight PAHs were detected in serum samples at an average concentration of 43.8 ± 8.8 ng·g−1 of lipid (mean of ∑ individual congeners). In contrast, a high-molecular-weight PAH, benzo[a]pyrene (BaP), was detected in one participant. Pyrene (PYR) and fluoranthene (FLU) were the predominant congeners, suggesting combustion-related sources, primarily vehicular emissions. Serum PAH levels showed a correlation with the frequency of consumption of canned fish and meat, but not with short-term airborne PAH measurements. These exploratory findings suggest that dietary intake is a primary pathway of bioaccumulation during acute exposure and plays a key role in determining the parental PAHs burden during pregnancy in polluted environments. However, additional data on parent PAHs and their metabolites are needed to provide a more comprehensive assessment of cumulative exposure arising from dietary sources and chronic inhalation of airborne PAHs.

Journal of Xenobiotics doi: 10.3390/jox16040123

Authors: Zvonimir Užarević Martina Šrajer Gajdošik Elvira Kovač-Andrić Lidija Kalinić Mihaela Vranješ Delać Dinko Puntarić Eda Puntarić Domagoj Vidosavljević Mario Begović Vlatka Gvozdić

Selenium is a trace element of vital importance for ecosystem functioning and human health mainly due to its antioxidant and protective properties. Since both selenium deficiency and excess can have harmful effects on living organisms, monitoring its distribution in biological systems and the environment is of significant scientific and public health interest. This study systematically assessed selenium concentrations in biological (urine, serum, and hair) and environmental (soil and dandelion) samples in eastern Croatia. Selenium concentrations were determined using the inductively coupled plasma mass spectrometry method. In biological samples, median selenium concentrations were from 15.02 to 37.15 μg·L−1 in urine, from 80.76 to 114.05 μg·L−1 in serum, and from 0.21 to 0.46 µg·g−1 in hair. In environmental samples, median selenium concentrations varied depending on location, ranging from 0.32 to 0.51 mg·kg−1 in soil and from 6.55 to 84.41 µg·kg−1 in dandelion. PCA was applied to identify overall patterns and groupings among biological and environmental samples, showing that dandelion samples from urban locations exhibited the highest selenium concentrations, indicating the influences of urban environmental conditions on selenium accumulation.

Journal of Xenobiotics doi: 10.3390/jox16040122

Authors: Chrysi A. Papadimitriou Christina Emmanouil Amalia Moriki Vasileios Bartzis

Tetracyclines (TCs) are antibiotics widely used in human and veterinary medicine as well as in agricultural practices. They may be retained in soil or drift into freshwater, thereby exerting effects on non-target organisms and deteriorating ecological quality. In this study, tetracycline (T), oxytetracycline (OT), chlortetracycline (CT), and their binary and ternary mixtures were evaluated using a battery of bioassays including terrestrial plants, aquatic crustaceans, a ciliate protist and a bacterial species. Results showed a concentration-dependent effect for parameter immobilization in Daphnia magna and Artemia salina, seed germination in the terrestrial plants, and bioluminescence inhibition and growth inhibition in Aliivibrio fischeri and Tetrahymena thermophila, respectively. For A. fischeri, statistically significant interactions were observed between dose and exposure time. A. salina demonstrated greater sensitivity than D. magna in all cases. Both A. salina and A. fischeri showed increased toxicity to OT and the ternary mixture. Dicots presented greater sensitivity than the monocot species in all cases. In the combined exposures, there was a deviation from the concentration addition (CA) model, with possible synergism for CT + T and the ternary mixture for A. fischeri. The concurrent environmental exposure of non-target organisms to TCs should be investigated further.

Journal of Xenobiotics doi: 10.3390/jox16040121

Authors: Ahmad A. Alresheedi Omnia A. Nour Dalia H. El-Kashef Manar A. Nader

Background/Objectives: Glucocorticoid therapy remains clinically indispensable, yet its long-term use is profoundly constrained by insulin resistance (IR), hepatic steatosis, and progressive metabolic dysfunction. Methylsulfonylmethane (MSM), a naturally occurring sulfur-containing nutraceutical with established antioxidant and anti-inflammatory activities, has emerged as a promising metabolic modulator; however, its therapeutic relevance in glucocorticoid-induced hepatic IR has not previously been explored. Methods: Male Wistar rats received MSM (200 or 400 mg/kg/day, p.o.) for 14 days, while dexamethasone (DEX) (8 mg/kg/day, i.p.) was administered during the final 7 days to induce severe metabolic dysfunction. Results: DEX provoked profound IR, dyslipidemia, oxidative stress, hepatocellular injury, and steatotic degeneration accompanied by marked ultrastructural abnormalities. Remarkably, MSM conferred dose-dependent metabolic and hepatoprotective effects, significantly restoring glucose homeostasis, insulin responsiveness, lipid metabolism, and hepatic structural integrity. Mechanistically, MSM exerted a pleiotropic regulatory effect through suppression of the glucocorticoid-responsive kinase SGK1, restoration of AMPK/mTOR signaling balance, and normalization of insulin signaling pathways and metabolic transcriptional regulators. Furthermore, MSM effectively attenuated oxidative stress and inflammatory amplification consistent with modulation of the NLRP3/NF-κB/IL-6 axis. Importantly, the current work identifies angiogenic remodeling demonstrated by DEX-induced upregulation of VEGF and CD34, both of which were substantially suppressed by MSM treatment. Conclusions: This study provides novel evidence that MSM mitigates glucocorticoid-induced hepatic IR through coordinated modulation of glucocorticoid-responsive kinases, metabolic signaling networks, redox–inflammatory cascades, and pathological angiogenesis. Consequently, MSM may represent a promising candidate for further preclinical and clinical evaluation regarding its capacity to limit glucocorticoid-associated metabolic burdens.

Journal of Xenobiotics doi: 10.3390/jox16040120

Authors: Azza Fahmy Amany Mohammed Mohmmed Hegab Hanan S. Mossalem Samah Sulaiman Abuzahrah Saud Omar Alafghani Alaaeldin Ahmed Hamza Nouf Juaid Amr Amin

Although praziquantel (PZQ) is the main antischistosomal drug currently in use, concerns remain regarding incomplete reversal of schistosomiasis-induced pathology and the emergence of drug resistance. This study evaluates the combined effect of PZQ with safranal, a bioactive saffron constituent, on Schistosoma mansoni-induced pathology in mice. Male CD1 Swiss albino mice were exposed to 60 S. mansoni cercariae and, at week 9 post-infection, were treated with PZQ (500 mg/kg orally for two consecutive days), safranal (50 mg/kg/day), or both, for three weeks. The animals were sacrificed at week 11 post-infection. Worm and egg burdens, liver histopathology, fibrotic markers, oxidative stress, and inflammatory cytokines were assessed. Combined PZQ + safranal therapy significantly reduced adult worm counts and hepatic and intestinal egg loads compared to PZQ alone. All treatments decreased liver index (hepatomegaly), with the combination treatment providing the best intervention. Histological analyses revealed significantly reduced granuloma size and hepatic necrosis post-treatment, particularly in the combination group. The levels of proinflammatory cytokines (TNF-α, IL-1β) and Th2 cytokines (IL-4, IL-5, IL-6, IL-10) were significantly lowered in treated mice, most notably with the combination treatment. Oxidative stress was also markedly attenuated, and infected mice exhibited elevated malondialdehyde and depleted antioxidant enzymes (SOD, CAT, GSH). Interestingly, PZQ and/or safranal restored antioxidant status and reduced lipid peroxidation, with the combination being most effective. Furthermore, collagen deposition and expression of hepatic fibrotic markers α-smooth muscle actin (α-SMA), TGF-β1, and matrix metalloproteinase-9 were most effectively suppressed by combined therapy. To conclude, safranal enhances PZQ’s antischistosomal efficacy and confers additive protection against Schistosoma-induced liver fibrosis.

Journal of Xenobiotics doi: 10.3390/jox16040119

Authors: Jeremy J. Ramsden

Most airline passengers and crew assume that the air in the cabin is free from harmful or hazardous substances, as is mandated by airworthiness regulations. While fresh air entering the cabin is sterile (and if recirculated is usually efficiently filtered to remove microorganisms), if the fresh air is bled off the turbine compressors (as is the case in about 95% of airliners currently in service), it may be contaminated with traces of engine oil and ultrafine particles abraded from the turbine blades, and possibly traces of hydraulic fluid leaking from servo systems. Engine oil contains tricresyl phosphate (TCP) as an essential antiwear agent, but it is also a well-known neurotoxin, and it has been suggested that there may be no safe lower limit of exposure, not least because of considerable variation among individuals in sensitivity to tri-ortho-cresyl phosphate (ToCP) and other isomers with at least one ortho constituent. This paper reviews current knowledge about these hazards and discusses the medical and economic motivations for diminishing them. A calculation based on maintaining the life quality index shows that eliminating xenobiotic hazards in aircraft cabin air is likely to be affordable.

Journal of Xenobiotics doi: 10.3390/jox16030118

Authors: Fatouma Mohamed Abdoul-Latif Rohit Kumar Talal Mohamed Ali Merito N Chinmaya Kumar Ibrahim Houmed Aboubaker Pannaga Pavan Jutur

African soils face increasing levels of metal pollution due to industrialization, artisanal mining activities, improper waste management, and enhanced agricultural productivity. However, unlike many organic pollutants, heavy metals do not degrade naturally and therefore persist in environmental systems for prolonged periods. Heavy metals accumulate over many decades in the soil and bioaccumulate through the food chain causing severe health complications such as cancer, kidney problems, and neurological impairment. This paper reviews the current literature on the origin, prevalence, and behavior of the main pollutants Pb, Cd, Cr, As, Hg, and Cu. The major phytoremediation methods including phytoextraction, rhizofiltration, phytostabilization, and phytovolatilization are highlighted alongside in planta screening methods for hyperaccumulating plants including Berkheya coddii (Ni) and Haumaniastrum robertii (Co). The paper evaluates various enhancement techniques such as the use of chelators, Rhizobium inoculations, and genetic modifications. The significance of these approaches in tropical and subtropical climates is discussed. The paper suggests a holistic framework involving empirical kinetic modeling, geospatial machine learning (random forest, kriging), and molecular omics in prediction modeling. Major hurdles in such predictions include lack of field-based verification of the models, biotechnology safety of genetically modified (GM) organisms, and inadequate regulations. Future perspectives emphasize community-driven phytomining, biomass recycling, and resilient phytoremediation solutions.

Journal of Xenobiotics doi: 10.3390/jox16030117

Authors: Nuraeni Nuraeni Herdis Herdiansyah Fatmah Fatmah Haruki Agustina Rully Yusuf

This study analyzes the determinants of self-reported behaviours and perceptions associated with pesticide toxicity risk in children using the Social Practice Theory framework, linking individual factors and agricultural practices to understand vulnerability and prevention opportunities. This research was conducted in Pattapang Village, Tinggimoncong District, Gowa Regency, South Sulawesi Province, Indonesia. To examine the relationship between pesticide use patterns, social norms, competence, material, and individual aspects and the risk of sensitive toxicity in children, data were analyzed using structural equation modeling-partial least squares (SEM-PLS) with bootstrapping resampling. Pesticide use patterns had a significant negative effect on toxicity risk. Competence was the strongest predictor of pesticide use patterns, followed by materials and short-term goals. Personal values dominate personal norms and long-term goals, while social norms only influence personal norms. Self-efficacy, personal norms, and long-term goals showed no significant effects. The novelty of this research lies in the integration of a socio-ecological approach with individual psychological factors in a comprehensive structural model that explains the complex mechanisms of children’s protective behavior formation from pesticide toxicity, identifying that personal values—not personal norms or self-efficacy—are the most effective leverage points for farmer behavior change interventions.

Journal of Xenobiotics doi: 10.3390/jox16030116

Authors: Brankica Kartalović Rastko Tomanović Kristina Habschied Alma Mikuška Mirta Sudarić Bogojević Antonije Žunić Dora Bjedov

Atmospheric wet deposition represents a major pathway for the transfer of organic micropollutants into terrestrial and aquatic ecosystems. This study investigates the occurrence and spatial distribution of polycyclic aromatic hydrocarbons (PAHs), phthalate esters (PAEs), and BTEX compounds in rainwater across Northern Serbia (Vojvodina region). Rainwater samples were collected during the 2025–2026 heating season at three locations: a petrochemical site in Kikinda, a traffic- and residentially influenced site in Sremska Mitrovica, and an urban background site in Sombor. Total concentrations showed pronounced spatial variability, with the highest ΣBTEX and ΣPAE levels recorded in Kikinda (∑BTEX = 2.818 μg L∑1; ∑PAE = 0.930 μg L∑1). Diagnostic ratios identified a dominant petrogenic signature in Kikinda (LMW/HMW > 1), while pyrogenic sources prevailed in Sremska Mitrovica and Sombor ((Fla/Fla + Pyr) > 0.5). BTEX profiles across all sites were characterised by the absence of benzene and elevated toluene and xylene levels (B/T ≈ 0; T/X > 1). Health risk assessment indicated an acceptable but non-negligible carcinogenic risk from PAHs, particularly for children in industrial areas. These findings highlight the role of precipitation as an efficient scavenger of organic pollutants and emphasise the need for integrated atmospheric–hydrological monitoring frameworks in industrialised regions.

Journal of Xenobiotics doi: 10.3390/jox16030115

Authors: Katarina Baralić Katarina Vidić Đurđica Marić Jovana Živanović Aleksandra Buha Djordjevic Marijana Ćurčić Zorica Bulat Biljana Antonijević Danijela Đukić-Ćosić

Per- and polyfluoroalkyl substances (PFAS) are persistent, chemically stable compounds widely used in daily life. Perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorohexanesulfonic acid (PFHxS), and perfluorooctanesulfonic acid (PFOS) were identified as the most relevant PFAS due to their prevalence and toxicity. This study aimed to investigate the immunotoxic mechanisms of a mixture of these PFAS using an in silico approach. Comparative Toxicogenomic Database (CTD), GeneMANIA, CytoHubba (Cytoscape), ToppGene Suite, and Metascape were used for the analysis. A total of 65 immune-related genes were identified as common to all four PFAS, with IFNG, TNF, IL1B, IL6, TYK2, CD3E, CASP8, VAV1, ARHGAP4, and CARD11 emerging as key hub genes. CTD phenotype analysis indicated immune dysregulation, with decreased humoral and adaptive immune responses in humans and tissue-specific modulation of B- and T-cell activity in mice, while no immune-related phenotypes were observed for PFNA. Network analysis identified functional modules associated with apoptotic and immune signaling, endothelial cell migration and angiogenesis, and shared inflammatory and viral response pathways. Disease enrichment analysis associated PFAS with autoimmune disorders (rheumatoid arthritis, asthma), metabolic conditions, and cardiovascular diseases (experimental diabetes, hypertensive disease). These results highlight PFAS involvement in immune modulation, cytokine signaling, and disease susceptibility.

Journal of Xenobiotics doi: 10.3390/jox16030114

Authors: Sebastiano La Maestra Francesco D’Agostini Linda Ferrea

Air pollution is a major environmental determinant of premature mortality and population health burden. Liguria represents a vulnerable Mediterranean region due to intense urbanisation, port-related emissions, complex topography and an ageing population. This study quantified the mortality burden and Years of Life Lost (YLL) attributable to long-term exposure to PM2.5, NO2 and O3 in Liguria (Italy), and estimated the potentially avoidable burden under WHO guideline scenarios. A Health Impact Assessment (HIA) was conducted using ARPAL air quality data and ISTAT mortality data for the population aged ≥30 years during 2022–2024. Relative risks were derived from the European ELAPSE project and WHO meta-analyses. Attributable mortality was estimated using a log-linear Health Impact Function, while YLL were calculated using regional life tables and normalised per 100,000 inhabitants. PM2.5 was the main contributor to air pollution-related mortality, accounting for 1333 attributable deaths in 2022. Corresponding YLL ranged from 755 to 1012 per 100,000 inhabitants over the study period. NO2 showed a relevant but secondary contribution, while O3 effects were smaller and more uncertain. WHO guideline scenarios indicated a substantial potentially avoidable burden of deaths and YLL. These findings support targeted environmental and public health interventions in highly urbanised coastal regions.

Journal of Xenobiotics doi: 10.3390/jox16030113

Authors: Tarek M. Abdelghany Alaa A. Budastour Ahmed S. Kamel Sherehan M. Ibrahim Alex Charlton Simon Wilkinson Catherine Arden Noha F. Abdelkader Matthew C. Wright

M8OI is a cytotoxic methylimidazolium ionic liquid solvent through its binding to the ubiquinone binding site on complex I of the mitochondrial electron transport chain. Given the overlap in terms of toxic mechanism of action with the pesticide rotenone, the potential neurotoxic effects of M8OI were examined. In vitro, cytotoxicity and mitochondrial function were assessed in SH-SY5Y cells by measuring MTT reduction and oxygen consumption/extracellular acidification using a Seahorse analyser. SH-SY5Y cells were sensitised to M8OI toxicity by replacing medium glucose with galactose. Glucose protected the cells from M8OI toxicity, whereas galactose showed no clear dose–response protection. M8OI induced a dose-dependent reduction in oxygen consumption rate with a compensatory increase in extracellular acidification rate, consistent with inhibition of mitochondrial oxidative phosphorylation and a shift toward glycolysis. In vivo, rats were orally exposed via drinking water for 20 weeks and assessed using behavioural tests. In addition, the concentrations of M8OI and its metabolites were quantified by LC–MS in rat brain and other tissues. In rats, M8OI concentrations were ~30-fold higher in kidney than brain, and brain levels were at least 100-fold lower than the concentrations that affected SH-SY5Y cell viability in vitro. However, based on open field tests, M8OI exposure suppressed motor activity without any anxious behaviours. The cytotoxicity of M8OI in SH-SY5Y neuroblastoma cells was associated with metabolic mitochondrial dysfunction. However, the neurobehavioural changes observed in orally exposed rats occurred at significantly lower brain concentrations than would be predicted to lead to neural cell death. Nevertheless, direct comparisons between acute in vitro exposures and chronic in vivo outcomes should be interpreted cautiously.

Journal of Xenobiotics doi: 10.3390/jox16030112

Authors: Wonsik Shin Pil-Gon Kim Min-Ho Oak

The sorption behavior of antibiotics in wastewater treatment systems plays a critical role in determining their environmental fate and removal efficiency. In this study, the sorption of 15 antibiotics representing multiple classes was investigated using two sewage sludge samples with different physicochemical characteristics. Batch equilibrium experiments were conducted to evaluate time-dependent sorption behavior and to determine solid–water distribution coefficients (Kd). The results showed that sorption occurred rapidly, with most compounds approaching a stable concentration within 24 h. The Kd values varied widely depending on the compound, ranging from 74 to 737 L/kg. For 13 of the 15 investigated antibiotics, higher Kd values were observed in sludge B than in sludge A, with the largest difference observed for tiamulin (402 ± 53 and 737 ± 76 L/kg for sludge A and sludge B, respectively). Sludge B generally exhibited higher sorption capacity for most compounds than sludge A, despite having a lower specific surface area, indicating that sorption was governed primarily by chemical composition and pore structure rather than surface area alone. Elemental and morphological analyses suggested that differences in metal-associated components and pore structure may contribute to the higher sorption capacity observed in sludge B. However, the specific sorption mechanisms could not be directly confirmed by the present analyses. Comparison with previous studies confirmed that the measured Kd values fall within reported ranges but are generally higher for sulfonamides, suggesting enhanced sorption capacity of the investigated sludge matrices. Application of an equilibrium-based model demonstrated that sorption alone can account for approximately 20–70% of antibiotic removal under typical activated sludge conditions, depending on compound affinity. These findings highlight the importance of sludge-specific properties in controlling antibiotic partitioning and demonstrate that incorporating such characteristics into predictive models can improve the accuracy of environmental fate assessments in wastewater treatment systems.

Journal of Xenobiotics doi: 10.3390/jox16030111

Authors: Fatima Abrantes-Soares Mariana Marques Santos Melissa Mariana Margarida Lorigo Elisa Cairrao

In recent years, bisphenol F (BPF) and bisphenol S (BPS) have been used in several everyday products to replace bisphenol A (BPA), since exposure to BPA has been associated with the development of several pathologies. However, recent studies have also been associating exposure to BPA substitutes with the development of various pathologies, including cardiovascular diseases, and the safety of BPA substitutes for human health has been questioned. Thus, this study aimed to investigate and compare BPA, BPF and BPS effects on arterial tone and to explore the mechanisms involved. The results suggest that BPA, BPS and BPF exert non-genomic and endothelium-independent relaxant effects on arteries and smooth muscle cells from the umbilical cord. Regarding genomic effects, the results suggest that BPA, BPF, and BPS disrupted the primary mechanisms underlying HUA relaxation by interfering with the cGMP signaling pathway and modulating the Ca2+ channels activity. Moreover, these results suggest that BPF alters the vasorelaxant response more than BPA and BPS. Therefore, replacing BPA with its substitutes does not appear to be beneficial for human cardiovascular health. Thus, in the future, the vascular effects of these bisphenols should be further evaluated to clarify their modes of action and future implications for maternal-fetal health.

Journal of Xenobiotics doi: 10.3390/jox16030110

Authors: Anisma R. Gokoel Arti Shankar Simran F. Mokiem Ashna D. Hindori-Mohangoo Maureen Y. Lichtveld Jeffrey K. Wickliffe Wilco C. W. R. Zijlmans

Maternal anemia and prenatal exposure to neurotoxic metals are widespread in low- and middle-income countries and may affect early childhood development. In Suriname, where mercury contamination from artisanal gold mining and social disparities coexist, we examined associations between maternal anemia, prenatal exposure to mercury (Hg), lead (Pb), manganese (Mn), selenium (Se), and cadmium (Cd), and early neurodevelopment. The study included 755 pregnant women and 644 children (10–26 months) from the Caribbean Consortium for Research in Environmental and Occupational Health (CCREOH) cohort. Maternal anemia was defined using WHO criteria for Hb, metals were measured in maternal blood, and child development was assessed using the Bayley Scales of Infant and Toddler Development, third edition (BSID-III-NL). Analyses used non-parametric tests, correlations, and multivariable regression. Anemia, though common (34%), was not independently associated with cognitive, language, motor, or social–emotional outcomes. However, iron status was not directly measured; therefore, the absence of an observed association should not be interpreted as evidence that maternal iron deficiency is unrelated to early neurodevelopment. Pb showed the most consistent associations, with higher prenatal levels predicting poorer cognitive, motor, and language scores. Hg demonstrated weaker but significant negative associations with several domains, while Mn and Cd showed limited direct effects. Interaction analyses suggested a potential modifying role of Se in certain metal-neurodevelopment associations; however, these findings require confirmation in future studies. Overall, these results suggest that prenatal exposure to neurotoxic metals and sociodemographic disparities may be important contributors to variation in early neurodevelopment in this population, but causal inferences cannot be made from this cross-sectional analysis.

Journal of Xenobiotics doi: 10.3390/jox16030108

Authors: Alfredo Cruz-Gregorio Alejandro Silva-Palacios Javier A. Belmont-Díaz María del Pilar Ramos-Godinez Rebeca López-Marure

Food-grade titanium dioxide (E171) is widely used as a food additive and has raised concerns about its potential systemic toxicity. However, its impact on cardiac cellular metabolism and mitochondrial function remains incompletely understood. In the present study, we investigated the effects of E171 on intracellular labile iron, oxidative stress, mitochondrial ultrastructure, and mitochondrial bioenergetics in H9c2 cardiomyoblasts. Exposure to E171 leads to a significant increase in intracellular labile iron (Fe2+) levels. This alteration was accompanied by elevated reactive oxygen species (ROS) production and reduced intracellular glutathione levels, consistent with enhanced oxidative stress following E171 exposure. Ultrastructural analysis by transmission electron microscopy (TEM) revealed marked mitochondrial alterations, including reduced cristae density and structural damage. Functional assessment of mitochondrial bioenergetics demonstrated impaired oxidative phosphorylation and reduced maximal respiratory capacity in E171-treated cells. The potential protective role of quercetin (a powerful antioxidant and iron chelator) was explored in mitochondrial respiration assays; however, at the concentration and exposure conditions tested, quercetin treatment did not fully restore the bioenergetic parameters induced by E171. Collectively, these findings indicate that E171 increases intracellular labile iron levels and promotes oxidative stress, associated with alterations in mitochondrial ultrastructure and impaired bioenergetic function in H9c2 cardiomyoblasts, suggesting a potential mechanism by which food additives may affect cardiac cellular metabolism.

Journal of Xenobiotics doi: 10.3390/jox16030109

Authors: Sara Glorio Patrucco Roberta Giugliano Alessandra Griglione Giorgia Zicarelli Camilla Mossotto Leo Costa Giuseppe Esposito Alice Gabetti Serena Anselmi Tecla Bentivoglio Barbara Vivaldi Valentina Ciccotelli Bruno Aimone Marino Prearo Damià Barceló Monia Renzi Stefania Squadrone Paolo Pastorino

Invasive alien species (IAS) such as Silurus glanis and Procambarus clarkii represent major ecological threats but may also serve as effective bioindicators of environmental contamination; therefore, this study aimed to evaluate their potential for multi-contaminant monitoring and assess their suitability as alternative feed and food resources within a circular economy framework. Multi-contaminant profiles were investigated in S. glanis and P. clarkii from Avigliana Lakes (NW Italy), analyzing trace elements, rare earth elements (REEs), and organic contaminants in fish muscle, and microplastics (MPs) in intestinal tracts. In S. glanis, total trace element concentrations and ΣREEs were markedly higher in Small Lake than in Great Lake, with ΣREEs reaching 0.445 and 0.056 mg/kg w.w., respectively. Mean concentrations of the regulated elements in Great Lake were 0.017 mg/kg w.w. (As), 0.003 mg/kg w.w. (Cd), and 0.16 mg/kg w.w. (Pb), increasing in Small Lake to 0.19, 0.03, and 1.86 mg/kg w.w., respectively. In P. clarkii, contamination levels were lower, with ΣREEs averaging 0.074 mg/kg w.w. and mean concentrations of As, Cd, and Pb of 0.25, 0.006, and 0.21 mg/kg w.w., respectively. Organic contaminants, including polycyclic aromatic hydrocarbons (PAHs), non-dioxin-like polychlorinated biphenyls (NDL-PCBs), and pesticides, were generally below limits of quantification. MPs were detected in 100% of specimens, with mean concentrations of 4.2 ± 2.15 and 4.4 ± 2.70 MPs per intestinal tract in S. glanis (Great and Small Lake, respectively) and 2.7 ± 2.39 MPs/intestinal tract in P. clarkii. Permutational multivariate analysis of variance (PERMANOVA) indicated significant site-related differences in S. glanis and species-related differences between S. glanis and P. clarkii within Great Lake. Most regulated contaminants were below applicable EU thresholds; however, Pb in S. glanis from Small Lake exceeded the maximum level established for fish muscle intended for human consumption.

Journal of Xenobiotics doi: 10.3390/jox16030107

Authors: Luzmila Burbano Pilar Rodríguez-Viso Manuel Zúñiga Vicente Monedero Vicenta Devesa Dinoraz Vélez

The aim of this study was to characterize the toxicokinetics of dietary Hg and to identify lactic acid bacteria (LAB) strains capable of reducing methylmercury (MeHg) accumulation in organs. BALB/c mice and Wistar rats were exposed via drinking water [Hg(II) MeHg] and feed (swordfish, mushrooms). The data showed that the feed matrix did not modify MeHg accumulation, though it reduced Hg(II) accumulation compared to the data observed with drinking water. Two in vitro LAB strains (L. intestinalis LE1 and L. johnsonii LE2) were selected to determine their efficacy in reducing tissue accumulation in mice exposed for 40 days to dietary MeHg. Daily dosing of both strains produced a relevant reduction in Hg content in the organs of animals exposed through drinking water (18–64%). The LE1 strain also reduced Hg content in animals exposed via feed (13–40%). LAB could be a useful strategy in populations which are chronically exposed to Hg through their diet.

Journal of Xenobiotics doi: 10.3390/jox16030106

Authors: Doonyah Alucozai Elizabeth Kwo

Chronic opioid use disrupts the female endocrine system, affecting the hypothalamic–pituitary–gonadal (HPG), hypothalamic–pituitary–adrenal (HPA), and hypothalamic–pituitary–thyroid (HPT) axes. In women, these disruptions manifest as menstrual irregularity, infertility, early menopause, reduced bone mineral density, adrenal insufficiency, and altered mood and sexual function. Despite the magnitude of the opioid epidemic and its impact on women of reproductive age, the existing evidence base is overwhelmingly male: across the major studies of opioid-induced endocrinopathy reviewed here, 99.5 percent of participants were male. Sex-specific mechanisms, prevalence estimates, and clinical thresholds therefore remain poorly defined, and current guidelines do not adequately address the reproductive, skeletal, and adrenal consequences of chronic opioid exposure in women. This review synthesizes available human and preclinical evidence on opioid-induced endocrine dysfunction in women across the lifespan, distinguishing established findings from hypotheses extrapolated from male or animal data. We propose a practical framework for routine endocrine screening of HPG, HPA, and HPT axis function, bone health, and fertility, and outline the roles of relevant specialties in multidisciplinary care. Available evidence suggests a substantial risk of endocrine dysfunction in women on chronic opioid therapy, but the precise prevalence remains unknown. Sex-sensitive research, guidelines, and routine screening are urgently needed to close this gap.

Journal of Xenobiotics doi: 10.3390/jox16030104

Authors: Maria Assunta Iovine Mariacristina Filice Luisa Albarano Alessia Caferro Sandra Imbrogno Rosa Mazza Francesca Esposito Maria Costantini Valerio Zupo Alfonsina Gattuso Giovanni Libralato Maria Carmela Cerra

Coastal ecosystems are increasingly threatened by human activities, highlighting the need for sensitive tools to assess environmental risk. An active biomonitoring approach, using the Mediterranean mussel (Mytilus galloprovincialis), was employed to evaluate anthropogenic chemical contamination in the North Ionian Sea, a still poorly studied area, by comparing mussel health status before (PrePT) and after (PostPT) the peak tourist season. Bioaccumulation of metal(loid)s was quantified in whole organisms. Oxidative stress was assessed in the gills and digestive gland through catalase (CAT), superoxide dismutase (SOD), lipid peroxidation (LPO), and oxidized carbonyl proteins (OMP). Neurotoxicity was evaluated via acetylcholinesterase (AChE) activity, while gene expression of stress-related biomarkers was analysed for metallothioneins (mt10, mt20), sod, cat, Glutathione S-transferase (gst), and Heat Shock Protein 70 (hsp70). Results suggest a progressive contaminant accumulation likely associated with intensified summer anthropogenic activity. Biomarker responses revealed clear activation of oxidative stress, with tissue-specific patterns. The findings confirm the effectiveness of active biomonitoring and multibiomarker approach in assessing coastal water quality and provide valuable baseline data for the management of marine ecosystems.

Journal of Xenobiotics doi: 10.3390/jox16030105

Authors: Chongxi Liao Luxin Ren Lulu Xu Renjie Zhao Baocong Zhao Sihao Lin Ting Zhang Yijie Zhuang Yanpeng Gao Yuemeng Ji

Toluene, as a common organic solvent in academic laboratories in university campuses, poses potential exposure concerns to students and staff in university campuses. Hence, by using a computational fluid dynamics simulation, we investigated the dispersion characteristics of toluene at a campus in Guangzhou under meteorological conditions and the impact of newly constructed buildings on toluene concentrations. The numerical simulation results reveal that toluene is readily accumulated in the free movement area under the prevailing east wind, in the administrative area under the prevailing north-northeast wind, and in the teaching area under the prevailing south wind. Therein, the teaching buildings (TB3–TB6) possess the highest average concentration of toluene compared with other functional areas. In the presence of newly constructed buildings, the toluene concentrations are decreased under the south-southeast wind but are aggravated under the southeast wind. As the height increases, under south-southeast winds, the merging of vortex structures continuously reduces toluene concentrations at TB3 and TB4 and the expansion of the wake region rebounds the toluene pollution at TB5 and TB6; under southeast winds, the expanding vertical vortex structures aggravate toluene pollution at TB3 and TB5 but attenuate toluene pollution at TB4 and TB6. Our results reveal that the teaching areas of the target campus represent a critical zone for potential student exposure during summer and require particular attention. This study provides new insights into the coupled effects of prevailing wind conditions and campus morphology on VOC dispersion characteristics and improves the understanding of airflow pollutant interactions in complex campus environments.

Journal of Xenobiotics doi: 10.3390/jox16030103

Authors: Imen Hammami Fatma Arrari Rahma Mahjoub Ridha Ben Ali Haifa El Hentati Afef Nahdi Eduardo Alberto López-Maldonado Emna Talbi

Cadmium is a widespread environmental xenobiotic that poses serious risks to hepatic, renal, and male reproductive functions. Natural compounds such as silymarin, a bioactive extract from Silybum marianum, have gained attention for their protective potential against xenobiotic-induced toxicity. This study investigated whether subchronic oral administration of silymarin (30 mg/kg) mitigates cadmium-induced toxicity (5 mg/kg) in adult rats over six weeks. Twenty-four rats were assigned to four groups: control, cadmium-exposed, silymarin-treated, and co-treated. Biochemical, hematological, oxidative stress, and reproductive parameters were assessed. Sperm quality was evaluated using CASA, and testicular tissues were examined histologically. Cadmium exposure significantly reduced body weight (−30.8%), elevated transaminases (AST, ALT; p < 0.01), increased serum creatinine and total cholesterol, and induced multi-organ oxidative stress, as reflected by elevated malondialdehyde and markedly reduced SOD, CAT, and thiol group levels in testicular, hepatic, and renal tissues (p < 0.01). Sperm concentration dropped from 75.2 to 21.8 × 106/mL, with total motility falling to 35% and progressive motility to 18%, accompanied by severe seminiferous tubule degeneration (Score III in 5 rats). Co-administration of silymarin partially restored these parameters, sperm concentration recovered to 38.5 × 106/mL, total motility improved to 50.2%, and antioxidant enzyme activities and liver/kidney biomarkers showed significant but incomplete recovery (p < 0.05). Molecular docking revealed favorable binding affinities of silybin toward GPx (−8.4 kcal/mol), CAT (−8.3 kcal/mol), and SOD (−6.4 kcal/mol), offering a preliminary computational hypothesis suggesting possible interactions between silybin and antioxidant enzymes, pending experimental validation. Silymarin alone exerted no adverse effects. These findings establish silymarin as a partial but promising multi-organ cytoprotectant against cadmium toxicity, and highlight the need for future studies optimizing dosing strategies, exploring longer treatment durations, and investigating combination approaches with metal chelators or Nrf2-activating agents to achieve complete tissue recovery.

Journal of Xenobiotics doi: 10.3390/jox16030102

Authors: Marko Šolić Nina Đukanović Tamara Apostolović Jelena Beljin Irina Jevrosimov Dragana Tamindžija Ivana Bajić Stanko Milić Tijana Zeremski Marijana Kragulj Isakovski Snežana Maletić

Metal-contaminated dredged sediments represent heterogeneous environmental matrices in which remediation responses are frequently constrained by elevated background metal loads and complex geochemical conditions. Within such systems, phytoremediation has been discussed as a nature-based management approach whose outcomes depend on plant biomass, internal metal allocation, and context-dependent interactions between plants and sediment. The present study evaluated whether bacterial and fungal plant growth-promoting microorganisms (PGPMs) were associated with changes in plant metal uptake and internal allocation in Sorghum bicolor L. and Cannabis sativa L. grown in dredged sediment collected from the Bega Canal. An outdoor pot experiment was conducted under environmentally relevant conditions, including bacterial and fungal inoculation treatments alongside non-inoculated controls, with plant responses to Cr, Ni, Cu, Zn, As, Cd, and Pb characterized using concentration- and mass-based uptake metrics, root–shoot partitioning, and sediment geochemical assessment based on pseudo-total concentrations and BCR sequential extraction fractions. Across treatments, plant responses were largely governed by intrinsic species traits and biomass production, while PGPM-associated effects remained modest and variable. Root-dominated metal retention and limited translocation were evident irrespective of species, consistent with a phytostabilization-type response rather than systematic extraction. Absolute metal uptake accounted for only a minor fraction of total sediment metal pools, underscoring the importance of interpreting concentration-based indices jointly with mass-based metrics when evaluating system-scale responses. Altogether, the findings indicate that under the investigated outdoor dredged sediment pot conditions, PGPM inoculation acts primarily as a context-specific modulator of plant responses rather than a driver of enhanced phytoremediation performance, reflecting the central role of intrinsic plant traits and stabilization-oriented processes in complex sediment systems.

Journal of Xenobiotics doi: 10.3390/jox16030101

Authors: Daniela Arias-Camacho Brian Antonio Rochin-Peraza Miguel Betancourt-Lozano Selene Abad-Rosales José Basilio Heredia Nayely Leyva-López Samuel Calderón-Liévanos Alejandra García-Gasca

The widespread use of pesticides in agriculture has increased their levels in water bodies, threatening aquatic ecosystems. Among these, glyphosate and chlorpyrifos are widely used in Mexico and can cause toxic effects even at low doses. In aquatic organisms, early exposure to these pollutants can disrupt vital processes, such as sex determination, through oxidative stress. This study assessed the effects of exposure to environmental concentrations of glyphosate (100 μg/L), chlorpyrifos (1.5 μg/L), and their combination on zebrafish (Danio rerio) from early stages to 90 days post fertilisation (dpf). Survival was measured using Kaplan–Meier curves; lipid peroxidation was assessed by malondialdehyde (MDA); sex-related gene expression was measured by qPCR of selected genes at 30 dpf; and gonadal development was assessed by histology at 65 dpf. The results showed increased MDA levels in exposed fish. Glyphosate caused early toxicity and a higher proportion of undifferentiated fish, implying delayed sex determination. Chlorpyrifos induced oxidative stress and affected amh gene expression linked to masculinisation. Combined exposure reduced survival and altered gene expression and gonadal development. Exposure shifted the sex ratio toward males, suggesting that pesticide-induced oxidative stress may alter the expression of sex determination genes during early development.

Journal of Xenobiotics doi: 10.3390/jox16030100

Authors: Zaina Kalaji Ibrahim Hachim Marwa Almazrouei Hanaa Habbal Vidya Bijosh Mohan Mohammad G. Mohammad Rifat Hamoudi Rabih Halwani

Larixyl acetate, a primary component of Larch turpentine, is a naturally occurring compound with a broad spectrum of medicinal properties, including anti-inflammatory effects. It is a potent and selective inhibitor of TRPC6, a widely expressed Ca2+ channel that is involved in many respiratory diseases. Despite its demonstrated efficacy, it lacks a well-defined preclinical and phenotypic safety profile, which limits its therapeutic potential and implementation. In this study, female BALB/c mice were used to assess the toxicity of intranasally administered Larixyl acetate through a subacute model based on OECD Test Guideline 412, followed by a detailed analysis of physical, blood, biochemical, and tissue changes at the administration sites and beyond. Within the study’s 30-day timeframe, our results show no statistically significant differences (p > 0.05) in any of the examined toxicity parameters between the controls or three treatment groups (0.5, 1, and 2 mg/kg). While no pharmacokinetic data were obtained to confirm local or systemic exposure of Larixyl acetate, these findings are crucial for establishing a solid foundation for future therapeutic endeavors, especially in the context of TRPC6-driven respiratory diseases.

Journal of Xenobiotics doi: 10.3390/jox16030099

Authors: Alessandro Polizzi Rossella Rotondo Sabrina Donati Zeppa Monia Cecati Magdalena Smolik Valentina Schiavoni Angelo Montana Valentina Pozzi Davide Sartini Roberto Campagna Gaetano Isola

Head and neck cancers (HNCs) represent a major global health burden and remain associated with substantial morbidity and limited therapeutic options, particularly in advanced or recurrent disease. Increasing interest has focused on naturally derived bioactive compounds with potential chemopreventive and therapeutic properties. Sulforaphane, a dietary xenobiotic isothiocyanate derived from glucoraphanin in cruciferous vegetables, has attracted attention due to its ability to modulate redox balance, epigenetic regulation, and multiple oncogenic signaling pathways. This manuscript reviews current evidence regarding the biological effects of sulforaphane in HNCs. Particular attention is given to the molecular mechanisms underlying its modulation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, a key regulator of cellular antioxidant and detoxification responses that can be activated by sulforaphane. Several studies indicate that sulforaphane can inhibit tumor growth through several mechanisms beyond Nrf2 activation, including induction of apoptosis, cell cycle arrest, epigenetic modulation, and suppression of oncogenic signaling pathways. In addition, sulforaphane has been shown to enhance the efficacy of conventional treatments, including chemotherapy, radiotherapy, and photodynamic therapy. Overall, the literature suggests that sulforaphane may represent a promising chemopreventive or therapeutic adjunct in HNC, although further clinical investigation is required to clarify its translational potential.

Journal of Xenobiotics doi: 10.3390/jox16030098

Authors: Khadija Ramadhan Makame Moustafa Sherif Le Vinh Hoi Thong Balázs Ádám Károly Nagy

Pesticide formulations may influence toxicological outcomes beyond the intrinsic properties of active ingredients; however, these differences are often overlooked in regulatory risk assessment. Using human liver HepG2 cells, this study compared the cytotoxic and genotoxic effects of two commercial lambda-cyhalothrin formulations that differ in formulation type and in composition: an emulsifiable concentrate (Lambda-Cyhalothrin 5% EC) and a suspension concentrate co-formulated with thiamethoxam (Duer SC). Cytotoxicity was assessed using propidium iodide exclusion, while genotoxicity was evaluated using the cytokinesis-block micronucleus (CBMN) and alkaline comet assays. Lambda-Cyhalothrin 5% EC showed significant cytotoxicity from 500 μM onward, whereas Duer SC induced cytotoxic effects even at lower concentrations beginning at 200 μM, indicating greater cytotoxic potency of the combined formulation. Lambda-Cyhalothrin 5% EC induced a concentration-dependent increase in the number of binucleated cells containing micronuclei, with a significant effect at 50 μM. Duer SC also increased micronucleus frequency but did not differ significantly from the negative control. Proliferation indices remained comparable to controls for both formulations. The comet assay showed that Lambda-Cyhalothrin 5% EC produced significant DNA damage from 10 μM onward, while Duer SC exhibited minimal genotoxic effects. These findings demonstrate that formulation type modifies the toxicological profile of lambda-cyhalothrin.

Journal of Xenobiotics doi: 10.3390/jox16030097

Authors: Stanila Stoeva-Grigorova Ivanesa Yarabanova Maya Radeva-Ilieva Galina Uzunova Georgi Bonchev Ivanka Gospodinova Violeta Borisova Elmira Vicheva Simeon Marinov Petko Marinov Snezha Zlateva

Introduction: Scopolamine is a tropane alkaloid and non-selective muscarinic receptor antagonist capable of inducing a central anticholinergic toxidrome with pronounced neuropsychiatric and autonomic manifestations. Despite its well-characterized pharmacology, diagnosis remains clinically challenging due to variable presentation and often absent or unreliable exposure history, particularly in non-medical contexts. Methods: We report a case of a 34-year-old female presenting with an acute onset of altered mental status following a social event. Clinical findings included delirium, hallucinations, psychomotor agitation, dysarthria, bilateral mydriasis with preserved light reflex, sinus tachycardia, and marked urinary retention. Routine laboratory investigations were largely within normal limits, and standard urine toxicology screening was negative except for iatrogenic benzodiazepines. Given the characteristic toxidrome, extended toxicological analysis using gas chromatography–mass spectrometry (GC–MS) was performed, confirming the presence of scopolamine. Supportive treatment, including benzodiazepines and off-label intravenous galantamine, was administered, resulting in progressive clinical improvement and complete recovery without sequelae. Results: This case underscores the diagnostic limitations of routine toxicology screening for tropane alkaloids and highlights the importance of a pattern-based toxidrome approach. Given the absence of reliable exposure history and the presence of marked anterograde amnesia, an exogenous exposure cannot be excluded; however, no direct forensic evidence supports intentional administration. Furthermore, galantamine may represent a pharmacologically plausible alternative to physostigmine in settings where it is unavailable, although evidence remains limited. Conclusions: Scopolamine intoxication should be considered in cases of acute anticholinergic syndrome with negative standard screening, and confirmation requires advanced analytical methods such as GC–MS.

Journal of Xenobiotics doi: 10.3390/jox16030096

Authors: Jacopo Fabrello Giovanni Martino Rigodanza Francesco Boldrin Federico Caicci Marco Munari Valerio Matozzo

This study investigated the individual and combined effects of mercury (as HgCl2) and Bisphenol S (BPS) on the blue crab Callinectes sapidus, focusing on biomarkers measured in hemolymph, gills, and hepatopancreas and ultrastructure (transmission electron microscopy, TEM) observations. Adult males were exposed for seven days to environmentally relevant concentrations of each contaminant, alone and in mixture. Results revealed clear tissue-specific responses. In hemolymph, both contaminants reduced total hemocyte count and affected immune-related parameters, including hemocyte proliferation and enzymatic activities. In gills, mercury significantly decreased total antioxidant capacity, while both contaminants increased lipid peroxidation, indicating oxidative stress. BPS and the mixture stimulated catalase activity, whereas electron transport system activity increased only under combined exposure. In contrast, the hepatopancreas showed limited biochemical alterations, suggesting a higher resilience to medium-term exposure. TEM observations revealed a general decrease in granularity of hemocytes from crabs exposed to BPS and mixture, suggesting cell degranulation. In addition, infiltration of hemocytes into gills was observed in crabs exposed to experimental conditions, suggesting migration of hemocytes from hemolymph to peripheral tissues, while alterations of the microvilli arrangement were detected in digestive cells of BPS-treated crabs. Overall, the findings suggested that hemolymph and gills were more sensitive to contaminant exposure, while the hepatopancreas appears more resistant, at least under the experimental conditions tested. The study also suggests potential non-additive interactions between mercury and BPS.

Journal of Xenobiotics doi: 10.3390/jox16030095

Authors: Jagjeet Singh Annu Phogat Reena Sheoran Arun Hasanpuri Vijay Kumar Manoj Kumar Yadav Vinay Malik

Background: Acetamiprid (ACMP) exposure mediates a variety of pathological complications, including testicular toxicity. Berberine (BBR) is a plant-derived alkaloid with potential pharmacological properties. This study sought to evaluate the ameliorative effects of BBR against ACMP-induced testicular toxicity. Methods: Male Wistar rats were divided into four groups: control, BBR-treated, ACMP-exposed, and BBR+ACMP co-treated, and were administered with BBR (150 mg/kg b.wt) and ACMP (21.7 mg/kg b.wt) for 21 days. Biochemical and FTIR analyses, RT-PCR, computational analyses, and histopathological examination were conducted to assess alterations in lipid and protein profiles, as well as apoptotic and structural changes. Results: ACMP exposure was associated with oxidative injury, functional alterations (stretching of -OH, -CH2, -NH, C=O, C-N, -COO-, -PO2−), and compositional changes in proteins and lipids. Pre-treatment of BBR (2 h prior) was associated with attenuation of the functional and compositional alterations in proteins and lipids in co-treated rats. RT-PCR and computational analysis showed increased Bax and caspase-3 and decreased Bcl-2 mRNA expression, suggesting a potential modulation of ACMP-induced apoptosis by BBR. Histological examination showed that pre-treatment with BBR prevented ACMP-induced structural alterations, including cellular disorganization and alteration in seminiferous tubules. Conclusions: The study suggested that the BBR may exert ameliorative effects against ACMP-induced testicular toxicity by modulating lipid and protein changes and the anti-apoptotic pathway. Thus, BBR could be used as a potential ameliorative agent against oxidative stress. However, more mechanistic studies are needed for broader biological relevance and validity.

Journal of Xenobiotics doi: 10.3390/jox16030094

Authors: María del Rocío Duarte-Martínez Aldo Amaro-Reyes Juan Campos-Guillen Miguel Angel Ramos-López Eloy Rodríguez-de León Monserrat Escamilla-García Vanessa Vallejo-Becerra Alejandra Álvarez-López Yesenia Mendoza-Burguete Mónica López Velarde-Santos Héctor Pool Luisa Ramírez-Granados Ricardo Chaparro-Sánchez José Alberto Rodríguez-Morales

Antimicrobial resistance (AMR) constitutes a growing global threat, facilitated by the dissemination of antibiotic resistance genes (ARGs) through wastewater treatment plants (WWTPs). This systematic review, conducted following the PRISMA guidelines, compiles the risks associated with ARGs, as well as the factors that promote horizontal gene transfer (HGT) and the technologies applied for their removal. The literature shows that WWTPs act as reservoirs, where biological treatment conditions and the presence of sub-inhibitory contaminants (antibiotics, metals, and pharmaceuticals) accelerate HGT. Although conventional methods (chlorination, ozonation, UV) are effective at eliminating antibiotic-resistant bacteria (ARB), their ability to degrade persistent genetic material is insufficient. Therefore, advanced oxidation processes (AOPs) emerge as a key solution, with the photo-Fenton process standing out due to efficiently generating hydroxyl radicals, achieving the degradation of ARGs, an essential step to mitigate the spread of AMR into the environment.

Journal of Xenobiotics doi: 10.3390/jox16030093

Authors: Maryam Mallek Damià Barceló

Emerging contaminants (ECs) and microplastics (MPs) are increasingly detected in surface waters, wastewaters, and drinking water, often as complex mixtures, transformation products, and particle-associated burdens that challenge routine monitoring. This critical review examines current analytical strategies for the detection and characterization of both molecular and particulate emerging contaminants in aquatic systems, with particular emphasis on their relevance to environmental and human health risk assessment. For molecular ECs, targeted LC–MS/MS and GC–MS and GC–MS/MS approaches are evaluated alongside high-resolution mass spectrometry (HRMS)-based suspect and non-target screening, retrospective data mining, and transformation-product elucidation. For MPs, particle-resolved vibrational spectroscopy including µ-FTIR and µ-Raman is critically assessed in comparison with complementary thermal analysis methods, such as pyrolysis–GC–MS and thermal extraction–desorption GC–MS (TED–GC–MS). Particular attention is given to the influence of sampling design, matrix-adapted sample preparation, analytical confidence, and method-dependent size and polymer coverage on data quality and interstudy comparability. The review further highlights the risks of ECs in relation to exposure pathways, mixture effects, and the potential carrier role of MPs for ECs, additives, and microorganisms. Finally, key priorities are identified for next-generation monitoring frameworks, including harmonized workflows, transparent confidence reporting, and stronger integration of analytical evidence with fate, exposure, and risk assessment.

Journal of Xenobiotics doi: 10.3390/jox16030092

Authors: Umberto Cornelli Giovanni Belcaro Claudio Casella

Microplastics (MPs) are emerging contaminants that have been detected in human blood and tissues, raising concerns regarding systemic exposure and potential health effects. Internal MP burden mitigation techniques, nevertheless, are yet largely unexplored. We evaluated whether oral administration of chitosan derived from Procambarus clarkii (PCC) could reduce circulating MPs in humans via gastrointestinal sequestration in this pilot-controlled study. 11 healthy adults received PCC supplementation (0.8 g/day) for 15 days, while 10 matched controls received a placebo. Using stereomicroscopy, scanning electron microscopy (SEM), and micro-Fourier transform infrared spectroscopy (µFTIR), blood MP concentrations were quantified and characterised according to size, shape, and polymer type. At baseline, MPs were found in every subject. Following PCC supplementation, mean MP concentrations decreased from 1.84 ± 0.28 µg/mL to 1.34 ± 0.20 µg/mL (−26.3%, p < 0.01, paired analysis). The control group observed no significant differences. While polymer-resolved analysis consistently indicated reductions across major polymer classes, size-resolved analysis indicated preferential reductions in intermediate particle fractions (11–50 µm). The circulating MPs’ estimated mean residence time (MRT) was 58 ± 28 days. These findings provide preliminary evidence that chitosan-based gastrointestinal sequestration could potentially reduce the systemic MP burden in humans.

Journal of Xenobiotics doi: 10.3390/jox16030091

Authors: Michal Sima Helena Libalova Zuzana Simova Kristyna Vrbova Antonin Ambroz Jiri Klema Lubos Dittrich Michal Vojtisek-Lom Pavel Rossner

Exposure to air pollution is linked to adverse health outcomes. To better reflect real-world conditions, we employed a mobile exposure system enabling direct field exposure of the human airway epithelial model MucilAir™ to ambient air in a traffic-burdened locality. This study represents a follow-up to our previous work, in which a 5-day exposure period under extreme traffic-related pollution conditions resulted in premature cell loss. Under different meteorological conditions characterized by increased precipitation and lower particle number concentrations, MucilAir™ cultures were exposed to traffic-polluted air for 2 days. The exposure resulted in a mild but significant increase in cytotoxicity markers, including lactate dehydrogenase release and elevated levels of 15-F2t-isoprostane, indicating induction of the cellular stress response rather than severe cytotoxicity. A transcriptomic analysis revealed extensive gene expression changes; the enrichment of the pathways related to polycyclic aromatic hydrocarbon detoxification and amino acid biosynthesis suggests adaptive metabolic responses to oxidative and genotoxic stress. In parallel, the pathways associated with epithelial proliferation and repair, extracellular matrix organization, focal adhesion, and immune signaling were suppressed, indicating potential disruption of the epithelial homeostasis. Overall, these findings demonstrate that 2 days of exposure to real-world traffic-polluted air elicits adaptive stress responses in airway epithelial cells while simultaneously impairing the processes essential for epithelial integrity, potentially leading to airway dysfunction.

Journal of Xenobiotics doi: 10.3390/jox16030090

Authors: Syed Sayyam Abbas Syed Ali Musstjab Akber Shah Eqani Ismat Nawaz Mansoor A. Alghamdi Ahmed S. Summan Abdul Qadir Shabbar Abbas Iqra Rasheed Syeda Maria Ali Mustafa Nawaz Shafqat Mohammed I. Orif Heqing Shen Nadeem Ali

Data on arsenic (As) and other potentially toxic elements (PTEs) in Pakistani retail meats are limited, constraining evidence-based dietary risk assessment and management. This study aimed to determine the concentrations and profiles of As and seven other PTEs (Cr, Ni, Mn, Pb, Cd, Cu, Zn) in commonly consumed meats and to evaluate the associated non-carcinogenic health risks. Ninety-two paired liver and muscle samples from broiler chicken, goat (mutton), and beef cattle were collected from four cities across the Indus Plain and analyzed using inductively coupled plasma mass spectrometry (ICP-MS). Dietary exposure was evaluated using estimated daily intake (EDI), target hazard quotient (THQ), and hazardous index (HI) under typical and high-consumption scenarios. Overall, Zn and Cu exhibited the highest concentrations, followed by Mn and Cr, whereas As, Pb, Ni, and Cd occurred at comparatively lower but environmentally relevant levels. Beef liver exhibited the highest contamination levels, exceeding FAO/WHO permissible limits for Pb, Cu, and Cd in up to 40% of samples. In contrast, mutton and beef muscle contained the highest As and Zn concentrations, while chicken muscle showed elevated Cr levels. Multivariate statistical analysis revealed three dominant co-variation patterns, suggesting potential contamination pathways: (i) geogenic groundwater sources enriched with As, Cr, and Ni; (ii) atmospheric and industrial dust inputs linked with Pb, Cd, and Mn; (iii) mineral-enriched feed additives potentially contributing to elevated Zn and Cu, particularly in poultry. Under high-consumption scenarios, THQ values for As, Cr, Cu, and Zn exceeded the safety threshold (THQ > 1), highlighting beef products as the dominant source of chronic dietary risk. Overall, the findings highlight pronounced tissue- and species-specific accumulation trends, and emphasizes the urgent need for stricter feed and water quality control measures to minimize dietary exposure to PTEs.

Journal of Xenobiotics doi: 10.3390/jox16030089

Authors: Muhammad Salman Zeb Mansour A. Alghamdi Ahmed Summan Javed Nawab Muhammad Imtiaz Rashid Nadeem Ali

Brominated flame retardants (BFRs) are widely used in automotive polymers and electronic components, yet vehicles remain an under-characterized and potentially high-exposure microenvironment, particularly in hot climates. This study provides the first comprehensive assessment of BFR occurrence, sources, and exposure risks in vehicle dust from Saudi Arabia, addressing a critical regional data gap. This study systematically investigates the occurrence, compositional patterns, sources, and human exposure risks of polybrominated diphenyl ethers (PBDEs) and selected alternative BFRs in dust from 80 vehicles (domestic cars and taxis; model years 2015–2022) operating in Jeddah, Saudi Arabia. Dust samples were collected using a standardized vacuuming protocol, extracted and cleaned using solvent extraction and silica SPE, and analyzed via GC–NCI–MS. Both legacy PBDE congeners and emerging alternatives (including DBDPE and TBB) were consistently detected, with BDE-209 dominating the overall BFR burden with mean concentrations of 6560 ng/g in domestic vehicles and 5454 ng/g in taxis, with maximum values reaching 220,860 ng/g. Lower-brominated PBDEs occurred at substantially lower concentrations, reflecting the ongoing global transition away from Penta- and Octa-BDE formulations. Taxis exhibited generally higher concentrations than domestic vehicles, likely due to prolonged occupancy, increased usage intensity, and enhanced dust resuspension dynamics. Multivariate analysis (PCA and correlation) revealed two distinct source categories: (i) legacy Penta-BDE-related congeners associated with polyurethane foam and textile materials and (ii) high-brominated PBDEs and DBDPE linked to hard plastics and electronic components. Human exposure assessment demonstrated that dust ingestion is the dominant exposure pathway, while dermal and inhalation routes contribute minimally. Non-carcinogenic hazard indices (HI) were well below unity for all compounds (HI < 1.67 × 10−6), and incremental lifetime cancer risks (ILCR) for BDE-209 remained within or near accepted risk thresholds (7.52 × 10−6–1.04 × 10−5), although occupational exposure among taxi drivers was consistently higher. Overall, the results demonstrate that modern vehicle cabins act as significant microenvironments for chronic BFR exposure, particularly under high-temperature conditions. Despite generally low estimated risks, the combined effects of chemical persistence, bioaccumulation potential, and mixture toxicity—amplified by extreme in-cabin temperatures—highlight vehicles as overlooked yet significant exposure environments. These findings provide the first comprehensive dataset for the Arabian Peninsula and emphasize the need for climate-sensitive exposure assessment, safer material design, and targeted mitigation strategies in vehicle interiors.

Journal of Xenobiotics doi: 10.3390/jox16030088

Authors: Anita Ferri Sandro Sacchi Chiara Losi Martina Amico Nicola Franchi Davide Malagoli

The increasing fragmentation of plastic debris into nanosized particles represents a threat to freshwater ecosystems, yet the biological effects of nanoplastics (NPs) on freshwater invertebrates remain poorly understood. This study investigated tissue distribution, cellular effects and immune responses following acute exposure to polyethylene terephthalate nanoplastics (PET-NPs) in the freshwater gastropod Pomacea canaliculata, a species of high ecological relevance and physiological resilience. Adult snails were injected with PET-NPs at 5 or 10 mg/L and sampled after 24 and 72 h. PET-NPs accumulation in the anterior and posterior kidneys was assessed by fluorescence imaging and tissue morphology was evaluated. Stress- and inflammation-related genes (Pc-Heat Shock Protein (HSP)70, Pc-HSP90 and Pc-Allograft inflammatory factor 1) expression was quantified by RT-qPCR. PET-NPs uptake and phagocytic activity were analyzed in circulating hemocytes in vivo and ex vivo. PET-NPs were accumulated in renal tissues, persisting up to 72 h without histopathological alterations. Gene expression analyses revealed non-linear and dose/time-dependent responses. Hemocytes of different morphologies internalized PET-NPs in a dose-dependent manner and showed intercellular particle transfer. Overall, acute PET-NP exposure determines rapid immune handling and tissue sequestration with limited short-term physiological impact, underscoring the potential involvement of immune processes in NPs fate and highlighting the need for chronic exposure studies.

Journal of Xenobiotics doi: 10.3390/jox16030087

Authors: Janiel Jawahar Samuel James

The interplay between the environmental exposome and the cancer genome remains a critical gap in precision oncology. While somatic mutational signatures—genomic fossils imprinted by exposures such as ultraviolet radiation; tobacco smoke; and industrial pollutants—are well characterised for their etiological significance; their functional impact on therapeutic efficacy remains largely unexplored. We hypothesised that these environmental genomic scars induce distinct pharmacogenomic vulnerabilities and resistance mechanisms that vary by geographical exposure patterns. This study employs two complementary analytical frameworks. First, a linear regression-based pharmacogenomic screen across four datasets (GDSC1, GDSC2, CTRP, CCLE; 1001 cell lines, 31 cancer types) identified 608 statistically significant (p < 0.01) mutational signature–drug interactions, revealing that UV-associated signature SBS7a is associated with broad-spectrum therapeutic resistance, including to BRAF inhibitors (PLX-4720, p < 10−4), while pollution-driven oxidative stress (SBS18) is associated with sensitivity to p38 MAPK inhibition (VX-702, r = −0.45, p < 10−9). Second, an XGBoost predictive model trained exclusively on 33,679 GDSC2 records using a 1265-feature matrix integrating 40 SBS signatures, drug chemistry descriptors, proteomic features, and two satellite-derived environmental variables (NASA PM2.5 and UV)—achieved R2 = 0.7973 on a 20% holdout set (grouped cross-validation R2 = 0.7296). SHAP analysis revealed that satellite-derived PM2.5 (Zone_PM25) ranked 7th of 1265 features, exceeding all 40 individual SBS mutational signatures. Synthesising these findings with satellite-derived atmospheric data, we constructed an exploratory spatially interpolated risk surface spanning 122 nations, generating the hypothesis that uniform drug efficacy assumptions may not apply globally. These findings suggest that a patient’s environmental exposure history may constitute a measurable pharmacogenomic variable. This exploratory framework warrants validation in independent datasets and with individual-level geographic data before clinical application.

Journal of Xenobiotics doi: 10.3390/jox16030086

Authors: Natalia Ferrando Elisa Langa Laura Botello-Morte Pedro Rodríguez-López Diego Ballestero María Rosa Pino-Otín

Trans-cinnamaldehyde (CIN), the main component of cinnamon essential oil, is a promising sustainable alternative to synthetic pesticides. Despite its use, ecotoxicological data on non-target species remain fragmented. This study systematically evaluates CIN’s acute toxicity across multiple trophic levels to characterize the biological sensitivity and environmental response of key organisms. Aquatic assays measured bioluminescence inhibition in Aliivibrio fischeri and immobilization in Daphnia magna. Terrestrial evaluations included lethality tests on Eisenia fetida and root elongation in Allium cepa. Additionally, the impact on soil and river microbial communities was analyzed via Biolog EcoPlates™. Significant dose–response relationships were observed across all bioindicators (p < 0.0001). A. fischeri was the most sensitive species (EC50 = 1.428 mg·L−1), followed by D. magna (EC50 = 4.533 mg·L−1). In terrestrial models, A. cepa (EC50 = 11.644 mg·L−1) exhibited higher sensitivity than E. fetida (LC50 = 412.519 mg·kg−1). Microbial metabolic activity showed dose-dependent inhibition, particularly affecting carbohydrate and polymer degradation at high concentrations. These findings define the first ecotoxicological benchmarks for CIN, establishing EC10 and EC50 values under standardized conditions. These data provide the necessary toxicological constraints to ensure environmental safety in future field-scale applications of this natural compound.

Journal of Xenobiotics doi: 10.3390/jox16030085

Authors: Victoria Yurchenko Alexey Morozov

Glyphosate and its primary metabolite aminomethylphosphonic acid (AMPA) are widely detected in aquatic environments, yet their combined effects on fish remain insufficiently understood. This study used label-free blood plasma proteomic profiling to explore molecular patterns associated with 14-day exposure of juvenile common carp (Cyprinus carpio) to environmentally relevant concentrations of glyphosate (100 µg/L), AMPA (100 µg/L), and their mixture (50 + 50 µg/L). Across the three exposure groups, 41 proteins of interest showed pronounced abundance differences relative to the control based on fold-change selection criteria. These proteins were mainly associated with immune recognition, innate immune and complement-associated functions, coagulation and extracellular protease regulation, lipid/sterol transport, and extracellular matrix organization. In the mixture group, proteins of interest spanned several functional categories, suggesting that combined exposure deserves further attention in future studies of plasma-level responses to glyphosate and AMPA. Overall, these findings provide preliminary insights into blood plasma protein patterns associated with systemic responses of fish to glyphosate, AMPA, and their mixture at environmentally relevant concentrations and highlight the importance of considering parent compounds, metabolites, and their co-occurrence when assessing the potential biological effects of herbicide contamination in aquatic ecosystems.

Journal of Xenobiotics doi: 10.3390/jox16030084

Authors: Iman Al-Saleh Ghofran Al-Qudaihi Yara Aljerayed Kafa Abuhdeeb Rola Elkhatib Hissah Alnuwaysir Mashael Alsubaie Norah Alotaibi

Styrene, a constituent of polystyrene food-contact materials, can migrate into hot beverages, but data on short-term consumer exposure and associated biological responses remain limited. In this single-arm longitudinal human biomonitoring pilot study, 40 healthy adults consumed tea or coffee daily in Styrofoam cups for approximately two weeks. Biomarkers were measured at baseline, day 6, and day 11, including urinary mandelic acid (MA) and phenylglyoxylic acid (PGA), salivary malondialdehyde (MDA), comet assay parameters in peripheral blood lymphocytes, and micronucleus (MN) frequency in buccal cells. Measured styrene migration into beverages ranged from 3.3 to 7.1 μg/L, below the World Health Organization guideline value. Urinary metabolites and salivary MDA showed substantial interindividual variability and no consistent temporal pattern. In contrast, generalized estimating equation models showed progressive increases in comet assay indicators over the exposure period. Tail intensity and tail moment increased over time, with stronger changes among participants consuming two cups daily. MN frequency did not change significantly. These findings suggest that repeated short-term consumption of hot beverages in polystyrene cups was associated with modest changes in selected early biomarkers of biological effect under consumer-use conditions. The results should be interpreted cautiously in light of the modest sample size, short follow-up, and absence of more specific mechanistic endpoints, but they support further study of repeated low-level exposure to food-contact materials.

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.