Abstract: Background: Medicinal plants used in traditional Mexican medicine represent a valu-able source of bioactive compounds with potential anticancer activity. Beyond cyto-toxic potency, selectivity toward cancer cells over normal cells is a critical toxicologi-cal parameter for identifying safer therapeutic candidates. This study aimed to evalu-ate the selective cytotoxic and antiproliferative effects of extracts from four Mexican medicinal plants across human cancerous and non-cancerous cell lines. Methods: Hexane, acetone, and methanolic extracts from Semialarium mexicanum, Eryngium heterophyllum, Piper auritum, and Cochlospermum vitifolium were evaluated in a panel of human cancer cell lines and non-tumoral models, including primary human uterine fibroblasts (HUF). Cytotoxicity was assessed after 48 h of treatment using in-creasing extract concentrations, and selectivity indices were calculated. Cell cycle dis-tribution and nuclear morphology analyses were performed to explore antiprolifera-tive effects. Additionally, GC–MS-based chemical profiling was conducted on selected extracts to obtain a tentative characterization of major bioactive constituents. Results: The extracts exhibited differential cytotoxic profiles depending on plant spe-cies and solvent polarity. Semialarium mexicanum, particularly its hexane extract, showed the highest cytotoxic potency and selectivity toward HeLa cervical cancer cells, achieving selectivity indices higher than those observed for paclitaxel when compared with HUF cells. Hexane extracts of Semialarium mexicanum and Eryngium heterophyllum, were generally more active than polar extracts, whereas Piper auritum displayed limited cytotoxicity and Cochlospermum vitifolium showed moderate, sol-vent-dependent effects. Cell cycle perturbations and nuclear alterations supported an antiproliferative response. Chemical profiling suggested the presence of lipophilic triterpenoid-related compounds in non-polar extracts and phenolic constituents in polar fractions. Conclusions: These findings provide in vitro evidence of selective anticancer activity of Mexican medicinal plant extracts and establish a basis for future mechanistic stud-ies medicinal plant extracts and lay the groundwork for future mechanistic investigations.

Abstract:

Chronic kidney disease (CKD) is characterized by progressive glomerulosclerosis and interstitial fibrosis, ultimately leading to irreversible renal dysfunction. Tacrolimus, a calcineurin inhibitor widely used after kidney transplantation, is known to accelerate chronic renal injury, whereas everolimus has been introduced as a calcineurin inhibitor–sparing agent. However, their direct effects on renal fibrogenesis remain incompletely defined. In this study, we established a chronic renal failure model using 5/6 nephrectomized rats to evaluate tacrolimus-accelerated renal interstitial fibrosis and the impact of concomitant everolimus treatment. Low-dose tacrolimus (1 mg/kg every other day) administered for two weeks starting four weeks after nephrectomy markedly accelerated interstitial fibrotic progression, accompanied by histological deterioration and increased albuminuria. Co-administration of low-dose everolimus (1 mg/kg every other day) significantly attenuated tacrolimus-accelerated fibrosis and improved functional and structural renal parameters. Histological analyses demonstrated reduced interstitial fibrosis and glomerular alterations in the everolimus-treated group. Although α-smooth muscle actin expression was not significantly suppressed, everolimus restored autophagic activity, as indicated by normalization of the LC3-II/LC3-I ratio in both in vivo and in vitro experiments. These findings indicate that tacrolimus accelerates renal fibrogenesis in a remnant kidney model and that concomitant low-dose everolimus effectively mitigates this progression, providing a time- and cost-efficient experimental platform for evaluating antifibrotic strategies in CKD.

Abstract:

Microplastics (MPs) are synthetic solid polymers (1µm – 5mm) which are non-biodegradable. The toxicological effects of MPs have been well investigated, but research on how these particles affect PBMCs leaves much to be explored. Different concentrations 0.5 µg/ml, 5 µg/ml, 50 µg/ml, 500 µg/ml of PEG and manually grinded natural MPs were exposed to PBMCs in RPMI medium for 24 hours. Cell viability assay, Neutral Red phagocytosis assay, Griess colorimetric assay, Nitroblue Tetrazolium test was done to examine the cytotoxic effect of MPs on PBMCs. The present study results indicated that both natural MPs and Polyethylene Glycol (PEG) significantly reduced cell viability in a concentration-dependent manner. At highest concentrations, Natural MPs induced phagocytic activity of PBMCs. These MPs may act as stimulants to increase phagocytic activity. Regarding oxidative stress, Natural MPs exposure with PBMCs showed a significant increase in ROS production, whereas PEG exposure didn’t induce notable ROS production. NO production levels remained unchanged in PBMCs after exposure to both PEG and Natural MPs, showing that under the tested conditions, neither treatment significantly influenced the NO-mediated inflammatory pathways. In summary, this present study showed that MPs exposure to humans can impair cell viability, induce phagocytosis and induce ROS production without altering the NO mediated inflammatory pathways.

Abstract: Lipid nanoparticles (LNPs) are a critical structural element of modern mRNA therapeutics, including COVID‑19 modRNA vaccines. Each formulation is a multicomponent system in which the LNP serves not as a passive carrier but as an active, biointeractive entity whose ionizable lipids engage directly with cellular membranes. Current evidence from cellular, transcriptomic, and proteomic analyses indicates that LNPs, with or without active mRNA cargo, alter transcriptomic programs and protein expression. This suggests that, even during uptake and interaction with the membrane (transfection), the membrane serves as an initial site for inflammatory, detoxifying, and stress responses. Simultaneously, pathways involved in fat metabolism and detoxification are affected, such as the peroxisome proliferator-activated receptor γ (PPARγ) and cytochrome P450 (CYP) enzyme systems. We believe that the phosphatidylinositol (PI) cycle is the initial point for these disorders. This cycle regulates both organelle trafficking and membrane restructuring following endocytic processes, including macropinocytosis. When this cycle is disrupted, membrane restructuring and organelle dysfunction occur, triggering downstream signaling cascades such as nuclear factor kappa-B (NF- κB), mitogen-activated protein kinases (MAPKs), Janus kinase–signal transducer (JAK-STAT) pathways, and mechanistic target of rapamycin (mTOR) complexes. Transfection with LNPs may induce a systemic condition we call lipid-nanoparticle-driven membrane dysfunction (L‑DMD), where transfection results in broader dysregulation of cellular communication, stress response, and energy balance. This hypothesis-driven review offers a mechanistic foundation for understanding the diffuse, often enduring, biological effects observed after exposure to messenger RNA LNP formulations. It highlights a needed perspective at the intracellular level and within systems biology.

Abstract: External causes of death contribute to over four million annual global fatalities, with drug use representing a significant risk factor. However, the true national impact and regional variations of psychoactive substance use in these deaths remains unde-fined in Brazil. To address this critical knowledge gap, this pioneering four-region study sought to elucidate the influence of alcohol and drug use on external cause mor-tality. We collected post-mortem blood from 3,577 victims of violent death across four distinct Brazilian regions using a standardized protocol to identify alcohol, illicit drugs, and psychoactive medicines. Analysis revealed a predominantly male cohort (89.7%; 56.0% aged 30 years or more), with homicide as the primary manner of death (67.3%). Critically, over half of the victims (53.0%) were positive for at least one psy-choactive substance prior to death, most commonly cocaine (29.6%) and alcohol (27.7%). Substance consumption was highest among homicide victims (55.7%), pre-dominantly cocaine (36.0%), and in self-harm cases (26.4%), which showed high ben-zodiazepine rates (20.0%). Consumption patterns varied regionally: alcohol-related deaths were more common in the Northeast, drug-only deaths concentrated in the Southeast and North, and the South showed a higher prevalence of alcohol use versus drug use. This widespread, regionally heterogeneous prevalence underscores the ur-gent need for targeted, region-specific interventions. By critically linking psychoactive substance use to various modes of violent death, these data provide crucial forensic and public health insights to inform tailored preventive strategies.

Abstract: Small plastics polymers have the potential to bioaccumulate in tissues and initiate toxicity, raising concerns about the long-term impacts towards filter-feeders such as freshwater mussels. The purpose of this study was to examine the toxicity of two common plastic polymers (polyvinyl chloride-PVC; polyethylene terephthalate-PET) and the plasticizer dibutylphthalate (DBP) to quagga mussels Dreissena bugensis. Mussels were exposed to 5, 50 and 100 µg/L of the above compounds for 96h at 15oC. They were then analyzed for total plastic accumulation, esterase, peroxidase, lipids and protein aggregation in the soft tissues. The data revealed that DBP reduced survival at concentrations > 5 µg/L and all died at 100 µg/L. An estimated bioavailability factor of 90, 40 and 1580 for PVC, PET and DBP was determined. These differences could be explained by particle size and form for PVC (0.5 µm fiber) and PET (2 µm diameter). DBP tissue levels were also detected in mussels exposed to PVC and PET suggesting its presence in plastics. Esterase activity was strongly increased in DBP (ester)-treated mussels and was slightly decreased in mussels exposed to ester-containing PET. The biomarker data also revealed an increase in lipids, peroxidase and protein aggregation in a concentration-dependent manner. It is concluded that these compounds are bioavailable to mussels and the changes in esterase activity could be a factor leading to oxidative stress and protein aggregation in mussel tissues.

Abstract:

Persistent environmental pollutants such as per- and poly-fluoroalkyl substances (PFAS) have been associated with a wide range of toxic effects, including cancer. There are over 12,000 PFAS compounds, which may act as carcinogens individually or in combinations. Therefore, efficient in vivo models of carcinogenicity are needed for evaluating environmental contaminant compounds and chemical mixtures. Here, we use the larval zebrafish xenograft assay to identify tumor growth activity of perfluorooctanesulfonic acid (PFOS), a known carcinogenic PFAS. Dose response curves for PFOS exposure were used to identify the Maximum Tolerated Concentration (MTC) and Lethal Concentration causing 50% death (LC50) under xenograft conditions. Zebrafish xenografts were established by injecting fluorescently labeled kidney cancer cells into the embryonic body cavity near the developing kidney, followed by treatment with PFOS at a concentration of 5%, 10% and 20% of the MTC. When treated with PFOS, zebrafish xenografts using renal cell carcinoma (ACHN) cells and clear renal cell carcinoma (Caki-1) cells show dose-dependent changes in tumor area. This study is the first to directly show cancer-promoting activity of a PFAS, using a rapid in vivo zebrafish xenograft assay, and demonstrates the utility of this model for validation of predicted cancer-promoting properties of environmental contaminants.

Abstract: Niclosamide has been the primary molluscicide for schistosomiasis control for over 50 years, but it poses significant threats to non-target large freshwater mollusks. Cipangopaludina cahayensis, a dominant species in East Asian schistosomiasis-endemic regions, was chronically exposed under environmentally relevant concentrations of niclosamide using a mesocosm approach to assess its toxic effects. Digestive glands accumulated more niclosamide than foot tissues. Prolonged exposure disrupted metabolic processes in the digestive glands, leading to tubular atrophy, inflammatory responses, and nutrient depletion. In the foot, niclosamide caused structural damage to both tissue and muscle fibers, resulting in reduced mobility. Furthermore, niclosamide impaired nutritional coupling between the digestive glands and foot, exacerbating motor dysfunction in the foot. Persistent niclosamide exposure destabilized molluscan trophic-level population dynamics, ultimately triggering multi-faceted ecological disturbances, including a decline in ecosystem resilience and disruption of key ecological functions. Our findings underscore the significant toxicological risks that niclosamide poses to freshwater mollusks and highlight the urgent need for the development of ecologically sound usage guidelines to minimize off-target effects.

Abstract: Cystic fibrosis (CF) airway mucus is thick and sticky, which makes it hard for nanoparticles to move and deliver drugs effectively. In this study, degradable PLGA nanoparticles were coated with polyethylene glycol (PEG) of different molecular weights (2, 5, and 10 kDa) to test their movement and depth of penetration in CF-like mucus. The nanoparticles had a uniform size of 150 ± 10 nm and were measured using fluorescence recovery after photobleaching (FRAP) and confocal imaging. The PEG (5 kDa) coating showed the best results, with a diffusion rate of 1.45 ± 0.12 µm²/s, about 3.8 times higher than that of uncoated particles. In a 6-hour test, the particles moved over 70% deeper into the mucus and showed much higher uptake by airway cells (P < 0.01). These results show that medium-length PEG chains reduce mucus adhesion and help nanoparticles move more easily through dense airway mucus. This work provides a practical way to improve drug carriers for CF and other lung diseases with mucus blockage.

Abstract: The effects of 4-phenylbutyrate (4-PBA) on particle-induced macrophage apoptosis in vitro have not been previously documented. This study aims to investigate the thera-peutic potential of 4-PBA in alleviating titanium nanoparticle (TiNP)-induced in-flammatory injury mediated by macrophages. Our findings demonstrate that TiNPs exhibit specific properties capable of inciting inflammatory injury to macrophages. Furthermore, the extent of inflammatory injury inflicted by TiNPs on macrophages progressively increases in correlation with both the concentration of particles and du-ration of exposure. Exposure to TiNPs significantly reduces macrophage viability, promotes apoptosis, enhances the expression of inflammatory cytokines, and exacer-bates the production of reactive oxygen species (ROS). Co-treatment with 4-PBA sig-nificantly alleviates the severity of particle-induced inflammatory injury while simul-taneously facilitating apoptosis in stressed macrophages. Pharmacological intervention with 4-PBA facilitates TiNP-induced macrophage apoptosis by increasing the expres-sion of pro-apoptotic proteins such as Caspase-3 and Bax, while decreasing the levels of anti-apoptotic protein Bcl-2. When used as an independent treatment, 4-PBA does not provoke a pronounced inflammatory response nor induce significant changes in mac-rophage apoptosis, underscoring its potential as a safe therapeutic option. These find-ings substantiate the hypothesis that employing a specific approach to facilitate mac-rophage apoptosis could serve as an effective therapeutic strategy for managing aseptic loosening. Significantly, this approach demonstrates a favorable safety profile, posi-tioning it as a promising avenue for future clinical applications in this domain.

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

Abstract:

Pesticides remain among the most significant threats to biodiversity and natural ecosystems. Non-invasive methods, such as the analysis of bird faeces, have shown great potential for detecting pesticide exposure. In this study with a new approach, we analysed faecal sacs from nestlings of Blue tits (Cyanistes caeruleus) and Great tits (Parus major) to gain deeper insights into pesticide contamination during the breeding period. Samples were collected from three distinct sites near Münster, Germany. In total, we detected 65 substances from 57 different pesticides, as well as caffeine, with pesticides present in 16.07% of the 168 samples. Concentrations varied between species and sites and were higher for fungicides and insecticides in nests located closer to agricultural fields. While no direct effects on reproductive success were found, our results underscore the potential of faecal sac analysis as a valuable tool for spatially resolved pesticide monitoring. Importantly, we show that pesticide exposure also occurs in in nestlings and birds breeding outside of intensive farmland. To better understand the ecological consequences, future studies should incorporate environmental variables and conduct a separate analysis of urate and faeces of feacal sacs to precisely determine concentrations.

Abstract: Beans are widely consumed worldwide and are a good source of amino acids and micronutrients; however, they contain anti-nutrients, such as lectins, tannins, protein inhibitors, saponins, and phytic acid, among others, which can reduce the food’s quality and cause adverse health effects. In this study, we analyzed the genotoxic and cytotoxic effects of a protein extract from Phaseolus acutifolius (TBE) seeds. The extract contained some antinutritional compounds, with a higher lectin content, with an activity of 2701.85 HU. The acute toxicity test in mice showed that the extract was non-toxic at the concentrations tested, as it did not cause any mortality. The in vitro cytotoxicity study on small intestinal epithelial cells indicated that the lectin-rich extract was cytotoxic in both assays, with IC50 values of 10.08 µg/mL and 108.93 µg/mL for the free cell and intestinal fragment assays, respectively. In the in vivo study, an erythropoiesis-stimulatory effect was observed, with significant genotoxic damage noted at 48 h, evidenced by 11 micronucleated erythrocytes at 1000 mg/kg TBE. However, no genotoxicity was detected with prolonged treatment times. These results indicate that TBE is cytotoxic within the tested concentration range, and genotoxic damage is influenced by both concentration and exposure time.

Abstract: Bortezomib is a 26S proteasome inhibitor used to treat multiple myeloma and systemic amyloidosis. While effective in prolonging survival, bortezomib has been increasingly associated with cardiovascular adverse events (CVAEs), such as cardiac failure and arrhythmias. However, a comprehensive post-marketing safety profile has not been fully established. We analyzed cardiovascular adverse events reported between May 2003 and May 2025 using the U.S. Food and Drug Administration Adverse Event Reporting System (FAERS) via the Open Vigil 2.1 platform. Disproportionality analysis was conducted using the reporting odds ratio (ROR) with 95% confidence intervals (CI) to detect statistically significant adverse reports. Of over 9 million drug-related adverse events in the FAERS database, 552 Cardiac events were linked to bortezomib. Cardiac amyloidosis showed the strongest association (ROR: 35.58; 95% CI: 28.16–44.95), followed by atrial flutter (ROR: 4.34; 95% CI: 3.41–5.52), left ventricular dysfunction (ROR: 4.25; 95% CI: 3.26–5.55), cardiac failure, acute (ROR: 3.66; 95% CI: 2.75–4.86), congestive cardiomyopathy (ROR: 3.23; 95% CI: 2.33–4.49), hypertrophic cardiomyopathy (ROR: 3.44; 95% CI: 1.95–6.08), cardiac failure (ROR: 3.18; 95% CI: 2.93–3.46), cardiomyopathy (ROR: 2.81; 95% CI: 2.30–3.43), atrial fibrillation (ROR: 2.65; 95% CI: 2.43–2.89), right ventricular failure (ROR: 2.30; 95% CI: 1.63–3.24), myocarditis (ROR: 2.25; 95% CI: 1.73–2.91), and supraventricular tachycardia (ROR: 2.17; 95% CI: 1.61–2.93). cardiac failure led to the highest number of hospitalizations (301) and deaths (208), followed by atrial fibrillation and cardiac amyloidosis. Older adults (≥65 years) and patients with amyloidosis or myeloma were the most vulnerable groups. Bortezomib is associated with a wide spectrum of serious cardiac adverse events, with cardiac amyloidosis and cardiac failure demonstrating particularly high disproportionality and mortality. These findings emphasize the need for routine cardiovascular risk assessment and proactive monitoring in high-risk patient populations receiving bortezomib.

Abstract: Biochar, a carbon-rich material traditionally used to improve soil health and as a feed additive, has recently attracted attention for its potential biological activity. This study investigated the effects of an aqueous biochar extract (BC-AE) on human intestinal epithelial cells (Caco-2), specifically examining its impact on cell viability and apoptosis. The metabolic activity of Caco-2 cells exposed to BC-AE was first evaluated using an MTS assay. A concentration of 3 mg/mL, which promoted Caco-2 metabolic activity, was selected for further testing at 24 and 72 hours. The effect of BC-AE on cell viability was assessed by epifluorescence microscopy (morphology) and flow cytometry (apoptosis profiling). The transcriptional response of cell viability-related genes (BAX, BAD, BCL-2, BCL-xL, MCL-1, P21, and P53) and microRNAs (miR-15b, miR-19, miR-21, miR-33a, miR-155, and miR-486) was analyzed by RT-qPCR. In parallel, selected proteins (BAD, BAX, BCL-2, and MCL-1) were examined by Western blotting. BC-AE decreased cell viability after 24 hours via late apoptosis, while 72-hour exposure increased necrosis without further viability loss. Both BAX and MCL-1 protein levels increased in Caco-2 cells after 72 hours of BC-AE treatment, and miR-15b and miR-21 were upregulated, suggesting the involvement of a regulatory mechanism controlling cell survival. The obtained findings highlight the importance of considering both concentration and exposure duration when assessing biochar bioactivity.

Abstract: Cytotoxicity testing remains a cornerstone of modern toxicology, providing essential insight into how chemicals and drugs affect cell viability and function. Classical colourimetric assays such as MTT, LDH release, and neutral red uptake established the methodological basis of in vitro toxicology and continue to serve as regulatory benchmarks. However, their limited mechanistic depth and physiological relevance have prompted the field to evolve towards more predictive and human-centred approaches. Recent advances in high-content imaging, flow cytometry, and real-time impedance analysis have transformed cytotoxicity testing into a multiparametric discipline capable of detecting subtle, adaptive, and sub-lethal cellular responses. Large-scale initiatives such as Tox21 and ToxCast, supported by computational pipelines like tcpl, have standardised data interpretation and improved reproducibility. In parallel, three-dimensional organoid cultures, organ-on-chip platforms, and bioprinted constructs now replicate the architec-ture, perfusion, and metabolic activity of human tissues, enhancing translational accuracy. Stem cell–based models using human embryonic and induced pluripotent stem cells provide ethically sustainable systems for organ-specific and developmental toxicity testing, while in silico tools - QSAR, machine learning, and physiologically based pharmacokinetic (PBPK) modelling - enable quantitative in vitro–in vivo extrapolation (QIVIVE). Together, these developments underpin New Approach Methodologies (NAMs) and Integrated Approaches to Testing and Assessment (IATA), marking the transition from descriptive assays to predictive, mechanism-anchored frameworks that advance both biomedical research and regulatory science.

Abstract: Ensuring access to safe drinking water is a fundamental public health priority, yet the growing diversity of contaminants demands more human-relevant toxicity assessment frameworks. Conventional models based on immortalized cell lines or sentinel species, while informative, lack the tissue complexity and inter-individual variability required to capture realistic human responses. Organoids, three-dimensional epithelial structures derived from adult or pluripotent stem cells, retain the genomic, histological, and functional characteristics of their original tissue, enabling assessment of contaminant-induced toxicity, short-term peak exposures, and inter-donor variability within a single system. This study examined whether current international drinking water guidelines remain protective or if recent organoid-based findings reveal toxicity at differing concentrations. Comparative synthesis indicates that PFAS often display organoid toxicity at concentrations above current thresholds, suggesting conservative guidelines, whereas most metals are properly regulated. However, some metals exhibit toxicity at concentrations that include levels below guideline values, highlighting the need for further investigation. Emerging contaminants, including pesticides, nanoparticles, microplastics, and endocrine disruptors, induce adverse effects at environmentally relevant concentrations, despite limited or absent regulatory limits. Integrating organoid-based toxicology with high-frequency monitoring and dynamic exposure modeling could refine water quality guidelines and support adaptive regulatory frameworks that better reflect real-world exposure patterns and human diversity.

Abstract:

Background: Hexavalent chromium [Cr(VI)] and gamma radiation are environmental toxicants that cause DNA damage, oxidative stress, and endocrine disruption, with transgenerational impacts on offspring health. Preventive strategies against inherited toxic effects are lacking, prompting research into protective interventions such as phytopreparations. Methods: Adult male rats were exposed to Cr(VI) (180 mg/L in drinking water for 14 days) and/or a single gamma irradiation (0.2 Gy), with subgroups receiving stinging nettle (Urtica dioica) or burdock (Arctium lappa) seed oil (0.5 mL/day) prophylaxis before irradiation. After exposure, rats were bred and male first-generation (F1) offspring were evaluated at 16 months for serum testosterone and thyroxine (T4), oxidative stress markers (MDA, catalase, SOD), sperm concentration/morphology, and testicular histology. Group differences were analyzed via one-way ANOVA (p<0.05). Results: Parental exposure to Cr(VI) and gamma radiation caused significant reproductive and endocrine impairment in F1 males: testosterone and sperm concentration decreased, abnormal sperm morphology increased, and T4 levels were disrupted compared to controls. However, parental supplementation with nettle or burdock oil significantly mitigated these effects, improving offspring hormone levels and sperm quality. Notably, burdock oil co-treatment restored testosterone and T4 toward control values and reduced sperm abnormalities in the combined Cr(VI)+γ group, indicating preserved spermatogenesis. Conclusions: Phytopreparation prophylaxis (nettle and burdock oils) in exposed parents partially normalized hormonal and reproductive parameters in F1 offspring and preserved testicular structure. These findings highlight the potential of phytoprotective interventions to attenuate transgenerational toxicity from Cr(VI) and radiation, thereby safeguarding future generations.

Abstract: Micro- and nanoplastics (MNPs) are emerging environmental immunotoxins with widespread human exposure through ingestion, inhalation, and dermal contact. Detected in the placenta, lungs, blood, bone marrow, and brain, MNPs accumulate in immune organs where they disrupt innate and adaptive cell functions. Experimental evidence shows that MNPs impair macrophage phagocytosis, skew dendritic cell maturation, trigger neutrophil extracellular traps, and alter T and B cell responses. Mechanistically, these effects are driven by oxidative stress, mitochondrial dysfunction, and activation of NF-κB, MAPK, and NLRP3 inflammasome pathways, leading to apoptosis, pyroptosis, and chronic low-grade inflammation. Barrier disruption and microbiome dysbiosis further compromise immune tolerance, linking MNPs exposure to autoimmune risk. Preclinical models demonstrate exacerbation of systemic lupus erythematosus, inflammatory bowel disease, and rheumatoid arthritis, though human causal data remain limited. Standardized protocols, advanced imaging and omics, and longitudinal cohorts are urgently needed to establish causal links and inform public health strategies.

Abstract:

Background: Developmental neurotoxicity (DNT) arises from disruption of key neurodevelopmental processes, including proliferation, differentiation, migration, and synaptogenesis. Traditional in vivo testing is costly and lacks mechanistic resolution, prompting OECD and EFSA to endorse new approach methodologies (NAMs) such as the neurosphere assay (NSA). Methods: We validated a mouse-derived NSA over a three-week differentiation period using multiparametric endpoints—proliferation, neuronal and glial differentiation, radial migration, synaptogenesis, and astrocytic maturation. Baseline characterization was established by flow cytometry, confocal immunofluorescence, and qPCR. Toxicants were applied under two exposure scenarios: (i) chronically during the 7-day proliferation phase, and (ii) chronically after three days of proliferation and maintained throughout the three-week differentiation period. Chlorpyrifos (CPF) served as a DNT-positive reference, and a biomonitoring-relevant mixture of PFAS (PFOS, PFOA, PFUnDA, PFHxS) was designed from the French Esteban study. Results: Baseline analyses confirmed progressive neuronal and glial differentiation, synaptic maturation, and growth factor responsiveness. CPF induced biphasic effects: early enlargement, subsequent growth arrest, migration impairment, overt cytotoxicity at 250 µM, and GFAP downregulation. PFAS mixtures produced modest viability effects but consistently reduced migration and downregulated GFAP and SYP at low-nM levels, consistent with epidemiological and experimental evidence linking PFAS to neurodevelopmental disorders. Migration emerged as a more sensitive endpoint than bulk viability, revealing functional impairments below overt toxicity thresholds. Conclusions: The NSA faithfully recapitulates corticogenesis and detects toxicant-specific liabilities, with CPF impairing astrocytic maturation and PFAS mixtures disrupting astrocytic and synaptic programs at environmentally relevant levels. By integrating complementary endpoints under chronic exposure conditions across proliferation and differentiation, this study advances the NSA as a mechanistic, human-relevant, and regulatory-ready NAM for DNT assessment.