BACKGROUND: Preclinical toxicity assessment is a critical step in drug development, despite advances in in vitro screening, these models remain insufficient to predict complex systemic toxicity, necessitating in vivo eva...BACKGROUND: Preclinical toxicity assessment is a critical step in drug development, despite advances in in vitro screening, these models remain insufficient to predict complex systemic toxicity, necessitating in vivo evaluation for novel compounds. 7-Methoxy-3-Phenyl-4H-Chromen-4-one, a structurally relevant chromen derivative, lacks prior in vivo toxicological data despite its potential biological activity. METHODS: Acute and sub-acute oral toxicity studies were conducted in in female Wistar albino rats following OECD guidelines. Acute toxicity was assessed using single-dose administration, while sub-acute toxicity involved repeated dosing over 28 days at graded dose levels. Safety evaluation included clinical observations, body weight, hematological and biochemical parameters, cardiac biomarkers and histopathological examination of major organs. RESULTS: Acute exposure did not produce significant systemic toxicity. Sub-acute administration at lower doses maintained normal physiological and biochemical profiles, whereas higher doses induced dose-dependent alterations in hepatic, renal and cardiac biomarkers along with histopathological changes. CONCLUSION: The compound demonstrates a defined safety margin, with a no observed adverse effect level (NOAEL) of 50 mg/kg and lowest observed adverse effect level (LOAEL) of 300 mg/kg. This study was specifically designed to address a critical gap in safety evaluation, providing essential in vivo toxicological data to support future pharmacological and translational investigations.
Mancozeb (MZB), a widely used fungicide, induces oxidative stress-mediated cytotoxicity through excessive reactive oxygen species (ROS) generation. Present study evaluated the cytoprotective potential of zerumbone (Zer),...Mancozeb (MZB), a widely used fungicide, induces oxidative stress-mediated cytotoxicity through excessive reactive oxygen species (ROS) generation. Present study evaluated the cytoprotective potential of zerumbone (Zer), a bioactive sesquiterpenoid, against MZB-induced toxicity in Vero cells. Dose-response and time-course assays determined the EC of MZB and a non-cytotoxic dose of Zer. Pre-treatment with Zer (10 μM, 18 h) significantly improved cell viability (66 ± 7%) following MZB exposure (22 μM, 6 h) and markedly reduced intracellular ROS levels. Zer attenuated MZB-induced DNA damage, lowering genotoxicity from 44 ± 6% to 23 ± 12% (p < 0.05), and significantly reduced apoptotic and necrotic cell death. Expression of apoptosis (Bax, Bcl-2, caspase-3, -8, -9), antioxidant (Nrf2, Keap1, catalase, HO-1, NQO1, SOD, GPx, PHD2) and inflammation-related (p53, Akt) markers was analysed and validated by western blotting to characterize molecular responses to Zer pre-treatment. Dysregulated expression of apoptosis and antioxidant markers, including caspase-3, catalase and GPx, was restored by Zer. Additionally, Zer normalized MZB-elevated Nrf2 levels while inducing its regulatory partners Keap1, NQO1 and HO-1. Overall, zerumbone confers substantial protection against MZB-induced oxidative injury by restoring redox balance, reducing genotoxic stress and preventing cell death, highlighting its therapeutic potential against fungicide-induced toxicity.
The global prevalence of metabolic diseases, including obesity, type 2 diabetes mellitus (T2DM), and metabolic dysfunction-associated steatotic liver disease (MASLD), continues to rise, representing a major global health...The global prevalence of metabolic diseases, including obesity, type 2 diabetes mellitus (T2DM), and metabolic dysfunction-associated steatotic liver disease (MASLD), continues to rise, representing a major global health threat and economic burden. Dihydroberberine (DHB), a reduced derivative of berberine (BBR), has recently garnered attention due to its superior lipophilicity and intestinal absorption. Pharmacokinetic studies suggested that DHB achieves significantly higher blood concentrations compared to BBR at equivalent doses. This review systematically synthesized the current preclinical evidence regarding the metabolic regulatory mechanisms of DHB. Key pharmacological targets identified in cell and animal models included the activation of AMP-activated protein kinase (AMPK) and glucokinase (GCK), modulation of lipid metabolism, and attenuation of inflammatory and oxidative stress pathways. Furthermore, DHB interacted extensively with the gut microbiota, acting both as a microbial metabolite of BBR and a modulator of microbial composition. Toxicological assessments indicated a favorable safety profile, although potential risks such as hERG channel inhibition required careful evaluation. Importantly, while in vitro and animal studies demonstrated significant metabolic benefits, human clinical trials assessing direct disease outcomes remained highly limited. This review highlighted the pharmacokinetic advantages of DHB and outlined the critical translational gaps that must be addressed in future research.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
The responses underlying heightened sensitivity of young populations to deoxynivalenol (DON) toxicity remain uncharacterized in translational murine models. This study aimed to fill this gap by providing the first system...The responses underlying heightened sensitivity of young populations to deoxynivalenol (DON) toxicity remain uncharacterized in translational murine models. This study aimed to fill this gap by providing the first systematic, multi-organ (intestine, liver, and kidney) comparison of DON toxicity, contrasting the responses of prepubertal and adult mice. Prepubertal (21-day-old) and adult (65-day-old) Swiss mice (n = 10/group) were fed control or 10 mg/kg DON-contaminated diets for 15 or 28 days, respectively. Growth performance was monitored, and intestine, liver, and kidney were subsequently collected for histopathological, morphometric, and biochemical analyses. DON induced a severe, acute weight gain reduction (∼59.6%) in prepubertal mice, with no effect on adults or systemic biochemical profiles. DON toxicity was highly organ-specific: the prepubertal intestine mounted an adaptive response, coupling oxidative damage with compensatory antioxidant production and enterocyte hypertrophy, but without structural lesions. In contrast, the liver and kidney sustained direct histopathological damage in both age groups. However, their biochemical responses diverged: the liver showed a young-specific inflammatory signal, whereas the kidney remained biochemically silent despite the damage it sustained. This multi-organ comparison in mice shows that DON toxicity is age- and organ-specific, demonstrating that at the exposure dose used, prepubertal mice exhibit more pronounced toxic responses.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
The existing information supports the use of this material as described in this safety assessment. β-Naphthyl isobutyl ether was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respirator...The existing information supports the use of this material as described in this safety assessment. β-Naphthyl isobutyl ether was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Target data and data from read-across analog β-naphthyl methyl ether (CAS # 93-04-9) show that β-naphthyl isobutyl ether is not expected to be genotoxic. Data on β-naphthyl isobutyl ether provide a calculated Margin of Exposure (MOE) >100 for the repeated dose toxicity endpoint. Data on read-across analog β-naphthyl methyl ether (CAS # 93-04-9) provide a calculated MOE >100 for the reproductive toxicity endpoint. Data show that there are no safety concerns for β-naphthyl isobutyl ether for skin sensitization under the current declared levels of use. The photoirritation/photoallergenicity endpoints were evaluated based on ultraviolet/visible (UV/Vis) spectra; β-naphthyl isobutyl ether is not expected to be photoirritating/photoallergenic. The local respiratory toxicity endpoint was evaluated using the Threshold of Toxicological Concern (TTC) for a Cramer Class III material, and the exposure to β-naphthyl isobutyl ether is below the TTC (0.47 mg/day). The environmental endpoints were evaluated; β-naphthyl isobutyl ether was found not to be Persistent, Bioaccumulative, and Toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards, and its risk quotients (RQs), based on its current volume of use (VoU) in Europe (EU), North America (NA), Asia-Pacific (AP), and South America (SA) (i.e., Predicted Environmental Concentration/Predicted No Effect Concentration [PEC/PNEC]), are <1.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a highly toxic persistent organic pollutant (POP), is associated with musculoskeletal disorders, yet its molecular mechanisms in osteoarthritis (OA)-related cartilage damage re...2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a highly toxic persistent organic pollutant (POP), is associated with musculoskeletal disorders, yet its molecular mechanisms in osteoarthritis (OA)-related cartilage damage remain unclear. This study aimed to explore this mechanism and identify key functional targets. Core genes were screened by integrating network toxicology and OA cartilage transcriptome data, validated via independent cohorts and machine learning, and further identified using SHAP combined with WGCNA. RT-qPCR verified the core target's gene expression in chondrocytes; zebrafish jaw cartilage transcriptome data (GSE11893) confirmed its expression changes after TCDD exposure; Western Blot (WB) validated protein expression and enriched pathways; and a rat OA model with proteomics further verified the target. Molecular docking and dynamics simulations assessed TCDD-target binding affinity. Bioinformatics analysis identified MMP9 as a key candidate gene, and experimental validation confirmed elevated MMP9 expression and increased p38 MAPK phosphorylation under OA-like cartilage injury conditions. Supplementary Western blot analysis in the IL-1β-induced OA-like injury model further showed increased phosphorylation of ERK and JNK, suggesting broader MAPK branch activation under inflammatory OA-like injury conditions. TCDD exposure time-dependently upregulated MMP9 in the zebrafish jaw cartilage transcriptome dataset, and molecular docking/dynamics simulation suggested a potential TCDD-MMP9 interaction. These findings indicate that MMP9 may be associated with TCDD-related OA cartilage damage and may participate in cartilage ECM dysregulation through MAPK signaling, particularly the p38 MAPK branch. Further human biomonitoring and low-dose chronic exposure studies are required to validate the clinical relevance of this mechanism.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Methcathinone (MCAT), a synthetic cathinone structurally analogous to amphetamine, poses substantial public health concerns due to its high addictive liability and pronounced neurotoxicity. In the present study, rat mode...Methcathinone (MCAT), a synthetic cathinone structurally analogous to amphetamine, poses substantial public health concerns due to its high addictive liability and pronounced neurotoxicity. In the present study, rat models of MCAT-induced neurotoxicity were established using low (0.5 mg/kg), medium (5 mg/kg), and high (20 mg/kg) doses. Cognitive function was assessed using the Morris water maze, while hippocampal synaptic morphology and ultrastructure were examined via Golgi staining and transmission electron microscopy. To elucidate the underlying molecular mechanisms, whole-transcriptome sequencing was performed to profile mRNAs, miRNAs, circRNAs, and lncRNAs in the hippocampus across exposure groups relative to controls. Differential expression analysis identified extensive transcriptional alterations, including 1646, 1539, and 1477 DEmRNAs; 32, 28, and 23 DEmiRNAs; 749, 728, and 753 DEcircRNAs; and 391, 369, and 371 DElncRNAs in the low-, medium-, and high-dose groups, respectively. Functional enrichment analyses consistently implicated synapse-related processes and neurodegeneration-associated pathways. Notably, activity-dependent immediate-early genes (c-Fos, Nr4a1, Arc, Egr1, Egr2, and Npas4) were uniformly downregulated across all exposure levels, indicating impaired neuronal activity-dependent transcriptional responses. Integration of multi-layered transcriptomic data enabled the construction of circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks, revealing extensive post-transcriptional regulatory interactions. A core ceRNA network was identified, comprising 6 hub mRNAs, 9 miRNAs, 95 lncRNAs, and 146 circRNAs. Quantitative RT-PCR validation demonstrated high concordance with RNA-seq results, supporting the robustness of the dataset. These findings demonstrate that MCAT induces cognitive deficits and synaptic structural impairments by disrupting activity-dependent gene expression and neurotrophic signaling through complex ceRNA-mediated regulatory networks. This study provides novel mechanistic insights into MCAT-induced neurotoxicity and identifies potential molecular targets for therapeutic intervention in psychostimulant-related cognitive dysfunction.
Nicotine pouches are marketed as a smokeless alternative to traditional tobacco products, delivering nicotine through a powder blend of water, salts, aromatic oils, and other additives. Building on previous work examinin...Nicotine pouches are marketed as a smokeless alternative to traditional tobacco products, delivering nicotine through a powder blend of water, salts, aromatic oils, and other additives. Building on previous work examining nicotine release under simulated use, this study assessed additional substances transferred into saliva during pouch exposure. GC-HRMS analysis revealed a broad range of compounds, including terpenes, benzene derivatives, phenols, ethers, and alcohols. Of sixty-five detected terpenes and terpenoids, only fourteen are authorized under Regulation (EC) No. 1334/2008, and several constituents are known allergens or suspected carcinogens. Phthalates such as dibutyl and bis(2-ethylhexyl) phthalate-both banned in cosmetics and recognized environmental pollutants-were also present. Two chlorinated compounds, tentatively identified as 2-(1-chloroethoxy)acetaldehyde and ((chloromethoxy)methyl)benzene, were detected, though their toxicological relevance remains unknown. Quantification of myosmine, a tobacco alkaloid not permitted as a food additive, detected 0.04 mg in pouch types F and 0.03 mg in pouch S, but nothing in V. Nicotine release after one hour in artificial saliva was highest in F (3.11 mg), followed by V (2.75 mg) and S (1.24 mg). A 50 kg child may be at risk from 25-50 mg nicotine, and in a worst-case scenario, consuming about ten strong pouches in an hour could approach a potentially lethal dose. These findings underscore the need for stricter regulation and comprehensive toxicological evaluation of nicotine pouch ingredients.
Colitis-associated liver injury rarely causes acute mortality, but whether alcohol and a high-fat diet (HFD) aggravate hepatic injury during colitis remains unclear. Here, dextran sulfate sodium (DSS)-induced colitic mic...Colitis-associated liver injury rarely causes acute mortality, but whether alcohol and a high-fat diet (HFD) aggravate hepatic injury during colitis remains unclear. Here, dextran sulfate sodium (DSS)-induced colitic mice were exposed to alcohol, HFD, or both. Compared with DSS alone, combined HFD and alcohol exposure (DSSHA) caused greater body-weight loss, poorer body condition, and reduced survival. These effects were linked mainly to liver rather than intestinal deterioration, as shown by an increased liver/body-weight ratio, elevated serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and severe hepatic steatosis, whereas colon length, colon weight, and intestinal histological injury were not further aggravated. Mechanistically, alcohol-HFD co-exposure impaired the gut-liver axis, reduced intestinal tight-junction proteins, increased gut permeability and circulating lipopolysaccharide (LPS), and promoted hepatic macrophage and neutrophil infiltration with increased interleukin-6 (IL-6). Transcriptomics identified serum amyloid A1 (SAA1) as a highly upregulated acute-phase gene and revealed Toll-like receptor 4 (TLR4) pathway enrichment. TLR4 inhibition TAK-242 significantly improved survival and attenuated liver injury. Multi-omics further showed increased hepatic cortisol and corticosterone, which cooperated with IL-6 to induce SAA1 expression in HepG2 cells. Thus, an IL-6/glucocorticoid-SAA1-TLR4 axis drives alcohol-HFD-aggravated liver injury in colitis and may represent a therapeutic target.
Exposure to bisphenol A (BPA) and long-term high-fat diet (HFD) have been proven to have adverse effects on human health. This study demonstrated that the sequential exposure to BPA in early life and high-fat diet after...Exposure to bisphenol A (BPA) and long-term high-fat diet (HFD) have been proven to have adverse effects on human health. This study demonstrated that the sequential exposure to BPA in early life and high-fat diet after weaning jointly impaired intestinal health. We found that BPA altered the gut microbiota composition of offspring, reducing microbial diversity and increasing Proteobacteria abundance. While short-term HFD intake partially restored microbial diversity, prolonged HFD feeding exacerbated dysbiosis. Early-life BPA exposure and subsequent HFD synergistically increased intestinal permeability, elevated serum zonulin and endotoxin (ET), and induced colonic inflammation. Notably, serum ET levels were significantly influenced by sex. Transcriptomic analysis revealed that the T cell receptor (TCR) and cell adhesion molecule (CAMs) signaling pathways were the key pathways associated with intestinal barrier impairment. Immunohistochemical analysis further showed increased CD3 T cell infiltration and ICAM-1 positive area in the colon of 13-week-old male offspring, particularly in the BPA + HFD group. These findings highlight a critical developmental window during which environmental and dietary factors interact to program long-term intestinal vulnerability.
Dietary ingestion of nanoplastics through contaminated food and drinking water has become a pressing human health concern. Despite this pressing health concern, the developmental reproductive impacts of gestational nanop...Dietary ingestion of nanoplastics through contaminated food and drinking water has become a pressing human health concern. Despite this pressing health concern, the developmental reproductive impacts of gestational nanoplastics exposure on female offspring remains poorly characterized. Using ICR mouse models subjected to prenatal exposure to 60-nm polystyrene nanoplastics (PS-NPs: 5, 50, or 500 mg/kg/day), this study demonstrated that maternal perinatal PS-NPs exposure significantly disrupted ovarian development, induced precocious puberty, and caused ovarian follicular depletion in female offspring, evidenced by reduced ovarian coefficients, advanced vaginal opening, depleted primordial follicles, and elevated atretic follicles. Mechanistically, ovarian development dysregulations arise from suppressed FOXL2-CYP19A1 signaling and downregulated RSPO1/WNT4/β-catenin pathways, while precocious puberty is driven by earlier elevation of serum GnRH levels and hypothalamic reprogramming. Quantitative proteomics reveals dose-dependent pathway alterations: low-dose PS-NPs dysregulate immune/estrogen responses, whereas high-dose exposure causes steroidogenic disruption and apoptotic dysregulation. Serum hormone profiling showed markedly reduced AMH/E2 and elevated LH, corroborated by transcriptional downregulation of key receptors (amh, ERα, and AR). These findings highlight a potential risk to fetal ovarian development posed by maternal nanoplastics exposure, urging the incorporation of developmental and reproductive toxicity into the comprehensive risk assessment of human exposure to micro/nano-plastics.
Alternariol (AOH), a mycotoxin produced by Alternaria species, has been associated with oxidative and inflammatory toxicity, although its effects on astrocytes remain poorly understood. This study investigated AOH-induce...Alternariol (AOH), a mycotoxin produced by Alternaria species, has been associated with oxidative and inflammatory toxicity, although its effects on astrocytes remain poorly understood. This study investigated AOH-induced cellular responses and the protective effects of eicosapentaenoic acid (EPA) in human astrocytes. Exposure to AOH (25-125 μM) reduced cell viability, increased intracellular reactive oxygen species (ROS) production, depleted glutathione (GSH), and elevated the expression of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6. AOH also induced apoptosis-related molecular alterations, characterized by increased Bax, cleaved caspase-9, and cleaved caspase-3 expression together with reduced Bcl-2 levels. In addition, AOH modulated the expression of Nrf2 and its downstream antioxidant targets, heme oxygenase-1 (HO-1) and NAD(P)H oxidoreductase 1 (NQO1), suggesting involvement of antioxidant defense-related signaling. Pretreatment with EPA (5 μM) significantly attenuated AOH-induced oxidative stress, inflammatory responses, and apoptosis-related molecular changes. These findings further identify astrocytes as a sensitive cellular target of Alternaria mycotoxins. Collectively, the findings indicate that AOH disrupts redox homeostasis and promotes cellular stress responses in human astrocytes, whereas EPA mitigates these effects and helps maintain cellular homeostasis.
Alternariol (AOH) is a mycotoxin produced by fungi belonging to the genus Alternaria, detected in a large variety of commodities such as cereals, fruits, vegetables, beverages, nuts, and other food products. The present...Alternariol (AOH) is a mycotoxin produced by fungi belonging to the genus Alternaria, detected in a large variety of commodities such as cereals, fruits, vegetables, beverages, nuts, and other food products. The present study investigates the effect of the mycotoxin alternariol on gut integrity and permeability using an intestinal porcine epithelial cell (IPEC-1) monolayer as a model for intestinal barrier and transport function. Exposure of IPEC-1 cells to higher concentrations of AOH (20 and 50 μg/mL) induced a dose-dependent decrease in cell proliferation, an increase in DNA fragmentation, and a significant increase in LDH activity in porcine intestinal epithelial IPEC-1 cells, indicating an important alteration in the capacity of intestinal epithelial renewal induced by AOH. Higher concentrations of AOH (5 and 10 μg/mL) decreased transepithelial electrical resistance and increased the passage of a fluorophore-conjugated macromolecule, indicating an alteration of the IPEC-1 monolayer integrity and permeability. These alterations are due to a decrease in tight junction proteins and mucins and are associated with an increase in nitric oxide synthesis and a decrease in the levels of pro-inflammatory cytokines IL-6 and IL-8. Taken together, these results showed an important alteration of the gut epithelium with important consequences for pig health. Our results should be confirmed by in vivo trials in swine in order to validate these in vitro findings.
The Adverse Outcome Pathway (AOP) framework has become a powerful tool in mechanistic toxicology, linking molecular initiating events through key biological changes to adverse health outcomes. In food safety, where chron...The Adverse Outcome Pathway (AOP) framework has become a powerful tool in mechanistic toxicology, linking molecular initiating events through key biological changes to adverse health outcomes. In food safety, where chronic exposure to diverse contaminants is common, AOPs offer a promising alternative to traditional animal testing by supporting in vitro and in silico approaches. This review evaluates current AOPs developed for major dietary contaminants such as mycotoxins, heavy metals, acrylamide, dioxins, and packaging-related chemicals. It critically assesses the quality of AOPs in the AOP-Wiki, focusing on the strength and biological relevance of key event relationships. The study also examines how regulatory agencies incorporate AOPs into risk assessment, revealing limited formal adoption despite their potential. Key gaps are identified, including insufficient quantitative data, challenges in addressing mixture toxicity, and limited cross-species applicability. Additionally, the role of computational tools in AOP development and validation is analyzed. Overall, the review highlights a disconnect between AOP research and regulatory implementation and provides recommendations to enhance their integration into food safety assessment frameworks, ultimately supporting more efficient, ethical, and mechanism-based risk evaluation.
Syzygium cumini seed preparations are clinically used during pregnancy for gestational hyperglycaemia, yet the developmental safety of any defined flavonoid glycoside of the species has not been examined in a mammalian m...Syzygium cumini seed preparations are clinically used during pregnancy for gestational hyperglycaemia, yet the developmental safety of any defined flavonoid glycoside of the species has not been examined in a mammalian model. We examined whether oral exposure to a chemically standardised, syringetin-3-O-glucoside (S3G)-enriched isolate at 0, 20, 40, or 80 mg/kg/day from gestational day 5 through 19 perturbs the maternal-fetal thyroid axis in Sprague Dawley (SD) rats. Maternal serum TSH rose dose-dependently; total T4, T3, and rT3 declined; thyroglobulin rose six-fold at the high dose; and thyroid histopathology progressed through follicular hyperplasia and colloid depletion to solid cellular sheets. Reproductive indices and fetal body weight were unaffected, and fetal findings were limited to minor variations. RNA-sequencing of maternal thyroid identified 930 differentially expressed genes, with compensatory upregulation of Tpo, Slc5a5/NIS, Tg, and Iyd. The isolate inhibited iodide uptake at sub-cytotoxic concentrations in rat FRTL-5 thyrocytes (IC 18.42 μg/mL) and was ∼3.4-fold less potent in human Nthy-ori 3-1 cells (IC 62.15 μg/mL). Intrathyroidal syringetin, the bioavailable aglycone, reached 440.0 ± 36.7 nM, and molecular docking prioritised NIS among favourably scored targets. These convergent findings localise the lesion to post-transcriptional NIS inhibition and distinguish S3G from the classic flavonoid-thyroid peroxidase inhibitor class.