Search Results (681)

N-nitrosamines and acrylamide are food mutagens classified as “probably carcinogenic to humans (Group 2A)” by the International Agency for Research on Cancer (IARC) from evidence of carcinogenicity. One of the main objectives of food safety is to reduce the presence of these substances in food. Therefore, the present study aimed to evaluate the effect of the addition of propolis, royal jelly or a combination of both bee products (2–10%) to chestnut and thyme honey on their protective properties against food mutagen-induced genotoxicity. DNA damage was evaluated by the alkaline comet assay. N-nitrosamines (N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR)) and acrylamide (AA) induced genotoxicity in human hepatoma HepG2 cells. All tested samples at all concentrations used (0.1–10 µg/mL) decreased genotoxic effects of the three food mutagens. The protective effects of honey samples and mixtures towards DNA damage induced by food mutagens were in the following order: NDMA > AA > NPYR, being more effective against NDMA compared to AA and NPYR. The mixtures of chestnut honey with 10% propolis, or 10% royal jelly, and 10% propolis showed a greater protective effect against NDMA, NPYR and AA compared to the honey sample alone. This protective activity may be attributable to the phenolic compound content and antioxidant capacity exhibited by the analyzed samples. Overall, the results suggest that thyme and chestnut honey supplemented with bee-derived products could represent potential natural chemopreventive candidates against food-borne mutagens. Full article

Background: Obesity is a chronic endocrine–metabolic disorder. The risk of comorbidities increases with a higher body mass index (BMI), particularly when BMI ≥ 35.0 kg/m². Common complications include insulin resistance, type 2 diabetes, dyslipidemia, and chronic low-grade inflammation, which collectively impair DNA stability by promoting the formation of genotoxic species. Methods: This non-randomised, quasi-experimental clinical intervention study included 53 participants (both sexes) with a BMI ≥ 35.0 kg/m², who were assigned to parallel experimental or control streams based on clinical needs and institutional eligibility. During a three-week intervention, the experimental group received a hospital-supervised very-low-calorie diet (VLCD; ~600 kcal/day) under continuous medical monitoring. Conversely, the control group followed a standard reduced-calorie diet (SRD) of 1500 kcal/day in a free-living home environment. Before and after the intervention, primary, oxidative, and permanent DNA damage were measured using alkaline, FPG-modified comet (peripheral blood mononuclear cells), and cytokinesis-block micronucleus cytome assays (phytohaemagglutinin-stimulated binucleated lymphocytes), alongside anthropometric and biochemical tracking. Results: Within-group evaluations revealed that both dietary regimens improved several metabolic health indicators, notably modulating insulin resistance, lipid profiles, and leukocyte counts. However, participants in the VLCD stream experienced significantly greater downward changes in body weight, BMI, and absolute lipid values. Crucially, the VLCD intervention was associated with a highly significant within-group reduction in parameters of permanent chromosomal damage, effectively halving the frequencies of micronuclei and nuclear buds, independent of baseline variations, in adjusted multivariate regression models. Conversely, the home-based SRD regimen demonstrated no measurable impact on permanent genomic damage. Neither diet induced a significant change in repairable primary or oxidative DNA lesions over this short timeframe. Conclusions: These exploratory findings suggest that strict calorie restriction can rapidly stabilise genome stability in advanced clinical settings, warranting future randomised controlled trials with long-term longitudinal follow-up to assess permanent risk reductions. Due to structural baseline variations in age, chronic comorbidities, and compliance environments between the cohorts, direct comparative superiority cannot be definitively established. Full article

Commercially available graphene quantum dots (GQDs) are promising nanomaterials for applications in research and preclinical diagnostics, drug delivery, and bioimaging. Their bioactivity is highly dependent on dose, route of exposure, duration, cell type, uptake mechanisms, tissue and cellular distribution, and physicochemical properties. This study aimed to evaluate genotoxic, cytotoxic, and cytostatic endpoints of blue- (B-GQDs) and green-emitting (G-GQDs) GQDs in human blood and salivary leukocytes. GQDs were tested at concentrations ranging from 2.5 to 100 µg/mL using distinct treatment periods. Fourier transform infrared spectroscopy (FTIR), trypan blue exclusion, comet, and cytokinesis-block micronucleus cytome (CBMN cyt) assays were performed. FTIR analysis revealed that G-GQDs, unlike B-GQDs, exhibit an absorption band typically associated with amine functional groups, which may contribute to their pronounced genotoxic effects. Peripheral blood mononuclear cells and salivary leukocytes showed higher sensitivity to G-GQDs compared to whole blood samples. Although no cytotoxic effects were observed, both GQDs induced significant DNA damage, with G-GQDs demonstrating greater genotoxic potential. These findings demonstrate that GQDs can induce DNA damage in the absence of detectable cytotoxic effects under the conditions tested, highlighting the importance of considering both physicochemical properties and cellular models in the safety assessment of nanomaterials. Full article

Aspergillus cristatus is a key microorganism involved in the fermentation of Fu brick tea, but systematic strain-level safety data remain limited despite its long history of use in dark tea production. In this study, the toxicological safety of A. cristatus CCNH008 was assessed through acute oral toxicity assay, a 90-day repeated oral toxicity study with a recovery period and genotoxicity assays. In acute oral toxicity tests, CCNH008 caused no mortality, no adverse clinical signs, and no treatment-related effects on bodyweight or pathology in mice or rats, with an LD₅₀ greater than 10 g/kg bodyweight. In the 90-day repeated oral toxicity study in rats, the oral administration of CCNH008 at doses up to 1.67 g/kg bodyweight/day produced no treatment-related changes in clinical signs, bodyweight, food consumption, hematological or biochemical parameters, urinalysis, organ weights, or histopathology, including during a 28-day recovery period. Genotoxicity was evaluated using a bacterial reverse mutation assay, an in vitro mammalian chromosome aberration test, and an in vivo mammalian erythrocyte micronucleus test; no mutagenic or clastogenic effects were observed. Collectively, these findings demonstrate that CCNH008 shows no evidence of acute, subchronic oral toxicity, or genotoxicity, providing important strain-level safety evidence to support its potential application in food-related products. Full article

The potential effects of microplastics (MPs) on humans and ecosystems are of great concern, and it has been reported that the ingestion of contaminated food is the main route of exposure. In the present study, Perca fluviatilis was selected as a vertebrate model to evaluate the possible cellular effects induced by five different plastic polymers and car tire debris (CT) after 4- and 7-month exposure periods. The Cytome assay was chosen to check chromatin alteration in perch’s peripheral blood. The results indicated an increase in micronuclei and cytotoxic effect in specimens co-exposed to MPs + CT for 7 months. Increases in dicentric chromosomes were observed in specimens exposed both to MPs alone and to the mixture of MPs + CT, indicating for the first time a genotoxic effect induced by CT debris in fish in terms of structural aberrations. Increases in micronucleated erythrocyte frequency assessed after 7 months only after the addition of CT debris to the mixture of MPs might suggest an aneugenic action of CT in fish. In the same groups, the higher values of frequency in 8-shaped erythrocytes also indicate possible cell cycle toxicity exerted by CT exposure. An association between total erythrocyte nuclear morphology abnormalities (ENA) and glutathione reductase activity was also found, indicating a potential involvement of oxidative processes in modulating the genotoxicity observed. The present experimental model is a useful tool to study cellular mechanisms related to both MP- and CT-induced chromatin structure alterations indicating possible interference with human health as well. Full article

Chronic inhalation exposure to nanoplastics, specifically polyethylene terephthalate (PET) nanoplastics (PET-NPLs) is an emerging health concern, yet the long-term consequences for genomic stability and DNA damage response (DDR) capacity in bronchial epithelial cells remain poorly characterized. For this study, human bronchial epithelial BEAS-2B cells were continuously exposed to PET-NPLs for over 20 weeks, after which elevated basal DNA genotoxic damage was observed, as assessed by the alkaline comet assay. In addition, a broad transcriptional suppression of the DDR, with 27 of 84 profiled genes involved in DDR showing reduced expression relative to passage-matched control was observed. The suppressed genes span ATM/ATR checkpoint signaling, homologous recombination (HR), base excision repair (BER), nucleotide excision repair (NER), and apoptotic pathways. To determine whether chronic PET-NPL exposure altered susceptibility to acute genotoxic challenge in a damage-type-specific manner, cells were treated with methyl methanesulfonate (MMS), ultraviolet-C (UV-C) radiation, or bleomycin. While MMS and UV-C induced comparable levels of DNA damage in control and PET-exposed cells, bleomycin produced significantly greater damage in PET-exposed cells, indicating selective sensitization to doble-strand breaks (DSB)-type and oxidative genotoxic insults. Transcriptional profiling during bleomycin challenge identified 18 DDR genes with relatively higher expression in PET-exposed cells compared to passage-matched controls, encompassing HR, BER, ATM/ATR signaling, the Fanconi anemia pathway, and apoptosis. Furthermore, PET-exposed cells retained significantly higher residual DNA damage after 3 h of bleomycin challenge, indicating a persistent early repair deficit. Together, these findings suggest that chronic PET-NPL exposure specifically compromises the bronchial epithelial DDR, with potential implications for long-term genomic stability in respiratory epithelia subjected to nanoplastic inhalation. Full article

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. Full article

Background: Nosocomial infections caused by multidrug-resistant microorganisms are a significant public health concern. Antimicrobial resistance (AMR) is closely linked to the excessive and indiscriminate use of antibiotics, which creates selective pressure and promotes the emergence of resistant pathogens. Objectives: This study evaluates the synergistic potential of intragenic antimicrobial peptides (IAPs) combined with plant alkaloids against susceptible and multidrug-resistant human pathogenic bacteria, assessing antimicrobial activity, biofilm inhibition, and hemocompatibility. Methods: The tested molecules included berberine, tomatidine, sinomenine, and the IAPs Hs02 and Gr01. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) assays were performed against both ATCC (E. coli ATCC 25922 and S. aureus ATCC 25923) and clinical strains (E. coli KPC+ HRAN 1812446 and S. aureus MDR LACEN 3730529). Synergistic interactions were evaluated by checkerboard assay, followed by biofilm inhibition and hemolysis assays using human red blood cells. Results: Berberine exhibited a MIC of 1024 µM when tested individually, while tomatidine and sinomenine showed no significant activity. As expected, the IAPs showed strong antimicrobial properties at 8 µM (Hs02) and 4 µM (Gr01). When tested in synergy, alkaloids and IAPs reduced the MIC by up to 128-fold. The combination of IAPs and alkaloids reduced the biofilm biomass of S. aureus and E. coli by 50%, by the crystal violet assay (p < 0.05). Notably, sinomenine had not previously been reported to have antimicrobial activity. Conclusions: These results highlight the importance of further exploring combinations of natural and synthetic bioactive molecules as promising antimicrobial candidates. This approach may help to extend the useful life of conventional antibiotics. However, further studies are needed to assess safety, cytotoxicity, genotoxicity, inflammation, and in vivo effects. Full article

This study evaluates the cytotoxic and genotoxic-like potential of an ipfencarbazone-based herbicide formulation (IPF-BH; commercial product Hokuto, containing 250 g/L of the triazolinone herbicide ipfencarbazone) in human peripheral lymphocytes in vitro across a concentration range of 62.5–1000 µg/mL. Cytotoxicity was monitored via the mitotic index (MI), while cytogenetic damage was assessed using the cytokinesis-block micronucleus (MN) assay and the alkaline comet assays. Comparisons were performed using one-way ANOVA, followed by Dunnett’s post hoc test, against the negative control. Results indicated a concentration-dependent cytotoxic effect, with a marked reduction in MI observed at all tested concentrations (p < 0.001). MN frequency was significantly elevated at concentrations ≥125 µg/mL, whereas the 62.5 µg/mL concentration did not induce significant micronuclei formation. The comet assay revealed increased DNA damage parameters (tail length, tail intensity (%), and tail moment) across the tested concentration range, albeit with a non-monotonic profile for tail length and tail intensity. These findings suggest that IPF-BH exposure is associated with marked cytotoxicity and a genotoxic response in this in vitro model at concentrations within the OECD 487-acceptable cytotoxicity window, together with cytotoxicity-associated genotoxic-like effects at strongly cytotoxic concentrations in human peripheral lymphocytes under in vitro conditions. Because IPF-BH is a commercial formulation, and no direct mechanistic endpoints (e.g., reactive oxygen species, mitochondrial transmembrane potential, lipid peroxidation, glutathione) were measured, and because the present design was performed without exogenous metabolic activation (no S9 supplementation), the observed effects cannot be unambiguously attributed to ipfencarbazone alone or to a defined mechanism of action; extrapolation to in vivo genotoxicity requires complementary +S9 and rodent in vivo follow-up studies. Full article

Mesenchymal stromal cells derived from adipose tissue (ASCs) are widely used in regenerative medicine, requiring scalable expansion strategies that preserve both cellular function and biological quality. However, current bioprocess optimization approaches are primarily guided by proliferation and phenotypic stability, often overlooking genomic integrity as a critical attribute. In this study, we developed a stirred-tank bioreactor system for ASC expansion on microcarriers and applied a genotoxicity-informed optimization strategy by integrating growth kinetics, metabolic profiling, and DNA damage assessment across multiple operational conditions (B₁–B₅), including variations in dissolved oxygen, agitation, inoculum density, and medium renewal. Optimized culture conditions (B5) enabled high cell productivity within a reduced cultivation period (9 days), while maintaining high viability (>90%), mesenchymal immunophenotype, and differentiation capacity. Distinct metabolic profiles were associated with enhanced proliferation, with increased glycolytic activity observed under optimized conditions. Despite these favorable outcomes, genotoxic analyses revealed a progressive, time-dependent accumulation of DNA damage and increased micronucleus frequency during expansion. Notably, these alterations did not impair cell proliferation, phenotype, or differentiation potential, indicating that conventional optimization metrics may not fully capture underlying genomic changes. Collectively, our findings demonstrate that bioprocess optimization based solely on classical performance parameters may overlook relevant biological alterations. By incorporating genotoxic endpoints into the evaluation framework, this study provides a refined approach for assessing large-scale stem cell expansion and contributes to improving the robustness and reliability of biomanufacturing strategies for therapeutic applications. Full article

Sumizyme PEG, a glucanase/cellulase enzyme preparation produced by Talaromyces versatilis PF8, was investigated to characterize its systemic and genotoxic toxicity profile to support its intended use in food processing applications. A comprehensive toxicological program was conducted in accordance with OECD guidelines, comprising a bacterial reverse mutation (Ames) test, an in vitro chromosomal aberration assay, an in vivo micronucleus test, and a 90-day repeated-dose oral toxicity study in male and female Crl:CD(SD) rats. In the subchronic study, Sumizyme PEG was administered by oral gavage at doses of 107, 1070, and 10,700 U/kg/day. No treatment-related adverse effects were observed across clinical, hematological, biochemical, urinalysis, organ weight, or histopathological endpoints, and the highest dose was identified as the NOAEL. Genotoxic testing showed no consistent mutagenic or clastogenic response across the test battery. A positive in vitro signal was observed in CHL/IU cells; however, this was not reproduced in a human TK6 cell assay or in vivo micronucleus testing, indicating assay-dependent sensitivity within a weight-of-evidence framework. Overall, the integrated dataset does not indicate a consistent treatment-related systemic or genotoxic effect under the conditions of the studies conducted. Full article

The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B₁ levels were below regulatory limits. Acute oral toxicity studies established LD₅₀ values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability. Full article

Humans spend a substantial proportion of their time indoors, where exposure to environmental pollutants such as radon gas and particulate contaminants in household dust is common. While radon is a well-established risk factor for lung cancer, household dust may serve as a reservoir for a complex mixture of indoor and outdoor pollutants. However, the biological effects of such exposures, particularly under sequential conditions, remain incompletely understood. This study aimed to investigate the cytotoxic and genotoxic effects of sequential exposure to household dust extract followed by indoor radon using human lung adenocarcinoma (A549) cells as an in vitro model. Household dust samples from upper northern Thailand were extracted and applied to cells, followed by controlled radon exposure. Cellular responses were evaluated using cell viability assays, cytokinesis-block micronucleus (MN) formation assays, and Western blot analysis of oxidative stress-related (Nrf2/HO-1), DNA damage-related (γ-H2AX), autophagy-related (LC3), and inflammatory-related (IL-6) protein expression. Exposure to household dust extract was associated with reduced cell viability and increased MN formation, while radon exposure alone produced relatively modest effects under the present conditions. Sequential exposure to household dust extract followed by indoor radon was associated with increased oxidative stress-related responses and elevated DNA damage than either treatment alone under the present experimental conditions. A trend toward autophagy-related responses was also observed, and the overall findings may indicate possible combined biological responses under sequential exposure conditions. These findings suggest that sequential exposure may be associated with changes in oxidative stress-related pathways, DNA damage responses, and autophagy-related processes in this in vitro model. However, the results should be interpreted with caution as they are derived from a single cancer cell line and there are limitations to the in vitro exposure model. Further studies using additional cell models and in vivo systems are warranted to further clarify the potential biological and human health relevance of these findings. Full article

Cenostigma pluviosum var. peltophoroides, known as “sibipiruna,” is a plant rich in polyphenols used in traditional medicine for gastrointestinal disorders. The study aimed to investigate the chemical composition of the crude ethanolic extract of the stem bark (CEECP), evaluating its in vivo toxicity, genotoxicity, mutagenicity and anti-inflammatory activity. The plant material was macerated in 95% ethanol for 72 h, and the solvent was removed by rotary evaporation to obtain CEECP. Chemical characterization was performed by HPLC-ESI-MS/MS in negative mode. In vivo approaches were performed using male/female Swiss albino mice. Acute toxicity was assessed at a single high dose of 2000 mg/kg. Mutagenicity was investigated by the micronucleus test and genotoxicity by the comet assay, both at a dose of 2000 mg/kg. Anti-inflammatory activity was evaluated in carrageenan-induced paw edema and peritonitis models, at doses of 50, 100, and 200 mg/kg. HPLC-ESI-MS/MS analysis showed the presence of hydrolyzable tannins, phenolic acid heterosides, and biflavonoids. The safety profile of the CEECP was demonstrated for the first time, with no evidence of acute toxicity, mortality, mutagenicity, or genotoxicity at the tested doses. The extract significantly reduced paw edema in a dose-dependent manner at doses of 100 and 200 mg/kg, with inhibition rates of 65.78% and 73.12%, respectively, and also decreased leukocyte migration in the peritonitis model by 61.81% and 72.79% at the same doses. These findings indicate the CEECP as a source of pharmacologically relevant phytocompounds and, most notably, demonstrate its pronounced anti-inflammatory activity. Furthermore, the extract exhibited a favorable safety profile in the toxicological evaluations, highlighting the extract as a promising anti-inflammatory agent. Full article

Background/Objectives: Calcium phosphate cement (CPC) is extensively utilised in bone repair owing to its biocompatibility, osteoconductivity, and compositional similarity to native bone. Functionalisation of CPC with palm tocotrienol may enhance its regenerative potential. However, the incorporation of phytochemicals requires safety evaluation to exclude potential genotoxic risks. This study investigated the mutagenic potential of CPC and tocotrienol-enriched CPC (CPC-T3) using the bacterial reverse mutation assay. Methods: Mutagenicity was evaluated in five bacterial strains, including Salmonella typhimurium TA100, TA98, TA1535, TA1537, and Escherichia coli WP2 trp uvrA, under both non-metabolic and metabolic activation conditions. Revertant colonies were quantified at multiple concentrations and mutagenicity ratios were calculated relative to the negative control. Results: Across all strains and metabolic conditions, neither CPC nor CPC-T3 induced reproducible or concentration-dependent increases in revertant colony numbers. Although isolated elevations were detected at certain concentrations, these findings lacked dose–response relationships and did not meet the criteria for a positive mutagenic response according to Organisation for Economic Co-operation and Development (OECD) Test Guideline No. 471. The performance of negative and positive controls confirmed the validity and sensitivity of the assay. Notably, the inclusion of palm tocotrienol did not alter the overall mutagenicity profile of CPC. Conclusions: CPC and CPC-T3 demonstrated no evidence of mutagenic activity under the conditions of the bacterial reverse mutation assay. These findings represent preliminary genotoxicity screening. Further mammalian genotoxicity and in vivo studies are warranted to support future translational development as implantable medical devices. Full article