Search Results (38)

Waste tires (WTs) constitute a potentially significant source of pollution, and the large quantities that are disposed of require proper handling. Pyrolysis has emerged as an environmentally friendly and effective method for WT treatment. In the present study, the cyto-genotoxic and toxic effects of untreated and acid-treated pyrolytic tire char (PTC_UN and PTC_AT, respectively) were investigated. The cytokinesis block micronucleus (CBMN) assay, using human lymphocytes, and the Aliivibrio fischeri bioluminescence assay were used for the assessment of cyto-genotoxicity and ecotoxicity, respectively. According to the results, both PTC_UN and PTC_AT exhibited genotoxicity at all concentrations tested (2.5, 5, and 10 μg mL⁻¹), which was more pronounced in the case of PTC_AT. Cytotoxicity induction was reported for PTC_UN and PTC_AT at all concentrations. Both demonstrated a relatively low potential for ecotoxicity induction against A. fischeri. Since the cyto-genotoxic and toxic effects of PTC_AT seemed to be more pronounced, the toxic profile of tire char should be investigated in depth before selecting the appropriate applications, thereby avoiding detrimental effects in the environment and humans alike. Full article

Exposure to arsenic, a known environmental and occupational genotoxicant, poses significant health risks. Identifying agents capable of mitigating its effects is crucial for public health. This study evaluates the protective potential of Argovit™ silver nanoparticles (AgNPs) against cytotoxic and genotoxic damage induced by sodium arsenite in ex vivo cultured human lymphocytes obtained from the whole blood of healthy donors. Lymphocytes were exposed to sodium arsenite (3.7 × 10⁻³ µg/mL) and Argovit™ AgNPs (3.6 × 10⁻³ µg/mL). The cytokinesis-block micronucleus (CBMN) assay was performed using a modified 144 h protocol to assess delayed effects across two cell cycles. Four groups were analyzed: untreated control, sodium arsenite only, AgNPs only, and sodium arsenite followed by AgNPs. Arsenite exposure increased cytotoxic and genotoxic biomarkers. In contrast, post-treatment with AgNPs significantly reduced these effects. All treatments were performed in duplicate, and data were analyzed using the Kruskal–Wallis test with Dunn’s post hoc comparison (p < 0.05). Statistical analysis confirmed the antigenotoxic and cytoprotective properties of Argovit™. These findings support its potential application as a mitigating agent in scenarios of environmental or occupational exposure to genotoxic compounds. Full article

Fludioxonil is a widely used fungicide that is frequently used to combat fungal plant diseases. Consequently, excessive concentrations of fludioxonil may enter and accumulate over time in aquatic systems, harming (micro) organisms in several ways. Thus, it is of great importance to evaluate the potential toxic effects of fludioxonil using bioassays. In the present study, various in vitro assays were used to assess the possible effects of fludioxonil in human cells and aquatic microorganisms. For the investigation of the toxic effects of fludioxonil on freshwater microalgae, Scenedesmus rubescens and Dunaliella tertiolecta were exposed to various environmentally relevant concentrations of the fungicide for a period of 96 h. Fludioxonil at 50–200 μg L⁻¹ significantly limited the growth of both microalgae, especially in the first 24 h of the exposure, where inhibitions up to 82.34% were calculated. The toxicity of fludioxonil was further evaluated via the Microtox test, and the studied fungicide was found to be less toxic for the bacteria Aliivibrio fischeri. Regarding human cells, the fludioxonil’s toxic and cyto-genotoxic effects were assessed using the Trypan blue exclusion test and the Cytokinesis Block MicroNucleus (CBMN) assay. Cell viability in all fludioxonil-treated concentrations was similar to control values according to the results of the Trypan blue exclusion test. However, the CBMN assay was used and revealed that fludioxonil had genotoxic potential in higher concentrations and exerted cytotoxic activity against human lymphocytes. Specifically, only the highest dose of fludioxonil, i.e., 10 μg mL⁻¹, exerted genotoxic effects against human lymphocytes, whereas treatment with 0.5, 1, and 5 μg mL⁻¹ did not lead to statistically significant induction of micronuclei (MN) frequencies compared with the control culture. However, fludioxonil-mediated cytotoxicity was statistically significant, which was demonstrated by the decreased CBPI (cytokinesis block proliferation index) values in all cases except for the lowest dose, i.e., 0.5 μg mL⁻¹. Full article

This study evaluates DNA damage and multi-element exposure in populations from La Mojana, a region of North Colombia heavily impacted by artisanal and small-scale gold mining (ASGM). DNA damage markers from the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay, including micronucleated binucleated cells (MNBN), nuclear buds (NBUDs) and nucleoplasmic bridges (NPB), were assessed in 71 exposed individuals and 37 unexposed participants. Exposed individuals had significantly higher MNBN frequencies (PR = 1.26, 95% CI: 1.02–1.57, p = 0.039). Principal Component Analysis (PCA) identified the “Soil-Derived Mining-Associated Elements” (PC1), including V, Fe, Al, Co, Ba, Se and Mn, as being strongly associated with high MNBN frequencies in the exposed population (PR = 10.45, 95% CI: 9.75–12.18, p < 0.001). GAMLSS modeling revealed non-linear effects of PC1, with greater increases in MNBN at higher concentrations, especially in exposed individuals. These results highlight the dual role of essential and toxic elements, with low concentrations being potentially protective but higher concentrations increasing genotoxicity. Women consistently exhibited higher MNBN frequencies than men, suggesting sex-specific susceptibilities. This study highlights the compounded risks of chronic metal exposure in mining-impacted regions and underscores the urgent need for targeted interventions to mitigate genotoxic risks in vulnerable populations. Full article

Cistus monspeliensis L. (C. monspeliensis) is used in Italian folk medicine. This study was performed to determine genotoxic and antigenotoxic effects of C. monspeliensis leaf extract against mitomycin C (MMC) using an in vitro cytokinesis-block micronucleus assay (CBMN) in the Chinese Hamster Ovarian K1 (CHO-K1) cell line. The phytochemical composition of C. monspeliensis extract was evaluated using an untargeted metabolomic approach by employing UPLC-PDA-ESI/MS. The automated in vitro CBMN assay was carried out using image analysis systems with a widefield fluorescence microscope and the ImageStreamX imaging flow cytometer. The phytochemical profile of C. monspeliensis extract showed, as the most abundant metabolites, punicalagin, myricetin, gallocathechin, and a labdane-type diterpene. C. monspeliensis, at the tested concentrations of 50, 100, and 200 μg/mL, did not induce significant micronuclei frequency, thus indicating the absence of a genotoxic potential. When testing the C. monspeliensis extract for antigenotoxicity in the presence of MMC, we observed a hormetic concentration-dependent effect, where low concentrations resulted in a significant protective effect against MMC-induced micronuclei frequency, and higher concentrations resulted in no effect. In conclusion, our findings demonstrate that C. monspeliensis extract is not genotoxic and, at low concentration, exhibits an antigenotoxic effect. In relation to this final point, C. monspeliensis may act as a potential chemo-preventive against genotoxic agents. Full article

The increasing use of Cannabis sativa products for medicinal, dietary, and recreational purposes has raised concerns about mycotoxin contamination in cannabis and hemp. Mycotoxins persist in these products’ post-processing, posing health risks via multiple exposure routes. This study investigated cytotoxic and genotoxic interactions between cannabidiol (CBD) and the mycotoxin citrinin (CIT) using human cell models: SH-SY5Y, HepG2, HEK293, and peripheral blood lymphocytes. IC₅₀ values and membrane disruption were initially assessed, followed by an evaluation of genotoxicity in lymphocytes using the Comet Assay and Cytokinesis Blocked Micronucleus Cytome Assay. Obtained findings demonstrate that cell-type sensitivity varied across treatments, with combined CBD and CIT exposure exhibiting distinct interactions. Lactate dehydrogenase (LDH) release remained minimal, suggesting cytotoxicity did not stem from membrane disruption but likely involved intracellular pathways. In lymphocytes, CBD alone produced negligible cyto/genotoxic effects and weak antiproliferative responses, whereas CIT displayed clear toxic impacts. DNA damage indicates that CIT may induce genome instability through indirect mechanisms rather than direct DNA interaction, with evidence of potential aneuploidic effects from the CBMN Cyt Assay. Combined exposure led to a reduction in CIT-induced DNA and cytogenetic damage, suggesting CIT’s potential interference with the beneficial properties of CBD. These results provide a foundation for further toxicological assessments and highlight the necessity of standardized mycotoxin monitoring in cannabis-derived products. Full article

This study evaluates the cytotoxic and genotoxic effects of PM_2.5 collected from an open-cast coal mining area in northern Colombia. Cyclohexane (CH), dichloromethane (DCM), and acetone (ACE) extracts were obtained using Soxhlet extraction to isolate compounds of different polarities. Human lymphocytes were exposed to the extracted compounds, and cytotoxicity and genotoxicity were assessed using the cytokinesis block micronucleus (CBMN) and comet assays, incorporating FPG and ENDO III enzymes to detect oxidative DNA damage. Chemical analysis revealed that the organic fractions consisted mainly of modified hydrocarbons and volatile organic compounds. The CBMN assay showed a significant increase in micronuclei in binucleated (MNBN) and mononucleated (MNMONO) cells and nucleoplasmic bridges (NPB) in exposed lymphocytes. The comet assay revealed substantial oxidative DNA damage, particularly with the ACE extract, which significantly increased oxidized purines and pyrimidines. DCM induced similar effects, while CH showed moderate effects. CREST immunostaining revealed aneugenic activity, particularly in cells exposed to ACE and DCM extracts. These results suggest that polar fractions of PM_2.5, likely containing metals and modified PAHs, contribute to DNA damage and chromosomal instability. The study highlights the need to monitor the composition of PM_2.5 in mining regions to implement stricter environmental policies to reduce exposure and health risks. Full article

This study comprises the phytochemical characterization, the evaluation of the total phenolic content (TPC) and antioxidant activity (AA), and the investigation of the cyto-genotoxic and antigenotoxic potential of hydromethanolic extract derived from Salvia verticillata L. leaves. HPLC–DAD–ESI-MS and HPLC–DAD were used for the characterization of the extract and determination of the major ingredients. Afterwards, the TPC and AA were determined. The cytotoxic and genotoxic effect of the extract on cultured human lymphocytes at concentrations of 10, 25, and 50 μg mL⁻¹ was investigated via the Cytokinesis Block MicroNucleus (CBMN) assay. Moreover, its antigenotoxic potential against the mutagenic agent mitomycin C (MMC) was assessed using the same assay. The hydromethanolic extract comprises numerous metabolites, with rosmarinic acid being the major compound. It had a high value of TPC and exerted significant AA as shown by the results of the Ferric Reducing Antioxidant Power (FRAP) and Radical Scavenging Activity by DPPH• assays. A dose-dependent cytotoxic potential was recorded, with the highest dose (50 μg mL⁻¹) exhibiting statistically significant cytotoxicity. None of the tested concentrations induced significant micronuclei (MN) frequencies, indicating a lack of genotoxicity. All tested concentrations reduced the MMC-mediated genotoxic effects, with the two lowest showing statistically significant antigenotoxic potential. Full article

Major strides have been made in the development of FLASH radiotherapy (FLASH RT) in the last ten years, but there are still many obstacles to overcome for transfer to the clinic to become a reality. Although preclinical and first-in-human clinical evidence suggests that ultra-high dose rates (UHDRs) induce a sparing effect in normal tissue without modifying the therapeutic effect on the tumor, successful clinical translation of FLASH-RT depends on a better understanding of the biological mechanisms underpinning the sparing effect. Suitable in vitro studies are required to fully understand the radiobiological mechanisms associated with UHDRs. From a technical point of view, it is also crucial to develop optimal technologies in terms of beam irradiation parameters for producing FLASH conditions. This review provides an overview of the research progress of FLASH RT and discusses the potential challenges to be faced before its clinical application. We critically summarize the preclinical evidence and in vitro studies on DNA damage following UHDR irradiation. We also highlight the ongoing developments of technologies for delivering FLASH-compliant beams, with a focus on laser-driven plasma accelerators suitable for performing basic radiobiological research on the UHDR effects. Full article

The continuous biomonitoring of a population directly or indirectly exposed to pesticides could be an additional tool for decision makers to improve their health conditions. In this work, we performed biomonitoring on two groups of people from the Mexicali Valley who were continuously exposed to pesticides using the cytokinesis-block micronucleus cytome assay (L-CBMN) to evaluate cytotoxic and genotoxic damage in human peripheral blood lymphocytes. The study groups comprised 14 indigenous Cucapah with non-vegetarian habits (NV group) from Ejido el Mayor (32.12594°, −115.27265°) and 21 lacto-ovo vegetarian (LOV) persons from the Seventh-day Adventist Church of Ejido Vicente Guerrero (32.3961°, −115.14023°). The L-CBMN assay determines the nuclear division index (NDI), apoptosis, necrosis, micronuclei (MNs), nuclear buds (NBUDs), and nucleoplasmic bridges (NPBs). Our results show that, regardless of diet or daily habits, both the studied groups presented with cytogenotoxic damage compared with non-exposed pesticide individuals, without modifications to the nuclear division index. In the rest of the evaluated biomarkers, the NV group exhibited greater cytotoxic and genotoxic damage than the LOV group. Nevertheless, individuals practicing a lacto-ovo vegetarian diet (LOV) showed lower damage than those with non-vegetarian habits (NV), suggesting a better antioxidant response that helps decrease the genotoxic damage due to the enhanced intake of folates and antioxidants from a plant-based diet. Full article

This study investigated the beneficial properties of prickly pear peel (PPP) extracts from Opuntia ficus-indica (L.) Mill. Extracts were obtained via the Soxhlet extraction method using methanol (P1), ethanol (P2) and ethanol-water (P3) as extraction solvents. Their total phenolic and flavonoid content (TPC and TFC, respectively) and their antioxidant activity (AA) were determined. The PPP extracts were characterized in detail using mass spectrometry techniques. Their cyto-genotoxic effect and antigenotoxic potential against mitomycin C were evaluated via the cytokinesis block micronucleus (CBMN) assay on human lymphocytes. Enhanced TPC, TFC and AA values were recorded for all the extracts. Moreover, P1 and P2 were cytotoxic only at the highest concentrations, whereas P3 was found to be cytotoxic in all cases. No significant micronucleus induction was observed in the tested extracts. The PPP extracts contain bioactive compounds such as flavonoids, carboxylic acids, alkaloids, fatty acids and minerals (mainly K, Si, Mg, Ca, P and Zn). The results showed that all three extracts exerted high antigenotoxic activity. Our findings confirm the beneficial and genoprotective properties of PPP extracts and further studies on the bioactive compounds of Opuntia ficus-indica (L.) Mill. are recommended, as it constitutes a promising plant in pharmaceutical applications. Full article

The bioavailability of glucoside flavonoids is influenced by the nature of the sugar, glucosides being absorbed faster than rhamnoglucosides, for example. One strategy to enhance the bioavailability is enzymatic hydrolysis. In this study, some kinetic parameters of hesperidinase-mediated hydrolysis of rutin were evaluated using an UHPLC/QTOF-MS^E analysis of the products of a bioconversion reaction. The resulting hydrolyzed rutins (after 4, 8 and 12 h of reaction) were submitted to anti-proliferative and Cytokinesis-Block Micronucleus (CBMN) assays in CHO-K1 cells. In the hesperidinase-mediated hydrolysis, the final concentration of quercetin-3-O-glucoside (Q3G) was directly proportional to the rutin concentration and inversely proportional to the reaction time. At an anti-proliferative concentration (2.5 μg/mL), hydrolyzed rutin derivatives did not show a mutagenic effect, except for the sample with a higher content of Q3G (after 4 h of the enzymatic hydrolysis of rutin). Moreover, the higher Q3G content in hydrolyzed rutin protected the CHO-K1 cells 92% of the time against methyl methanesulfonate-induced mutagenic damage. These results suggested that the anti-mutagenic effect of hydrolyzed rutin might be related to antioxidant and cell death induction. Presenting a good lipophilicity/hydrophilicity ratio, together with antioxidant and anti-mutagenic activities, the hesperidinase-mediated hydrolyzed rutin seemed to be a promisor raw material for the development of food supplements. Full article

Over the past 20 years, numerous tyrosine kinase inhibitors (TKIs) have been introduced for targeted therapy of various types of malignancies. Due to frequent and increasing use, leading to eventual excretion with body fluids, their residues have been found in hospital and household wastewaters as well as surface water. However, the effects of TKI residues in the environment on aquatic organisms are poorly described. In the present study, we investigated the cytotoxic and genotoxic effects of five selected TKIs, namely erlotinib (ERL), dasatinib (DAS), nilotinib (NIL), regorafenib (REG), and sorafenib (SOR), using the in vitro zebrafish liver cell (ZFL) model. Cytotoxicity was determined using the MTS assay and propidium iodide (PI) live/dead staining by flow cytometry. DAS, SOR, and REG decreased ZFL cell viability dose- and time-dependently, with DAS being the most cytotoxic TKI studied. ERL and NIL did not affect viability at concentrations up to their maximum solubility; however, NIL was the only TKI that significantly decreased the proportion of PI negative cells as determined by the flow cytometry. Cell cycle progression analyses showed that DAS, ERL, REG, and SOR caused the cell cycle arrest of ZFL cells in the G0/G1 phase, with a concomitant decrease of cells in the S-phase fraction. No data could be obtained for NIL due to severe DNA fragmentation. The genotoxic activity of the investigated TKIs was evaluated using comet and cytokinesis block micronucleus (CBMN) assays. The dose-dependent induction of DNA single strand breaks was induced by NIL (≥2 μM), DAS (≥0.006 μM), and REG (≥0.8 μM), with DAS being the most potent. None of the TKIs studied induced micronuclei formation. These results suggest that normal non-target fish liver cells are sensitive to the TKIs studied in a concentration range similar to those previously reported for human cancer cell lines. Although the TKI concentrations that induced adverse effects in exposed ZFL cells are several orders of magnitude higher than those currently expected in the aquatic environment, the observed DNA damage and cell cycle effects suggest that residues of TKIs in the environment may pose a hazard to non-intentionally exposed organisms living in environments contaminated with TKIs. Full article

Turpentine is a fluid used mainly as a solvent for thinning oil-based paints, obtained by distilling the resin of coniferous trees. Fine art painters use turpentine on a daily basis. The aim of this study was to investigate the genotoxic effect of turpentine and to determine the lymphocyte proliferation index in the peripheral blood of individuals occupationally exposed to turpentine. For this purpose, the cytokinesis-block micronucleus assay (CBMN) was used to determine the total number of micronuclei (MNi), nucleoplasmic bridges (NPB), and nuclear buds (NBUD), as well as the cell proliferation index (CBPI) in the peripheral blood lymphocytes of the subjects. Twenty-two subjects exposed to turpentine daily through their work participated in the study and were compared to twenty subjects in the control group. The results showed a significant increase in the number of micronuclei and other genotoxicity parameters, as well as significant cytotoxicity based on CBPI values. In addition, the genotoxic and cytotoxic effects of turpentine were found to be time-dependent, i.e., the deleterious effects of turpentine on genetic material increase with prolonged exposure. These results strongly suggest that exposure to turpentine vapors may affect genome stability and that occupational safety measures should be taken when using turpentine. Full article

Nuclear imaging is a highly sensitive and noninvasive imaging technique that has become essential for medical diagnosis. The use of radiolabeled nanomaterials capable of acting as imaging probes has shown rapid development in recent years as a powerful, highly sensitive, and noninvasive tool. In addition, quantitative single-photon emission computed tomography (SPECT) images performed by incorporating radioisotopes into nanoparticles (NPs) might improve the evaluation and the validation of potential clinical treatments. In this work, we present a direct method for [^99mTc]Tc-radiolabeling of FITC-tagged silk fibroin nanoparticles (SFN). NPs were characterized by means of dynamic light scattering and scanning electron microscopy. In vitro studies were carried out, including the evaluation of stability in biological media and the evaluation of hemocompatibility and genotoxicity using the cytokinesis block micronucleus (CBMN) assay. The radiolabeling method was reproducible and robust with high radiolabeling efficiency (∼95%) and high stability in biological media. Hydrodynamic properties of the radiolabeled NPs remain stable after dual labeling. The interaction of SFN with blood elicits a mild host response, as expected. Furthermore, CBMN assay did not show genotoxicity induced by [^99mTc]Tc-FITC-SFN under the described conditions. In conclusion, a feasible and robust dual-labeling method has been developed whose applicability has been demonstrated in vitro, showing its value for further investigations of silk fibroin NPs biodistribution in vivo. Full article