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Keywords = cytokinesis-block micronucleus cytome assay

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28 pages, 2377 KiB  
Article
Genetic Damage and Multi-Elemental Exposure in Populations in Proximity to Artisanal and Small-Scale Gold (ASGM) Mining Areas in North Colombia
by Pedro Espitia-Pérez, Lyda Espitia-Pérez, Ana Peñata-Taborda, Hugo Brango, Karina Pastor-Sierra, Claudia Galeano-Páez, Gean Arteaga-Arroyo, Alicia Humanez-Alvarez, Ruber Rodríguez Díaz, Javier Salas Osorio, Luís Armando Valderrama and Tatiana Dillenburg Saint’Pierre
Toxics 2025, 13(3), 202; https://doi.org/10.3390/toxics13030202 - 11 Mar 2025
Viewed by 1312
Abstract
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 [...] Read more.
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
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24 pages, 4040 KiB  
Article
Preliminary Insights into the Cyto/Genoprotective Properties of Propolis and Its Constituent Galangin In Vitro
by Mateo Jakac, Irena Brčić Karačonji, Andreja Jurič, Dražen Lušić, Danijel Milinčić, Aleksandra Dramićanin, Mirjana Pešić, Nediljko Landeka and Nevenka Kopjar
Toxics 2025, 13(3), 194; https://doi.org/10.3390/toxics13030194 - 8 Mar 2025
Cited by 1 | Viewed by 804
Abstract
Propolis has been well known for centuries as a natural preventive and therapeutic agent. Its numerous health benefits are mainly attributed to its high content of phenolic compounds that have a remarkable antioxidant activity. Since phenolics may exert a dual nature (pro-oxidant and [...] Read more.
Propolis has been well known for centuries as a natural preventive and therapeutic agent. Its numerous health benefits are mainly attributed to its high content of phenolic compounds that have a remarkable antioxidant activity. Since phenolics may exert a dual nature (pro-oxidant and antioxidant) the aim of this study was to investigate the safety profile of the ethanolic extract of propolis and the related flavonoid galangin and their ability to protect lymphocytes from irinotecan-induced cyto/genotoxicity in vitro. Isolated human peripheral blood lymphocytes were exposed for 3 h to three concentrations of propolis extract and galangin corresponding to the average daily dose of 0.25 mL of extract [propolis in 70% ethanol (3:7, w/w)], as well as a five- and ten-fold higher concentration. Cyto- and genoprotective effects were tested using a cytokinesis-block micronucleus cytome assay. Treatment with propolis and galangin in the selected concentrations exerted high biocompatibility with lymphocytes and diminished the level of cytogenetic damage caused by irinotecan. Propolis at the same concentration offered a stronger protective effect than single galangin. Also, apoptosis was the prevailing mechanism of cell death in our experimental conditions. These preliminary results speak in favour of future investigations of propolis using other available cytogenetic methods and cell models. Full article
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21 pages, 4145 KiB  
Article
Assessing the Potential Synergistic/Antagonistic Effects of Citrinin and Cannabidiol on SH-SY5Y, HepG2, HEK293 Cell Lines, and Human Lymphocytes
by Dubravka Rašić, Antonio Zandona, Maja Katalinić, Martin Češi and Nevenka Kopjar
Toxins 2024, 16(12), 534; https://doi.org/10.3390/toxins16120534 - 11 Dec 2024
Cited by 1 | Viewed by 1393
Abstract
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 [...] Read more.
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 article belongs to the Special Issue Occurrence, Toxicity, Metabolism, Analysis and Control of Mycotoxins)
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14 pages, 3082 KiB  
Article
Genomic Instability and Cytotoxicity Evaluation of Two Communities Exposed to Pesticides in the Mexicali Valley by the L-CBMN Assay
by Balam Ruiz-Ruiz, Olivia Torres-Bugarin, Erika Zúñiga-Violante, Francisco Casillas-Figueroa, Roberto Luna-Vázquez-Gómez, Verónica Campos Gallegos, Ana Erika Ruiz-Arellano and María Evarista Arellano-García
Toxics 2023, 11(10), 807; https://doi.org/10.3390/toxics11100807 - 25 Sep 2023
Cited by 2 | Viewed by 2566
Abstract
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 [...] Read more.
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 article belongs to the Section Agrochemicals and Food Toxicology)
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12 pages, 1351 KiB  
Article
The Relationship between Telomere Length and Nucleoplasmic Bridges and Severity of Disease in Prostate Cancer Patients
by Varinderpal S. Dhillon, Permal Deo and Michael Fenech
Cancers 2023, 15(13), 3351; https://doi.org/10.3390/cancers15133351 - 26 Jun 2023
Cited by 4 | Viewed by 1740
Abstract
Telomeres are repetitive nucleotide (TTAGGG) sequences that stabilize the chromosome ends and play an important role in the prevention of cancer initiation and progression. Nucleoplasmic bridges (NPBs) are formed when chromatids remain joined together during mitotic anaphase either due to mis-repair of DNA [...] Read more.
Telomeres are repetitive nucleotide (TTAGGG) sequences that stabilize the chromosome ends and play an important role in the prevention of cancer initiation and progression. Nucleoplasmic bridges (NPBs) are formed when chromatids remain joined together during mitotic anaphase either due to mis-repair of DNA breaks or due to chromatid end fusion as a result of telomere loss or telomere dysfunction. We tested the hypotheses that (i) telomere length (TL) is shorter in prostate cancer (PC) patients relative to healthy age-matched individuals, (ii) TL differs in different stages of PC and (iii) shorter TL is significantly correlated with NPBs formation in PC cases. TL was measured in whole blood by well-established quantitative PCR method and the frequency of NPBs was measured in lymphocytes using cytokinesis-block micronucleus cytome (CBMNcyt) assay. Our results indicate that TL is shorter and NPBs are increased in PC patients relative to age-matched healthy controls. Furthermore, TL was significantly shorter (p = 0.03) in patients with a Gleason score more than 7 and there was also a significant trend of decreasing TL across all three stages (p trend = 0.01; Gleason score <7, 7 and >7). Furthermore, TL was significantly inversely correlated with NPB frequency in PC patients (r = −0.316; p = 0.001) but not in controls (r = 0.163; p = 0.06) and their relationships became stronger with higher Gleason scores. More studies are required that can confirm our observations and explore mechanistic differences in the role of telomeres in NPB formation in PC cases relative to non-cancer cases. Full article
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19 pages, 2944 KiB  
Article
Protective Effects of Arbutus unedo L. Honey in the Alleviation of Irinotecan-Induced Cytogenetic Damage in Human Lymphocytes—An In Vitro Study
by Andreja Jurič, Irena Brčić Karačonji, Uroš Gašić, Dušanka Milojković Opsenica, Saša Prđun, Dragan Bubalo, Dražen Lušić, Nada Vahčić and Nevenka Kopjar
Int. J. Mol. Sci. 2023, 24(3), 1903; https://doi.org/10.3390/ijms24031903 - 18 Jan 2023
Cited by 6 | Viewed by 2921
Abstract
Strawberry tree (Arbutus unedo L.) honey (STH) has been used since ancient times as a folk medicine remedy, especially in certain Mediterranean countries. This honey, rich in phenolic content, is well recognized for its antioxidant, anti-inflammatory, and antimicrobial activities, and is used [...] Read more.
Strawberry tree (Arbutus unedo L.) honey (STH) has been used since ancient times as a folk medicine remedy, especially in certain Mediterranean countries. This honey, rich in phenolic content, is well recognized for its antioxidant, anti-inflammatory, and antimicrobial activities, and is used for the treatment of skin lesions as well as gastrointestinal and respiratory disorders. This study investigated whether STH alleviates genome damage in human peripheral blood lymphocytes produced by the cytotoxic drug irinotecan. The phenolic profile of STH was previously estimated by ultra-high-performance liquid chromatography coupled to a linear ion trap–Orbitrap hybrid mass spectrometer. The effects of STH were evaluated at three concentrations (1×, 5×, and 10×), based on the daily consumption of the honey by an adult person. After 2 h of in vitro exposure, standard lymphocyte cultures for the analysis of chromosome aberrations and the cytokinesis-block micronucleus cytome assay were established. Our results demonstrate that STH offered remarkable geno- and cytoprotection when administered with irinotecan. These findings are relevant for drawing preliminary conclusions regarding the in vitro safety of the tested honey. However, further studies are needed with the application of more complex experimental models. Full article
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17 pages, 419 KiB  
Article
Assessment of Cytogenetic Damage and Cholinesterases’ Activity in Workers Occupationally Exposed to Pesticides in Zamora-Jacona, Michoacan, Mexico
by Rafael Valencia-Quintana, Rosa María López-Durán, Mirta Milić, Stefano Bonassi, Ma. Antonieta Ochoa-Ocaña, Mayrut Osdely Uriostegui-Acosta, Guillermo Alejandro Pérez-Flores, José Luis Gómez-Olivares and Juana Sánchez-Alarcón
Int. J. Environ. Res. Public Health 2021, 18(12), 6269; https://doi.org/10.3390/ijerph18126269 - 10 Jun 2021
Cited by 18 | Viewed by 3690
Abstract
Pesticides have been considered as potential chemical mutagens; however, little is known about toxic and genotoxic effects during pesticide application in Zamora-Jacona, Michoacan State in Mexico. This study sought to determine DNA damage and cholinesterase activities inhibitions in 54 agricultural workers exposed to [...] Read more.
Pesticides have been considered as potential chemical mutagens; however, little is known about toxic and genotoxic effects during pesticide application in Zamora-Jacona, Michoacan State in Mexico. This study sought to determine DNA damage and cholinesterase activities inhibitions in 54 agricultural workers exposed to complex mixtures of pesticides vs. control group (26 individuals) using Comet assay in peripheral whole blood, micronucleus (MN) test in oral mucosa cells, Cytokinesis-blocked MN assay in lymphocytes (L-CBMNcyt) and measuring AChE and BChE activities in whole blood and plasma samples, respectively. Exposed subjects demonstrated significantly elevated levels of primary (Comet assay: tail intensity, tail length, tail moment, Olive tail moment) and permanent DNA damage (MN assay: in blood/buccal cells; frequencies of nuclear buds, binucleated cells, cells with condensed chromatin, karyorrhexis, pyknosis, and karyolysis). However, inhibition of cholinesterase activities (AChE and BChE) was not observed in the workers. Confounding factors including sex, age, BMI, working exposure period, protection level, smoking habit (cigarettes per day units), alcohol consumption (weekly), medication, were considered in the analysis. These combined techniques demonstrated usefulness in the health hazards risks pesticide exposure assessment and suggested the need for periodic monitoring together with the education and the training of occupational workers for the safe application of potentially harmful pesticides. Full article
(This article belongs to the Special Issue Prevention of Occupational Risks)
15 pages, 1608 KiB  
Article
Cytotoxic and Genotoxic Effects of Cyanobacterial and Algal Extracts—Microcystin and Retinoic Acid Content
by Michal Bittner, Alja Štern, Marie Smutná, Klára Hilscherová and Bojana Žegura
Toxins 2021, 13(2), 107; https://doi.org/10.3390/toxins13020107 - 2 Feb 2021
Cited by 23 | Viewed by 4487
Abstract
In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures [...] Read more.
In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04–2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2–2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs. Full article
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13 pages, 2118 KiB  
Review
Cytokinesis-Block Micronucleus Cytome Assay Evolution into a More Comprehensive Method to Measure Chromosomal Instability
by Michael Fenech
Genes 2020, 11(10), 1203; https://doi.org/10.3390/genes11101203 - 15 Oct 2020
Cited by 105 | Viewed by 10362
Abstract
This review describes the cytokinesis-block micronucleus (CBMN) cytome assay and its evolution into a molecular cytogenetic method of chromosomal instability (CIN). Micronuclei (MNi) originate from whole chromosomes or chromosome fragments that fail to segregate to the poles of the cell during mitosis. These [...] Read more.
This review describes the cytokinesis-block micronucleus (CBMN) cytome assay and its evolution into a molecular cytogenetic method of chromosomal instability (CIN). Micronuclei (MNi) originate from whole chromosomes or chromosome fragments that fail to segregate to the poles of the cell during mitosis. These lagging chromosomes are excluded from the daughter nuclei and are enveloped in their own membrane to form MNi. The CBMN assay was developed to allow MNi to be scored exclusively in once-divided binucleated cells, which enables accurate measurement of chromosome breakage or loss without confounding by non-dividing cells that cannot express MNi. The CBMN assay can be applied to cell lines in vitro and cells such as lymphocytes that can be stimulated to divide ex vivo. In the CBMN assay, other CIN biomarkers such as nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) are also measured. Use of centromere, telomere, and chromosome painting probes provides further insights into the mechanisms through which MNi, NPBs and NBUDs originate. Measurement of MNi is also important because entrapment within a micronucleus may cause chromosomes to shatter and, after nuclear reintegration, become rearranged. Additionally, leakage of DNA from MNi can stimulate inflammation via the cyclic GMP-AMP Synthase—Stimulator of Interferon Genes (cGAS-STING) DNA sensing mechanism of the innate immune system. Full article
(This article belongs to the Special Issue Triggering Forces of Chromosome Instability)
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