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Journal articles on the topic 'Micronuclei assay'
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Lepage, Chloe C., Laura L. Thompson, Bradley Larson, and Kirk J. McManus. "An Automated, Single Cell Quantitative Imaging Microscopy Approach to Assess Micronucleus Formation, Genotoxicity and Chromosome Instability." Cells 9, no. 2 (2020): 344. http://dx.doi.org/10.3390/cells9020344.
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Micronuclei are small, extranuclear bodies that are distinct from the primary cell nucleus. Micronucleus formation is an aberrant event that suggests a history of genotoxic stress or chromosome mis-segregation events. Accordingly, assays evaluating micronucleus formation serve as useful tools within the fields of toxicology and oncology. Here, we describe a novel micronucleus formation assay that utilizes a high-throughput imaging platform and automated image analysis software for accurate detection and rapid quantification of micronuclei at the single cell level. We show that our image analysis parameters are capable of identifying dose-dependent increases in micronucleus formation within three distinct cell lines following treatment with two established genotoxic agents, etoposide or bleomycin. We further show that this assay detects micronuclei induced through silencing of the established chromosome instability gene, SMC1A. Thus, the micronucleus formation assay described here is a versatile and efficient alternative to more laborious cytological approaches, and greatly increases throughput, which will be particularly beneficial for large-scale chemical or genetic screens.
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Wojcik, A., M. Kowalska, E. Bouzyk, et al. "Validation of the micronucleus-centromere assay for biological dosimetry." Genetics and Molecular Biology 23, no. 4 (2000): 1083–85. http://dx.doi.org/10.1590/s1415-47572000000400055.
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The micronucleus assay is frequently used for purposes of biological dosimetry. Due to high interindividual variability in the spontaneous frequency of micronuclei, its sensitivity in the low dose region is poor. It has been suggested that this problem can be mitigated by selectively analyzing the frequency of those micronuclei which contain only acentric fragments. Using a pan-centromeric FISH probe we have studied the dose dependence of micronuclei with centromeres in peripheral lymphocytes of human donors. In contrast to previous publications, our approach is based on determining the relative frequency of micronuclei with and without centromeric signals. Our results confirm previous observations that in the low dose range of ionizing radiation, the micronucleus-centromere assay is more sensitive than the conventional micronucleus test.
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Udroiu, Ion. "A Double Fluorescent Staining Method Increases the Sensitivity of the Cytokinesis-Block Micronucleus Assay." Methods and Protocols 8, no. 1 (2025): 3. https://doi.org/10.3390/mps8010003.
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The micronucleus test is one of the most popular genotoxicity assays. In order to avoid underestimation of micronuclei frequencies by counting non-replicating cells, the cytokinesis-blocked micronucleus test has been developed. In this technique, only binucleated cells are scored. One underestimated problem is the potential difficulty in discriminating binucleated from mononucleated cells when using DAPI staining, i.e., the possibility that two neighboring mononucleated cells could be mistaken for a binucleated one. The new protocol presented here comprises the addition of acridine orange in order to stain the cytoplasm (in addition to DAPI to stain nuclei and micronuclei). This new technique can increase the sensitivity of the cytokinesis-blocked micronucleus test and avoid underestimation of micronuclei frequencies, an important issue when high doses are employed.
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Guogytė, Kamilė, Aista Plieskienė, Olga Sevriukova, Rima Ladygienė, Julius Žiliukas, and Vinsas Janušonis. "Micronuclei And G2 Assays For Assessment Of Chromosomal Radiosensitivity As Assistant Tool In Radiotherapy: Method-Comparison Study." Sveikatos mokslai 26, no. 5 (2016): 63–68. http://dx.doi.org/10.5200/sm-hs.2016.073.
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Radiation therapy is widely used for cancer treatment. Medical application of ionizing radiation can cause different responses in human depending on individual radiosensitivity. Therefore, assessment of individual radiosensitivity could be proposed as assistant tool in optimizing radiotherapy. The cytokinesis- block micronucleus and G2 chromosomal radiosensitivity assays were proposed as appropriate methods for assessment of individual radiosensitivity. In current study we carried out a pilot cytokinesis-block micronucleus and G2 chromosomal radiosensitivity assays comparison by evaluating specificity of chromatid breaks yield and micronuclei frequency in peripheral blood lymphocytes as biomarkers of individual radiosensitivity in three cancer patients treated with radiotherapy. Our study revealed positive correlation between higher increase in frequency of micronuclei and chromatid breaks after in vitro irradiation in radiotherapy patients peripheral blood lymphocytes with occurrence of adverse radiation effects in tissue which are not being targeted. G2 assay appeared to be more sensitive than micronuclei assay for assessment of irradiation-induced alterations in individual radiosensitivity during the radiotherapy that could affect development of treatment side effects. Therefore, further investigations involving more radiotherapy patients as well as healthy donors are required to select the most sensitive method and reveal the possible correlation between individual radiosensitivity and adverse effect of radiotherapy.
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Rincón, Guido, and Claudia Sánchez. "Short Assay Design for Micronucleus Detection in Human Lymphocytes." BioMed Research International 2021 (September 11, 2021): 1–6. http://dx.doi.org/10.1155/2021/2322257.
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There has been a constant need to develop new and faster cytogenetic assays to measure the instability induced by genotoxic agents in the field of cytogenetic research, an example of which is the micronucleus assay. Micronuclei are fragments or complete chromosomes that remain in the cytoplasm during mitosis. With their high sensitivity and specificity detection, their presence can indicate environmental and occupational genotoxic effects. However, the prolonged periods of cell incubation this assay necessitates are costly and extensive. Hence, it is essential to develop an improved assay that can achieve standardization by being reproducible in practice. The standard protocol for the detection of micronuclei in lymphocytes uses a total assay time of 72 hours. Theoretically, it is possible to reduce the incubation period, and consequently, the total assay time, considering a lymphocyte, completes its mitosis in 24 hours. This study, after careful review of literature, proposes an experimental design to reduce the incubation period and demonstrates its usefulness in practice through the design of a collaborative trial.
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CRISPIM, BRUNO A., JULIANA C. V. SPÓSITO, ROSILDA M. MUSSURY, LEONARDO O. SENO, and ALEXÉIA B. GRISOLIA. "Effects of atmospheric pollutants on somatic and germ cells of Tradescantia pallida (Rose) D.R. HUNT cv. purpurea." Anais da Academia Brasileira de Ciências 86, no. 4 (2014): 1899–906. http://dx.doi.org/10.1590/0001-3765201420140338.
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Anatomical alterations in leaves and DNA damage in cells caused by the accumulation of atmospheric pollutants can be measured by epidermal leaf analyses and Tradescantia micronuclei assay with early pollen tetrad cells. The present study examined the feasibility of using somatic and germ cells of Tradescantia pallida for biomonitoring purposes in the city of Dourados, state of Mato Grosso do Sul (MS), Brazil. Stomatal, micronucleus and epidermal leaf analyses were performed, using standard methodologies, on plants growing at three locations during six different time periods. Tradescantia micronuclei data were analyzed using SAS 9.2 software package and stomatal data were analyzed using SANEST software. Analyses of stomatal characteristics and micronuclei examination in T. pallida were found to be an efficient tool for monitoring atmospheric pollution. The micronucleus assay suggested that the number of micronuclei in early pollen tetrad cells was related to the intensity of vehicular traffic. Increased number of epidermal cells and stomata and increased stomatal density observed at locations with greater vehicular traffic are likely physiological responses of those plants to the increased gas exchange in highly polluted environments.
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Michalová, V., M. Galdíková, B. Holečková, S. Koleničová, and V. Schwarzbacherová. "Micronucleus Assay in Environmental Biomonitoring." Folia Veterinaria 64, no. 2 (2020): 20–28. http://dx.doi.org/10.2478/fv-2020-0013.
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AbstractNowadays many chemicals are widely used in agriculture to ensure high crop yields or in veterinary/human medicine to cure diseases. After their improper usage they may contaminate the environment, persist in it and adversely affect both the target and/or the non-target organisms. One of the ways to detect the occurrence of chemicals in the environment is to assess their impact on aquatic and farm animals; both are directly or indirectly exposed via their feed and water. The micronucleus assay is a standardly used cytogenetic test for the simultaneous detection of clastogenic and aneugenic agents. Additionally, cytotoxic effects are also assessed by analysing the proliferation changes using the cytokinesis-blocked proliferation index. The occurrence of micronuclei is analysed in many types of cells like the peripheral blood cells, bone marrow or cell lines according to standards for micronuclei detection. The analysis of published results has shown that the micronucleus assay is, together with the chromosomal aberration test, one of the most often used test in genotoxicity assessment. Its results have contributed to reassessing the use of multiple chemicals available on the market. Moreover, it is a compulsory test before approving the chemical/ pesticide for the market.
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Nefić, Hilada, Jasmin Mušanović, Kemajl Kurteshi, Enida Prutina, and Elvira Turcalo. "The effects of sex, age and cigarette smoking on micronucleus and degenerative nuclear alteration frequencies in human buccal cells of healthy Bosnian subjects." Journal of Health Sciences 3, no. 3 (2013): 196–204. http://dx.doi.org/10.17532/jhsci.2013.107.
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Introduction: This study was performed to establish a baseline value of micronucleus frequency in buccal cells and to estimate the impact of the most common factors (sex and age, and smoking) on micronucleus and degenerative nuclear alteration frequencies in the sample of healthy Bosnian subjects.Methods: The Buccal Micronucleus Cytome (BMCyt) assay, based on scoring not only micronucleus frequency but also other genome damage markers, dead or degenerated cells, provides a measure of cytotoxic and genotoxic effects.Results: Our results showed the baseline buccal micronucleus frequency was 0.135% or 1.35‰, as well as positive correlations between micronucleus frequencies and formations of degenerative nuclear alterations (nuclear buds, karyolytic and karyorrhectic cells). The number of micronuclei in buccal cells was significantly higher in females than in males. There was positive association between the age and frequency of analysed cytogenetic biomarkers. Buccal cell micronuclei and degenerative nuclear alternations were more frequent among cigarette smokers than non-smokers and significantly higher in female smokers than in male smokers. Cytogenetic damages showed significantly positive correlation between intensity of smoking and the number of nuclear alterations. The years of smoking had a significant influence not only on the number of nuclear alterations but also in micronuclei and nuclear buds in buccal cells.Conclusions: The sex influences the number of micronuclei in human buccal cells. The ageing increased the number of micronuclei and other biomarkers of DNA damage. The cigarette smoking significantly increases the frequencies of micronuclei and nuclear buds, pyknotic, karyolytic and karyorrhectic cells.
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Anand, Prem Rajan, and Shamundeeswari.Anandan. "INVESTIGATION OF CARCINOGENIC AND MUTAGENIC PROPERTY OF FOOD COLOR USING CAT FISHCLARIAS BATRACHUSBY USING ALKALINE SINGLE-CELL GEL ELECTROPHORESIS (COMET) ASSAY AND MICRONUCLEUS ASSAY." International Journal of Medical Research and Pharmaceutical Sciences 4, no. 7 (2017): 29–34. https://doi.org/10.5281/zenodo.831536.
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In recent years, humans are exposed to various carcinogenic agents, artificial colors like Green #3, Red#3, Citrus red #2, Saccharin, Etc.The present study emphasis is made to know the effect of these common agents in particular with reference to artificial food color Allura red (FD & C # 40)- orange red food dye on DNA damage using Cat FishClarias batrachus as an model organism. The methodology will be to understand direct damage to nucleus by Micronuclei test and Chromosomal damage by alkaline single-cell gel electrophoresis (COMET) ASSAY. The freshwater fish catfish Clarias batrachus was used for specificity genotoxic indicators micronucleus assay and COMET assays. The fish was exposed to 1g/L of Allura red (FD & C # 40) for a period of 7, 14, 28, and 35 days ectodermally. The blood sample was tested for genotoxicity. The results revealed DNA damage through alkaline single-cell gel electrophoresis (comet) and micronuclei assays. Hence it is concluded that the usage of food color containing Allurared(FD & C # 40) –orange red food dye may be toxic at genetic level if the usage is prolonged
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KUCHTA-GŁADYSZ, MARTA, ANNA GRZESIAKOWSKA, AGNIESZKA OTWINOWSKA-MINDUR, PRZEMYSŁAW BARAN, and OLGA SZELESZCZUK. "Development of the optimal dose of cytochalasine B in the MN assay in a domestic cat." Medycyna Weterynaryjna 78, no. 12 (2022): 6708–2022. http://dx.doi.org/10.21521/mw.6708.
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Cytochalasine B is an indicator in the CBMN assay, but can also be geno- or cytotoxic. The aim of the study was to determine the optimal dose of cytochalasine B for the Cytokinesis-Block Micronucleus (CBMN) assay for the domestic cat. The results revealed a surprisingly high number of micronuclei (MN) in relation to the doses used. It was found that a concentration of 2.5 μg/ml is enough to obtain binuclear cells, with a minimal toxic effect on the lymphocytes of the domestic cat. In the material analysed, three forms of chromatin structure abnormalities were found. The most prevalent abnormalities were isolated micronuclei, with the mean number of binucleated cells with one micronucleus (BNCs + 1 MN) equal to 39.96 ± 9.02. Moreover, the numbers of binucleated cells with one micronucleus, nuclear buds and nucleoplasmic bridges did not differ significantly according to the sex or age of European cats.
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Balasem, Abbas N. "Micronucleus Assay as a Biological Indicator for In Vitro Exposure of Human Lymphocytes to Gamma Rays." Journal of Biotechnology Research Center 6, no. 1 (2012): 51–55. http://dx.doi.org/10.24126/jobrc.2012.6.1.200.
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Micronucleus Assay was employed to detect the effects of acute exposure of human peripheral blood lymphocytes in vitro to Cs -137 gamma rays. Human whole blood samples were irradiated with different doses of gamma rays namely ) 0.1, 0.2, 0.3, 0.4, 0.5, 1.00) Gy, respectively in addition to a control non-irradiated sample. The samples were tissue cultured and cytokinesis blocked method was used to investigate the frequency of micronuclei. In vitro exposure of lymphocytes to this doses led to elevation of micronuclei in comparison with non –irradiated samples However, inclusion of mono-, tri-,and quadrinucleated cells in micronucleus assay probably gives more satisfying result than restriction the test on binucleated cells. Computed programmed were employed to establish dose – response relationships to be used as biological dosimeter during radiation accidents.
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Depuydt, Julie, Ans Baeyens, Stephen Barnard, et al. "RENEB intercomparison exercises analyzing micronuclei (Cytokinesis-block Micronucleus Assay)." International Journal of Radiation Biology 93, no. 1 (2016): 36–47. http://dx.doi.org/10.1080/09553002.2016.1206231.
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Yang Jee Kim, Jun Yeol Choi, Yoon Hee Cho, Hae Dong Woo, and Hai Won Chung. "Micronucleus-centromere assay in workers occupationally exposed to low level of benzene." Human & Experimental Toxicology 29, no. 5 (2010): 343–50. http://dx.doi.org/10.1177/0960327110361500.
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Benzene is a well-known carcinogen that induces chromosomal instability, including chromosome aberration and aneuploidy. In order to assess aneugenic effect of low-level benzene, micronucleus-centromere assay using specific probes for chromosomes 7 and 9 was performed in workers occupationally exposed to low-dose benzene at a petroleum refinery. A micronucleus-centromere assay using a pan-centromeric probe was also performed to determine the origin of benzene-induced micronucleus (MN). Frequency of aneuploidy of chromosomes 7 and 9 was significantly higher among workers compared to the unexposed control group. Poisson regression analysis revealed that aneuploidy frequency of chromosome 7 or 9 was significantly associated with benzene level after adjusting for confounding variables such as age, smoking, alcohol intake, and duration of work (p = .042). Additionally, frequencies of MN and centromere-negative micronuclei (MNC—) were significantly higher in benzene-exposed workers compared to controls, while frequency of centromere-positive micronuclei (MNC+) was similar in both groups. In conclusion, aneuploidy of chromosomes 7 and 9 could be a useful biomarker to assess the effect of low-level benzene exposure, and benzene-induced MN originates from chromosome breaks rather than chromosome non-disjunction.
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Ibrulj, Slavka, Sanin Haverić, Anja Haverić, Adaleta Durmić-Pašić, and Damir Marjanović. "Effect of War and Postwar Genotoxins on Micronuclei Frequency in Sarajevo Study Group." Bosnian Journal of Basic Medical Sciences 6, no. 4 (2006): 54–57. http://dx.doi.org/10.17305/bjbms.2006.3121.
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During the 1992-1995 siege, as well as after the war activities, citizens of Sarajevo were most probably exposed to various potential genotoxic agents. The effects of those potential genotoxins were evaluated by micronucleus-cytokinesis blocked assay. The study included 30 individuals who resided in the area of Sarajevo during the war and the postwar period. Point bi-serial coefficient analysis did not reveal any relationship between the frequencies of binuclear cells with micronuclei as well as total number of micronuclei and smoking habits or gender. Simple linear regression revealed statistically significant positive correlation between the age and micronuclei formation. Due to the war related environmental contamination more extensive study is recommended.
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Minhas, Rashid, Huma Naz, Sajid Abdullah, Khalid Abbas, Tanveer Ahmed, and Nimra Zahid. "Evaluation of Genotoxicity induced by Cobalt to Freshwater Fish, Cirrhina mrigala using Micronuclei Assay." Journal of Zoo Biology 5, no. 1 (2022): 19–25. http://dx.doi.org/10.33687/zoobiol.005.01.4511.
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Due to industrial advancement, a variety of toxic chemicals including metals are released into the aquatic environment which not only disturbs the physico-chemical properties of the water bodies but also influences the aquatic food chain to cause physiological and cytogenetic alternations in the aquatic animals. Metals have the ability to produce reactive oxygen species (ROS) that would cause the oxidative of nucleic acid. Micronucleus test has been commonly used for the estimation of biological impacts of water pollutants on genotoxic damage in fish. Therefore, the present research work was designed to check the genotoxic potential of cobalt for fish Cirrhinus mrigala by using a micronuclei assay. Fish were exposed to the various sub-lethal concentrations of cobalt metal such as 2/3rd, 1/3rd, 1/4th, and 1/5th of LC50 concentration for one month and sampling was done after 10 days intervals. Blood sample from the caudal vein of fish was collected to see the micronuclei and binucleated nuclei. Results showed that all test concentrations induced micronuclei and binucleated nuclei in peripheral erythrocytes of C. mrigala. Maximum nuclear abnormalities in peripheral erythrocytes of C. mrigala were observed in 2/3rd concentration followed by the orders: 1/3rd 1/4th 1/5th.Due to industrial advancement, a variety of toxic chemicals including metals are released into the aquatic environment which not only disturbs the physico-chemical properties of the water bodies but also influences the aquatic food chain to cause physiological and cytogenetic alternations in the aquatic animals. Metals have the ability to produce reactive oxygen species (ROS) that would cause the oxidative of nucleic acid. Micronucleus test has been commonly used for the estimation of biological impacts of water pollutants on genotoxic damage in fish. Therefore, the present research work was designed to check the genotoxic potential of cobalt for fish Cirrhinus mrigala by using a micronuclei assay. Fish were exposed to the various sub-lethal concentrations of cobalt metal such as 2/3rd, 1/3rd, 1/4th, and 1/5th of LC50 concentration for one month and sampling was done after 10 days intervals. Blood sample from the caudal vein of fish was collected to see the micronuclei and binucleated nuclei. Results showed that all test concentrations induced micronuclei and binucleated nuclei in peripheral erythrocytes of C. mrigala. Maximum nuclear abnormalities in peripheral erythrocytes of C. mrigala were observed in 2/3rd concentration followed by the orders: 1/3rd 1/4th 1/5th.
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Matthopoulos, Demetrios P. "Dynamic Analysis of DNA Damage by Flow Cytometry and FISH." Scientific World JOURNAL 6 (2006): 563–70. http://dx.doi.org/10.1100/tsw.2006.112.
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The micronucleus assay, developed to assess DNA damage induced by noxious agents, supplies information on whether the damage is due to clastogenic or aneugenic action. Although it is the test that can be used to assess agents' toxicity, it cannot provide information on the molecular events that result in the induction of micronuclei. To study the molecular events, the combination of both microscopic and analytical techniques is required. Flow-sorting induced micronuclei, based on their DNA content, in combination with chromosomal FISH and other molecular techniques, may provide information on these events.
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Velickova, Nevenka, Misko Milev, Tatjana Ruskovska, Biljana Petrova, Bojana Nedeljkovik, and Pale Gorgieva. "Cytogenetic abnormalities in lymphocytes evaluated with micronucleus assay in medical personnel occupationally exposed to ionizing radiation." Genetika 47, no. 3 (2015): 927–39. http://dx.doi.org/10.2298/gensr1503927v.
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The aim of this study was to evaluate the genotoxicity of ionizing radiation on medical personnel using the micronucleus assay and to determine the human health risk. Paired Student?s t-test shows significant statistical difference between the total number of binucleated (BN) cells with micronuclei within the two groups (exposed and control) (t=6,812; p<0,05). The mean of MN frequencies in the exposed group increased in comparison with the mean of MN frequencies in the control group. The formation of small and large micronuclei indicates that medical personnel who are exposed on radiation in their work place, have a chromosomal instability and a risk of cancer.
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Nito, Shinji, Fumio Ariyuki, and Azusa Okaniwa. "Spontaneous expulsion of micronuclei by enucleation in the micronucleus assay." Mutation Research Letters 207, no. 3-4 (1988): 185–92. http://dx.doi.org/10.1016/0165-7992(88)90085-1.
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Khondkaryan, Lusine, Ani Tevosyan, Hayk Navasardyan, et al. "Datasets Construction and Development of QSAR Models for Predicting Micronucleus In Vitro and In Vivo Assay Outcomes." Toxics 11, no. 9 (2023): 785. http://dx.doi.org/10.3390/toxics11090785.
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In silico (quantitative) structure–activity relationship modeling is an approach that provides a fast and cost-effective alternative to assess the genotoxic potential of chemicals. However, one of the limiting factors for model development is the availability of consolidated experimental datasets. In the present study, we collected experimental data on micronuclei in vitro and in vivo, utilizing databases and conducting a PubMed search, aided by text mining using the BioBERT large language model. Chemotype enrichment analysis on the updated datasets was performed to identify enriched substructures. Additionally, chemotypes common for both endpoints were found. Five machine learning models in combination with molecular descriptors, twelve fingerprints and two data balancing techniques were applied to construct individual models. The best-performing individual models were selected for the ensemble construction. The curated final dataset consists of 981 chemicals for micronuclei in vitro and 1309 for mouse micronuclei in vivo, respectively. Out of 18 chemotypes enriched in micronuclei in vitro, only 7 were found to be relevant for in vivo prediction. The ensemble model exhibited high accuracy and sensitivity when applied to an external test set of in vitro data. A good balanced predictive performance was also achieved for the micronucleus in vivo endpoint.
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Petrovic, Sandra, Andreja Leskovac, and Gordana Joksic. "Positive correlation between micronuclei and necrosis of lymphocytes in medical personnel occupationally exposed to ionizing radiation." Archive of Oncology 13, no. 2 (2005): 65–68. http://dx.doi.org/10.2298/aoo0502065p.
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BACKGROUND: Current radiation protection standards are based on premise that any radiation dose may result in detrimental health effects. The aim of this study was to evaluate extent of the DNA damages (measured by induction of micronuclei) and interphase cell death in circulating lymphocytes of medical personnel exposed to ionizing radiation. METHODS: Baseline micronuclei were assessed using the cytokinesis-block micronucleus test. Cytotoxicity was analyzed by flow cytometry for human white blood cells to identify cells that displayed apoptosis-associated DNA condensation. Necrotic cells were analyzed simultaneously. All parameters were compared with corresponding controls. RESULTS: A statistically significant difference (t = 4.54, p = 0.002) was found between exposed and control group in the yield of baseline micronuclei. The level of baseline micronuclei correlated positively with necrosis of leucocytes (r=0.09, p=0.68 in exposed group, r=0.02, p=0.97 in controls). An inverse correlation between baseline micronuclei and apoptosis was noted in both groups of examinees (r = -0.26, p = 0.27 in exposed group, r = -0.09, p=0.80 in controls). The data obtained also suggested an inverse correlation between necrosis and apoptosis (r = -0.37, p = 0.11 in exposed group, r = -0.89, p = 0.001 in controls). CONCLUSION: Flow cytometry being a rapid, fast, and accurate method is strongly recommended in evaluation of radiation injuries. The integration of apoptosis and necrosis into micronucleus assay could be very important in the assessment of cumulative effects of ionizing radiation in occupationally exposed medical personnel.
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Güez, Camila Martins, Emily Pansera Waczuk, Karina Braccini Pereira, Marcus Vinícius Morini Querol, João Batista Teixeira da Rocha, and Luís Flávio Souza de Oliveira. "In vivo and in vitro genotoxicity studies of aqueous extract of Xanthium spinosum." Brazilian Journal of Pharmaceutical Sciences 48, no. 3 (2012): 461–67. http://dx.doi.org/10.1590/s1984-82502012000300013.
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The use of plants as a source of palliative or cure for pathological conditions is quite common worldwide. Xanthium spinosum (Asteraceae), popularly known in Brazil as 'espinho de carneiro', is an annual weed from South America, which has been used by empiric medicine to treat neoplasias. Owing to the extensive use of the above-mentioned plant and to the lack of reports about the real effects of its infusion, current study evaluated the genotoxic potential of its aqueous extract at concentrations 0.02 g L-1, 0.1 g L-1 and 0.2 g L-1 by fish micronucleus test and by comet human leukocytes assay. The micronucleus test featured at least 50 cells with micronuclei to every 2,000 cells scored, as a mutagenic parameter. The comet assay was used as a parameter for assessing the level of cell damage and the damage index. Since no significant changes in strain cells exposed to the aqueous extract in the comet and micronucleus assays were reported, it seems that no genotoxicity evidence is extant at the concentrations and in the assays performed.
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da Costa, Bruna Filipa Tavares, Alexandra Teixeira, Joana C. Prata, and Daniel Pérez-Mongiovi. "Application of the Buccal Micronucleus Cytome Assay for Genotoxicity Detection in Dogs." Animals 15, no. 3 (2025): 382. https://doi.org/10.3390/ani15030382.
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In Europe, there is a growing concern for animal welfare, encompassing both their rights and health. Consequently, identifying biomarkers that predict serious pathological conditions has become crucial in veterinary medicine. The Buccal Micronucleus Cytome (BMCyt) assay is a minimally invasive method that uses biomarkers to evaluate DNA damage and chromosomal instability, using exfoliated buccal cells. A rising frequency of anomalies, such as micronuclei formation, strongly indicates an elevated risk of cancer, neurodegenerative diseases, or accelerated aging, potentially originating from exposure to genotoxins and cytotoxins. This method has been validated in humans, but very little research has been conducted on animals. This work aims to provide a detailed description of an optimized method for collecting buccal exfoliated cells in dogs and to characterize a biomarker related to genomic damage using optical and fluorescent microscopy. Samples from dogs in breeding kennels, including pregnant animals, were tested for chromosomal instability. By following procedures similar to those used in humans, we were able to detect and count major nuclear abnormalities. The percentage of micronuclei was higher compared to other studies. Technical aspects, such as avoiding artifacts and ensuring prior training of the operator, must be taken into account. This work validated the BMCyt method for collecting and processing samples in dogs, potentially enhancing the understanding of micronuclei as biomarkers for pre-pathological states in canines.
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Mitra, Debanjana, and Abilash Vg. "GENOTOXIC EFFECT OF PESTICIDES ON HUMAN LEUKOCYTE CULTURE: A REVIEW." Asian Journal of Pharmaceutical and Clinical Research 9, no. 5 (2016): 29. http://dx.doi.org/10.22159/ajpcr.2016.v9i5.13080.
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ABSTRACTPesticides have long been in use in the agricultural fields, plantation firms, and also for household purposes. However, there are a considerablenumber of studies which prove the detrimental effects of the pesticides that include biochemical, histopathological, and genetic effects. The aim ofthis article is to present a review on the effects of pesticides on leukocytes which have been analyzed through various assays including chromosomeanalysis, cytokinesis-block micronuclei assay, comet assay, semen, and sperm analysis. The studies have shown organophosphates and carbamates tobe the most potential sources of genotoxicity and the individuals exposed to these groups of pesticides are relatively much more prone to genotoxicity.Further investigation on molecular mechanism by which the pesticides affect the genome of cells needs to be carried out.Keywords: Pesticides, Genotoxicity, Chromosome analysis, Cytokinesis-block micronucleus cytome assay, Semen analysis.
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Sriambika, K. "Assessment of DNA damage through micronucleus studies in subjects exposed to formalin." Medpulse International Journal of Anatomy 19, no. 1 (2021): 01–05. http://dx.doi.org/10.26611/10011911.
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Background: Formaldehyde (FA) is the reactive and simplest of all the aldehydes. It is used as a preservative in anatomy, pathology and forensic laboratories. The international agency for research on cancer has classified FA as a carcinogen that can cause nasopharyngeal carcinoma, Leukaemia, Liver and pancreatic cancer. Objective And Method: The aim of the study was to assess the DNA damage in peripheral blood lymphocytes and in buccal cells by Micronucleus assay in Formalin exposed workers of Anatomy, Pathology and Forensic laboratories and compare with the control group, and also to analyze the relationship between frequency of Micronuclei and duration of exposure to formalin. Results: The mean and standard deviation (SD) of micronuclei in peripheral blood of exposed was 8.35 and in controls was 4.18. There was a significant increase in the frequency of MN in exposed group when compared with the comparison group (p<0. 5876). Pearson’s correlation test showed a positive correlation between the years of FA exposure and the number of micronuclei in buccal cells and peripheral blood indicating that DNA damage due to FA was directly proportional to the duration of exposure (r=0.8, 0.9). Conclusion: The present study was done to assess the DNA damage in people who were exposed to FA and a control group not exposed to FA by buccal cell and peripheral blood Micronucleus Assay. There was a significant increase in the MN in people exposed to FA which was directly proportional to the duration of exposure.
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Sommer, Sylwester, Iwona Buraczewska, and Marcin Kruszewski. "Micronucleus Assay: The State of Art, and Future Directions." International Journal of Molecular Sciences 21, no. 4 (2020): 1534. http://dx.doi.org/10.3390/ijms21041534.
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During almost 40 years of use, the micronucleus assay (MN) has become one of the most popular methods to assess genotoxicity of different chemical and physical factors, including ionizing radiation-induced DNA damage. In this minireview, we focus on the position of MN among the other genotoxicity tests, its usefulness in different applications and visibility by international organizations, such as International Atomic Energy Agency, Organization for Economic Co-operation and Development and International Organization for Standardization. In addition, the mechanism of micronuclei formation is discussed. Finally, foreseen directions of the MN development are pointed, such as automation, buccal cells MN and chromothripsis phenomenon.
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Burgum, Michael, Naouale Yamani, Eleonora Longhin, et al. "50 Human Hazard Assessment of Nanomaterials: Supporting Risk Decision Making Through Interlaboratory Trial Data." Annals of Work Exposures and Health 67, Supplement_1 (2023): i51—i52. http://dx.doi.org/10.1093/annweh/wxac087.125.
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Abstract The prevalent application of engineered nanomaterials (ENMs) has led to an extensive effort to develop tools facilitating their risk assessment and management. This interlaboratory trial (encompassing two laboratories) sought to investigate the genotoxic impact of ENMs using the hypoxanthine phosphoribosyltransferase (HPRT) forward mutation assay and in vitro cytokinesis-blocked micronucleus (CBMN) assay, utilising harmonised protocols. In the CBMN assay, human lymphoblast (TK6) cells were exposed to zinc oxide (ZnO), titanium dioxide (TiO2) and tungsten carbide-cobalt (WC/Co) for 24-hours after which 1000 binucleated cells were scored for micronuclei (n=2). For the HPRT assay, TK6 and mouse fibroblast (V79) cells were exposed to ZnO, TiO2, WC/Co, CuO and Nanocyl-CNTs for 24-hours after which 600 wells were scored for point mutations (n=2). Significant cytotoxicity and micronuclei frequency was reported following ZnO exposure (20µg/ml). Significant micronuclei induction was reported following WC/Co exposures (2-fold over control, 100µg/ml). No significant mutagenicity was detected with ZnO or TiO2; both laboratories observed significant cytotoxicity following exposure to ZnO (20µg/ml; 37% reduction in cell viability). In the second interlaboratory trial CuO induced significant cytotoxicity (36% reduction in viability: 0.5µg/ml), and a 32-fold increase in mutagenicity. The Nanocyl induced a 6-fold increase in mutagenicity. The data generated here has shown good statistical concordance between laboratories and has highlighted a promising route forward in supporting risk decision making for ENMs. The authors would like to acknowledge this research has received funding from the European Union’s Horizon 2020 research and innovation program for the RiskGONE project, grant agreement #814425.
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Smolewski, Piotr, Qin Ruan, Luciano Vellon, and Zbigniew Darzynkiewicz. "Micronuclei assay by laser scanning cytometry." Cytometry 45, no. 1 (2001): 19–26. http://dx.doi.org/10.1002/1097-0320(20010901)45:1<19::aid-cyto1140>3.0.co;2-g.
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Arnaoutoglou, Christos, Anastasia Keivanidou, Georgios Dragoutsos, et al. "Factors Affecting the Nuclei in Newborn and Children." International Journal of Environmental Research and Public Health 19, no. 7 (2022): 4226. http://dx.doi.org/10.3390/ijerph19074226.
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It is known that children are more sensitive to the effects of medical treatments and environment than adults. Today there is limited information regarding the differences in genotoxic effects in children. The micronucleus assay is a method that is used to monitor genotoxicity, and it was validated several years before. Today there is international interest for exfoliated buccal cells. Most of the micronuclei studies in children have been performed with the analyses of lymphocytes. However, there is vast interest in using exfoliated cells from the oral cavity. The reason is that other type of cells are acquired non-invasively, this is an important issue in paediatric cohorts. Unfortunately a limitation of measuring micronuclei frequency is that it has been observed to be low in newborns and on the other hand there are a large number of patients and cell sample counts. It has been observed that radiation exposure and environmental pollutants increase the micronuclei frequency in newborn and children. Regarding the medical treatments, there is little data and several studies are needed to optimise the doses. There is the need to observe if there is a relationship between micronuclei in lymphocytes and exfoliated cells and to identify the baseline of the micronuclei levels. Moreover, we evaluate the changes in response to the toxic agents. Prospective cohorts studies will clarify the predictive value of micronuclei for cancer and chronic diseases for both children and adults. Novel molecular technologies will assist in the elucidation of different biological pathways and molecular mechanisms connected with the micronulcei levels in newborn and children.
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Pavel, D. Kolesnichenko* Sergey V.Nadezhdin Yuri E. Burda Elena B. Artyushkova Natalya A. Bystrova Galina A. Lazareva Vladimir Y. Provotorov. "PRECLINICAL STUDY OF THE DNA-DAMAGING ACTIVITY AND GENOTOXICITY OF CARBAMYLATED DARBEPOETIN." Indo American Journal of Pharmaceutical Sciences 04, no. 10 (2017): 3792–97. https://doi.org/10.5281/zenodo.1020034.
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The article presents the results of a potential mutagenicity studying of a new drug from the group of erythropoietin, carbamylateddarbepoetin. The study was performed using DNA-comet assay in mice, and in micronucleus test. As a positive control there were used mice which had been injected with genotoxicantmethylmethane sulfonate at a dose of 40 mg/kg and as a negative control there were mice after administration of equivalent doses of a placebo. The results showed that the mutagenic effect of the drug in single (3.59±1.02) dosing and three times (4.0±0.78) dosing had not been different from the mutagenic effect of placebo (4.39±1.34). Micronucleus test was performed using a cytogenetic damage of bone marrow cells and checking the appearance of polychromatophil cells, containing micronuclei. It was established that carbamylateddarbepoetin within stated doses according to the used test, has no a potential carcinogenic effect. Key words: DNA-comet assay, mutagenicity, carbamylateddarbepoetin, cytogenetic damage.
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Clode, S. A., R. A. Riley, S. D. Blowers, T. C. Marrs, and D. Anderson. "Studies on the Mutagenicity of a Zinc Oxide-Hexachloroethane Smoke." Human & Experimental Toxicology 10, no. 1 (1991): 49–57. http://dx.doi.org/10.1177/096032719101000109.
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1 A suitable method has been developed for generating atmospheres of zinc oxide/ hexachloroethane smoke (ZnHCE). 2 The smoke was investigated using the Ames test and the micronucleus assay. 3 It was weakly mutagenic to the bacteria, but in the bone marrow no increases in micronuclei were detected up to toxic levels of the smoke. 4 The method used here could be applied to other pyrotechnic mixtures which give rise to complex mixtures of products.
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Tobólska, Sylwia, Sylwia Terpiłowska, Jerzy Jaroszewski, and Andrzej Krzysztof Siwicki. "Genotoxicity and mutagenicity of inosine pranobex." Journal of Veterinary Research 62, no. 2 (2018): 207–13. http://dx.doi.org/10.2478/jvetres-2018-0030.
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AbstractIntroductionInosine pranobex (Methisoprinol, ISO, Isoprinosine) is an immuno-modulatory antiviral drug that has been licensed since 1971 in several countries worldwide. In humans, the drug is approved for the treatment of viral infections, and it might also have therapeutic use in animals. The aims of the presented work were to investigate the genotoxicity of inosine pranobex on BALB/3T3 clone A1 and HepG2 cell lines and to elucidate its mutagenicity using the Ames test.Material and MethodsThe BALB/3T3 clone A1 and HepG2 cells were incubated with inosine pranobex at concentrations from 0.1 to 1,000 μg/mL. The genotoxicity was determined by comet and micronucleus assays, and the mutagenicity was determined by Ames assay.ResultsInosine pranobex did not induce a significant dose-related increase in the number of comets or micronuclei in BALB/3T3 clone A1 and HepG2 cells. Moreover, based on the results of the Ames test, it was concluded that inosine pranobex is not mutagenic in theSalmonella typhimuriumreverse mutation assay.ConclusionBased on the results of a comet assay, micronucleus assay, and Ames test, it was concluded that inosine pranobex is neither genotoxic nor mutagenic.
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Ansu, Ann Abraham, Denny E. Ceena, Manaktala Nidhi, Natarajan Srikant, Ahmed Junaid, and Yadav Utkarsh. "Assessment of Genotoxic Effects of Various Forms of Smokeless Tobacco Using Micronuclei Assay." Transylvanian Review XXVI, no. 25 (2018): 6739–44. https://doi.org/10.5281/zenodo.6237318.
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Aim: To assess the micronuclei assay based on age and gender variability from the buccal mucosal cells in patients using various forms of smokeless tobacco. Materials and Methods: A cross sectional study was conducted among sixty eight individuals who had a history of chewing various forms of smokeless tobacco. The study subjects were divided according to their habits into four groups. The groups were arecanut, betel quid, gutkha and control group. Buccal smears were obtained and the exfoliated cells were stained using Feulgen stain. The micronuclei count was done under 40X magnification and compared among the various groups. Results: One way ANOVA analysis was done to compare the difference in ages of the various groups in the present study. The mean age in years were: control group (21 years), gutkha (30 years), betel quid (50 years) and arecanut chewers (43 years), which was found to be significant (p< 0.01). One way ANOVA analysis was also done to compare the micronuclei values in the various groups and it was found that it was the highest in betel quid chewers (P< 0.05). The comparison of the micronuclei value between control and each of the habits was found to be significant (p< 0.001). Similarly, while comparing the micronuclei value between betel quid and the arecanut was found to be significant (p= 0.003). Conclusion: The frequency of Micronuclei assay formation could be used as markers for early identification of the genotoxic effects of chewing tobacco. Policy implications: Awareness of the toxic effects should be carried out with these target groups in mind.
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Shabina, Lal Nand, and Shilpa. "Effect of Years of Exposure on Micronucleus Frequency in Buccle Epithelium Cell through Micronucleus Assay in Subjects Exposed to Formaldehyde." International Journal of Pharmaceutical and Clinical Research 15, no. 1 (2023): 293–99. https://doi.org/10.5281/zenodo.13132082.
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<strong>Background:</strong> The stratified squamous cell epithelium continuously loses superficial buccal cells, which are replaced by cell division. When these cells split, chromosomal fragments or full chromosomes may be left behind during mitotic anaphase and manifest as tiny nuclear particles, or micronuclei, in the cytoplasm of daughter cells. The cytogenetic changes can only be examined from the exfoliated cells after the cells have matured and migrated to the surface. A novel genotoxicity method that shows promise for the investigation of epithelial carcinogens is the micronuclei assay (MA) on exfoliated buccal cells. For detecting tissue-specific genotoxic damage in people exposed to carcinogenic combinations, micronuclei (MN) are ideal internal dosimeters. <strong>Aim:</strong> To analyse effect of years of exposure on micronucleus frequency in buccle epithelium cell through micronucleus assay in subjects exposed to formaldehyde. <strong>Methods and Materials:</strong> After obtaining informed consent, 50 male and female participants who are exposed to formaldehyde were chosen for the study. The duration of formaldehyde exposure in days, months, and years was recorded. After properly washing the mouth, buccal cells were scraped off the inside of the cheek. The cells were moved into centrifuge tubes with 5ml of 0.9% saline, and they underwent two centrifugal separations. 2 drops of fixative (3:1 Methanol and acetic acid) and the supernatant trash were added. The cells were dropped onto a cooled slide using a Pasteur pipette before being air dried. Per sample, two slides were created. The slide was air dried, labelled, and fixed in fixative for 20 minutes before being stored for 24 hours before staining. Giemsa staining solution was used to mark the slides. The criteria provided by Tolbert <em>et al</em> that were used to identify the MN. <strong>Results:</strong> There was positive correlation of various parameters of degree of exposure to formaldehyde with MN count /500 cells but statistically significant correlation was with year of exposure and total exposure only. (p=0.004 & p=0.003 respectively). <strong>Conclusion:</strong> It was concluded that years of exposure to formaldehyde were significantly correlated to the frequency of micronucleus. Micronucleus assay in human epithelial buccal cells is most reliable and attainable method to assess occupational menace resulting in genetic abnormality due to environmental and chemical factors.
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Padilla-Camberos, Eduardo, Moisés Martínez-Velázquez, José Miguel Flores-Fernández, and Socorro Villanueva-Rodríguez. "Acute Toxicity and Genotoxic Activity of Avocado Seed Extract (Persea americanaMill., c.v.Hass)." Scientific World Journal 2013 (2013): 1–4. http://dx.doi.org/10.1155/2013/245828.
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The use of vegetal extracts requires toxicological and genotoxic evaluations to establish and verify safety before being added to human cosmetic, pharmaceutical medicine, or alimentary products.Persea americanaseeds have been used in traditional medicine as treatment for several diseases. In this work, the ethanolic seed extract ofPersea americanawas evaluated with respect to its genotoxic potential through micronucleus assay in rodents. The frequency of micronuclei in groups of animals treated with avocado seed extract showed no differences compared to the negative control (vehicle); therefore, it is considered that the avocado seed extract showed no genotoxic activity in the micronucleus test.
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Mary, Vincy T., Ramya Vengatesan, and Vallabi Rajasekaran. "A research investigation into the micronuclei assay within urothelial cells as an innovative biomarker for the carcinogenesis of cervical cancer conducted at a tertiary care facility." MGM Journal of Medical Sciences 11, no. 1 (2024): 8–14. http://dx.doi.org/10.4103/mgmj.mgmj_218_23.
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Abstract Background: Cervical carcinoma ranks as the fourth most prevalent cancer in women worldwide, following breast, colorectal, and lung cancers. Screening is a crucial secondary prevention strategy, particularly in developing nations such as India, ensuring effective prevention and treatment. This study investigates the micronuclei assay in urothelial cells as a potential biomarker for cervical carcinoma screening, focusing on its correlation with colposcopic and histopathological observations. Materials and Methods: The prospective study was conducted from September 2016 to September 2017 at the Department of Pathology, Institute of Obstetrics and Gynecology, Madras Medical College, Chennai, Tamil Nadu, India. It involved 108 women who visited the colposcopic clinic. Urine samples were collected from 50 women with normal colposcopic findings and 58 with abnormal colposcopy results. Micronuclei scoring was performed on 1000 epithelial cells from the smears. Additionally, cervical biopsies were conducted for the 58 cases with abnormal colposcopic findings, and the resulting histopathological data were juxtaposed with the findings of the micronuclei assay. Results: The study’s findings demonstrated a sensitivity of 82.76% and a specificity of 90%, with positive predictive values of 90.75% and negative predictive values of 81.82%. Additionally, a statistically significant linear correlation was observed between the micronuclei assay and histopathological diagnosis, with a P value of less than 0.001. Conclusion: Therefore, the micronuclei assay offers a means to detect genotoxicity, monitor cervical carcinogenesis, and identify high-risk groups using simple, reliable, and cost-effective techniques suitable for large-scale screening programs.
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Wikanthi, Layung Sekar Sih, Nindi Wulandari, Yuni Fajar Esti, Nur Fitra Sari, and Ratna Asmah Susidarti. "Cinnamomum Essential Oil Prevents DNA Damage-Induced by Doxorubicin on CHO-K1 Cells." Indonesian Journal of Cancer Chemoprevention 8, no. 1 (2017): 27. http://dx.doi.org/10.14499/indonesianjcanchemoprev8iss1pp27-31.
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DNA damage usually happens due to the several chemical materials that induce genotoxic effect in normal cells. Cinnamon essential oil (CEO), which contains cinnamaldehyde as its major compound, has been reported to possess antioxidant activity to prevent DNA damage. The aim of this study is to evaluate the genotoxic and cytotoxic effect of CEO on doxorubicin-induced Chinese Hamster Ovary (CHO-K1) cells. The cytotoxic effect of CEO was determined by MTT assay with the parameter of IC50 while the genotoxic effect was carried out by micronucleus (MN) assay by using acridine orange fluorescent staining with the parameter of MN/1000 cells reduction number. Based on MTT assay, CEO showed cytotoxic activity with the IC50 value of 30 μg/ml and for MN assay, 3 μg/ml (1/10 IC50) of CEO decreased the percentage of micronucleus per 1000 cells up to 94,55%. Thus, the result can be summarized that CEO does not induce genotoxic and has the potency to prevent DNA damage caused by doxorubicin on CHO-K1 cells.Keywords: genotoxic, cinnamomum essential oil (CEO), micronuclei assay, in vitro
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Vidya, Puthan Variyam, and Kumari Chidambaran Chitra. "Evaluation of Genetic Damage in Oreochromis mossambicus Exposed to Selected Nanoparticles by Using Micronucleus and Comet Bioassays." Croatian Journal of Fisheries 76, no. 3 (2018): 115–24. http://dx.doi.org/10.2478/cjf-2018-0015.
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Abstract The purpose of the present study is to extend knowledge on the adverse effects of nanoparticles by evaluating genotoxicity as environmental risk assessment in Oreochromis mossambicus. Fish were exposed to sublethal concentrations of the selected nanoparticles, namely silicon dioxide (SiO2NPs-12mg/L), aluminium oxide (Al2O3NPs-4mg/L), titanium dioxide (TiO2NPs-16.4mg/L) and iron oxide (Fe3O4NPs-15mg/L) for short-term (24, 72 and 96 h) and long-term durations (15, 30 and 60 days). Genetic damages such as cytoplasmic, nuclear and DNA damage were measured in the erythrocytes of fish by using standard genotoxicity tests such as micronucleus test and comet assay. The frequencies of micronuclei along with nuclear and cytoplasmic abnormalities were scored and compared with the control group. The intensity of micronuclei along with other nuclear and cytoplasmic anomalies are found to be increased significantly (p<0.05) in time-dependent manner in all exposure groups when compared to the control group, thereby indicating chromosomal damage as a result of contact with nanoparticles. The tail length and percent of tail DNA within the comet significantly (p<0.05) increased in time-dependant manner after exposure to all nanoparticles, demonstrating an increase in DNA damage. Taken together, by using micronucleus test and comet assay, it is evident that the selected nanoparticles at sublethal concentrations induced genetic damage in Oreochromis mossambicus.
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Lusiyanti, Yanti, Harry Nugroho Eko Surniyantoro, Nastiti Rahajeng, Viria Agesti Suvifan, Sofiati Purnami, and Lina Choridah. "The Cytogenetic Effects on Peripheral Blood Lymphocytes in Cancer Patients After Radiation Therapy: Chromosome Aberrations and Micronuclei." Biosaintifika: Journal of Biology & Biology Education 13, no. 1 (2021): 9–15. http://dx.doi.org/10.15294/biosaintifika.v13i1.25264.
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Individual responses to radiotherapy are often observed whether or not regimes with identical treatments were applied. Patient-related factors, the therapeutic process, and therefore the intrinsic factors of individual radiosensitivity are considered to be causing the variability of side effects. A preliminary evaluation was done on cytogenetic biomarkers found in cancer patients. The purpose of this present study was to assess the individual response of patients with cancers after radiation therapy. The sample obtained from 11 patients with different types of cancer as a case group and 12 people as a control group from a healthy volunteer. Blood samples were stimulated by an in vitro culture using phytohemagglutinin, and the cultures were assessed by using the Dicentric and Cytokinesis- Block Micronucleus (CBMN-) assay. These two methods were compared. The results showed that the overall dicentric chromosome and micronuclei in binucleate cells (MN/BNC) have a significantly higher frequency in the breast, head, and neck compared to extremity cancer. A high frequency of micronuclei in lymphocyte patients was seen after radiotherapy treatment but relatively not much higher compared to the range of micronuclei backgrounds in healthy people The CBMN is the most effective assay for evaluation of the cytogenetic studies in cancer patients because it is more radiosensitive to study individual responses. By evaluating the effects of radiotherapy based on DNA damage, the severity of radiation exposure can be studied. This study can be useful for researchers and related stakeholders in the application of radiotherapy.
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Jagetia, Ganesh Chandra. "Grapefruit flavonoid naringin protects v79 cells against the vinblastine-induced DNA damage in vitr." MOJ Anatomy & Physiology 10, no. 1 (2023): 35–40. http://dx.doi.org/10.15406/mojap.2023.10.00336.
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Vinblastine an antimitotic agent is used to treat hematological malignancies and in combination cancer chemotherapy. Vinblastine produces second malignancies in the survivors which can be reduced/prevented in cancer patients by intervention with natural products. Naringin a grapefruit bioflavonoid was tested for its chemoprotective activity in cultured V79 cells. The V79 cells were treated with 1, 2, 5, 10, 25, 50, 100, and 200 µg/mL of naringin 1 h before exposure to 10 µg/mL of vinblastine. The DNA damage was evaluated by micronucleus assay at 22 h after vinblastine exposure in mononucleate, binucleate, and trinucleate cells. Vinblastine treatment increased the frequency of micronuclei significantly in both the mononucleate and binucleate V79 cells whereas different concentrations of naringin significantly reduced the formation of vinblastine-induced micronuclei. A maximum reduction in micronuclei formation was recorded at 200 µg/mL naringin in both mononucleate and binucleate cells. The determination of cell proliferation indicates that naringin treatment impacted the cell proliferation, especially at 2 to 25 µg/mL which increased at 200 µg/mL in both the naringin and Naringin+Vinblastine groupsgroup as indicated by a rise in binucleate and trinucleate cell numbers. The present study indicates that naringin protects the V79 cells against vinblastine-induced DNA damage indicated by significant attrition in micronuclei formation and 200 µg/mL naringin was highly effective.
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Luqman, Master, Syed Sadatullah, Ali Azhar Dawasaz, Ahmed A. Almeshari, and Rafi Ahmad Togoo. "Radiation Risk Assessment in Professionals Working in Dental Radiology Area using Buccal Micronucleus Cytome Assay." Journal of Contemporary Dental Practice 14, no. 6 (2013): 1024–27. http://dx.doi.org/10.5005/jp-journals-10024-1444.
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ABSTRACT Objective The aim of this study was to assess the incidence of micronuclei (MN) in buccal mucosal cells of professionals working in radiology area to determine the risk of stochastic effects of radiation. Materials and methods All the professionals and students working in King Khalid University - College of Dentistry radiology area were included in the Risk Group (RG = 27). The Control Group (CG = 27) comprised of healthy individual matching the gender and age of the RG. Buccal mucosal scraping from all the 54 subjects of RG and CG were stained with Papanicolaou stain and observed under oil immersion lens (×100) for the presence of micronuclei (MN) in the exfoliated epithelial cells. Results There was no significant difference between the incidence of MN in RG and CG (p = >0.05) using t-test. Conclusion Routine radiation protection protocol does minimize the risk of radiation induced cytotoxicity, however, screening of professionals should be carried out at regular intervals. How to cite this article Sadatullah S, Dawasaz AA, Luqman M, Assiry AA, Almeshari AA, Togoo RA. Radiation Risk Assessment in Professionals Working in Dental Radiology Area using Buccal Micronucleus Cytome Assay. J Contemp Dent Pract 2013;14(6):1024-1027.
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Swati Parhar and Amani Mahajan. "Assessment of Micronuclei Frequency in Individuals with a Habit of Tobacco Chewing by Means of Exfoliated Oral Buccal Cells." International Healthcare Research Journal 4, no. 7 (2020): OR15—OR18. http://dx.doi.org/10.26440/ihrj/0407.10277.
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INTRODUCTION: Majority of cancers are diagnosed at an advanced stage resulting in poor prognosis and survival rates among patients. Hence early diagnosis of oral cancers seems to be the need of the hour. Analysis of exfoliated buccal cell micronuclei (MN) is a sensitive method of monitoring genetic damage. AIM: The present study has been carried out with an objective to evaluate the genotoxic effects of tobacco chewing by means of micronucleus assay in exfoliated cells of buccal mucosa.MATERIALS AND METHOD: This cross sectional study was carried out in Department of Oral Pathology, Swami Devi Dyal Hospital And Dental College, Golpura, Barwala, Panchkula. The study population comprised of a total of 50 subjects, divided into five groups: Group 1comprising of 10 age and sex matched healthy subjects without any habits as controls, Group 2 comprising of 10 subjects with a history of chewing tobacco. Group 3 comprising of 10 subjects with a history of chewing tobacco and cigarette smoking, Group 4 comprising of 10 subjects with a history of chewing tobacco and drinking and Group 5 comprising of 10 subjects with a history of chewing tobacco, smoking and drinking. Oral exfoliated cells were obtained from buccal mucosa of the subjects, slides were prepared from each subject stained with stain respectively.RESULTS: The mean numbers of micronuclei in group 1 were 7.86±6.7, Group 2 were 63.37±10.01, Group 3 were 65.49±12.32, Group 4 were 68.22±11.11 and Group 5 were 69.43±10.71. On comparison we observed that the difference in Mean micronuclei frequency among all the 5 study groups came to be statistically also highly significant (p<0.0001*)CONCLUSION: Micronuclei assay is an effective tool that reflects severity of disease. Even though tobacco induced cancers are preventable, banning the use of tobacco has not been possible for social and political reasons.
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Santiago-Manzano, Dayna. "Staining criteria, collection and number of cells evaluated to identify micronuclei in buccal mucosa cells of agricultural workers exposed to pesticides." Mexican Journal of Medical Research ICSA 9, no. 18 (2021): 34–38. http://dx.doi.org/10.29057/mjmr.v9i18.5717.
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Although there are several publications that refer to the basic criteria established for the micronucleus (MN) test in exfoliated buccal cells, there is still a difference in the quantity, method of obtention and staining of the cells for the evaluation of the presence of micronuclei. Objective. Identify the criteria for evaluation of MN in oral mucosa cells exposed to pesticide in investigations carried out. Material and methods. A systematic review was carried out on the internet, based on articles published in Crossref, JCR, Scopus, PubMed, Google academic, using keywords such as: micronuclei, buccal mucosa cells, genotoxic damage, pesticides and biomarker. Results. In the six selected articles, four presented statistically significant values with the presence of MN when comparing the exposed groups with respect to the control groups, and the criteria for staining, collection and number of cells evaluated to identify micronuclei were very varied. Conclusions. It is important to follow validated and standardized protocols for the MN oral cytoma assay. Being considered in all the parameters suggested in the protocol will increase the reliability of the studies and will give the possibility of comparing the results obtained.
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Colombelli, Renato Alberto Weber, Gabriela Cristina Uebel, Diene da Silva Schlickmann, et al. "DNA DAMAGE AND OTHER NUCLEAR ANOMALIES AMONG GYM USERS: A COMPARATIVE STUDY BETWEEN BRAZIL AND SPAIN." Revista Jovens Pesquisadores 12, no. 2 (2022): 10–18. http://dx.doi.org/10.17058/rjp.v12i2.17487.
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Populations from different countries may present different cellular modifications among themselves, and the Buccal Micronucleus Cytome (BMCyt) assay in human buccal mucosal tissue may be a marker to evaluate these modifications. This study evaluated and compared DNA damage and other nuclear anomalies between Brazilian and Spanish gym users. This is a cross-sectional study carried out with gym users of Santa Cruz do Sul/Brazil and Madrid/Spain. The BMCyt assay was performed for biomarkers of DNA damage (micronuclei and/or nuclear buds), cytokinetic defects (binucleated cells), proliferative potential (basal cell frequency) and/or cell death (condensed chromatin, karyorrhexis, pyknotic and karyolytic cells) in human buccal mucosal. Of the 228 individuals evaluated, 163 were Brazilian, and 65 were Spanish. Gym users of both countries differed between weight, body mass index, body fat, and muscle mass. The Brazilians presented a significantly higher frequency of micronuclei, nuclear buds, cells with condensed chromatin and karyorrhexis. Spaniards, however presented a significantly higher frequency of karyolytic cells. In conclusion, Brazilian gym users presented significantly higher rates of DNA damage and cell death, while the Spanish presented a higher frequency of advanced stage cell death.
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Sharma, Dinesh C. "Micronuclei assay to detect arsenic-induced cancer." Lancet Oncology 5, no. 7 (2004): 393. http://dx.doi.org/10.1016/s1470-2045(04)01517-7.
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Minina, V. I., and V. Yu Buslaev. "Estimating genotoxic effects of anticancer drugs using cytokinesis-block micronucleus assay on human lymphocytes." Fundamental and Clinical Medicine 4, no. 3 (2019): 95–101. http://dx.doi.org/10.23946/2500-0764-2019-4-3-95-101.
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Here we review the current experience of using cytokinesis-block micronucleus (CBMN) assay on cultures of human lymphocytes to evaluate genotoxic effects of anticancer drugs. Having performed search in PubMed, Scopus, Web of Science, TOXLINE, and the Cochrane Library, we identified a total of 172 relevant studies. Out of them, 89 were conducted in vitro, and 41 were published within the last decade. The mentioned studies concordantly demonstrated a significant increase in micronuclei, protrusions, nucleoplasmic bridges, and a decrease in proliferation in cells treated with anticancer drugs in a time- and dose-dependent manner. Notably, the results of CBMN assay are consistent with the data obtained from other cytogenetic techniques (comet assay, chromosomal aberration analysis, analysis of mutations in housekeeping genes, and fluorescence in situ hybridisation). Conclusion. CBMN assay permits a reliable evaluation of the mutagenic effects related to anticancer drugs.
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Akca, Cetin, Ozgur Vatan, Dilek Yilmaz, Huzeyfe HURIYET, Nilüfer Cinkilic, and Tolga Cavas. "In vitro cytotoxic and genotoxic effects of donkey milk on lung cancer and normal cells lines." Czech Journal of Food Sciences 37, No. 1 (2019): 29–35. http://dx.doi.org/10.17221/221/2018-cjfs.
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In vitro cytotoxic and genotoxic effects of donkey milk on cancer (A549) and normal (BEAS-2B) lung cell lines were investigated. The XTT and WST-1 tests as well as clonogenic assays were used to evaluate cytotoxicity. The comet assay and micronucleus test were used as genotoxicity endpoints. Donkey milk showed lower cytotoxic effects against normal lung cell line BEAS-2B in comparison to the tumor cell line A549. Genotoxicity experiments revealed dose dependent increases in the frequencies of micronuclei and single stranded DNA breaks in A549 cells whereas no significant damage was observed in BEAS-2B cells. The results indicate that donkey milk has anti-proliferative and genotoxic effects on lung cancer cells at concentrations which are non-toxic to normal lung cells.
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Maes, A., R. Anthonissen, and Luc Verschaeve. "Testing Chemical Agents with the Cytokinesis-Block Micronucleus Cytome Assay." Folia Biologica 58, no. 5 (2012): 215–20. http://dx.doi.org/10.14712/fb2012058050215.
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In order to evaluate the applicability of the cytokinesis-block micronucleus cytome assay in routine mutagenicity testing we investigated with this method different chemicals having different mechanisms of action: non-mutagens, direct-acting base-altering mutagens, direct-acting cross-linking mutagens, clastogens including a radiomimetic chemical, indirect-acting spindle poisons and indirect-acting enzyme inhibitors. We looked at the presence of micronuclei as biomarkers for either the loss of chromosome fragments (clastogen) or the loss of a whole chromosome (aneugen), nucleoplasmic bridges as biomarkers for complex rearrangements (e.g., dicentric chromosomes) and nuclear buds as biomarkers for gene amplification. The cytome assay proved to be a suitable tool to investigate genetic effects of environmental agents and to provide insight into their working mechanisms as all chemicals tested showed the expected response.
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Rudd, N. L., D. I. Hoar, S. E. Williams, and U. G. G. Hennig. "Genotype and the cryopreservation process affect the levels of aneuploidy and chromosome breakage in cultured human fibroblasts." Genome 32, no. 2 (1989): 196–202. http://dx.doi.org/10.1139/g89-429.
Full textAbstract:
Spontaneous micronucleus frequencies were measured in 11 human fibroblast strains, with early-passage cells that had never been frozen and with cells of comparable population doublings that had been cryopreserved in liquid nitrogen. The mean micronucleus frequency of the 11 strains increased from 14.0 ± 0.7 to 20.4 ± 1.8/1250 mononucleated cells (P = 0.002) after the freeze – thaw process. The nature of this increase in micronucleus frequency was examined using an immunodetection assay for the in situ identification of kinetochores in micronuclei. The increase in micronucleus frequency occurred primarily in the kinetochore-positive fraction, which is indicative of aneuploidy, but also by an increase in chromosome breakage in several strains. The findings were reproducible in repeat biopsies from two donors. Plating efficiencies of the 11 strains were studied during 1 – 9 and 10 – 20 population doublings from primary outgrowth, before freezing and again after freeze – thaw. The mean plating efficiency of frozen – thawed cells before nine doublings was significantly lower than that of cells of similar ages that had never been frozen (P = 0.004). The four strains that had a greater than 25% decrease in plating efficiency post freeze-thaw also had the highest aneuploidy index post freeze-thaw, suggesting that chromosomal imbalance contributes to the observed reduction in growth. We conclude that the genotype and culture manipulations of a fibroblast strain influence the outcome of the micronucleus assay.Key words: micronuclei, kinetochore immunofluorescence, freeze – thaw, aneuploidy, chromosome breakage.
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Slowinski, Jerzy, Grazyna Bierzynska-Macyszyn, Urszula Mazurek, et al. "CYTOKINESIS-BLOCK MICRONUCLEUS ASSAY IN HUMAN GLIOMA CELLS EXPOSED TO RADIATION." Image Analysis & Stereology 23, no. 3 (2011): 159. http://dx.doi.org/10.5566/ias.v23.p159-165.
Full textAbstract:
Biological tests are efficient in reflecting the biological influences of several types of generally harmful exposures. The micronucleus assay is widely used in genotoxicity studies or studies on genomic damage in general. We present methodological aspects of cytokinesis-block micronucleus assay performed in human gliomas irradiated in vitro. Eight human glioblastoma cell lines obtained from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany) were gamma-irradiated (60Co) over a dose range of 0-10 Gy. Cytokinesis-block micronucleus assay was performed to quantitate cytogenetic damage. The cells were fixed directly on dishes, stained with fluorochrome DAPI and evaluated under fluorescent and phase contrast microscope. The micronucleus frequency was expressed as a micronuclei (MN) per binucleated cell (BNC) ratio, calculated after scoring at least 100 BNC per dish. The frequency of spontaneous MN ranged from 0.17 to 0.613 (mean: 0.29 ± 0.14). After irradiation increase of MN frequency in the range of 0.312 - 2.241 (mean: 0.98 ± 0.68) was found at 10 Gy. Gliomas are extremely heterogenous in regard to cytogenetic effects of irradiation, as shown in this study by cytokinesis-block micronucleus assay. This test is easily performed on irradiated glioma cell lines and can assist in determining their radiosensitivity. However, in order to obtain reliable and reproducible results, precise criteria for MN scoring must be strictly followed. Simultaneous use of fluorescent and phase contrast equipment improves imaging of morphological details and can further optimize MN scoring.
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Santos, Jean L., Eliana A. Varanda, Priscila L. Bosquesi, Adélia E. Almeida, and Chung Man Chin. "Mutagenic and Genotoxic Effect of Hydroxyurea." International Journal of Biomedical Science 7, no. 4 (2011): 263–67. http://dx.doi.org/10.59566/ijbs.2011.7263.
Full textAbstract:
The hydroxyurea, a cytotoxic drug, is the mainly available therapeutical strategy for the treatment of sickle cell disease. This study aimed to evaluate the mutagenic and genotoxic potential of the hydroxyurea through the Salmonella/Microsome assay and micronucleus test in peripheral blood of mice.The doses were evaluated at 29.25-468 μmol/plate in Salmonella/Microsome assay in presence and absence of metabolic activation the drug. In the micronucleus test the doses were evaluated at 12.5; 25; 50; 75 and 100 mg/kg. The results show that hydroxyurea present mutagenic activity in TA98 and TA100 in doses above 117 μmol/plate and 234 μmol/plate respectively. The drug induced a significant increase in the frequency of micronuclei in reticulocytes of mice at concentrations of 50, 75 and 100 mg/kg, compared to negative control (water). These results demonstrated the mutagenic and genotoxic potential of hydroxyurea.