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      Emergence of micronuclei as a genomic biomarker

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          Abstract

          The presence of micronuclei (MN) in mammalian cells is related to several mutagenetic stresses. MN are formed as a result of chromosome damage and can be readily identified in exfoliated epithelial cells. MN is chromatin particles derived from acentric chromosomal fragments, which are not incorporated into the daughter nucleus after mitosis. It can be visualized by chromatin stains. A variety of factors influences the formation of MN in cells such as age, sex, genetic constitution, physical and chemical agents, adverse habits such as tobacco, areca nut chewing, smoking, and alcohol consumption. Micronucleation has important implications in the genomic plasticity of tumor cells. The present paper reviews the origin, fate and scoring criteria of MN that serves as a biomarker of exposure to genetic toxins, and for the risk of cancer.

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          Most cited references26

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          Buccal micronucleus cytome assay.

          The Buccal Micronucleus Cytome (BMCyt) assay is a minimally invasive method for studying DNA damage, chromosomal instability, cell death and the regenerative potential of human buccal mucosal tissue. This method is increasingly used in molecular epidemiological studies for investigating the impact of nutrition, lifestyle factors, genotoxin exposure and genotype on DNA damage, chromosome malsegregation and cell death. The biomarkers measured in this assay have been associated with increased risk of accelerated ageing, cancer and neurodegenerative diseases. This protocol describes one of the current established methods for buccal cell collection using a small-headed toothbrush, the generation of a single-cell suspension, slide preparation using cytocentrifugation, fixation and staining using Feulgen and Light Green for both bright field and fluorescence microscopic analysis. The scoring criteria for micronuclei and other nuclear anomalies are also described in detail. The protocol in its current form takes approximately 4 h to complete from the time of buccal cell collection to the generation of stained slides for microscopic analysis.
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            The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage: the HUMN project perspective on current status and knowledge gaps.

            The micronucleus (MN) assay in exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. This overview has concluded that although MN assay in buccal cells has been used since the 1980s to demonstrate cytogenetic effects of environmental and occupational exposures, lifestyle factors, dietary deficiencies, and different diseases, important knowledge gaps remain about the characteristics of micronuclei and other nuclear abnormalities, the basic biology explaining the appearance of various cell types in buccal mucosa samples and effects of diverse staining procedures and scoring criteria in laboratories around the world. To address these uncertainties, the human micronucleus project (HUMN; see http://www.humn.org) has initiated a new international validation project for the buccal cell MN assay similar to that previously performed using human lymphocytes. Future research should explore sources of variability in the assay (e.g. between laboratories and scorers, as well as inter- and intra-individual differences in subjects), and resolve key technical issues, such as the method of buccal cell staining, optimal criteria for classification of normal and degenerated cells and for scoring micronuclei and other abnormalities. The harmonization and standardization of the buccal MN assay will allow more reliable comparison of the data among human populations and laboratories, evaluation of the assay's performance, and consolidation of its world-wide use for biomonitoring of DNA damage.
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              The HUman MicroNucleus Project--An international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans.

              The International Collaborative Project on Micronucleus Frequency in Human Populations (HUMN) was organized to collect data on micronucleus (MN) frequencies in different human populations and different cell types. The test procedures considered by this project are assays using human lymphocytes (cytokinesis-block method), exfoliated epithelial cells, and other cell types. Data (including descriptions of the populations monitored, detailed test protocols, and test results) are being obtained from a large number of laboratories throughout the world and are being entered into a unified database. The information will be used to: (1) determine the extent of variation of 'normal' values for different laboratories and the influence of other factors potentially affecting baseline MN frequency, e.g., age, gender and life-style; (2) provide information on the effect of experimental protocol variations on MN frequency measurements; (3) design and test optimal protocols for the different cell types; and (4) determine the extent to which MN frequency is a valid biomarker of ageing and risk for diseases such as cancer.
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                Author and article information

                Journal
                Indian J Med Paediatr Oncol
                Indian J Med Paediatr Oncol
                IJMPO
                Indian Journal of Medical and Paediatric Oncology : Official Journal of Indian Society of Medical & Paediatric Oncology
                Medknow Publications & Media Pvt Ltd (India )
                0971-5851
                0975-2129
                Oct-Dec 2015
                : 36
                : 4
                : 212-218
                Affiliations
                [1] Department of Oral and Maxillofacial Pathology, Bhojia Dental College and Hospital, Baddi, Himachal Pradesh, India
                [1 ] Department of Endodontics, Himachal Dental College, Sundernagar, Himachal Pradesh, India
                [2 ] Department of Oral Medicine and Radiology, Darshan Dental College & Hospital, Udaipur, India
                [3 ] Department of Oral Pathology and Microbiology, Himachal Dental College, Sundernagar, Himachal Pradesh, India
                [4 ] Department of Oral and Maxillofacial Surgery, Institute of Dental Sciences, Bhubneshwar, India
                [5 ] Department of Endodontics, IDS, Uttar Pradesh, India
                [6 ] Department of Periodontics, DJ College of Dental Sciences and Research, Uttar Pradesh, India
                Author notes
                Address for correspondence: Dr. Robin Sabharwal, Bhojia Dental College and Hospital, Baddi, Himachal Pradesh, India. E-mail: robin.sabharwal@ 123456yahoo.co.in
                Article
                IJMPO-36-212
                10.4103/0971-5851.171541
                4711219
                26811590
                e20ec5de-1224-429c-b662-c2dd91a96329
                Copyright: © Indian Journal of Medical and Paediatric Oncology

                This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

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                Categories
                Review Article

                Oncology & Radiotherapy
                aneugens,clastogens,micronuclei
                Oncology & Radiotherapy
                aneugens, clastogens, micronuclei

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