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Scoring criteria for the evaluation of micronuclei in oral exfoliated cells

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      HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures.

      Criteria for scoring micronuclei and nucleoplasmic bridges in binucleated cells in the cytokinesis-block micronucleus assay for isolated human lymphocyte cultures are described in detail. Morphological characteristics of mononucleated cells, binucleated cells, and multinucleated cells as well as necrotic and apoptotic cells and nuclear buds are also described. These criteria are illustrated by a series of schematic diagrams as well as a comprehensive set of colour photographs that are of practical assistance during the scoring of slides. These scoring criteria, diagrams and photographs have been used in a HUman MicronNucleus (HUMN) project inter-laboratory slide-scoring exercise to evaluate the extent of variability that can be attributable to individual scorers and individual laboratories when measuring the frequency of micronuclei and nucleoplasmic bridges in binucleated cells as well as the nuclear division index. The results of the latter study are described in an accompanying paper. It is expected that these scoring criteria will assist in the development of a procedure for calibrating scorers and laboratories so that results from different laboratories for the cytokinesis-block micronucleus assay may be more comparable in the future.
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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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          An in vivo cytogenetic analysis of human oral squamous cell carcinoma

          Background: Oral cancer ranks in the top three of all cancers in India, which accounts for over 30% of all cancers reported in the country. The micronucleus test (MNT) is one of the most widely applied short term tests used in genetic toxicology to evaluate the mutagenicity and carcinogenicity. Aims: The present study aims at an in vivo cytogenetic analysis of human oral squamous cell carcinoma and to assess the applicability of MNT in diagnosing early detection of oral carcinoma. Materials and Methods: Exfoliated scrape smears were collected from the clinically diagnosed 136 patients suffering from oral precancerous and cancerous lesions. The wet fixed smears were stained by adopting Papanicolaou's staining protocol and counter-stained with Giemsa's solution. Results: The frequency of micronucleated cells has been observed to be in increasing order with the increase of the age-groups and from control to precancerous to cancerous cases significantly in both sexes. Conclusion: Micronucleus formation in the oral mucosa could be a biomarker of genetic damage and also a potential onco-indicator in the long run of oral carcinogenesis. Therefore, MNT can be applied for the early detection of oral carcinoma in the human being.
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            Author and article information

            Affiliations
            Department of Oral and Maxillofacial Pathology, Christian Dental College, CMC, Ludhiana, Punjab, India
            [1 ]Department of Oral and Maxillofacial Pathology, Bapuji Dental College and Hospital, Davangere, Karnataka, India
            Author notes
            Correspondence to: Dr. Sonal Grover, E-mail: sonalgrvr@ 123456yahoo.com
            Journal
            South Asian J Cancer
            South Asian J Cancer
            SAJC
            South Asian Journal of Cancer
            Medknow Publications & Media Pvt Ltd (India )
            2278-330X
            2278-4306
            Apr-Jun 2017
            : 6
            : 2
            : 89
            5506822 SAJC-6-89a 10.4103/2278-330X.208842
            Copyright: © 2017 The South Asian Journal of Cancer

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