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      Genotoxicity in Oral Mucosal Epithelial Cells of Petrol Station Attendants: A Micronucleus Study

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          Abstract

          Introduction:

          Occupational exposure to petrol derivatives possesses an increased risk of various cancers including that of the oral mucosa. Scientific studies have shown the correlation of micronuclei assay (MN) with the cytogenotoxic changes in petrol station attendants. However, very few have reported the use of MN assay as a promising tool for assessing the impact of smoking in these workers.

          Aim:

          To explore the cytogenotoxic damage in exfoliated buccal cells obtained from petrol station attendants and control subjects using the MN assay along with additional effects due to smoking.

          Materials and Methods:

          The study comprised 60 males who were divided into Group I–IV with each having 15 subjects. These subjects were categorized as exposed smokers, exposed nonsmokers, unexposed smoker group, and unexposed nonsmokers. The MN and additional nuclear abnormalities (karyorrhexis [KH], binucleation [BN], pyknosis [P], and karyolysis [KL]) were calculated in PAP-stained slides.

          Results:

          Statistically higher mean frequencies of overall nuclear anomalies were observed in petrol pump workers in comparison with the control group. Petrol pump smokers carry the highest nuclear anomalies followed by non-exposed smokers than exposed non-smokers and the count was the least among unexposed non-smoker workers.

          Discussion and Conclusion:

          The present study indicated that the petrol pump workers are under higher cytogenotoxic damage. Also, smoking added to the frequency of damage. Thus, MN and other nuclear anomalies are in-vitro reliable biomarker assays available and should be routinely employed as a screening tool in their periodic medical evaluation.

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          Most cited references24

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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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            Cytogenetic biomonitoring in petrol station attendants: micronucleus test in exfoliated buccal cells.

            To study the effects of occupational exposure to petroleum derivates such as benzene, exfoliated buccal cells from 50 petrol station attendants and 50 age- and sex-matched control subjects were examined for micronucleus (MN) frequency. Frequencies of nuclear abnormalities (NA) other than micronuclei, such as binucleates, karyorrhexis and karyolysis, were also evaluated. Benzene exposure was ascertained by measuring urinary phenol levels. The mean urinary phenol level of station workers was found to be significantly higher than that of control subjects (P < 0.05). Analysis of buccal cells revealed that MN and NA frequencies in petrol station workers were significantly higher than in control subjects (P < 0.01) and also significantly related to smoking habit (P < 0.01). Our findings indicate that the petrol station workers are under risk of significant cytogenetic damage.
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              Combining the in vivo comet and micronucleus assays: a practical approach to genotoxicity testing and data interpretation

              Despite regulatory directives requiring the reduction of animal use in safety testing, recent modifications to genotoxicity testing guidelines now propose the use of two in vivo genotoxicity assays as a follow-up to an in vitro positive (International Conference on Harmonization Consensus Draft Guidance S2[R1] released March, 2008). To address both goals, the in vivo comet and micronucleus (MN) assays can be successfully combined into one informative study. Combining these two assays with such differences in sensitivity, endpoints measured and the type of data generated significantly improves upon the current standard capabilities for detecting genotoxicity without requiring additional animals. But to take full advantage of the benefits of incorporating the comet assay in safety testing, these same differences must be recognized and considered. Developed from over 15 years experience using the in vivo comet and MN assays in genotoxicity testing of chemicals and pharmaceuticals, this paper presents guidelines for the appropriate experimental design, dose selection and data interpretation for combined in vivo comet/MN assay studies. To illustrate the approach, data from combined assay studies are presented and discussed.
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                Author and article information

                Journal
                J Cytol
                J Cytol
                JCytol
                Journal of Cytology
                Wolters Kluwer - Medknow (India )
                0970-9371
                0974-5165
                Oct-Dec 2021
                22 November 2021
                : 38
                : 4
                : 225-230
                Affiliations
                [1 ]Department of Oral Pathology, Sudha Rustagi College of Dental Sciences and Research, Faridabad, Haryana, India
                [2 ]Department of Oral Pathology, Government College of Dentistry, Indore, Madhya Pradesh, India
                [3 ]Department of Public Health Dentistry, Sudha Rustagi College of Dental Sciences and Research, Faridabad, Haryana, India
                Author notes
                Address for correspondence: Dr. Shweta Rehani, Department of Oral Pathology, Sudha Rustagi College of Dental Sciences, Faridabad, Haryana - 121003, India. E-mail: rehanishweta@ 123456gmail.com
                Article
                JCytol-38-225
                10.4103/JOC.JOC_44_21
                8670452
                35002116
                442ef005-7db8-496c-b83f-cb162a6b8f06
                Copyright: © 2021 Journal of Cytology

                This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.

                History
                : 05 March 2021
                : 03 October 2021
                : 28 October 2021
                Categories
                Original Article

                Pathology
                buccal smear,cytogenotoxicity,exfoliative cytology,micronuclei,petrol fumes,smoking
                Pathology
                buccal smear, cytogenotoxicity, exfoliative cytology, micronuclei, petrol fumes, smoking

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