Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Dr Bhanu K Bhakhri

"The Journal of Clinical and Diagnostic Research (JCDR) has been in operation since almost a decade. It has contributed a huge number of peer reviewed articles, across a spectrum of medical disciplines, to the medical literature.
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Faculty, Pediatric Medicine
Super Speciality Paediatric Hospital and Post Graduate Teaching Institute, Noida
On Sep 2018




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Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
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I wish JCDR a great success and I hope that journal will soar higher with the passing time."



Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




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Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




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Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help ones reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journalsNo manuscriptsNo authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011

Important Notice

Dentistry
Year : 2010 | Month : December | Volume : 4 | Issue : 6 | Page : 3607 - 3613

Evaluation Of Micronuclei Using Papanicolaou And May Grunwald Giemsa Stain In Individuals With Different Tobacco Habits – A Comparative Study

PALASKAR S*, JINDAL C**

*Professor, Dept of oral pathology, Sinhgad dental college and hospital, Pune, Maharastra, India, 411041, palaskarsangeeta@gmail.com, 00918806791307; **Senior lecturer, Dept of oral pathology, Derabassi, Chandigarh, India, 00919466949753, ichhavi25@gmail.com

Correspondence Address :
*Professor, Dept of oral pathology, Sinhgad dental college and hospital, Pune, Maharastra, India, 411041, palaskarsangeeta@gmail.com.

Abstract

Aims: To compare the Papanicolaou’s (Pap) and May Grunwald’s Giemsa (MGG) staining techniques which are done to detect micronuclei (MN) in exfoliated buccal mucosal cells in individuals with different tobacco habits. To determine the severity of human buccal cell changes which are associated with smoking and smokeless (‘‘chewing’’) tobacco (SLT).
Methods and Material: A total of 45 male subjects (15 smokers, 15 smokeless tobacco users and 15 non users/ non smokers) were examined. Two cytological smears were taken from the apparently normal buccal mucosa from each individual. 45 smears (1 per individual) were wet fixed and stained with Pap and the remaining 45 smears were air dried and stained with the MGG stain. All the smears were assessed for cellularity and were scored for MN. Statistical analysis used: The ANOVA (one way analysis of variance) was used to analyse the frequency of cells with micronuclei. Bonferroni multiple comparisons were done to determine the significance (p<0.05) of the mean difference.
Results: MNs were easily seen in the clear cytoplasm in the Pap smears. Regarding the MGG smears, bacteria and cell debris masked the effect of the MNs as compared to the Pap smears, where the fixative destroyed the bacteria and clearly demarcated the cell boundaries. The score of the MN frequency decreased as we moved from the smokeless tobacco chewers to the smokers and then to the non users and the non smokers.
Conclusions: Pap is a better stain as compared to MGG for counting micronuclei. Smokeless tobacco chewers showed an increased number of MNs as compared to the smokers, thus laying emphasis on the greater carcinogenic potential of tobacco which was used in the chewable form.

Keywords

Micronuclei, Smokers, Smokeless tobacco users, Papanicolaou, May Grunwald’s Giemsa

Introduction
The buccal cell micronucleus (MN) assay was first proposed in 1983 (1) and it continues to gain popularity as a biomarker of genetic damage in numerous applications. MN assays provide information on the cytogenetic damage in the tissues, that are targets of human carcinogens and from which carcinomas can develop. Oral squamous cell carcinomas are characterized by complex karyotypes that involve many chromosomal deletions, translocations and structural abnormalities. Cells from these type of tumours often have errors in chromosome segregation that lead to the formation of a lagging chromosome or chromosome parts that become lost during the anaphase stage of cell separation and are excluded from the reforming nuclei. The laggards are observed in the cytoplasm as micronuclei (2),(3).
Significantly higher frequencies of MNs have been observed in exfoliated buccal cells, from people who are exposed to organic solvents, antineoplastic agents, diesel derivatives, polycyclic aromatic hydrocarbons, lead-containing paints and solvents and drinking water which is contaminated with arsenic (4)-(23). Recent studies have also suggested the genotoxicity and the cytotoxicity of the urban air pollution and ozone during the summer season, particularly in places with high ambient levels (24)-(26).

The lifestyle factors that are associated with genetic damage include smoking, alcohol consumption and diet, especially vitamin deficiencies and supplementation (2),(27). A majority of the studies which reported a significant increase in MNs in the buccal mucosa cells, which were related to a risk of oral cancer, were performed in subgroups of subjects with specific lifestyle habits, i.e. chewers of betel quids (areca nut, betel leaves, slaked lime and tobacco) from India, Taiwan and Philippines; reverse smokers from India and Philippines; snuff dippers from Canada; users of Khaini tobacco (tobacco mixed with slaked lime) from India, and other similar practices (28)-(31). But comparative studies on individuals who consumed tobacco in different forms are scarce.

Also, only little attention has been given, until now, to the effect of different staining procedures on the results of micronuclei assays. An evaluation of the literature shows that a variety of different stains is used in micronuclei studies. Among the DNA-specific stains, the ones which are most widely used are Feulgen and acridine orange; in some experiments, 4¶,6-diamidino-2-phenylindole (DAPI) and propidium iodide were also used. About 30% of the studies on epithelial cells were conducted by using nonspecific stains (Giemsa, May- Grunwald’s Giemsa, and less frequently, Orcein) (32)-(34). Hence, the present study was done to fulfill the following study objectives:
1. To compare Papanicolaou’s (Pap) and May Grunwald’s Giemsa (MGG) staining techniques to detect micronuclei (MN) in exfoliated buccal mucosal cells in individuals with different tobacco habits.
2.To determine the severity of human buccal cell changes which are associated with smoking and smokeless (‘‘chewing’’) tobacco (SLT).

Material and Methods

Subjects
A total of 45 male subjects (15 smokers, 15 smokeless tobacco users and 15 non users/non smokers) were selected from among the outpatients who attended the Department of Oral and Maxillofacial Pathology, M M College of Dental Sciences and Research, Mullana, Ambala, Haryana, from January 2009 to October 2009. The smokers who smoked every day for at least five years and consumed >80 packs/year and smokeless tobacco chewers who chewed four or more packets daily for at least five years, were included in their respective groups. All were occasional drinkers who consumed alcohol once a week. The control group persons were not habituated to any form of tobacco consumption or pan chewing and were occasional drinkers who consumed alcohol once a week. The majority of them were living in rural areas.
Before collecting the samples, the written consent of each individual was taken. Each subject was asked about his lifestyle, food consumption, infectious diseases, X-ray exposure, medication, etc. Individuals who had had a recent viral infection or had been exposed to X-rays or those who had been under medication were excluded from the study. The lifestyle (except the habit of tobacco) and dietary habits of the controls were similar to those of the users.
Cytological preparations and examination
Before sampling, each individual rinsed his/her mouth thoroughly with tap water. The exfoliated cells were obtained by scraping the buccal mucosa with a moistened wooden spatula. The scraped cells were placed onto pre-cleaned slides. Two slides were made from each subject. One was air dried and stained with the MGG stain (Table/Fig 1), while the other was wet fixed and stained with PAP.

(Table/Fig 1): Modified MGG staining method
The slides were randomized and scored by a single observer. From each slide, ~ 1000 cells were examined under the 400X magnification and when MN cells were located, they were examined under the 1000X magnification. The criterion which was developed by Tolbert et al (Table/Fig 2) was used for counting the micronuclei.

(Table/Fig 2): Tolbert et al criteria

Results

The individual data is presented in (Table/Fig 3). The frequency of cells with micronuclei was analysed by means of ANOVA, the one way analysis of variance.

(Table/Fig 3): Frequency of distribution of micronuclei in buccal mucosal cells in both Pap and MGG stained smears.

The mean values which were obtained from the smokeless tobacco chewers, the smokers, the tobacco non users and the non smokers were compared in both the PAP and the MGG staining techniques (Table/Fig 4). Bonferroni multiple comparisons were done amongst the different groups to determine the significance of the mean difference (Table/Fig 5).

(Table/Fig 4): Mean number of micronucleated cells in different groups under different stains.

(Table/Fig 5): Multiple comparisons bonferroni*The mean difference is significant at the .05 level
The mean number of the micronuclei which was found among the control group by using the PAP stain was 6.13 +/- 2.29, whereas that which was obtained by using the MGG stain was 3.53 +/- 1.407. Similarly, the mean number of micronuclei among the smokers which was obtained by using the PAP stain was higher than that which was obtained by using the MGG stain i.e. 22.07 +/- 5.88 and 17.67 +/- 5.76, respectively. Again, among the smokeless tobacco chewers, the Pap stain smears showed more number of micronuclei ie. 45 +/- 6.18 than the MGG stained smears, where the mean number of micronuclei was 38.6 +/- 6.51. As evident, the mean number of micronuclei cells was higher in the PAP stained smears in all the three groups than in the MGG stained smears. But the difference in the number of micronuclei in the MGG and PAP stained smears among the groups was not statistically significant. The mean number of micronuclei was significantly higher in smokeless tobacco users than in the smokers, non smokers and the non chewers in both the Pap and the MGG stained smears. Mutiple comparisons showed the mean difference to be highly statistically significant amongst the three groups in both the PAP and MGG stained smears (Table/Fig 5).

Discussion

The usefulness of a micronucleus test to detect and quantitate the genotoxic action of carcinogens and mutagens has been well established in vitro as well as in vivo studies (35). The sensitivity of the micronucleus test is comparable to that of scoring chromatid breaks and exchanges. A reasonable relationship between the carcinogenicity of chemicals and their capacity to induce micronuclei, as well as the ease of scoring, stimulated the application of the micronucleus test to exfoliated human cells. The MN assay can be used for exfoliated cells, which offers the advantage of conducting a genotoxicity test on material from an intact organism with its multitude of defense systems (35).

Our findings showed that the results of the micronuclei assay in exfoliated oral mucosal cells of tobacco users and non tobacco users depended strongly on the staining method. According to the results of the present study, for the routine micronuclei assay, PAP, which is the most commonly used cytological stain, was found to show better staining results as compared to the MGG, a Romanowsky’s stain which is used widely in field studies. As for the PAP stain, the smear was wet fixed in alcohol which gave a clear background when compared to MGG, where the smear was air dried and resulted in a background which was full of cell debris and salivary proteins, thus masking the counting of the micronuclei (Table/Fig 6) and (Table/Fig 7).

(Table/Fig 6): PAP stained smear showing micronuclei in exfoliated buccal mucosal cells.


(Table/Fig 7): MGG stained smear showing micronuclei in exfoliated buccal mucosal cells

These findings were consistent with the findings of Sohair etal (36) who concluded that Pap stain was the preferred method in field studies for scoring and detecting MN in the cells of the buccal mucosa.

Hence, only Pap stained micronuclei values were used for multiple comparisons among the different groups.

As seen in the results section, the percentage of MN cells was significantly higher in smokeless tobacco users than in the non smokers/ non users and the mean difference between the two was statistically highly significant (Table/Fig 4) and (Table/Fig 5). These findings are consistent with the recent studies of Sellapa et al (37) and Patel et al (38) where the MN count in smokeless tobacco users were higher than that in the control group. Carcinogenic and mutagenic compounds, including tobacco-specific nitrosamines, which are present in smokeless tobacco forms (39), are believed to be responsible for the induction of micronuclei. These compounds are produced from nicotine by bacterial or enzymatic activity. The same formation occurs in the mouth under the influence of saliva (40).

In a study by Suhas et al (41) on buccal cell changes which are associated with smoking by using the micronucleus assay, there was found to be a significant correlation between the habit of smoking and the frequency of the micronucleated oral mucosal cells. The results of present study are in accordance with this study.

Ozkul etal (42) compared micronuclei formation in the buccal mucosal cells of habitual Maras powder (a form of smokeless tobacco) users with those of the smokers and found that the mean percentage of the micronuclated cells was 1.86+/- 0.26 in the users and 1.99+/-0.33 in the smokers. But there was no difference between themean percentages of the micronucleated cells in these two groups (p>0.05). In the present study, the mean percentage of the micronucleated cells was 4.5 +/- 0.61 in smokeless tobacco users while it was 2.20+/- 0.5 in the smokers. The mean difference between the two groups was highly statistically significant.

The risk of cancer in smokeless tobacco users has been attributed to the presence of tobacco specific nitrosamines (TSNAs) (43). There are four principal compounds: N-nitrosonornicotine (NNN), 4-methyl-N-nitrosamino-1-(3-pyridyl)-1-butanone (NNK), N-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB). Only two TSNAs, NNN and NNK, are considered to be potential carcinogens (IARC, 1985a). On the other hand, both NAT and NAB which are designated by IARC are not classifiable with regards to their carcinogenicity (IARC, 1985b). In India, SLT processing is performed by individual farmers and small companies with little control over fermentation and curing, which increases the production of TSNAs (43). Also, SLT is not homogeneous in India, since the tobacco is often combined with betel leaf (Piper betle) and sliced areca nut (Areca catechu) and/or powdered slaked lime, which are additives that enhance the toxicity as well as the psychotropic effect of tobacco (44).
Also, studies have proven that the nicotine content of the Indian brands of smoking tobacco is slightly high as compared to that of the international brands (45). The nicotine content in commercially available chewing tobacco products was found to be much lower than that in the smoking form of tobacco, but the average daily consumption has made it comparable to the smoking form (45).
According to the centre for disease control (CDC), chewing tobacco which was used 7-8 times a day could be equivalent to smoking 30-40 cigarettes per day. Other factors such as the use of slaked lime and continuous contact with the oral mucosa, led to more absorption of nicotine through smokeless tobacco use. Additionally, in contrast to the smokers who absorbed nicotine primarily through the pulmonary vasculature, chewing tobacco users were found to absorb nicotine through the buccal mucosa and the gastrointestinal tract mucosa.

Chewing tobacco is said to have increased the carcinogenic and genotoxic potential. The odds for oral cancer are estimated to be 7.3 in smokers, 1.3 in alcoholics and 11.4 in those who are habituated to chewing tobacco (46).

Conclusion

This study concludes that Pap is a better stain over MGG for the micronucleus assay screening of the buccal cells. The severity of the buccal changes which were associated with smokeless tobacco use was more than that in smokers, thus indicating the more genotoxic effect of smokeless tobacco. The limitation of this study was the sample size which could have been larger. This technique is primitive and further research by using fluorescent dyes and molecular markers is recommended.

References

1.
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