The effect of Maras powder and smoking on the microRNA deregulation of oral mucosa

Abstract Objective This study aimed to investigate the effects of Maras powder (a type of smokeless tobacco obtained from Nicotiana rustica Linn and mixed with the ashes of wood, especially from oak, walnut or grapevine) on the microRNA (miRNA) deregulation of oral mucosa, and it compares these effects with those of smoking. Methodology Oral mucosal samples were collected from 74 patients, consisting of 16 nonusers, 26 smokers, and 32 Maras powder users. Genes associated with oral cancer were selected and 90 microRNAs targeting these genes were identified. MicroRNA were isolated and purified using the microRNA isolation kit. MicroRNA were expressed using Fluidigm RT-PCR. Results A positive correlation between the duration of Maras powder use with miR-31 expression levels, and a negative correlation between the Maras powder chewing time and miR-372 expression levels was found. In addition, there is a negative correlation between the amount of Maras powder consumed and expression levels of miR-375, miR-378a, miR-145, and miR-10b; moreover, another negative correlation is observed between the number of cigarettes consumed and the expression levels of miR-23a, miR-23b, miR-203a, miR-200b, and miR-375. However, miR-200b and miR-92a levels were downregulated significantly more in Maras powder users when compared with smokers and nonusers (p<0.05). Conclusion The results show both chewing Maras powder and smoking have an effect on deregulation of miR-200b and miR-92a expressions. This leads to the belief that assessing the expression of these two miRNAs is a promising noninvasive method of analysis, especially in mutagen exposures. Finally, large-scale and high-throughput studies may help to identify an extensive miRNA expression profile associated with tobacco use and improve the understanding of oral malignancies.

Study of Some Cardiovascular Risk Factors in Tobacco Chewers and Non Tobacco Chewers.

Tobacco chewing is known causative factor of cardiovascular disorders and cancer. In India tobacco and gutkha chewing is very common. In present study effect of tobacco chewing was studied on other cardiovascular risk factors. This is comparative study done to see the association of smokeless tobacco consumption with blood pressure, heart rate, BMI and serum cotinine in adult male tobacco chewers. Methods: In present study 175 apparently healthy males were selected. Out of these 80 were tobacco non chewers and 95 were tobacco chewers. Parameters such as BMI, B.P., heart rate and serum cotinine were measured in both the groups. Result: No significant difference were observed among tobacco chewers and controls for heart rate and BMI, however, significant increase was observed in systolic and diastolic blood pressure (p > 0.001) in tobacco chewers as compared to tobacco non chewers. Conclusion: Significantly increased values of systolic and diastolic blood pressure indicated that tobacco consumption in any form increases the risk for cardiovascular diseases.

Smokeless tobacco use: A meta-analysis of risk and attributable mortality estimates for India.

BACKGROUND: Use of smokeless tobacco (SLT) is widely prevalent in India and Indian subcontinent. Cohort and case–control studies in India and elsewhere report excess mortality due to its use. OBJECTIVE: The aim was to estimate the SLT use‑attributable deaths in males and females, aged 35 years and older, in India. MATERIALS AND METHODS: Prevalence of SLT use in persons aged 35 years and older was obtained from the Global Adult Tobacco Survey in India and population size and deaths in the relevant age‑sex groups were obtained from UN estimates (2010 revision) for 2008. A meta‑relative risk (RR) based population attributable fraction was used to estimate attributable deaths in persons aged 35 years and older. A random effects model was used in the meta‑analysis on all‑cause mortality from SLT use in India including four cohort and one case–control study. The studies included in the meta‑analysis were adjusted for smoking, age and education. RESULTS: The prevalence of SLT use in India was 25.2% for men and 24.5% for women aged 35 years and older. RRs for females and males were 1.34 (1.27–1.42) and 1.17 (1.05–1.42), respectively. The number of deaths attributable to SLT use in India is estimated to be 368127 (217,076 women and 151,051 men), with nearly three‑fifth (60%) of these deaths occurring among women. CON CLUSION: SLT use caused over 350,000 deaths in India in 2010, and nearly three‑fifth of SLT use‑attributable deaths were among women in India. This calls for targeted public health intervention focusing on SLT products especially among women.

Rising incidence of oral cancer in Ahmedabad city.

CONTEXT: In 1999, an increase in mouth cancer incidence among young men (<50 years) in urban Ahmedabad was reported to be occurring along with decreasing mouth cancer incidence in older age groups and increasing oral submucous fibrosis incidence associated with areca nut consumption among young men in Gujarat. The aim was to investigate whether the increase in the incidence mouth cancer that had started among young men in the 1990s was continuing. SETTINGS AND DESIGN: Ahmedabad urban population, comparison of reported mouth cancer cases in the population across four time period. METHODS: Age‑specific incidence rates of mouth cancer (International Classification of Diseases [ICD]‑9:143–5; ICD‑10:C03–06) in five year age groups among men aged ≥15 years for the city of Ahmedabad for years 1985, 1995, 2007 and 2010 were extracted from published reports. For comparison, lung cancer (ICD‑9:169; ICD‑10:C33–C34) rates were also abstracted. STATISTICAL ANALYSIS USED: A cohort approach was used for further analysis of mouth cancer incidence. Age adjusted incidence rates of mouth and lung cancer for men aged ≥15 years were calculated and compared. RESULTS: The age specific incidence rates of mouth cancer among men increased over the 25‑year period while lung cancer rates showed a net decrease. Using a cohort approach for mouth cancer, a rapid increase in younger age cohorts was found. CONCLUSIONS: Mouth cancer incidence increased markedly among men in urban Ahmedabad between 1985 and 2010, apparently due to increasing consumption of areca nut products, mawa and gutka. Gutka has now been banned all over India, but a more vigorous implementation is necessary.

A review of Indian literature for association of smokeless tobacco with malignant and premalignant diseases of head and neck region.

In India, about 60% of tobacco users use smokeless tobacco (ST) alone. Head and neck squamous cell carcinoma is one of the most common cancers in India. International Agency for Research on Cancer (IARC) monograph (Vol 89) found a significant association between ST use and oral cancer. However, only a few articles from India were included in this monograph. To overcome this lacuna, we have reviewed the articles published from India investigating the association between ST use and malignant and premalignant diseases of head and neck region. Data collection has been performed by computer-aided search of the MedLine and PubMed databases using different combinations of the key words. For malignant lesions, only cohort and case control studies were considered for review. For premalignant lesions and dental diseases other than case control studies, some cross-sectional studies have also been reviewed. Studies found a significant association between ST use and cancer of the oral cavity. The association was stronger for the buccal mucosa compared to tongue and for females compared to males. Significant association noted between cancer of the hypopharynx and oropharynx with ST use but no definitive association noted for cancer of the larynx and nasopharynx. Some dental disease and oral premalignant conditions were also associated with ST use. Indian studies suggest ST use is strongly associated with cancer of the oral cavity, oropharynx and hypopharynx.

Chemistry and toxicology of smokeless tobacco.

In most parts of the world, tobacco is used for smoking, whereas, in India, tobacco is used for smoking as well as in diverse smokeless forms. Absorption of toxic and carcinogenic chemicals in tobacco and other ingredients added to various products are causally associated with several non-communicable diseases including cancer, especially oral cancer, which is the leading cancer among men and the third most common cancer among women in India. This article highlights the toxicity, mutagenecity and carcinogenic effects of hazardous chemicals present in smokeless tobacco products. This endeavor was based on the extensive review of literature from various sources. The SLT products have influence on cellular metabolism, ability to cause DNA damage, and cancer in experimental animals. It is, therefore, essential to consider the collective role of chemical constituents of SLT products in the causation of adverse effect on human health.

Smokeless tobacco use among patients with tuberculosis in Karnataka: The need for cessation services.

Background. India is home to the largest population of patients with tuberculosis and tobacco users in the world. Smokeless tobacco use exceeds smoking and is increasing. There is no study to date that reports smokeless tobacco use before and after the diagnosis and treatment of tuberculosis. We assessed smokeless tobacco use among former patients of tuberculosis in Karnataka, India. Methods. We conducted a community-based, crosssectional study among 202 men, who had been diagnosed and treated for tuberculosis (mean age 48 years), selected by multistage, random sampling. Using a semi-structured interview schedule, retrospective smoking and smokeless tobacco use were captured at eight time-points before and after the diagnosis and treatment of tuberculosis. Results. Most patients suspended tobacco use during treatment. A high 44% prevalence of smokeless tobacco use 6 months before diagnosis was reduced to just 8% during the intensive phase of treatment and climbed to 27% 6 months after treatment. The tobacco use relapse rate 6 months after completion of treatment was higher for smokeless tobacco use (52%, 95% CI 41%–62%) than for smoking (36%, 95% CI 26%–45%). We also found that many patients who were advised to quit smoking continued using smokeless tobacco after completion of treatment. Additionally, new smokeless tobacco use was documented. Of the 11 new exclusive smokeless tobacco users, 10 shifted from smoking to smokeless tobacco use as a form of harm reduction. Conclusion. Patients with tuberculosis are advised by their doctors, at the time of diagnosis, to quit smoking. Several patients shift from smoking to smokeless tobacco use, which needs to be addressed while providing tobacco cessation services.

Relationship between ABO blood groups and oral cancer.

Background: Cancer is a unique disease characterized by abnormal growth of cells which have the ability to invade the adjacent tissues and sometimes even distant organs. Oral cancer has multifactorial etiology and is significantly associated with risk factors of the individual's lifestyle, particularly, chronic use of tobacco, spicy food, alcohol and smoking. Many studies have indicated that genetic factors also have an influence on the etiology of cancer. Aim: To evaluate if any of the ABO blood groups are associated with an increased risk for oral cancer. Materials and Methods: The present study was conducted at Kidwai Memorial Institute of Oncology (KMIO), Bangalore, after obtaining permission from the Director of the institute. The study sample comprised 235 oral cancer patients and 812 controls. For statistical analysis, Chi-square test and odds ratio were used to assess the relationship between ABO blood groups and oral cancer. Probability level was fixed at ≤0.05. Results: It was found that people with blood group A had 1.46 times higher risk of developing oral cancer compared to people of other blood groups. Conclusion: By employing a simple blood grouping test during community field programs, people with blood group A in the age group of 40-59 years having tobacco chewing habits can be apprised that they are more at risk to develop oral cancer than people with other blood groups.

Tobacco Consumption Practices among the School Students.

Research question: What is the prevalence of tobacco consumption in school students of Anand Taluka in Gujarat. Settings: Urban and rural areas of Anand taluka of Gujarat. Study design: A cross-sectional study. Participants: School students of class 11th and 12th of selected schools. Methodology: Sample size taken for the study purpose was 1200 in accordance with the prevalence of tobacco consumption in Kheda district of Gujarat. The study was conducted with the help of pre-tested, self-administered questionnaire. Urban and rural areas were selected by simple random sampling and school students were selected by systematic random sampling. Results: The prevalence of current tobacco users was found to be 8.2%. Chewing is the most preferable form of tobacco use. Maximum number of users started tobacco use at the age of 11-15. The study showed a lack of knowledge in the students regarding the consequences of tobacco use. Friend’s and teacher’s smoking behaviour is significantly associated with student’s tobacco consumption.

Evaluation of DNA damage in oral precancerous and squamous cell carcinoma patients by single cell gel electrophoresis.

Context: Single cell gel electrophoresis (SCGE) or "comet assay" is a rapid and very sensitive fluorescent microscopic method for detecting various forms of DNA damage at individual cell level. Aims: The aim of the present study was to detect the extent of DNA damage in oral cancer, oral submucous fibrosis (OSMF) and leukoplakia in comparison to normal individual. Settings and Design: A total of 44 consecutive patients with oral cancer (n=26), leukoplakia (n=12) and OSMF (n=6) and 10 healthy normal volunteers with normal oral epithelia (controls) were recruited from Dr. R. Ahmed Dental College and Hospital and were assessed for the extent of DNA damage using SCGE following clinical diagnosis. Materials and Methods: Peripheral blood was collected by venepuncture and comet assay was performed using SCGE. Mean tail length was compared between diagnostic groups and between different oral habit groups using t-tests and analysis of variance (ANOVA). Pearson's product moment correlation was used to examine the linear association between the extent of DNA damage and oral habit pack-years. Scheffe's pair-wise test was employed to adjust for multiple comparisons. Results: None of the controls were associated with any oral habits. Mean (±SD) tail lengths (in mm) for cancer (24.95 ± 5.09) and leukoplakia (12.96 ± 2.68) were significantly greater than in controls (8.54 ± 2.55, P<0.05). After adjustment, well-, moderately, and poorly differentiated carcinomas had significantly greater tail length than controls. Whereas the extent of DNA damage in cancer cases was significantly greater in leukoplakia than in compared to OSMF (11.03 ± 5.92), the DNA damage in latter was not different from controls. DNA damage for people with any oral habit (19.78 ± 7.77) was significantly greater than those with no habits (8.54 ± 2.55; P<0.0001). Conclusions: DNA damage measured by SCGE is greater in leukoplakia and squamous cell carcinoma, but not in OSMF. Deleterious oral habits are also associated with greater DNA damage.

Fas receptor (CD95) & Fas ligand (CD178) expression in patients with tobacco-related intraoral squamous cell carcinoma.

Background & objectives: Fas receptor and Fas Ligand (FasL) system has been implicated in the resistance to apoptosis, insensitivity to chemotherapy and in providing immune privileged status to most of the tumours. However, no reports are available on Fas and FasL expression in patients with tobacco-related oral carcinoma. Therefore, the present study was undertaken to observe Fas and FasL expression and their correlation with clinicopathological features as well as cell cycle parameters. Methods: Immunohistochemistry for Fas, FasL and DNA flow cytometry for cell cycle parameters was successfully done on 41 paraffin embedded tumour and 10 normal samples. The results were evaluated for possible association of Fas and FasL with clinicopathological features and cell cycle parameters. Results: Weak Fas expression was observed on the cell membrane only in 2 of 41 (5%) oral tumours while FasL immunoreactivity was seen in 26 of 41 (63.4%) tumours. In contrast, all ten normal oral tissues exhibited strong cytoplasmic and membrane Fas receptor immunoreactivity but absence of FasL staining. Older patients, greater tumour size and lymph node positivity were found to be associated with high expression of FasL. Significantly higher (P<0.01) expression of FasL was observed in oral tumours with aggressive DNA pattern like aneuploidy and high S-phase fraction. Interpretation & conclusions: Downregulation of Fas receptor and up-regulation of Fas ligand appear to be an important feature of tobacco-related intraoral carcinoma. Association of FasL expression with advanced clinical stage and aggressive DNA pattern suggests that the Fas and FasL system may be used as an important prognostic variable in patients with tobacco-related intraoral squamous cell carcinoma.

Verrucous carcinoma in association with oral submucous fibrosis.

Oral verrucous carcinoma is a form of well differentiated squamous cell carcinoma characterized by exophytic over growth. It is slow growing and locally invasive tumor occurring in 6 th and 7 th decade of life. Smoking and chewing tobacco is found to be the most common etiological factor of verrucous carcinoma although oral leukoplakia may act as a predisposing factor. This is a rare case of oral varrucous Carcinoma seen in association with oral submucous fibrosis in a younger patient with long standing history of chewing tobacco.

Correlation of cyclin D1 expression with aggressive DNA pattern in patients with tobacco-related intraoral squamous cell carcinoma.

Background & objectives: Cyclin D1 has been strongly implicated in cell proliferation particularly in the G1/S checkpoint of the cell cycle, and prognoses in human malignancies. We investigated the correlation between cyclin D1 overexpression and clinicopathological features as well as cell cycle parameters to understand its clinical significance in patients with tobacco-related oral squamous cell carcinoma (OSCC). Methods: Immunohistochemistry for cyclin D1 and DNA flowcytometry for cell cycle parameters was done on paraffin embedded tumour samples from 45 patients with OSCC Results: Higher expression of cyclin D1 was observed only in 30 (66.6%) of 45 cases that correlated with advanced age (P <0.02), higher tumour stage ((P<0.01), histological differentiation and lymph node metastasis (P <0.01). Analysis of nuclear DNA pattern revealed cyclin D1 immunoreactivity in tumours with aggressive DNA pattern such as aneuploidy ((P<0.05) and higher S phase fraction ((P<0.04). Interpretation & conclusions: Higher expression of cyclin D1 in oral cancer appears to be closely linked to cell proliferation, differentiation and lymph node invasion. Pre-operative evaluation of cyclin D1 in biopsy specimen may be useful in planning the most appropriate treatment strategies in patients with tobacco-related OSCC.

Trends in the epidemiology of oral squamous cell carcinoma in Western UP: An institutional study.

Objective: The purpose of the study was to identify trends in incidence rates of oral squamous cell carcinoma (OSCC) at specific anatomic sites or within specific age or sex groups in the Western Uttar Pradesh population. Materials and Methods: The study covers the period from January 2004 through April 2009. OSCC cases were retrospectively analysed for site, age, gender and habits and the findings were formulated to chart the trends in Western U.P. Results: The study revealed a male to female ratio of 2.2:1 with the largest number of OSCCs developing in the fourth and fifth decades of life. Overall, the most common site was the buccal mucosa (63.75%), followed by retromolar area (15%), floor of the mouth (11.25%), lateral border of the tongue (3.75%), labial mucosa (3.75%), and palate (2.5%). Smokeless tobacco habit was more prevalent than smoking tobacco in both men as well as women. Karl - Pearson's correlation coefficient was calculated to find the degree of association between the two variables i.e. between gender to buccal mucosa and gender to smokeless and smoking tobacco habits, which were found to be positively correlated with respect to the age. Conclusion: Oral cancer is an important cause of morbidity and mortality worldwide with an incidence rate that varies widely by geographic location. Even within one geographic location, the incidence varies among groups categorized by age, sex, site or habit.

Histological patterns and clinical presentation of oral squamous cell carcinoma

The objective of our study was to identify the different histological patterns of Oral Squamous cell carcinoma [OSCC] in relevance to demographic variables and to evaluate histological patterns according to various clinical presentations at the joint department of Oral Surgery, Karachi Medical and Dental College and Abbasi Shaheed Hospital Karachi. This descriptive study was conducted from December 2005 to November 2006 at Oral Surgery Department of Karachi Medical and Dental College and Abbasi Shaheed Hospital Karachi. A total of 60 cases presenting with OSCC were included in this study. Their demographic details, habit profile, clinical information of lesion were recorded on a proforma. All the patients underwent biopsy and their histological information was also noted on the same proforma. This study included 60 patients of OSCC. Fifty eight percent of patients were from age group between 41 -60 years. Approximately 63.3% males and 36.7% females were affected and male to female ratio was 1.7: 1. Approximately 48.3% of lesions involved alveolus. Nearly 86.7% of patients were indulged in habits of tobacco chewing. Clinical appearance of lesion showed that 66.7% of lesions were ulcerative type and histologically 48.3% were well-differentiated OSCC. Majority of patients were indulged in habit of tobacco chewing, Ulcerative type was the dominant clinical presentation whereas histologically well-differentiated OSCC was the most common presentation. Infiltration was a consistent finding in all cases

Oral snuff and carcinoma oesophagus

Carcinoma oesophagus is known to occur worldwide. Various dietary, environmental and genetic factors and certain pre-malignant conditions have been blamed in the causation of carcinoma oesophagus but the exact cause in unknown. This study was conducted to find out the possible role of oral snuff to carcinoma oesophagus. This descriptive analytical study was carried out in Medical, Surgical and ENT units of Khyber Teaching Hospital and Cardiothoracic unit of Lady Reading Hospital Peshawar from 1995 to 2005. Relation of oral snuff use to carcinoma oesophagus was studied in 100 patients confirmed on endoscopy and biopsy and histopathology. Oral snuff use was found to have very strong correlation [P<0.001] with carcinoma oesophagus. The effect of ingesting snuff also had very strong correlation [P<0.001] and so was the duration [more than 10 years] of use [P=0.004], however, the frequency of snuff use [i.e. number of time per day] did not have any significant effect [P=0.08]. A strong correlation of snuff use was also found with squamous cell carcinoma [P=0.001]. Oral snuff use is a risk factor for carcinoma oesophagus. Carcinoma oesophagus can be prevented by discouraging its use. Educating people against snuff use may prevent this harmful and lethal disease. This should be done on individual level by the doctors and on mass level through public media

Formas não habituais de uso do tabaco: [revisão] / Noncigarette forms of tobacco use: [review]

Existem diferentes produtos de tabaco, preparados de formas distintas, que podem ser divididos em produtores e não produtores de fumaça quanto a sua utilização. Dentre os que produzem fumaça, além do cigarro, citamos o charuto, cachimbo e o narguilé. O tabaco que não produz fumaça pode ser encontrado em preparações para ser mascado ou para ser absorvido pela mucosa oral ou nasal. Entretanto, todas as formas de utilização do tabaco liberam nicotina para o sistema nervoso central com risco potencial confirmado para causar dependência. Além disso, não existe forma segura de consumo do tabaco, e seus usuários têm, de forma significativa, risco aumentado para adoecimento e morte prematura por enfermidades relacionadas ao uso de tabaco.

There are many preparations for tobacco use, which can be classified as smoking or smokeless tobacco. Among the noncigarette preparations that produce smoke, we cite cigars, pipes and narghiles. Smokeless tobacco can be found in preparations for chewing or for being absorbed by nasal and oral mucosae (snuff). However, all tobacco products deliver nicotine to the central nervous system and there is a confirmed risk of dependence. In addition, there is no safe form of tobacco use, and tobacco users have a significantly increased risk of morbidity and premature mortality due to tobacco-related diseases.