Decoding the link associated with areca nut chewing and cardiovascular disease using hsCRP as a biomarker.

Background: Chewing of areca nut is associated with various oral and systemic ill effects. The deleterious oral effects of areca nut have been widely documented to range from reactive lesions, periodontal health compromise to oral pre-cancer and cancerous states and have been described widely in the literature. The reported systemic effects of areca nut chewing are relatively less documented but reported in literature. Areca nut chewing may predispose to cardiovascular disease due to a systemic inflammatory response from the elevated levels of circulating inflammatory mediators. High-sensitivity C-reactive protein (hsCRP) is a systemic inflammatory biomarker to assess the risk of cardiovascular disease. Materials and Methods: This cross-sectional, observational study was conducted among areca nut chewers (n = 50) and non-chewers (n = 50). Areca nut chewers were considered as chewers/cases. Blood samples were collected from the participants and hsCRP levels were studied. The data were analysed using SPSS software, version 21 for statistical significance. Chi-square test was used to compare categorical variables. Mann-Whitney U test was done to analyse continuous variables. The level of statistical significance was set at P value < 0.05. Results: hsCRP mean was higher among the chewers (2.3 ± 3.7) compared to non-chewers (0.9 ± 1.3). The difference in hsCRP levels between the study group was statistically significant (P = 0.002). Conclusion: This study demonstrates the link between areca nut and cardiovascular disease using hsCRP as biomarker.

Evaluation of stress among dental undergraduates and postgraduates using salivary cortisol as a biomarker.

Background: Stress is a non-specific response of the body to any demand. Professional education has a certain level of stress inherent in it. Dental students have been reported in the literature to experience more stress than medical students. Objective: To evaluate the stress levels of dental students: first-year undergraduates, house surgeons, and final-year postgraduates, using salivary cortisol as a stress biomarker. Materials and Methods: Dental students above 18 years of age and without any systemic illness were included in the study. Those students with a drug history of corticosteroids, anti-depressants, and oral contraceptives were excluded from the study. Female participants experiencing pre-menstrual symptoms, pregnancy, and familial stressor issues were excluded from the study. First-year undergraduates, house surgeons, and final-year postgraduates were enrolled in the study. Unstimulated whole saliva was collected in the morning and evening between 9:00 and 10:00 am and between 3:00 and 4:00 pm. Salivary cortisol levels were assessed by chemiluminescence immuno-assay (CLIA). Results: Paired sample t-test was used to compare the diurnal variation among morning and afternoon samples within the groups. The salivary cortisol levels were found to be significant within the groups. One-way ANOVA was used to compare the salivary cortisol levels between the groups. The salivary cortisol levels within the groups were not statistically significant. Conclusion: Salivary cortisol is a non-invasive biomarker of stress. Postgraduates had higher salivary cortisol levels than other study groups.

Study of Hypoxia-inducible factor-2α expression in the malignant transformation of Oral submucous fibrosis.

CONTEXT: Hypoxia-inducible factor (HIF)-2α is overexpressed in primary and metastatic human cancers, whose expression is correlated with tumor angiogenesis and patient mortality. HIF plays a role in the progression of fibrosis in oral submucous fibrosis (OSF). AIM AND OBJECTIVE: The aim is to study and compare the expression of HIF-2α in OSF (a), oral squamous cell carcinoma (OSCC) with areca nut usage (b), OSCC without areca nut usage (c) and normal mucosa (d) by immunohistochemistry. SUBJECTS AND METHODS: Immunohistochemical detection of HIF-2α was done on 51 samples, which included 11 cases (a), 15 cases (b), 15 cases (c) and the expression was compared with that of (d). STATISTICAL ANALYSIS: Data were analyzed using the SPSS™ software (ver. 21.0). Chi-square test and kappa analysis were performed to compare the intensity of staining between the groups and for inter-observer agreement, respectively. Value of P ≤ 0.05 was considered statistically significant. The mean labeling index between the groups was studied by the Kruskal-Wallis test. RESULTS: All the cases of (d), (a), (b) and (c) showed HIF-2α expression (P = 0.329). About 13% cases of (c) showed intense expression (P = 0.406) and 27% of (a) cases showed expression only in the connective tissue (P = 0.023). The number of positively stained nuclei in both (b and c) cases reduced as the tumor progression was from well to poorly differentiated. CONCLUSION: Areca nut initiates fibrosis and subsequent hypoxia in OSF which triggers HIF-2α expression in the epithelium. HIF-2α could be a surrogate marker for cancer initiation and progression.

Loricrin expression and its implication in oral submucous fibrosis, hyperkeratosis and normal mucosa with association to habits - An immunohistochemical study.

OBJECTIVE: The coarse fibres of areca nut and the continuous friction from occluding teeth are major causes of mechanical stress to the oral mucosa in conditions like oral submucous fibrosis and frictional keratosis. The continuous micro trauma provided in areca nut chewers, creates an environment where the keratinocytes exhibit alteration. Loricrin, is expressed abundantly in keratinizing epithelium in response to mechanical stress. Their expression or absence could play a role in malignant transformation. This study attempts to assess the potential of Loricrin as an early diagnostic marker in patients with chewing habit. METHODS: 73 archival samples of formalin fixed, paraffin embedded tissue specimens histopathologically confirmed, were segregated as normal mucosa 11, hyperkeratotic 32 and oral submucous fibrosis 30 and stained with antibodies to Loricrin and graded as negative, mild, moderate and intense based on the staining intensity. Pearson's chi square test was done for statistical analysis. RESULTS: Loricrin expression was observed in all groups with staining in the stratum granulosum showing a significant association to habits (P = 0.000). CONCLUSION: This prominent staining indicates a compensatory cytoskeletal rearrangement of surface epithelium during cell division in early oral submucous fibrosis showing potential as an early marker of the condition.

Identification of Helicobacter pylori in Saliva of Patients with and without Gastritis by Polymerase Chain Reaction.

AIMS AND OBJECTIVE: The aim of this study was to identify the presence of Helicobacter pylori in saliva of patients with and without gastritis by polymerase chain reaction (PCR) method. MATERIALS AND METHODS: The study comprised 20 patients in Group I presenting with various symptoms of gastritis and 10 asymptomatic subjects in Group II. The intestinal endoscopy antral biopsies were collected from 20 symptomatic patients with gastroduodenal disorders. The saliva specimens were taken from all patients before endoscopy. PCR was performed using genomic DNA, isolated from the saliva and the biopsies of the patients as the template to detect the presence of the 16S ribosomal RNA gene in H. pylori. RESULTS: In Group I, 10 (50%) cases of clinical gastritis were positive for H. pylori by endoscopy biopsy and 10 (50%) were negative. Of the 10 endoscopy biopsy positive cases for H. pylori, eight were PCR positive in saliva and two were negative. Of the 10 endoscopy biopsy negative cases, three were PCR positive for H. pylori in saliva and seven were negative. In Groups II, four were symptomatic for gastritis and six were negative. Of the six gastritis negative cases, three were PCR positive, four were gastritis positive, and three were PCR positive. Sensitivity and specificity of PCR were found to be 80% and 70%, respectively. The positive predictive and negative predictive values of PCR in saliva were 72.7% and 77.7%, respectively. CONCLUSION: PCR analysis of saliva may be handy in identification of H. pylori and serves as a noninvasive technique to diagnose and monitor the prognosis.

Awareness of Forensic Odontology among Legal Professionals, Chennai, India.

BACKGROUND: The forensic discipline of law is a multidisciplinary team comprising of specialists in forensic medicine, forensic odontology, security and law. AIM: The study was to find the awareness level of scope and utility of forensic odontology among lawyers in Chennai, South India. MATERIALS AND METHODS: A cross-sectional study using a self administered structured questionnaire was conducted in 200 lawyers between August and September of 2013. The data was analyzed depending on age, gender, type and years of practice. RESULTS: Lawyers above 40 years of experience were more aware of palatal rugae analysis (P = 0.02), and those with more than 20 years were aware of lip print (P = 0.001) and bite mark analysis (P = 0.001). Males were more aware of forensic odontology with respect to criminal identification (P = 0.001). The knowledge of bite mark analysis was higher among male lawyers (P = 0.001), civil and criminal practicing lawyers (P = 0.004). All participants were aware that loss or fracture of tooth constitutes a grievous injury under Indian Penal Code (IPC) 320 clause 7(5). CONCLUSION: This study highlighted the knowledge of forensic odontology among legal professionals and also identified the areas in which they need further appraisal.

On factors influencing the absorption capacity of surgical sponges.

BACKGROUND: Cotton balls and surgical gauzes (CSG) have been routinely used for mopping fluids and blood in surgeries. Polyurethane sheets (PUS) are increasingly used instead of CSG. The factors influencing the absorption of fluids by CSG and PUS have not been studied in detail. There are a variety of factors that could be modified by the manufacturer while a limited number of them could be manipulated by the operator. MATERIALS AND METHODS: Mathematical models and equations have been employed in this study. The impact of absorbency was performed by modulating various factors. RESULTS AND DISCUSSION: A variety of factors such as thickness, density of the web, radius of fiber, temperature, surface tension and viscosity of fluid, all influenced the absorption rate and capacity to varying degrees. The optimal variation by which the PUS can be effectively used is discussed. The suggested mathematical model will help the operator to choose the appropriate type of PUS based on the individual's needs. CONCLUSION: In an area of rapidly developing technology and change in biomaterials, where the choice of such materials are overwhelming, hopefully this basic knowledge will help the clinician to make a more educated and rational decision.

Immunohistochemical detection of p53 and p63 in oral squamous cell carcinoma, oral leukoplakia, and oral submucous fibrosis.

AIM: Oral squamous cell carcinoma (OSCC) may be preceded by potentially malignant disorders such as leukoplakia and oral submucous fibrosis (OSF). p63 in concert with p53 regulates cell proliferation and differentiation and may have a role in potentially malignant and malignant lesions of the oral cavity. The aim of the study was to evaluate and compare the expression of p53 and p63 proteins in OSCC, leukoplakia, and OSF by immunohistochemistry (IHC). METHODS: Tissue sections of OSCC (n = 20), leukoplakia (n = 20), OSF (n = 20) and normal oral mucosa (n = 10) were stained with p53 and p63 antibodies by IHC. Mean labeling index (LI) among the study groups were compared using the Kruskal-Wallis test. RESULTS: The mean LI of p53 for OSCC, leukoplakia, OSF, and normal mucosa were 56.9 ± 21.3, 37.6 ± 12.6, 34.6 ± 8.7 and 15.1 ± 9, while mean LI of p63 were 56.8 ± 19.6, 42.3 ± 10.5, 32.8 ± 12.1, and 26.4 ± 9.4. The mean LI of p53 and p63 were significantly higher in OSCC, leukoplakia and OSF compared to normal mucosa (P < 0.01; P < 0.01). CONCLUSION: The significant increase in expression of p53 and p63 proteins in OSCC, leukoplakia, and OSF suggests their role as surrogate markers of malignant transformation.

Impact of highly active antiretroviral therapy on salivary flow in patients with human-immuno deficiency virus disease in Southern India.

AIMS: To ascertain and compare between highly active antiretroviral therapy (HAART) and non-HAART patients, the stimulated salivary flow rates and unstimulated salivary flow rates (USFR and SSFR) and to correlate the salivary flow rates with immune suppression. MATERIALS AND METHODS: One hundred human-immuno deficiency virus seropositive patients attending RAGAS-YRG CARE were examined and divided into two groups, a HAART group (patients on combination antiretroviral therapy) comprising 50 patients and a non-HAART group comprising 50 patients. The HAART group was followed every 3 months after the baseline visit (0) for a period of 9 months, during which a clinical oral examination and collection of unstimulated and stimulated saliva was done. Their salivary gland function was assessed using a xerostomia inventory during each visit. The study on non-HAART group was cross-sectional. STATISTICAL ANALYSIS: Statistical analysis were performed with the aid of the Statistical Package for the Social Sciences (SPSS version 10.05) software. RESULTS: There was no significant difference in mean SSFR and USFR between the two groups at baseline. In the HAART group, the mean stimulated salivary flow rate increased from baseline to 3 months (P = 0.02), with the increase being maintained at 6 months and 9 months. When salivary flow rates were correlated with Cluster of Differentiation, CD4 counts, patients in the HAART group with a CD4 ≤ 200 at 6 months visit had a higher mean stimulated salivary flow rate when compared with patients with CD4 ≥ 200 (P = 0.02). The xerostomia inventory did not reveal any significant difference between the two groups and HAART was not significantly associated with xerostomia. CONCLUSION: In our study HAART was neither associated with xerostomia nor a reduction in salivary flow rate and immune suppression was not a significant factor for decreasing the salivary flow rate.

Expression of p53 and Cyclin D1 in oral squamous cell carcinoma and normal mucosa: An Immunohistochemical study.

AIM: To assess p53 and Cyclin D1 expression using Immunohistochemistry in normal mucosa and oral squamous cell carcinoma. MATERIALS AND METHODS: Twenty cases of Oral squamous cell carcinoma (OSCC) and 10 normal mucosa were used and the primary antibodies used were p53 (DAKO-DO7) and Cyclin D1 Mouse Anti human Cyclin D1 (DCS-6) 1: 100 dilution. STATISTICAL ANALYSIS: Labelling index was calculated and mean LI and SD were calculated, using Descriptive Statistics and t-test was used to compare mean LI between antibodies used in OSCC. Percentage positivity was done by Chi-Square test. Comparison of LI between p53 and Cyclin D1 was studied using t test. RESULTS: p53 was positive in 30% and Cyclin D1 in40% of normal cases and 65% and 95% of OSCC were positive for p53 and Cyclin D1 respectively. Mean LI of p53 and Cyclin D1 were found to be statistically significant between the normal mucosa and OSCC. The correlation of mean LI of p53 and Cyclin D1 in OSCC was found to be statistically significant. LI of p53 was found to be higher than Cyclin D1 in OSCC. CONCLUSION: In the present study, increased p53 and Cyclin D1 expression were seen in OSCC when compared to the normal mucosa and a positive correlation was seen between increased p53 and Cyclin D1 expression in OSCC.

Prevalence and correlates of tobacco use among urban adult men in India: a comparison of slum dwellers vs non-slum dwellers.

BACKGROUND: Tobacco use is reported to be rampant in urban slums in developing countries. Demographical variations in tobacco use between males living in urban slums vs those living in non-slum areas in India has not been reported, and this study was undertaken to address this issue. MATERIALS AND METHODS: Secondary data analysis of National Family Health Survey-3 (NFHS-3) was undertaken to study demographical variations in tobacco use between urban slum dwellers and non-slum dwellers in eight Indian cities. Demographic determinants for use of smoking and chewing forms of tobacco in the two groups were analyzed. SPSS version 16.0 (SPSS Inc., IL, USA) was used for statistical analysis. RESULT: The study population comprised 6887 (41.8%) males from slum areas and 9588 (58.2%) from non-slum areas of eight urban cities. Cigarette/beedi smoking was the commonest form of tobacco use among the study population. Pan masala use was the least common form of smokeless tobacco use, next only to snuff. There was a high statistical significance observed within the various demographic parameter studied in both the slum and non-slum dwelling males in study population. However, on studying the differences between the two groups, it was observed that statistical significance of P≤.001 was observed with age (15-49), secondary education, religion, household structure and marital status. The difference between the two groups in the mean number of cigarettes/beedis smoked was not statistically significant (P=.598). DISCUSSION AND CONCLUSION: Male slum dwellers are a distinct urban population, whose health needs assessment requires a different approach than that for non-slum dwellers who often can afford the services that an urban Indian city can offer.

Potential role of septins in oral carcinogenesis: An update and avenues for future research.

Septins belong to the GTPase superclass of conserved proteins and have been identified to play a role in diverse aspects of cell biology, from cytokinesis to the maintenance of cellular morphology. At least 14 septins have been identified in humans. With their complex patterns in gene expressions and interaction, it has been reported that alterations in septin expression are observed in human diseases. Although much is not known about the role of human septins in oral carcinogenesis, circumstantial evidence does indicate that it may play a major role. This review intends to summarize the basis of septin biology, with the focus being on the evidence for septin involvement in human oral cancer.

Concepts in sample size determination.

Investigators involved in clinical, epidemiological or translational research, have the drive to publish their results so that they can extrapolate their findings to the population. This begins with the preliminary step of deciding the topic to be studied, the subjects and the type of study design. In this context, the researcher must determine how many subjects would be required for the proposed study. Thus, the number of individuals to be included in the study, i.e., the sample size is an important consideration in the design of many clinical studies. The sample size determination should be based on the difference in the outcome between the two groups studied as in an analytical study, as well as on the accepted p value for statistical significance and the required statistical power to test a hypothesis. The accepted risk of type I error or alpha value, which by convention is set at the 0.05 level in biomedical research defines the cutoff point at which the p value obtained in the study is judged as significant or not. The power in clinical research is the likelihood of finding a statistically significant result when it exists and is typically set to >80%. This is necessary since the most rigorously executed studies may fail to answer the research question if the sample size is too small. Alternatively, a study with too large a sample size will be difficult and will result in waste of time and resources. Thus, the goal of sample size planning is to estimate an appropriate number of subjects for a given study design. This article describes the concepts in estimating the sample size.

Tooth decay in alcohol and tobacco abusers.

BACKGROUND: Alcohol and tobacco abuse are detrimental to general and oral health. Though the effects of these harmful habits on oral mucosa had been demonstrated, their independent and combined effect on the dental caries experience is unknown and worthy of investigation. MATERIALS AND METHODS: We compared 268 alcohol-only abusers with 2426 alcohol and tobacco abusers in chewing and smoking forms to test the hypothesis that various components of their dental caries experience are significantly different due to plausible sociobiological explanations. Clinical examination, Decay, Missing, Filled Teeth (DMFT) Index and Oral Hygiene Index - Simplified were measured in a predetermined format. Descriptive statistics, Chi-square test and one-way ANOVA analysis were done using SPSS Version 16.0. RESULT: The mean DMFT were 3.31, 3.24, 4.09, 2.89 for alcohol-only abusers, alcohol and chewing tobacco abusers, smoking tobacco and alcohol abusers, and those who abused tobacco in smoke and smokeless forms respectively. There was no significant difference between the oral hygiene care measures between the study groups. Presence of attrition among chewers and those with extrinsic stains experienced less caries than others. DISCUSSION AND CONCLUSION: The entire study population exhibited a higher incidence of caries experience. Use of tobacco in any form appears to substantially increase the risk for dental caries. Attrition with use of chewing tobacco and presence of extrinsic stains with tobacco use appear to provide a protective effect from caries. The changes in oral micro-flora owing to tobacco use and alcohol may play a critical role in the initiation and progression of dental caries.

Oral lichen planus: An update on pathogenesis and treatment.

Oral lichen planus (OLP) is a chronic inflammatory disease that affects the mucus membrane of the oral cavity. It is a T-cell mediated autoimmune disease in which the cytotoxic CD8+ T cells trigger apoptosis of the basal cells of the oral epithelium. Several antigen-specific and nonspecific inflammatory mechanisms have been put forward to explain the accumulation and homing of CD8+ T cells subepithelially and the subsequent keratinocyte apoptosis. A wide spectrum of treatment modalities is available, from topical corticosteroids to laser ablation of the lesion. In this review, we discuss the various concepts in the pathogenesis and current treatment modalities of OLP.

Rapidly enlarging mass following dental extraction.

Oral melanoma (OM) is a rare, malignant neoplasm of melanocytic origin. It accounts for 0.2 - 8% of all melanomas. In contrast to its cutaneous counterpart, OM has poor prognosis, possibly due to late clinical diagnosis, nonexistence of standardized clinical or histopathological grading and anatomical limitation in complete excision of the lesion. For better understanding of the lesion, case reports are still considered to be the source of information. Here we present a case of oral primary melanoma occurring in an uncommon site, the mandibular gingiva, with review of related literature.