Immunohistochemical Detection of Proliferative Marker Ki-67 in Benign and Malignant Salivary Gland Tumors.

Introduction: Salivary gland tumors are the most histologically heterogeneous group of tumors with the greatest diversity of morphologic features among their cells and tissues. The present study was aimed at assessing the validity of Ki-67, a cell proliferation marker, as a prognostic factor in benign and malignant salivary gland tumors and to study whether it is related to age, sex, anatomical site, and size of the lesion in salivary gland tumors. Materials and methods: A retrospective study consisted of benign salivary gland tumors (BSGTs) (n = 15), malignant salivary gland tumors (n = 18), and normal salivary gland parenchyma (n = 15). Results: There was a significant difference of Ki-67 labeling index (LI, %) in normal salivary gland parenchyma, BSGTs, and malignant salivary gland tumors. The Ki-67 LI (%) in normal salivary gland parenchyma is negligible (0.27 ± 0.31%), whereas malignant salivary gland tumors showed very high Ki-67 LI (%) of 18.79 ± 18.06% compared with BSGTs being 0.76 ± 2.02%. There was a significant correlation statistically of mean ± standard deviation (SD) of Ki-67 LI (%) with the age of the patients being the maximum (32.68 ± 15.87%) in the 50 to 59 years age group, whereas sex, site of the lesion, and size of the lesion in salivary gland tumors had no significant correlation. Conclusion: The Ki-67 is a useful marker for assessing prolif-erative potential of tumors. Clinical significance: The Ki-67 LI% can be used as a reliable adjuvant diagnostic tool to differentiate between the subtypes and grading of certain malignant tumors, such as mucoepi-dermoid carcinoma (MEC), adenoid cystic carcinoma (AdCC), and acinic cell carcinoma (AcCC), which are usually difficult to diagnose on histopathological criteria alone. Keywords: Immunohistochemistry, Ki-67, Salivary gland neoplasms.

Matrix metalloproteinases-3 gene-promoter polymorphism as a risk factor in oral submucous fibrosis in an Indian population: a pilot study.

AIM: Host-related factors could be important to the fundamental understanding of oral diseases. One such factor is matrix metalloproteinases (MMP). MMP3 (stromelysin 1) is crucial in connective tissue remodeling. Promoter regions are specific segments of DNA that control the rate of mRNA synthesis. Thus, polymorphism in the (5A/6A) promoter region of the MMP3 gene results in different transcriptional activities and is related to susceptibility in diseases. The aim of the present study was to detect polymorphism in the MMP3 gene-promoter region in oral submucous fibrosis patients (OSF), oral squamous cell carcinoma patients (OSCC), and normal individuals with tobacco and areca nut habit and without lesions, and to correlate the genotype as a risk factor in these diseases. METHOD: Genomic DNA from blood of OSF (n = 5), OSCC (n = 5), and normal individuals with tobacco and areca nut habit (n = 5) and without (n = 5) were subjected to polymerase chain reaction of the MMP3 gene-promoter region and DNA sequencing. RESULTS: The 5A allele in the MMP3 gene-promoter region was observed more frequently in the OSF group than the control groups. No significant difference was noted between OSCC and the control groups on the 5A allele. CONCLUSION: The results indicated that the 5A allele of the MMP3 gene-promoter region could be associated with OSF risk factor, but not OSCC, in an Indian population.

Detection of Porphyromonas gingivalis and Treponema denticola in chronic and aggressive periodontitis patients: A comparative polymerase chain reaction study.

BACKGROUND: The detection frequency of Porphyromonas gingivalis and Treponema denticola in chronic periodontitis (CP) and aggressive periodontitis (AgP) is not explored well in Indian population. AIM: The study was undertaken to detect P. gingivalis and T. denticola in CP as well as in AgP patients using polymerase chain reaction (PCR), and to determine the relationship between the frequency of these two microorganisms and the severity of clinical periodontal parameters. MATERIALS AND METHODS: Subgingival plaque samples were collected from ninety participants (thirty CP patients, thirty AgP patients, and thirty healthy participants) and the aforementioned two microorganisms were detected using PCR. RESULTS: However, when CP and AgP were compared for the detection frequency of two microorganisms, no statistically significant difference was noted. A statistically significant increase in the number of bacteria-positive sites increased as the score of plaque index (PI), gingival index (GI), and clinical attachment level of CP and AgP patients increased. Coexistence of P. gingivalis and T. denticola was frequently observed in deep periodontal pockets. CONCLUSIONS: Study findings suggest that P. gingivalis and T. denticola are significantly associated with the severity of periodontal tissue destruction. Statistically significant association exists between clinical periodontal parameters such as PI, GI, periodontal pocket depth (PPD), and clinical attachment loss and presence of both the microorganisms.

Role of podoplanin expression in deciding the invasive potential of ameloblastoma - A retrospective IHC study.

BACKGROUND: The aim of this study was to investigate the podoplanin expression in epithelial odontogenic tumors both non-aggressive and aggressive, tumors with and without ectomesenchyme and remnants of the odontogenic epithelium from dental follicles (DF) of unerupted teeth and to examine its role in progression and invasion of tumors. METHOD: Thirty paraffin embedded specimens AM (15 were non-aggressive ameloblastomas and 15 ameloblastoma showing aggressive behavior), 15 paraffin embedded specimens of AOT, 15 paraffin embedded specimens of CEOT, 15 paraffin embedded specimens of CCOT were obtained and were analyzed by immunohistochemistry using anti-human podoplanin. Podoplanin expression in odontogenic epithelial cells was evaluated using a scoring method, along with calculation of the percentage of positive odontogenic cells. RESULTS: Podoplanin was expressed strongly at the invasive front (in the peripheral odontogenic epithelial cells) of most tumors and dental follicles. Membranous expression of podoplanin in ameloblastomas was stronger in cases of ameloblastomas showing aggressive behavior than (NA) non-aggressive ameloblastomas. CONCLUSION: Expression of podoplanin at the invasive front (in peripheral cells) of odontogenic tumors considered to be associated with neoplastic odontogenic tissues. This molecule might play a role in progression and local invasion of odontogenic tumors. The migration and invasion mediated by podoplanin in odontogenic tumors could be related to cytoskeletal reorganization.

Efficacy of curcumin in the treatment for oral submucous fibrosis - A randomized clinical trial.

INTRODUCTION: Oral submucous fibrosis (OSF) is a chronic, insidious disease that is associated with significant functional morbidity and an increased risk for malignancy. Turmeric and its active ingredient "curcumin" are being studied upon as chemopreventive agents in various diseases. The present study aims to determine the efficacy of curcumin in the treatment of OSF. MATERIALS AND METHODS: Thirty clinically diagnosed OSF patients were divided into two groups, 15 patients in each group from the Outpatient Department. Test group patients were treated with Longvida (curcumin) lozenges and control group with Tenovate ointment (clobetasol propionate (0.05%). The treatment was given for 3 months duration and follow-up was done for 6 months. Both the groups were advised for physiotherapy exercises by mouth exercise device. The baseline and follow-up results were compared for IIO (interincisal distance on maximum mouth opening), Visual Analogue Scale (VAS) for normal food and VAS for spicy food. RESULTS: The test group showed 5.93 (±2.37) mm increase in mouth opening compared to 2.66 (±1.76) mm of the control group. In relation to VAS scale with spicy and normal food the average reduction was 64 (42-73) and 77 (70.5-82) as compared to 34 (14.5-64.5) and 64 (46-75.5) respectively in control group. The test group results achieved in the treatment span was sustained in the follow-up (P < 0.05) compared to control group which showed statistically significant (P < 0.05) relapse. CONCLUSION: It can be concluded that combination strategies for the management of OSF which include the stoppage of causative ill habits, appropriate medicinal and physiotherapy management is more efficient than single therapeutic modality. It is evident from the study that curcumin holds good promise in the treatment of OSF in future.

Estimation of vascular endothelial growth factor gene -460 C/T polymorphism as a biomarker in oral squamous cell carcinoma patients from the Indian subcontinent.

AIM: Oral squamous cell carcinoma (OSCC) is one of the most common malignancies in India. Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis, is known to play a key role in the pathophysiology of tumorigenesis. Polymorphisms of the VEGF gene have been shown to influence the risk of OSCC, but it remains to be elucidated in the Indian population. Thus, the present study was designed to detect the VEGF gene -460 C/T polymorphism in OSCC patients from the Indian population, which is genetically distinct from previously-studied populations. METHODS: Peripheral blood samples of 80 OSCC patients and 80 healthy individuals were collected for a case-control study design. The VEGF gene -460 C/T polymorphism was detected by polymerase chain reaction-based restriction analysis. RESULTS: The analysis revealed significant differences between normal individuals and patients with OSCC (P < 0.05). The distribution of the TT homozygote and the T allele in the patient group was significantly greater than in the control group. CONCLUSION: The present study provides novel evidence of the role of the VEGF gene -460 C/T polymorphism as a suitable genetic marker of OSCC in the Indian population.

Demonstration of mycobacterium tuberculosis in sputum and saliva smears of tuberculosis patients using ziehl neelsen and flurochrome staining- a comparative study.

BACKGROUND: Early detection of tuberculosis is important for reducing its morbidity and mortality especially in the patients with non-productive cough. To overcome the cumbersome process involved in collection and processing of the sputum specimen, the time consumed for reporting of sputum by Ziehl Neelsen (ZN) method and to introduce a routine screening test in suspected, symptomless tuberculosis patients, the present study was designed using saliva as diagnostic medium and Auramine Rhodamine (AR) as staining method. On review of literature, there was no study which has tried diagnosing tuberculosis using saliva with flurochrome stain; hence the present study was designed. AIM: To introduce a routine screening test for tuberculosis patient using saliva and to determine the diagnostic efficacy of routine ZN staining method and AR fluorescent staining method in sputum and saliva smears of pulmonary tuberculosis patients. SETTINGS AND DESIGN: Laboratory settings and Experimental design. MATERIALS AND METHOD: Fifty smears samples of sputum and saliva of known cases of pulmonary tuberculosis were stained with routine ZN stain and other with AR fluorescent stain. All the specimens were inoculated into Lowenstein-Jensen culture media. The smears were subjected for scanning of Mycobacterium tuberculous bacilli under X 1000 magnification for ZN stain and X 400 magnification for AR stain by grid pattern proposed by National tuberculosis institute and graded by RNTCP grading system. RESULTS: All 50 sputum samples showed 100% positivity by ZN and AR stain while only 76% positivity was seen by culture. Of the 50 saliva samples 10% cases were positive by ZN, 76% were positive by AR & 70% by culture method. Statistical analysis using chi square test was done, and the value was found to be statistically highly significant for AR staining technique. (p<0.001) CONCLUSION: Saliva can prove to be an important tool for the diagnosis as well as screening of the patients with pulmonary tuberculosis when aided with flurochrome staining method.

Relation of erythrocyte indices and serum iron level with clinical and histological progression of oral squamous cell carcinoma in Central India.

AIM: This study aims to evaluate the relation of erythrocyte indices and serum iron level with clinical and histopathological progression of oral squamous cell carcinoma (OSCC). METHODS: Eighty newly diagnosed OSCC patients recruited for the study were divided according to tumor size int three groups. Erythrocyte indices (e.g., hemoglobin [Hb], red blood cell [RBC] count, packed cell volume [PCV]) and serum iron level (S.Fe) were evaluated with normal reference values, control subject, primary tumor size and histopathological grading. Correlation of Hb to S.Fe was also analyzed. RESULTS: We found that 56.36, 61.81, and 83.63% of males and 76, 32, and 88% of female patients were anemic in terms of Hb%, RBC count and PCV respectively, whereas the remainding indices and S.Fe were within normal range (P < 0.05). Percent Hb, RBC count and PCV gradually decreased with increasing tumor size and histopathological grading (P > 0.05). Moderate to weak correlation was observed between Hb and S.Fe (P < 0.05). CONCLUSIONS: Tumor-induced hemolysis appeared to be responsible for anemia in OSCC and its severity increased with the progression of tumor. Moderate to weak correlation exists between Hb and S.Fe probably owing to the dual requirement of iron for bone marrow and the tumor.

An unusual presentation of clear cell odontogenic carcinoma in mandibular anterior region.

Clear cell odontogenic carcinoma (CCOC) is a rare, potentially aggressive odontogenic epithelial tumor with tendency for recurrence. It was first described as a clinicopathological entity in 1985 and to date only 73 cases has been reported in English literature. A case of CCOC in 64-year-old male patient in mandibular anterior region is presented which when recurred in soft tissue 5 years after wide surgical resection of mandible, revealed a biphasic pattern as against monophasic pattern of primary neoplasm and was unusually associated with primary squamous cell carcinoma, suggestive of hybrid tumor.

Oropharyngeal candidosis relative frequency in radiotherapy patient for head and neck cancer.

BACKGROUND: Radiation given during treatment of oral and pharyngeal malignancy frequently causes alteration of the oral environment predisposing to the colonization of the oral mucosa by yeast species most frequently Candida. OBJECTIVE: Thus, this study was undertaken in 107 patients to find out association between radiation therapy and frequency of oropharyngeal candidosis, to quantitate colony forming units (CFUs) to identify Candida at species level and to check the incidence of serotype A and B in C. albicans. MATERIALS AND METHODS: The study was done on patients suffering from oropharyngeal cancer who were advised radiotherapy. The oral rinse collection method was used to collect the sample. Sabourauds Dextrose Agar (SDA) was used as primary culture media and subsequently speciation was done using standard techniques. The strains of C. albicans were serotyped employing the method described by Hansclever and Mitchell (1961, J Bacteriol 1961;82:570-3). RESULTS: 26.16% patients were mycologically positive for candida before radiotherapy with CFUs 100. 14 ± 59.11 that increased to 60.74% patients during radiotherapy with an increase in CFUs to 490.15 ± 207.97. Clinically, grading of mucositis was done and also individual signs and symptoms were noted in each patient. The occurrence of erythmatous lesions, ulceration, and xerostomia were found to be statistically significant (P<0.05). C. albicans was the most frequently encountered species with higher prevalence of serotype A suggesting higher virulent species. CONCLUSION: It is proposed that in such patients taking radiotherapy prophylactic antifungal treatment should be given specially in patients showing development of oral mucosal lesions such as erythmatous lesions, ulcerations, and complaining about dryness of mouth, that is, xerostomia irrespective of presence or absence of clinical oral candidosis.

Oral manifestations of HIV infection and their correlation with CD4 count.

Human Immunodeficiency Virus (HIV)-related oral lesions can be used as markers of the immune status. The present cross-sectional study was conducted to identify the oral manifestations in HIV-infected individuals and their association with reduced Cluster of Differentiation 4 (CD4) count. The study population included known HIV-positive patients. A detailed case history of 399 HIV-positive patients was obtained and general examination was carried out. Diagnosis of oral lesions was done based on presumptive criteria of EEC Clearinghouse, 1993. The CD4 count was determined in 369 patients and correlated with oral manifestations. The prevalence of oral lesions was found to be 76.70% (n = 306). Oral candidiasis (157 (39.3%)) was the most common oral lesion associated with HIV infection. Amongst various forms of oral candidiasis, erythematous candidiasis (122 (39.3%)) outnumbered the other forms. The mean CD4 count of patients with oral lesions (207 cells/mm(3)) was less than in patients without oral lesions (291 cells/mm(3)) (P = 0.002). Oral candidiasis was found to be significantly correlated to a reduced CD4 cell count below 200 cells/mm(3) (P = 0.000; Odds ratio = 3.1; 95% Confidence interval 1.9-4.9) with good sensitivity, best specificity and positive predictive value. Oral manifestations may be used as an alternative to CD4 count at field-based settings to diagnose the immune compromised status of HIV-infected individuals.

Verrucous hyperplasia: A clinico-pathological study.

CONTEXT: Oral verrucous hyperplasia (OVH) is a premalignant lesion that may transform into an oral cancer. AIMS: The present retrospective study was carried out to analyze the clinico-pathological features of verrucous hyperplasia (VH). MATERIALS AND METHODS: Total 19 diagnosed cases of verrucous hyperplasia were retrospectively analyzed for demographic, clinical and histopathological features including dysplasia. RESULTS: Average age of occurrence of lesion was 4 (th) decade of life, with male predominance (2:1) and common site of occurrence being buccal mucosa. Clinically it present as verrucous exophytic growth with sharp or blunt projections on surface, which corresponds histologically. Tobacco lime quid placement in buccal vestibule was key etiologic factor. Histopathologicaly 68% cases showed dysplasia out of which moderate dysplasia predominates (42%). Moderately dysplastic cases were found to be associated with mixed habit pattern. Maximum cases were treated with excision. CONCLUSION: Biopsy specimen comprising of adjacent normal epithelium is key in distinguishing verrucous hyperplasia from verrucous carcinoma. Clinical behavior and recurrence potential needs to be assessed with long term follow up studies.

Detection of Candida by calcofluor white.

OBJECTIVE: To assess the efficacy of calcofluor white (CFW) is compared to Gram stain and periodic acid-Schiff in detection of Candida in oral precancer and cancer. STUDY DESIGN: The study group consisted of patients with precancer (n = 45), cancer (n = 45) and control group (n = 45). Presence of Candida was confirmed by culture inoculation along with a germ tube and carbohydrate fermentation test. The cytopathologic smears were analyzed by Papanicolaou-CFW and Gram staining, whereas, tissue sections were stained by periodic acid-Schiff and CFW staining. RESULTS: Candida albicans was the predominant species identified. A highly significant association of Candida was seen more often in cancer than in precancer. Both in cytology and histopathology Candida detection by CFW was higher. In precancer it was 48.88% in smears and 40% in tissue sections, whereas, in cancer 60% in smears and 55.55% in histopathology. CONCLUSION: Among the various diagnostic tools used in the present study, the use of CFW is seen to be a simple, effective, rapid and reliable method, both in cytopathology and histopathology.