The adverse effects of mining pollutants on oral mucosa in Bellary district: A clinical and genotoxicity study.

Background: The Bellary district in Karnataka, rich in mineral resources, is a major mining industry, but prolonged exposure to mining can lead to health hazards. The study aims to assess the genotoxic impact of mining pollutants on mine workers using the micro-nucleus (MN) assay. Setting and Design: Cross-sectional study. Materials and Methods: A total of 250 individuals (198 males and 52 females) working in mining areas were examined, and their oral findings were recorded in a proforma. For the micro-nucleus assay, buccal smears from 30 individuals working in mining areas with habits, 30 individuals working in mining areas without habits, and 30 individuals residing in non-mining areas (control group) were selected. Smears were stained with Giemsa stain to identity and quantify the MNs. Results: The frequency of oral mucosal problems among 250 persons working in mining regions was 170 (68.0%) with no oral mucosal conditions, 79 (32.6%) with oral mucosal conditions, 25 (10%) with leukoplakia, 1 (0.4%) with lichen planus, and 8 (3.2%) with ulcerations. Acute necrotising gingivitis was reported in one person (0.4%), candidiasis in two (0.8%), abscess in two (0.8%), OSMF in 39 (15.6%), and oral cancer in two (0.8%). The mean MN count was 2.40 + 1.57 in mine employees with habits, 2.18 + 1.25 in mine workers without habits, and 1.40 + 0.55 in normal healthy controls. Conclusion: Reduced occupational health risks brought on by exposure to mining contaminants require protective measures. After being exposed to mining pollutants, exfoliated buccal mucosal cells can be examined for genotoxicity.

Salivary L-fucose as a biomarker for oral potentially malignant disorders and oral cancer.

BACKGROUND: The objective of this study was to evaluate the serum and salivary L-fucose in oral potentially malignant disorders (OPMDs) and oral cancer (OC) in order to investigate the possibility of using this as biomarker for early diagnosis. MATERIALS AND METHODS: The study included 85 participants, who were grouped as control (30), OPMDs patients (25), and OC patients (30). Serum and unstimulated whole saliva were collected from participants of all groups and fucose estimation was done using spectrophotometry. The results were tabulated and analyzed statistically. RESULTS: The mean serum L-fucose levels in normal, OPMDs, and OC group were 3.49, 19.18, and 35.75 mg/dl, respectively, while the levels of salivary L-fucose were 3.18, 7.02, and 11.66 mg/dl, respectively. A highly significant rise (P < 0.001) in serum and salivary L-fucose was observed in the study participants compared to control. CONCLUSIONS: The present study showed a significant and gradual increase in serum and salivary L-fucose from control to OPMDs to OC. From this study, we suggest that L-fucose can be used as a reliable biomarker and saliva can be used as a diagnostic fluid for screening and early detection of OC.

Floret-like giant cells in intra-osseous schwannoma.

Floret-like giant cells are very commonly seen in neurofibroma and as an indication to be associated with neurofibromatosis type 1 gene. This particular case of intraosseous schwannoma of mandible showing such giant cells, which is rarely reported.

Omics data-driven analysis identifies laminin-integrin-mediated signaling pathway as a determinant for cell differentiation in oral squamous cell carcinoma.

BACKGROUND: In recent years, high-throughput omics technologies have been widely used globally to identify potential biomarkers and therapeutic targets in various cancers. However, apart from large consortiums such as The Cancer Genome Atlas, limited attempts have been made to mine existing datasets pertaining to cancers. METHODS AND RESULTS: In the current study, we used an omics data analysis approach wherein publicly available protein expression data were integrated to identify functionally important proteins that revealed consistent dysregulated expression in head and neck squamous cell carcinomas. Our analysis revealed members of the integrin family of proteins to be consistently altered in expression across disparate datasets. Additionally, through association evidence and network analysis, we also identified members of the laminin family to be significantly altered in head and neck cancers. Members of both integrin and laminin families are known to be involved in cell-extracellular matrix adhesion and have been implicated in tumor metastatic processes in several cancers. To this end, we carried out immunohistochemical analyses to validate the findings in a cohort (n = 50) of oral cancer cases. Laminin-111 expression (composed of LAMA1, LAMB1, and LAMC1) was found to correlate with cell differentiation in oral cancer, showing a gradual decrease from well differentiated to poorly differentiated cases. CONCLUSION: This study serves as a proof-of-principle for the mining of multiple omics datasets coupled with selection of functionally important group of molecules to provide novel insights into tumorigenesis and cancer progression.

BCL-X expression in oral cancer: Comparison between oral squamous cell carcinoma and verrucous carcinoma.

BACKGROUND: Verrucous carcinoma (VC) should be considered a distinct clinicopathologic entity different from the more common oral squamous cell carcinoma (OSCC) because of its unique biological behavior. Best way to understand the behavior of these carcinomas is to study them by means of molecular methods, especially in tumor progression tests and Bcl-X is an important antiapoptotic member of the Bcl-2 family and is one of the newest and most useful markers to determine the aggressiveness of many carcinomas. The relationship between this Bcl-X protein and carcinomatous behavior toward it is not studied extensively, which we attempted to evaluate using immunohistochemical analysis in selected carcinomas of the head and neck region. METHOD: We studied Bcl-X protein expression in sections of thirty OSCC and ten VC samples and correlated this with tumor differentiation. RESULTS: There was a significant difference in cytoplasmic staining of Bcl-X expression with statistical analysis (P < 0.005) for VC and OSCC when compared as a group. No significance was seen among the different histological grades of OSCC and when compared with VC individually. CONCLUSION: The significant result between OSCC and VC suggests that their biologic course is comparable and can be helpful in differentiating them with each other for establishment of a better treatment protocol.

Genotoxicity of non-alcoholic mouth rinses: A micronucleus and nuclear abnormalities study with fluorescent microscopy.

AIM: The aim of the present study was to evaluate the genotoxicity of non-alcoholic mouth rinses on buccal epithelial cells using a micronucleus test. METHODS: A total of 105 patients were selected and randomly divided into five groups. Four different mouth rinses and normal saline were given for 2 weeks' duration, and cytological smears were collected before and after exposure. These smears were subjected to micronucleus (MN) and other nuclear abnormalities (ONA) tests using acridine orange stain, and their frequencies were obtained in 500 buccal epithelial cells. The statistical analysis included mean, χ2 -test, analysis of variance, and post-hoc analysis by Bonferroni test. RESULTS: Micronucleated cells (P < .00) and MN (P < .00) were higher in individuals exposed to chlorhexidine (CHX), followed by chlorine dioxide (ClO2 ), potassium nitrate (KNO3 ), and sodium fluoride (NaF), amine fluoride (AmF), and normal saline. ONA were greater (P < .00) in individuals exposed to CHX, followed by ClO2 , AmF, KNO3 , and NaF and normal saline. Overall, the results showed that genotoxic damage was greater in the case of CHX, followed by ClO2 , KNO3 , and NaF, AmF, and normal saline. CONCLUSION: Chronic exposure to mouth rinses can cause genotoxic damage to buccal epithelial cells. Long-term injudicious and inadvertent use of mouth rinses should be discouraged.

Qualitative analysis of connective tissue stroma in different grades of oral squamous cell carcinoma: A histochemical study.

BACKGROUND: Detection of oral cancer at an early stage is of utmost importance to decrease morbidity and mortality. Tumor stroma plays a critical role during carcinogenesis. There is lack of information regarding the characteristics of the stroma in relation to the invading malignant epithelial cells and the interdependence between stroma and tumor cells in different grades of oral squamous cell carcinoma (OSCC). AIM: The present study was aimed to analyze and compare the nature of stroma in the vicinity of invading tumor islands in different grades of OSCC, using a histochemical technique picrosirius-polarization method. The present study also evaluated and correlated the possible role of inflammatory response in determining the nature of the stroma. SUBJECTS AND METHODS: The study included thirty cases of different grades of histologically diagnosed OSCC and ten sections of normal buccal mucosa as a control group. Nature of collagen was analyzed using picrosirius-polarization method, and intensity of inflammatory cell infiltrate was recorded using ImageJ software (1.42q, NIH, USA). The results were tabulated and analyzed statistically. RESULTS: Normal oral mucosa showed predominantly reddish birefringence. All cases of well-differentiated OSCC showed reddish-orange color. Nearly 70% moderately differentiated cases showed yellowish-orange (YO) and 60% of poorly differentiated cases, showed greenish-yellow (GY). The mean inflammatory cell count was highest in well-differentiated group. There was shift to YO and GY collagen when the cell differentiation and inflammatory cell count decreased in moderate and poorly differentiated cases. CONCLUSION: Both inflammatory cells and tumor cells have a role in determining the nature of the collagen fibers in tumor stroma of OSCC, probably with opposing effects on stromal behavior and hence both are significant in predicting prognosis.

Comparative immunohistochemical study of Bcl-X in ameloblastoma, keratocystic odontogenic tumor and adenomatoid odontogenic tumor.

OBJECTIVES: Since its recognition as a physiologic process associated with tumor, among molecular mechanisms involved in tumor progression, defects in regulation of apoptosis have generated an accelerating volume of research that has sought to elucidate the role of programed cell death in pathogenesis and treatment of various tumors. Therefore, this study was performed to understand better the diverse biological profile of epithelial odontogenic tumors with the help of immunohistochemical expression of Bcl-X protein. MATERIALS AND METHODS: We studied Bcl-X protein expression in 45 cases of epithelial odontogenic tumors which included 15 cases each of ameloblastomas, keratocystic odontogenic tumor (KCOT) and adenomatoid odontogenic tumor (AOT) and correlated the expression with their growth pattern. RESULTS: Cytoplasmic staining of Bcl-X revealed overexpression in ameloblastoma when compared to KCOT and AOT. Percentage of positive cells showed a statistically significant difference, P = 0.007 between ameloblastoma and KCOT, whereas P < 0.001 between ameloblastoma and AOT. However, no significance was observed between KCOT and AOT (P = 0.132). CONCLUSION: The present study supports the fact that epithelial odontogenic tumors show diverse growth profiles. An increased Bcl-X expression was seen in ameloblastoma compared to KCOT and least expression in case of AOT which could be indicative of more aggressive biological behavior and increased cell survival activity of ameloblastoma than KCOT and AOT. This signifies the diagnostic relevance of this biomarker and also could be a possible regulator of the proliferative compartment by contributing in tumor progression and cytodifferentiation of epithelial odontogenic tumors.

Estimation of Antimicrobial Properties of Aqueous and Alcoholic Extracts of Salvadora Persica (Miswak) on Oral Microbial Pathogens - An Invitro Study.

INTRODUCTION: Twigs of Salvadora persica (Miswak) plant are being used as a means of oral hygiene since ages for brushing teeth. Though clinical research and trials have shown promising results on effectiveness of Miswak, but some reports are conflicting. AIM: To evaluate the antimicrobial activity of crude aqueous and alcoholic extracts of Salvadora persica (Miswak) against the common microbial pathogens causing dental caries and periodontitis. MATERIALS AND METHODS: A prospective study of one year duration was conducted in Yenopoya dental and medical college, Mangalore. The twigs of Salvadora persica were collected and alcoholic and aqueous extracts were prepared using standard techniques. The antimicrobial properties of the extracts against common oral pathogens like Streptococcus mutans, Streptococcus mitis, Candida albicans, Lactobacillus acidophilus, Prevotella intermedia, & Peptostreptococcus were performed by agar well diffusion method and two fold broth dilution method. RESULTS: No significant results was obtained when water extracts of Salvadora persica was tested except for minimum inhibitory effect against Streptococcus mutans, Prevotella intermedia & Peptostreptococcus and Candida albicans. Relatively significant inhibitory effect was noted with respect to alcoholic extract of Salvadora persica. CONCLUSION: Although comparatively less than chlorhexidine which is a known antimicrobial agent, the alcoholic extracts of Salvadora persica showed antimicrobial effect against the common microbial pathogens causing dental caries and periodontitis indicating a potential beneficial effect of this plant. However, further research with more standardized extraction procedure and advanced techniques is required to find out the exact chemicals responsible for the antimicrobial properties of the plant extract.

Investigation of the Possibility of Using Serum Ischemia Modified Albumin (IMA) as a Novel and Early Marker of the Extent of Oxidative Stress Induced by Various Tobacco Products.

BACKGROUND: Ischemia Modified Albumin (IMA) is an altered serum albumin that forms under the conditions of oxidative stress and is considered as a biomarker of cardiac ischemia. The objective of this study was to evaluate the ischemia modified albumin (IMA) in the serum of the individuals with different types of tobacco habits in order to investigate the possibility of using this as a biomarker for the oxidative stress induced by the tobacco products. MATERIALS AND METHODS: The study included 90 subjects, who were Grouped as control (30), Group I (betel quid chewers), Group II (gutkha chewers), Group III (smokers) and Group IV (mixed). Serum was collected from subjects of all Groups and IMA estimation was done using Albumin Cobalt binding assay. The results were tabulated and analysed statistically. RESULTS: The mean serum IMA levels in control, Group I, Group II, Group III and Group IV were 0.52547 ABSU, 0.68767 ABSU, 0.47433 ABSU,0.36540 ABSU and 0.54593 ABSU respectively. CONCLUSION: The results show that serum IMA levels were increased in betel quid chewers and mixed Group compared to the controls. From the results noted in this study we suggest that IMA can be used as an early marker for tobacco related oxidative stress.

Estimation of argyrophilic nucleolar organizer regions in different grades of oral submucous fibrosis.

CONTEXT: Assessment of potential for malignant transformation of oral submucous fibrosis (OSF) through clinical or light microscopic examination of hematoxylin and eosin stained tissue sections is not totally satisfactory. The search is for such a tissue marker that will differentiate those cases of OSF, which carry a higher risk for malignant transformation. During the past few years, numerous workers have validated the usefulness of enumerating argyrophilic nucleolar organizer regions (AgNORs) in predicting the malignant potential of lesions. The present study was carried out to validate the diagnostic potential of this marker. OBJECTIVES OF THE STUDY: Quantitative and qualitative assessment of AgNORs in different grades of OSF and to compare the count of AgNORs in different grades of OSF and normal mucosa. MATERIALS AND METHODS: AgNORs were investigated in tissue specimens from 90 patients diagnosed with different histopathological grades of OSF. AgNORs were identified in tissue specimens stained with silver nitrate, using light microscope. AgNORs were counted as small, large and total count to analyze them both quantitatively and qualitatively. STATISTICAL ANALYSIS USED: Results were subjected to statistical analysis for obtaining significance value (P value) by unpaired Student's t-test. RESULTS: The mean total count of AgNORs was 2.464 ± 0.101, 4.358 ± 0.108, 3.704 ± 0.106 and 3.279 ± 0.161 in normal mucosa, Grades I, II and III of OSF, respectively. A qualitative difference was observed in the presentation of AgNORs in different grades of OSF. Mean value of small nucleolar organizer regions (NORs) decreased while the mean of large NORs increased as the grade of OSF increased. CONCLUSIONS: The study findings suggest that AgNORs are increased in OSF and they can serve as a reliable tool adjunct to histopathological diagnosis. Their ease of demonstration and high specificity to cellular proliferation make them the best available histopathological marker in the arsenal of an oral pathologist.

Histopathological, immunohistochemical and special stain unraveling the enigmatic carcinosarcoma--A case report.

Carcinosarcoma is an extremely rare, highly malignant tumor having a characteristic of both epithelial and mesenchymal components; the epithelial component exhibiting focal squamous cell carcinoma and mesenchymal component having sarcomatoid stroma. The occurrence of carcinosarcoma in the oral cavity is very rare and only few cases have been reported in the literature, although its occurrence in other parts of the body has been widely reported.

Estimation of copper in saliva and areca nut products and its correlation with histological grades of oral submucous fibrosis.

BACKGROUND: The purpose of this study was to estimate the copper levels in saliva of patients with oral submucous fibrosis (OSF) and different areca nut products and its correlation with different histological grades of OSF. METHODS: The study comprised 60 individuals, 30 OSF patients and 30 non-OSF individuals. Unstimulated whole saliva was collected, and copper analysis was performed using colorimetric method. The commercial areca nut products used by the patients were acquired and subjected to copper analysis through the atomic absorption spectrophotometer method. Oral biopsies were performed for OSF patients for histopathological correlation. RESULTS: The mean salivary copper level was 27.023 µg/dl in OSF patients when compared with 8.393 µg/dl in non-OSF individuals (P < 0.005). The mean copper content in different areca nut products was 13.313 ppm (P < 0.005). Comparison of copper content in different areca nut products with salivary copper levels of OSF patients showed negative correlation (P < 0.853). Comparison of salivary copper levels between different histological grades of OSF yielded a statistically significant association between grades I and III (P < 0.005) and grades II and III OSF (P < 0.019). Comparison of copper content in areca nut products and different histological grades of OSF yielded weak negative statistical correlation (r = -0.116). CONCLUSION: Despite high copper content in areca nut products, the observations yielded a negative correlation with different histological grades of OSF. This further raises a doubt about the copper content in areca nut as an etiological factor for this crippling disease.

Liposarcoma of the maxillary antrum: A case report.

Atypical Lipomatous Tumor/Well-Differentiated Liposarcoma. (ALT/WDL) is a soft tissue sarcoma of intermediate malignant behavior, which most frequently affects the retroperitoneum and lower extremities. Liposarcomas of head and neck are rare, representing only 2-8% of all sarcomas in this region. The majority of liposarcomas occur in middle-aged adults; however, very uncommonly cases have been reported in infancy and early childhood. We report a case of a 14-year-old girl diagnosed as ALT/WDL of the maxillary antrum.

Antimicrobial properties of Cocos nucifera (coconut) husk: An extrapolation to oral health.

BACKGROUND AND OBJECTIVES: Brushing the teeth with fibrous husk of Cocos nucifera (coconut) is a common oral hygiene practice among people of rural areas of South India. However, the probable antimicrobial properties of this plant material against common oral pathogens have not been proved scientifically. Therefore, the present study was designed. MATERIALS AND METHODS: Alcoholic extract of the husk of Cocos nucifera was prepared and the antimicrobial properties against common oral pathogens like cariogenic bacteria, periodontal pathogens, and candidal organisms were performed by the Agar Well Diffusion Method. The results obtained were then subjected to statistical analysis using One-Way Analysis of Variance (ANOVA) and the Tukey's Honestly Significant Difference (HSD). RESULTS: The alcoholic extract of Cocos nucifera showed a significant concentration-dependent antimicrobial activity, expressed as a zone of inhibition with respect to all tested organisms except Actinomyces species. The inhibitory effect was more significant, with a majority of cariogenic organisms and Candida, with a zone of inhibition ranging from 4.6 mm to 16.3 mm. However, the effect was lesser with Cocos nucifera compared to chlorhexidine. Minimum inhibitory concentration (MIC) ranged from 50 mg/ml to 75 mg/ml. CONCLUSION: Cocos nucifera has a significant inhibitory action against common oral pathogens, indicating the presence of highly effective antimicrobial compounds. Therefore, it is proved that its use can contribute to oral health to a great extent. Identification of these active compounds provides the scope for incorporating it into a modern oral care system, so as to control oral diseases.

Comparative Study of Genotoxicity in Different Tobacco Related Habits using Micronucleus Assay in Exfoliated Buccal Epithelial Cells.

BACKGROUND: Oral cancer is one of the most debilitating diseases afflicting mankind. Consumption of tobacco in various forms constitutes one of the most important etiological factors in initiation of oral cancer. When the focus of today's research is to determine early genotoxic changes in human cells, micronucleus (MN) assay provides a simple, yet reliable indicator of genotoxic damage. AIMS AND OBJECTIVES: To identify and quantify micronuclei in the exfoliated cells of oral mucosa in individuals with different tobacco related habits and control group, to compare the genotoxicity of different tobacco related habits between each group and also with that of control group. PATIENTS AND METHODS: In the present study buccal smears of 135 individuals with different tobacco related habits & buccal smears of 45 age and sex matched controls were obtained, stained using Giemsa stain and then observed under 100X magnification in order to identify and quantify micronuclei in the exfoliated cells of oral mucosa. RESULTS: The mean Micronucleus (MN) count in individuals having smoking habit were 3.11 while the count was 0.50, 2.13, and 1.67 in normal control, smoking with beetle quid and smokeless tobacco habit respectively. MN count in smokers group was 2.6 times more compared to normal controls. MN count was more even in other groups when compared to normal control but to a lesser extent. CONCLUSION: From our study we concluded that tobacco in any form is genotoxic especially smokers are of higher risk and micronucleus assay can be used as a simple yet reliable marker for genotoxic evaluation.

Cytomorphometric analysis of oral buccal mucosal smears in tobacco and arecanut chewers who abused with and without betel leaf.

BACKGROUND: Tobacco in any form (smoking or chewing), arecanut chewing, and alcohol are considered to be the major extrinsic etiological factors for potentially malignant disorders of the oral cavity and for squamous cell carcinoma, the most common oral malignancy in India. An increase in nuclear diameter (ND) and nucleus-cell ratio (NCR) with a reduction in cell diameter (CD) are early cytological indicators of dysplastic change. The authors sought to identify cytomorphometric changes in ND, CD, and NCR of oral buccal cells in tobacco and arecanut chewers who chewed with or without betel leaf. METHODS: Participants represented 3 groups. Group I consisted of 30 individuals who chewed tobacco and arecanut with betel leaf (BQT chewers). Group II consisted of 30 individuals who chewed tobacco and arecanut without betel leaf (Gutka chewers). Group III comprised 30 apparently healthy nonabusers. Cytological smears were prepared and stained with modified-Papanicolaou stain. RESULTS: Comparisons between Groups I and II and Groups II and III showed that ND was increased, with P values of .054 and .008, respectively, whereas a comparison of Groups I and III showed no statistical significance. Comparisons between Groups I and II and Groups II and III showed that CD was statistically reduced, with P values of .037 and <.000, respectively, whereas comparison of Groups I and III showed no statistical significance. Comparisons between Groups I and II and groups II and III showed that NCR was statistically increased, with P values of <.000, whereas a comparison of Groups I and III showed no statistical significance. CONCLUSIONS: CD, ND, and NCR showed statistically significant changes in Group II in comparison with Group I, which could indicate larger and earlier risk of carcinoma for Gutka chewers than in BQT chewers.

Quality and safety aspects in histopathology laboratory.

Histopathology is an art of analyzing and interpreting the shapes, sizes and architectural patterns of cells and tissues within a given specific clinical background and a science by which the image is placed in the context of knowledge of pathobiology, to arrive at an accurate diagnosis. To function effectively and safely, all the procedures and activities of histopathology laboratory should be evaluated and monitored accurately. In histopathology laboratory, the concept of quality control is applicable to pre-analytical, analytical and post-analytical activities. Ensuring safety of working personnel as well as environment is also highly important. Safety issues that may come up in a histopathology lab are primarily those related to potentially hazardous chemicals, biohazardous materials, accidents linked to the equipment and instrumentation employed and general risks from electrical and fire hazards. This article discusses quality management system which can ensure quality performance in histopathology laboratory. The hazards in pathology laboratories and practical safety measures aimed at controlling the dangers are also discussed with the objective of promoting safety consciousness and the practice of laboratory safety.