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.

Identifying Biomarkers and Therapeutic Targets by Multiomic Analysis for HNSCC: Precision Medicine and Healthcare Management.

Background: Head and neck squamous cell carcinoma (HNSCC) is one of the major types of cancer, with 900,000 cases and over 400,000 deaths annually. It constitutes 3-4% of all cancers in Europe and western countries. As early diagnosis is the key to treating the disease, reliable biomarkers play an important role in the precision medicine of HNSCC. Despite treatments, the survival rate of cancer patients remains unchanged, and this is mainly due to the failure to detect the disease early. Thus, the objective of this study is to identify reliable biomarkers for head and neck cancers for better healthcare management. Methods: In this study, all available, curated human genes were screened for their expression against HNSCC TCGA patient samples using genomic and proteomic data by various bioinformatic approaches and datamining. Docking studies were performed using AutoDock or online virtual screening tools for identifying potential ligands. Results: Sixty genes were short-listed, and most of them show a consistently higher expression in head and neck patient samples at both the mRNA and the protein level. Irrespective of human papillomavirus (HPV) status, all of them show a higher expression in cancer samples. The higher expression of 30 genes shows adverse effects on patient survival. Out of the 60 genes, 12 genes have crystal structures and druggable potential. We show that genes such as GTF2H4, HAUS7, MSN, and MNDA could be targets of Pembrolizumab and Nivolumab, which are approved monoclonal antibodies for HNSCC. Conclusion: Sixty genes are identified as potential biomarkers for head and neck cancers based on their consistent and statistically significantly higher expression in patient samples. Four proteins have been identified as potential drug targets based on their crystal structure. However, the utility of these candidate genes has to be further tested using patient samples.

Revisiting Vickers and Gorlin criteria in histopathological subtypes of ameloblastoma.

Background: Ameloblastoma is a benign tumour of odontogenic epithelial origin arising from enamel organ tissue that has not undergone differentiation to the point of hard tissue formation. Aims: This study was conducted with an aim to provide a baseline data to analyse whether various histopathological variants of ameloblastoma satisfies all the characteristic histopathological features of Vickers and Gorlin criteria. Settings and Design: A retrospective study of 25 cases of intraosseous ameloblastoma was carried out in the Department of Oral Pathology and Microbiology in accordance with the Institutional Ethics Committee. Methods and Materials: Histopathological slides of ameloblastoma subtypes were analysed microscopically to assess Vickers and Gorlin criteria. Statistical Analysis Used: Statistical analysis was done using the Chi-square test. A P- value of < 0.05 was set for statistical significance. Results: Presence of hyperchromatic nuclei was seen in all the variants (100%), except for the desmoplastic variant which showed only 60% positivity. Basal cell palisading, reverse polarity and subnuclear vacuolization were seen predominantly only in acanthomatous (100%), and follicular variants (83%). Conclusions: Vickers and Gorlin criteria have become an integral part of diagnosis of histopathological subtypes of ameloblastoma and should be applied vigilantly in the diagnosis as these may not always fulfill all the gold standard criteria when individual subtypes are assessed.

Age estimation using extracted teeth in coastal Karnataka population.

Introduction: Age estimation using regressive alterations such as root dentin translucency, PDL attachment, and attrition are easy and reliable way of predicting the age. However, extensive and population-specific formula has not been generated. This attempt was to assess the correlation of these alterations with age and to generate a Linear regressive formula, specific to this population. Methods and Material: Three alterations were assessed such as dental attrition, root dentin translucency, and periodontal attachment level from the extracted teeth. Dental attrition was measured using Johanson's and Li and Ji criteria. PDL attachment level and root dentin translucency was measured using the Johanson method and the Lamendin method. Statistical Analysis: SPSS software (Version 27), Pearson correlation test, and Linear regressive analysis were used. Results: Our results showed all three factors/parameters such as attrition, periodontal ligament, and translucency having a very good correlation with age and correlation coefficient r value ranging from 0.6 to 0.8. All the parameters were having statistically significant correlation with P value <0.005. Among them, root dentin translucency with Johanson G method showed excellent correlation with r = 0.83 followed by PDL attachment by Johanson G method with r = 0.702. Conclusions: Regressive changes such as Dentin translucency, PDL attachment and attrition on Coastal Karnataka showed a very good correlation with age. Among them, Dentin translucency by Johanson G method had the best correlation with the a standard error of estimate (SEE). Results of our study indicates that all these parameters [Translucency, PDL attachment, and attrition] can be utilized in age estimation.

Serum autoantibody profiling of oral squamous cell carcinoma patients reveals NUBP2 as a potential diagnostic marker.

Introduction: Oral Squamous Cell Carcinoma (OSCC), a common malignancy of the head and neck region, is frequently diagnosed at advanced stages, necessitating the development of efficient diagnostic methods. Profiling autoantibodies generated against tumor-associated antigens have lately demonstrated a promising role in diagnosis, predicting disease course, and response to therapeutics and relapse. Methods: In the current study, we, for the first time, aimed to identify and evaluate the diagnostic value of autoantibodies in serum samples of patients with OSCC using autoantibody profiling by an immunome protein array. The utility of anti-NUBP2 antibody and tissue positivity in OSCC was further evaluated. Results and discussion: We identified a total of 53 autoantibodies with significant differential levels between OSCC and control groups, including 25 that were increased in OSCC and 28 that were decreased. These included autoantibodies against Thymidine kinase 1 (TK1), nucleotide-binding protein 2 (NUBP2), and protein pyrroline-5-carboxylate reductase 1 (PYCR1), among others. Immunohistochemical validation indicated positive staining of NUBP2 in a large majority of cases (72%). Further, analysis of OSCC data available in TCGA revealed higher NUBP2 expression correlated with better disease-free patient survival. In conclusion, the differential serum autoantibodies identified in the current study, including those for NUBP2, could be used as potential biomarkers for early diagnosis or as screening biomarkers for OSCC pending investigation in a larger cohort.

Unveiling the arcanum of formalin-fixed paraffin-embedded archival tissue blocks: A valuable resource for genomic DNA extraction.

Background: Formalin-fixed paraffin-embedded (FFPE) tissue blocks are routinely preserved after pathological diagnosis and possess tremendous potential for biomarker discovery. These archival samples are prone to degradation on prolonged storage due to the formalin cross-linking. Aims: This study aimed to evaluate whether the storage period of the formalin-fixed paraffin-embedded tumor blocks had a significant impact on the yield and purity of the isolated DNA archived for 11 years. Settings and Design: A retrospective study was carried out in the Department of Oral Pathology and Microbiology in accordance with the Institutional Ethics Committee. Materials and Methods: Genomic DNA extraction was performed using TaKaRa DEXPAT Easy DNA kit from 40 FFPE tissue blocks of oral squamous cell carcinoma archived for 11 years (2006-2017). NanoDrop spectrophotometer was used to determine the DNA yield (A260) and purity (A260/A280 ratio). The quality of DNA fragments was validated using agarose gel electrophoresis. Statistical Analysis Used: Statistical analysis was obtained by SPSS 22, MS Excel and analyzed using the analysis of variance (ANOVA) test. P < 0.05 was set for statistical significance. Results: There was no statistically significant difference observed both in terms of DNA yield (P = 0.996) and purity (P = 0.997) of FFPE tumor blocks archived for 11 years among the study groups. Conclusions: It was concluded that, irrespective of years of storage of the FFPE, it is possible to extract genomic DNA and use it for molecular studies.

Predatory science: Unraveling a secret journey of fake journals and conferences.

The fraudulent behaviour of predatory journals/conferences through E-mail solicitations and author's perspective in unknowingly becoming victims of predatory publishing scheme, by being unaware of the fact that the journals in which they are involved are possibly predatory are highlighted here.

Molecular alterations in oral cancer between tobacco chewers and smokers using serum proteomics.

BACKGROUND: Tobacco exposure (through smoking or chewing) is one of the predominant risk factors associated with the development of oral squamous cell carcinoma (OSCC). Despite the growing number of patients diagnosed with OSCC, there are few circulating biomarkers for identifying individuals at a higher risk of developing the disease. Successful identification of candidate molecular markers for risk assessment could aid in the early detection of oral lesions and potentially be used for community screening of high-risk populations. OBJECTIVE: Identification of differentially expressed proteins in the serum of oral cancer patients which can serve as biomarkers for the diagnosis of the onset of oral cancer among tobacco users. METHODS: We employed a tandem mass tag (TMT)-based quantitative proteomics approach to study alterations in the serum proteomes of OSCC patients based on their tobacco exposure habits (chewing and smoking) compared to healthy individuals with no history of using any form of tobacco or any symptoms of the disease. RESULTS: Mass spectrometry-based analysis resulted in the identification of distinct signatures in the serum of OSCC patients who either chewed or smoked tobacco. Pathway analysis revealed opposing effects of dysregulated proteins enriched in the complement-coagulation signaling cascades with a high expression of the Serpin family of proteins observed in OSCC patients who chewed tobacco compared to healthy individuals whereas these proteins showed decreased levels in OSCC patients who smoked. ELISA-based validation further confirmed our findings revealing higher expression of SERPINA6 and SERPINF1 across serum of OSCC patients who chewed tobacco compared to healthy individuals. CONCLUSIONS: This study serves as a benchmark for the identification of serum-based protein markers that may aid in the identification of high-risk patients who either chew tobacco or smoke tobacco.

Molecular alterations in oral cancer using high-throughput proteomic analysis of formalin-fixed paraffin-embedded tissue.

Loss of cell differentiation is a hallmark for the progression of oral squamous cell carcinoma (OSCC). Archival Formalin-Fixed Paraffin-Embedded (FFPE) tissues constitute a valuable resource for studying the differentiation of OSCC and can offer valuable insights into the process of tumor progression. In the current study, we performed LC-MS/MS-based quantitative proteomics of FFPE specimens from pathologically-confirmed well-differentiated, moderately-differentiated, and poorly-differentiated OSCC cases. The data were analyzed in four technical replicates, resulting in the identification of 2376 proteins. Of these, 141 and 109 were differentially expressed in moderately-differentiated and poorly differentiated OSCC cases, respectively, compared to well-differentiated OSCC. The data revealed significant metabolic reprogramming with respect to lipid metabolism and glycolysis with proteins belonging to both these processes downregulated in moderately-differentiated OSCC when compared to well-differentiated OSCC. Signaling pathway analysis indicated the alteration of extracellular matrix organization, muscle contraction, and glucose metabolism pathways across tumor grades. The extracellular matrix organization pathway was upregulated in moderately-differentiated OSCC and downregulated in poorly differentiated OSCC, compared to well-differentiated OSCC. PADI4, an epigenetic enzyme transcriptional regulator, and its transcriptional target HIST1H1B were both found to be upregulated in moderately differentiated and poorly differentiated OSCC, indicating epigenetic events underlying tumor differentiation. In conclusion, the findings support the advantage of using high-resolution mass spectrometry-based FFPE archival blocks for clinical and translational research. The candidate signaling pathways identified in the study could be used to develop potential therapeutic targets for OSCC.

Demineralization of tooth enamel following radiation therapy; An in vitro microstructure and microhardness analysis.

OBJECTIVE: The objective of this study is to evaluate the effects of radiotherapy doses on mineral density and percentage mineral volume of human permanent tooth enamel. MATERIALS AND METHODS: Synchrotron radiation Xray microcomputed tomography (SRµCT) and microhardness testing were carried out on 8 and 20 tooth samples, respectively. Enamel mineral density was derived from SRµCT technique using ImageJ software. Microhardness samples were subjected to Vickers indentations followed by calculation of microhardness and percentage mineral volume values using respective mathematical measures. Data were analyzed using paired t-test at a significance level of 5%. Qualitative analysis of the enamel microstructure was done with two-dimensional projection images and scanned electron micrographs using µCT and field emission scanning electron microscopy, respectively. RESULTS: Vickers microhardness and SRµCT techniques showed a decrease in microhardness and an increase in mineral density, respectively, in postirradiated samples. These changes were related to mineral density variation and alteration of hydroxyapatite crystal lattice in enamel surface. Enamel microstructure showed key features such as microporosities and loss of smooth homogeneous surface. These indicate tribological loss and delamination of enamel which might lead to radiation caries. CONCLUSIONS: Tooth surface loss might be a major contributing factor for radiation caries in head-and-neck cancer patients prescribed to radiotherapy. Such direct effects of radiotherapy cause enamel abrasion, delamination, and damage to the dentinoenamel junction. Suitable measures should, therefore, be worked out to protect nontarget oral tissues such as teeth while delivering effective dosages to target regions.

Single-nucleotide polymorphisms of methylenetetrahydrofolate reductase gene in a South Indian cohort with nonsyndromic cleft lip with or without palate.

OBJECTIVE: Clefts of the lip, with or without cleft palate and cleft palate only, collectively called as orofacial clefts (OFCs) are one of the most common congenital malformations with varying degrees of penetrance and phenotype expressions. The aim of this study was to investigate the association between methylenetetrahydrofolate reductase (MTHFR) cytosine-to-thymine (c. 677 C>T), adenine-to-cytosine (c.1298 A>C) single- nucleotide polymorphisms (SNPs) and South Indian patients with the nonsyndromic cleft lip with or without palate (NSCL ± P). METHODS: A cohort consisting of 25 cases of NSCL ± P and 18 controls from a South Indian cohort were included in this case-control study. Genetic analysis of c.677C>T and c.1298A C polymorphisms in the MTHFR gene was carried out using Sanger sequencing and analyzed from chromatogram profiles. Data interpretation was done using statistical software MedCalc Statistical Software version 16.2 and the Statistical Package for the Social Sciences (SPSS version 22.0). RESULTS: DNA sequence analysis of the MTHFR gene revealed c. 677C>T and c. 1298A>C polymorphisms in 16% and 76% of NSCL ± P cases, respectively. Heterozygous variant in MTHFR c. 1298A>C polymorphism was found to be a significant risk factor (P = 0.0164) for NSCL ± P in South Indian ethnic population. c.677C>T polymorphism, in particular, was apparently dormant overall in the study population. These results offer certain novelty in terms of the distinctive pattern in SNPs of genotypes observed in the study. CONCLUSION: NSCL ± P is one of the most common and challenging congenital malformations with complex etiological basis. Common risk factors such as MTHFR SNPs, namely c.677C>T and c.1298A>C, are subjected to variations in terms of ethnic group, geographic region and micro/macro-environmental factors. Overall, our study has explored part of South Indian ethnic population and revealed a different and unique distribution of mutations in this sample population.

Spectroscopic analyses reveal radiotherapy-induced variations in elemental composition and crystallite properties of human permanent teeth enamel.

OBJECTIVE: To study the effect of radiation therapy on the structural and elemental composition of permanent teeth enamel in vitro. METHODS: Sections from 21 noncarious healthy human teeth were exposed to a cumulative radiation dose of 20-80 Gy. The sections were subjected to electron dispersive spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis to study the elemental composition, the ratio of inorganic and organic content, and the mineralization and crystalline properties of the hydroxyapatite crystal structure respectively. All measures were taken on specified areas of enamel surface before and after radiation exposure and compared. RESULTS: In FTIR and EDS studies, the calcium to phosphorus (Ca/P) and carbonate to phosphate (CO32-/PO3-4) ratios were significantly different (P < 0.05) in teeth sections exposed to 80 Gy, indicating the deterioration of inorganic calcium and phosphorous content. The XRD spectrum data showed loss of peaks at seven specific 2θ coordinate areas, flattened peaks and an increase in the crystallite size in the radiation-exposed groups due to mineralization loss and alteration of the hydroxyapatite crystal matrix in the tooth enamel. CONCLUSIONS: Radiotherapy can induce significant variations in the inorganic and organic functional groups constituting the tooth enamel surface; and these variations are dose dependent. The mechanism responsible for delamination and radiation caries needs to be explored by studying the protein lysis pattern, which might be a leading factor causing the enamel degradation and radiation caries.

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.

Clinicopathological analysis of oral squamous cell carcinoma among the younger age group in coastal Karnataka, India: A retrospective study.

AIMS: Oral squamous cell carcinoma (OSCC) primarily occurs in older age group. However, in the recent years, incidence of oral cancer in young people has been on rise worldwide. Towards this end, we sought to analyze the clinical and histopathological characteristics of OSCC in patients less than 45 years of age. MATERIALS AND METHODS: The clinical and histological features of patients diagnosed with squamous cell carcinoma of the oral cavity at two hospitals in the coastal Karnataka region of South India between 1996-2012 were reviewed. The tabulation and descriptive statistics of the study were carried out. RESULTS: A total of 420 patients were treated for OSCC in the 17-year period (1996-2012), of which 86 (20.5 %) patients were under 45 years of age. The most common site of involvement among the young was tongue (29.07%) and buccal mucosa (27.9%) respectively. A total of 47 (54.65%) patients were either habitual chewers, smokers, or alcoholics. Pathological grading of cases classified tumors into well differentiated (34.88%), moderately differentiated (46.51%) and poorly differentiated (4.65%). CONCLUSIONS: The data from this study reveals that a significant proportion of the OSCC cases are observed in patients of 45 years or younger. Additionally, our study also indicated an increase in the usage of tobacco and pan chewing in young adults in comparison to older individuals in the two hospitals of South India. The data obtained from this analysis emphasizes the need for screening programs that are tailor-made for individuals at high risk of developing oral cancer and warrants tobacco awareness programs in the community.

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.

Evaluation of homocysteine levels in individuals having nonsyndromic cleft lip with or without palate.

CONTEXT: Nonsyndromic cleft lip with or without palate (NSCL ± P) is a genetic predisposition involving defects in shape and makeup of the lip and palate. Elevation of homocysteine (Hcy) levels is seen in medical complications such as developmental anomalies causing neural tube defects, congenital vascular diseases, neurodegenerative and psychiatric conditions. Evaluation of serum Hcy levels forms an important feature to look further into molecular aspects. AIMS: The aim of this study was to evaluate the Hcy levels in NSCL ± P cases by comparing with control cases having no orofacial deformities. SETTINGS AND DESIGN: This study was performed with a biochemical assay in a research laboratory. MATERIALS AND METHODS: A cross-sectional prevalence study was done to compare the concentrations of Hcy between 25 NSCL ± P patients and 15 healthy controls. Blood samples were collected from both the patients and controls and assessed for serum Hcy level using competent chemiluminescent immunoassay technique. STATISTICAL ANALYSIS USED: Student's t-test was used for statistical analysis. RESULTS: The average Hcy concentration was 9.5 µmol/L in control group. There was an increase in Hcy concentration among the NSCL ± P cases with an average value of 18.4 µmol/L. The results were found to be statistically significant using Student's t-test. CONCLUSIONS: The results of this study indicate that Hcy concentration has a significant elevation in NSCL ± P patients when compared with that of control cases.