p53 & Cyclin D1 expression in surgically resected clear margins of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of the most well-known malignancies that affect the human population worldwide. The early diagnosis and early intervention of OSCC help improve the survival rate of the patients. The tumour free surgical margins are a positive prognostic factor for recurrence-free survival. The molecular markers can be used to detect the tumour free surgical margins. Aim: The aim of the study is to evaluate the expression of p53 & Cyclin D1 marker in resected surgical apparently clear margins and to correlate the p53 & Cyclin D1 expression with clinicopathological characteristics and patient outcome. Methods: The study population included retrospective cases of OSCC with apparently clear margins (2017-18) n=10 and Clinicopathological variables relevant to survival analysis were recorded. Finally, two margins were selected from each case, a total of 20 margins were included in this study. Paraffin-embedded wax blocks retrieved and tissue sections were made. Expression of cyclin D1 and p 53 was assessed by the immunohistochemical staining procedure Results: Positive expressions Cyclin D1 in 40% of mild dysplasia margins and 60% in clearance adequate margins were present. p53 expression was seen in 16% of mild dysplasia margins and 84% in clearance adequate margins. The expression of p53 and Cyclin D1 molecular markers are noted in the basal & parabasal layer of epithelium. Conclusion: Molecular markers could play a more reliable method for the assessment of dysplasia at the margins

Evaluation of muscle involvement & degeneration in advanced oral submucous fibrosis using three different stains ­ a retrospective cross-sectional study / Avaliação do envolvimento e degeneração muscular na fibrose submucosa oral avançada usando três colorações diferentes - um estudo transversal retrospectivo

Background: Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features could be identified in routine Hematoxylin and Eosin (H and E) staining in shades of pink. However, the use of special stains provides a contrast to various connective tissue components thereby aiding in better visualization of these connective tissue changes in advanced OSMF cases. Objective:To compare and evaluate muscle involvement and degeneration in advanced oral submucous fibrosis using three different stains namely, H&E, Van Gieson, and Masson's Trichrome. Material and Methods: 10 Formalin-fixed paraffin-embedded tissue sections of advanced OSMFwere stained using 3 different stains namely Hematoxylin and Eosin (H&E), Van-Gieson, and Masson trichrome. The results obtained were tabulated and statistically analyzed using Kruskal Wallis ANOVA. Results: The hyalinization and fibrosis involving the skeletal muscle were better visualized in Masson's Trichrome but were not statistically significant. The muscle degeneration in deeper areas was better visualized in Masson's trichrome when compared to the H&E and Van Gieson. Conclusion: Hematoxylin and eosin stains all the connective tissue components in various shades of pink, use of special stains bestows contrast between different components of connective tissue, thus improvising grading of OSMF. Masson's trichrome stain can be used as a single adjunct to hematoxylin and eosin stain as changes in the superficial and deeper connective tissue could be ascertained (AU)

Contexto: A fibrose submucosa oral (FSO) é classificada de acordo com vários fatores histológicos que incluem alterações epiteliais e do tecido conjuntivo. Essas características podem ser identificadas na coloração de rotina com Hematoxilina e Eosina (H&E) em tons de rosa. No entanto, o uso de colorações especiais fornece um contraste para vários componentes do tecido conjuntivo, auxiliando assim na melhor visualização dessas alterações do tecido conjuntivo em casos avançados de FSO. Objetivo: Comparar e avaliar o envolvimento e degeneração muscular da fibrose submucosa oral avançada usando três colorações diferentes: H&E, Van Gieson e Tricrômio de Masson. Material e Métodos: 10 seções de tecido fixadas em formalina e embebidas em parafina de FSO avançada foram coradas usando essas três colorações. Os resultados obtidos foram tabulados e analisados estatisticamente por meio do teste de Kruskal Wallis. Resultados: A hialinização e fibrose envolvendo o músculo esquelético foram melhor visualizadas no tricromo de Masson, mas não foram estatisticamente significativas. A degeneração muscular em áreas mais profundas foi melhor visualizada no tricrômico de Masson, quando comparado ao H&E e Van Gieson. Conclusão: Hematoxilina e Eosina coloram todos os componentes do tecido conjuntivo em vários tons de rosa. O uso de colorações especiais confere contraste entre os diferentes componentes do tecido conjuntivo, melhorando assim a avaliação da FSO. A coloração com tricrômio de Masson pode ser usada como um único adjunto para a coloração de Hematoxilina e Eosina, pois assim é possível verificar alterações superficiais e profundas no tecido conjuntivo. (AU)

Efficacy of Anhydrous Copper Sulphate as a Solid Dehydrant in Tissue Processing Procedure

Tissue processing in histotechnique is an important procedure after specimen collection involving three main procedures namely, dehydration using alcohol, clearing using xylene, and infiltration by paraffin wax. Isopropyl alcohol, the widely used dehydrating agent is toxic and when exposed to heated isopropyl alcohol fumes, it leads to numerous health hazards. Anhydrous copper sulphate is less toxic than isopropyl alcohol and requires less amount of exposure to the chemicals. The aim of the study was to investigate the dehydrating potential of anhydrous CuSO4 as an isopropyl alcohol substitute. METHODSA descriptive study of forty paired soft tissue specimens were obtained from the Department of Oral Pathology and Microbiology and were subjected to routine histopathological tissue processing with isopropyl alcohol and anhydrous CuSO4 as dehydrating agents. Histomorphological criteria namely nuclear staining, cytoplasmic staining, artefacts, and background staining were evaluated and the scores were tabulated, and statistical analysis was done using SPSS version 20 by IBM. RESULTSOur study results showed that 40 % of alcohol dehydrated tissue samples had better nuclear staining than tissue samples dehydrated by anhydrous copper sulphate and 15 % of alcohol dehydrated tissue samples had better cytoplasmic staining than tissues dehydrated by Anhydrous CuSO4. Background staining of more than 20 % was seen in anhydrous CuSO4 dehydrated tissue specimens than alcohol dehydrated tissue specimens. Artefacts were seen in similar ratios in both alcohol (80 %) and anhydrous copper sulphate (75 %) dehydrated tissue specimens. CONCLUSIONSAnhydrous CuSO4 has superior dehydrating properties than isopropyl alcohol which lead to over-dehydration of the tissue specimens. Further studies are required to validate the findings.

Quantitative analysis of copper levels in areca nut plantation area – A role in increasing prevalence of oral submucous fibrosis: An in vitro study

Background: Oral submucous fibrosis (OSMF) commonly seen in the South Asian countries is mostly associated with the chewing of areca nut (AN). Copper content in the AN has been implicated to play a major role in the pathogenesis of OSMF. It was found that most of the AN plantations in South India commonly use a copper-based fungicide, Bordeaux mixture (BM). Objective: To evaluate the level of copper in the AN, root, and soil of the AN plantation with and without the application of BM and to correlate the role of copper in the pathogenesis of OSMF. Materials and Methods: ANs, roots, and soil were obtained from plantations located in Tanniadi, Kerala. Four areas were selected from the plantation with and without BM application. The samples were collected twice with the interval of 6 months during January 2015 and July 2015, respectively. Statistical Analysis: Arithmetic mean and standard deviation were calculated. The differences between means were calculated by paired sample t-test. Results: There was statistically significant difference in the copper content of ANs, soil, and roots from both groups (P < 0.05). Samples treated with BM showed significantly higher copper levels as compared to their counterparts in January and July 2015. Conclusions: External copper from BM and increased processing for the commercial products could collectively increase the total copper content of the commercial AN products, and this high copper concentration may be implicated to the pathogenesis and the increasing prevalence of OSMF.

Evaluation of dermatoglyphic patterns using digital scanner technique in skeletal malocclusion: A descriptive study

Skin is the largest human organ, which performs a dynamic role in life. The ectodermal layers of the skin found on the palm with intricate lines are stable throughout life and have grabbed scientific attention. Any disturbances during their formation might possibly have the likelihood of a person developing malocclusion. Aims and Objectives: The aims of this study were to evaluate and compare the dermatoglyphic pattern with various skeletal malocclusions. Materials and Methods: The study was carried out on 40 outpatients reported with an age group of 18–20 years divided into four groups as follows: Group: I – 10 (Class I occlusion), Group: II – 10 (skeletal Class I malocclusion), Group: III – 10 (skeletal Class II malocclusion), and Group: IV – 10 (skeletal Class III malocclusion). The fingerprints were recorded using scanner exclusively designed for diagnostic purpose of the study. The collected data were analyzed using SPSS statistics software Version 23.0 to find which groups had significant differences. Results and Conclusion: Increased frequency of arch pattern was present in participants with Class I malocclusion, and loop patterns were observed in those with Class I occlusion and Class III malocclusion and whorl patterns in Class II malocclusion with P < 0.05. Dermatoglyphic pattern remains constant throughout life and it can be used as a noninvasive aid in determining the development of malocclusion at a very early age.

Expression of CD 68, CD 45 and human leukocyte antigen‑DR in central and peripheral giant cell granuloma, giant cell tumor of long bones, and tuberculous granuloma: An immunohistochemical study.

Background: Multinucleated giant cells (MNCs) form an integral part of numerous bone and soft tissue tumors, tumor‑like lesions and are often associated with granulomas of immunological and nonimmunological origin. The presence of various types of giant cells depends on the lesions in which they are present which are difficult to be diagnosed under routine histological techniques. Immunohistochemistry can be used for a better diagnosis and understanding of the origin of various giant cells using various markers of immune response like human leukocyte antigen‑DR (HLA‑DR) and those expressed on monocytes and macrophages like CD 68 and leukocyte common antigen (LCA). Materials and Methods: The study group consisted of 10 cases of giant cell tumor (GCT) of long bones, tuberculous granuloma, and giant cell granuloma to evaluate and analyze the expression pattern of LCA, CD 68, and HLA‑DR in various giant cell lesions. Results: Strong expression of CD 68 was observed in 80% of the lesions, strong and moderate expression of CD 45 observed in 70% of the lesions among and within the groups. In contrast, HLA‑DR demonstrated negative expression in 80% of cases except for tuberculous granuloma where all the 10 cases showed moderate to strong immunoreactivity. Conclusion: CD 68 and CD 45 expression was found in central giant cell granuloma, peripheral giant cell granuloma and GCT, suggesting the origin from mononuclear phagocyte system and considering their clinical behavior of osteoclast type. High expressivity of HLA‑DR in tuberculous granulomas which is an essential factor for presentation of the microbial antigen to CD 4 helper cells thus reassuring the fact that they are up‑regulated in response to infection.

Promoter hypermethylation patterns of P16, DAPK and MGMT in Oral Squamous Cell Carcinoma: A systematic review and meta-analysis.

Background: Oral squamous cell carcinoma (OSCC) is a common cancer world‑wide that is highly lethal due to its recurrence and metastasis. Methylation is a common epigenetic mechanism that leads to gene silencing in tumors and could be a useful biomarker in OSCC. The prevalence of P16, death‑associated protein kinase (DAPK) and O6‑methylguanine‑DNA‑methyltransferase (MGMT) promoter hypermethylation in OSCC has been evaluated for several years while the results remain controversial. Objective: The aim of this systematic review is to critically analyze and perform a meta‑analysis on the various studies in the literature that have reported the promoter hypermethylation of P16, DAPK and MGMT genes in OSCC. Search Strategy: Articles were searched and selected through PubMed. Hand search from the relevant journals was also performed. Articles were reviewed and analyzed. Results: The estimated prevalence of P16 methylation was 43%, DAPK methylation was 39.7% and MGMT methylation was 39.8%. Heterogeneity in methylation prevalences and correlations with the clinical outcomes of the disease prevailed in various studies. Conclusion: We can conclude from our systematic review that a higher prevalence of methylation of P16, DAPK and MGMT occur in OSCC. Further studies are required to substantiate the role of methylation of P16, DAPK and MGMT as a marker in OSCC.

Antioxidant and pro-oxidant activity of Vitamin C in oral environment.

Context: Antioxidant properties and Vitamin C. Background: Vitamin C is a naturally occurring organic compound and a potent antioxidant preventing oxidative damage to lipids and other macromolecules. It can also exhibit bimodal activity as a pro‑oxidant at a higher concentration. Vitamin C has a switch over role from being an antioxidant in physiologic conditions to a pro‑oxidant under pathologic conditions. A systematic review of this role would help to elucidate whether it is an antioxidant or a pro‑oxidant in the oral environment. Objective: To review studies reported in the literature elucidating the activity of Vitamin C and determine whether it is an antioxidant or a pro‑oxidant. Materials and Methods: Articles were searched in PubMed, MEDLINE using appropriate key words like “Vitamin C,” “antioxidant activity,” “pro‑oxidant activity,” “oral health” “oral disease.” Hand search of journals was also performed. Articles were reviewed and analyzed. Results: Search strategy reviewed 10 relevant articles which studied the dual role of Vitamin C. 65% of authors analyzed antioxidant action of ascorbic acid compared to 35% of the pro‑oxidant potential. Vitamin C acts as an antioxidant and a pro‑oxidant by a plethora of mechanisms. Factors determining its bimodal activity were studied, and the frequencies of their occurrence in the literature were depicted in percentage. Conclusion: The data validates the role of Vitamin C as an antioxidant under physiologic conditions exhibiting a cross over role as a pro‑oxidant in pathological conditions. Further studies are required to substantiate its pro‑oxidant activity to draw concrete conclusions.

Stromal changes in apparently normal mucosa of smokers and pan chewers: a multi-parametric approach

Aim: To compare and contrast the various changes in the connective tissue among patients with the habits of smoking, pan chewing and controls by using fluorescence spectroscopy, histopathology and transmission electron microscopy. Method: Thirty subjects were categorized into three groups: pan chewers, smokers and controls without any oral lesions. Fluorescence spectroscopy was carried out using FLUOROMAX-2. Excitation spectroscopy was performed at 280 and 320nm respectively and Emission spectroscopy was performed at 340 and 390nm excitation. Subsequently, histopathological evaluation and transmission electron microscopy was done for biopsies taken from test groups and controls. Results: The mean, standard deviation and test of significance of mean values between different groups for intensity-380nm, intensity–420nm and intensity – 460nm at 320 nm excitations, showed that the Mean values in group I and group II were significantly higher than the mean value in group III with a p-value of less than 0.001. Considering the histopathological parameters, the pattern of the sub-epithelial connective tissue, the presence of chronic inflammatory cells and lysis of connective tissue was significant. The ultrastructural analysis revealed the presence of epithelioid, spindle or elongated and stellate shaped fibroblasts in the connective tissue. Conclusion: In this pilot study, we could find variations in emission characteristics of various amino acids which correlated with histopathology and electron microscopy. Our study suggests that there are connective tissue changes in oral mucosa among smokers and pan chewers, though it is apparently normal in clinical presentation. These initial connective tissue changes could determine the progression of altered mucosa to a pre-cancer or cancer, which is further related to other complex interactions. Elaborate studies are required to evaluate the significance of our hypothesis.

Oral signs and salivary parameters as indicators of possible osteoporosis and osteopenia in postmenopausal women: a study of 45 subjects

Aim: To correlate the oral signs, salivary calcium, phosphorus, alkaline phosphatase levels and dental radiographic findings in postmenopausal osteoporotic, osteopenic and non-osteoporotic women. Materials and Methods: Forty-five subjects were selected based on bone mineral density (BMD) analyses and were assigned to 3 groups (n=15): Group 1 - established osteoporotic women; Group II - established osteopenic women; Group III (control) - non-osteoporotic women. Complete oral and radiographic examination, saliva collection and analysis of calcium, phosphorus and alkaline phosphatase were performed. Results: The results were tabulated and analyzed for statistical significance using the Mann-Whitney U-test. There was statistically significant difference (p<0.05) in salivary calcium, phosphorus and alkaline phosphatase levels when osteoporotic and osteopenic women were compared to the controls. Conclusion: Salivary parameters can be used as indicators to aid in the diagnosis of osteoporosis and osteopenia in postmenopausal women.