Comparison of Diagnostic Reliability of p63 and Smooth Muscle Actin in Salivary Gland Neoplasms.

Salivary gland neoplasms pose considerable diagnostic difficulty owing to their diverse histological features in individual lesions and the presence of a number of types and variants & similar histological features with other tumor entities. Myoepithelial and basal cells play a significant role in the pathogenesis of salivary gland neoplasm. p63 and smooth muscle actin are more reliable markers for identifying these cells and not studied much comparing their reliability in the diagnosis of salivary gland neoplasms. Hence, the aim of this study is to evaluate and compare the diagnostic reliability of immunohistochemical markers such as p63 and smooth muscle actin (SMA) in the diagnosis of various benign and malignant salivary gland neoplasms. The study comprises of 18 samples categorized into two groups: Group I comprised 9 cases, of which 4 cases were Pleomorphic adenoma, 2 cases were Myoepithelioma, 2 cases of Basal cell adenoma and 1 case was Warthin's tumor; and Group II consisted of 9cases, of which 3 was Mucoepidermoid carcinoma, 1 cases were Myoepithelial carcinoma and 5 cases were Adenoid cystic carcinoma. The selected cases were subjected to immunohistochemistry (IHC) procedure to assess the expression pattern of p63 and smooth muscle actin. The obtained data was analysed statistically by using Mann-Whitney test. In SMA, strong positivity for epithelial and connective tissue components of benign salivary neoplasm is about 22.2%respectively. In malignant salivary neoplasm, SMA was strongly positive for the epithelial and connective tissue component of about 77.7% and 88.8% cases respectively. The difference in the connective tissue components was found to be statistically significant (U = 24, P = 0.032). P63 was strongly positive for the epithelial and connective tissue component of benign salivary neoplasm of about 33.3% and 11.1% cases respectively.In malignant salivary neoplasm, p63 was strongly positive for the epithelial component of about 66.6% cases and connective tissue is completely negative. Alpha-SMA can be utilized as reliable IHC markers for salivary gland neoplasms due to its diagnostic importance in tumors with myoepithelial origin indicative of the histogenesis of salivary gland tumors and even p63 can be used as specific markers for differentiation of malignant salivary gland tumors.

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

Cedarwood oil as an alternative to xylene as a clearing agent in histopathological tissue processing - A comparative study.

BACKGROUND: Clearing in histopathological tissue processing should be able to make the tissues translucent and clear for the tissues to be visible under light microscopy and should render the clearing agent to be miscible with the dehydrant and the impregnation wax in the preceding and following processing steps. Xylene is a gold standard clearing agent but increasing concerns about the potential carcinogenicity, implementing eco-friendly agents in routine histopathology is necessary. AIM: The aim of the study is to assess the clearing ability of Cedarwood oil as an alternative to Xylene in routine tissue processing. MATERIALS & METHODS: The study was carried out in the Department of Oral Pathology and Microbiology. Formalin fixed 50 tissue samples of size 3-7mm were taken and subsequent dehydration done with acetone and alcohol. The dehydrated tissue is later processed using 90ml of Cedarwood oil with few drops of Xylene and Thymol. After clearing the tissues were subjected to impregnation and embedded in paraffin wax, later which sections were made and stained using H & E stain. RESULTS: The results of our study on comparison showed better outcome in tissues processed with cedarwood oil than xylene. Statistical Significant correlation was observed in nuclear staining (p value = 0.001) ; cytoplasmic staining (p value = 0.08) and background staining ( p value = 0.045) indicating a positive results on using cedarwood oil as clearing agent. CONCLUSION: The cedarwood oil can be considered as a safer natural alternative to xylene in laboratories. The cedarwood oil is eco - friendly and easily available with enhanced tissue processing qualities.

Inking of gross specimens: a systematic review.

BACKGROUND AND AIM: Histopathological Evaluation of surgical margins of a resected tumour specimen can give an insight about the extent of tumour spread. Errors in proper identification and orientation of resected tumour margins can lead to treatment failure and poor prognosis. Inking of resected margins is the most reliable and safe method. The aim of this study is to systematically review the studies which compares various materials used for inking of surgically resected specimen. MATERIALS AND METHODS: Articles searched from PubMed, Cochrane, Google search, manual search using key words - inking, tissue marking dyes, surgical margin, tumour margin, surgical pathology, grossing, gross specimens and back references of the articles, yielded three articles. Three articles with a total sample size of 1325 and compares properties of India ink, Acrylic colours and Tissue Marking Dyes were considered in this review. RESULTS: Both India ink and acrylic colours are good with respect to the Ease of application, visibility on paraffin wax blocks, Visibility on naked eye examination of slides and Visibility on microscope. Acrylic colours have less drying time than India ink. India ink do not result in contamination of tissue processing fluids, Interference with cellular and nuclear details and penetration in to tissues when compared with acrylic colours. CONCLUSION: India Ink will continue to dominate as the best surgical ink when comparing all the parameters till newer studies are available for acrylic colours or other dyes. Acrylic colours have the potential to be widely used as a tissue marking dyes except for the few disadvantages.

Evaluation of matrix metalloproteinase 9 gene expression in oral squamous cell carcinoma and its clinicopathological correlation.

BACKGROUND: Matrix metalloproteinases (MMPs) are calcium-dependent and zinc-containing endopeptidases which enhance cancer progression by regulating angiogenesis, migration, proliferation, and invasion. Oral squamous cell carcinoma (OSCC) is one of the most common malignancies in India, and it is observed over 90% of cases. In OSCC, MMP9 which belongs to the gelatinase group promotes tumor progression by angiogenesis, disturbing tissue morphology that allows tumor growth which breaks the basement membrane and enables metastasis, and its overexpression in OSCC is proven to have prognostic value. AIM AND OBJECTIVES: To assess the expression of MMP9 in OSCC and to correlate the MMP9 expression with pathological staging of the OSCC. MATERIALS AND METHODS: Ten OSCC tissue samples and normal tissue samples were collected. Total RNA was extracted and the complementary DNA was generated. The specific primers used in the primers were synthesized. Total reaction volume was 20 µl. The polymerase chain reaction condition included 95°C for 30 s followed by 40 cycles of two steps: 95°C for 5 s and 60°C for 30 s. The relative quantification of genes was evaluated. RESULTS: Upregulation of MMP9 gene regulation was observed in OSCC tissue samples when compared to the controls. Correlating with the pathological staging, we observed that 30% tumors were stage IVA with involvement of adjacent structures and 30% had lymph node involvement. CONCLUSION: This study is done to assess the possibility of MMP9 gene expression as a prognostic biomarker for OSCC.

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)

Diagnostic utility of touch imprint cytology for intraoperative assessment of surgical margins and sentinel lymph nodes in oral squamous cell carcinoma patients using four different cytological stains.

BACKGROUND: OSCC is most commonly associated with positive surgical margins. The important cause of loco regional recurrence is histologically positive or closed margins. Clear surgical margins might favor the patient with a better prognosis and prevent repetitive surgeries. The present study was designed to the diagnostic utility of touch imprint (TI) smears using H and E, Pap, Giemsa and Feulgen stains, so that they can be used on a routine basis. MATERIALS AND METHODS: A total of 720 smears from 130 margins resected from 32 patients who underwent surgical resection of OSCC were prospectively evaluated. The slides were fixed in alcohol and randomly divided into four different batches for staining with H&E, rapid Pap, Giemsa, Feulgen stain. TI of 30 sentinel lymph node were fixed in 95% alcohol, stained by (H&E) and evaluated by two independent observers. The results were compared with gold standard histopathology. RESULTS: H&E stain showed sensitivity 44%, rapid Pap 35%, Giemsa 29% and Feulgen stain 25%. Positive predictive value-100% for all the four stains. NPV-H&E 70%, Pap 66%, Giemsa 62%, Feulgen 59%. Diagnostic test accuracy of H&E stained smears was higher 72%, compared to Pap 67%. Giemsa 65%, and Feulgen 63%. In lymph nodes, Specificity was 94.74%, PPV 90.91%, NPV 94.74%, diagnostic accuracy 93.33%. CONCLUSION: TIC is effective in identifying an inadequate or severe dysplasia margin comparable to gold standard histopathology. It might be used to intraoperatively identify metastases in sentinel lymph nodes in clinically N0 Patients.

Saliva as a Diagnostic Tool in Oral Squamous Cell Carcinoma - a Systematic Review with Meta Analysis.

Whole saliva is mainly composed of fluid produced by major and minor salivary glands. Major salivary glands including parotid, submandibular, and sublingual glands, are known to secrete fluid transported from serum as well as surrounding glandular tissues [1]. Beside the secretions from salivary glands, oral mucosa, periodontium, as well as oral microflora also contribute to the final content of whole saliva [1]. Whole saliva therefore represents a complex balance among local and systemic sources [2]. This allows for the application of saliva in the diagnosis not only for salivary gland disorders but also for oral diseases and systemic conditions [2]. The role of saliva as a diagnostic tool in detecting Oral Squamous Cell Carcinoma. Articles published in PUBMED, EMBASE, COCHRANE, GOOGLE, manual search and back references of the articles for last 5 years extracted 77 articles. Studies which considered saliva as a diagnostic tool were included. Statistical analysis with Receivers Operating Curve to establish sensitivity and specificity of the salivary biomarkers as a diagnostic tool to detect Oral Squamous Cell Carcinoma were included for meta analysis. The measure of effect with 95% confidence interval were meta analysed for 9 articles in which 308 healthy individuals compared with 340 patients with Oral Squamous Cell Carcinoma. Highly sensitive salivary biomarkers for detecting Oral Squamous Cell Carcinoma were MMP-9, Chemerin, Choline + Betaine + Pipecolinic Acid + I - Carnitine(confidence interval ranges from 0.83-1.0). The narrow confidence interval of 0.95 + (0.88-1.00) was seen for MMP-9 followed by 1.00 + (0.78-1.00) for chemerin. Highly specific biomarkers for Oral Squamous Cell Carcinoma were MMP-9 (specificity -100%,), Chemerin(specificity-100%), over expressed mi RNA 136 with specificity of 0.88(0.69-0.97), under expressed mi RNA 27B with specificity of 1.0(0.66-1.00). Saliva can be used as a diagnostic tool with highly sensitive and specific markers namely MMP-9, Chemerin for early detection of Oral Squamous Cell Carcinoma.

Correlation between bizygomatic and maxillary central incisor width for gender identification / Correlação entre a distância bizigomática e a espessura de incisivo central maxilar para identificação de gênero

Objective: The aim of the present study is to analyze and evaluate the applicability of bizygomatic and maxillary central incisor width in identifying the sex of an individual for anthropological studies. Material and Methods: The study was conducted on 100 individuals in a private dental institution. The width of the central incisor was measured by requesting the subject to bite onto a sheet of modelling wax. The bizygomatic width was calculated with the help of a divider by taking the most prominent area of the zygomatic arch as the reference point bilaterally. Berry's formula was used to calculate the width of the maxillary central incisor from the bizygomatic width. Berry's Formula "Width of the maxillary central incisor = Bizygomatic width / 16." The data obtained was tabulated and analyzed statistically. Results: The results in our study indicated that the widths of both maxillary central incisors and the bizygomatic width were found to be higher in males when compared to females with a positive strong correlation. Conclusion: The Berry's index can be used for identifying the gender and can also be used for facial reconstruction. (AU)

Objetivo: O objetivo do presente estudo é analisar e avaliar a aplicabilidade da distância bizigomática e espessura de incisivos centrais maxilares na identificação do sexo de um indivíduo para estudos antropológicos. Material e métodos: O estudo foi conduzido com 100 indivíduos de uma instituição odontológica privada. A espessura do incisivo central foi medida pedindo ao sujeito que mordesse em uma folha de cera. A espessura bizigomática foi calculada com o auxílio de uma régua pegando a área mais proeminente do arco zigomático como ponto de referência bilateralmente. A fórmula de Berry foi usada para calcular a espessura do incisivo central maxilar da espessura zigomática. Fórmula de Berry: "Espessura do incisivo central maxial = Espessura bizigomática / 16". Os dados obtidos foram tabulados e analisados estatisticamente. Resultados: Os resultados em nosso estudo indicaram que as espessuras de ambos os incisivos centrais maxilares como as espessuras bizigomáticas foram maiores no sexo masculino do que no sexo feminino, com uma correlação positiva forte. Conclusão: O índice de Berry pode ser usado para identificação de gênero e também pode ser usado para reconstrução facial. (AU)

Surgical Specimen Handover from Operation Theater to Laboratory: A Survey.

INTRODUCTION: Essential communication between surgeons and pathologists is required when a specimen is transferred from operation theater to a laboratory. Any errors during transferring of specimen can lead to serious consequences such as wrong diagnosis, inappropriate treatment, reoperations, and physical and emotional disaster. AIM: To evaluate the incidence of mishaps and misses during the transfer of specimen from operation theater to pathology department. METHODOLOGY: This cross-sectional study was conducted among the oral and maxillofacial surgeons and postgraduate students of the Department of Oral and Maxillofacial Surgery. A self-administered questionnaire containing 15 questions pertaining to entry, collection, preservation, and transport of specimens to the laboratory was made. The questionnaire was validated and later distributed to the participants. RESULTS: Our study showed that there are misses and mishaps during the entry, collection, preservation, and transport of specimen to the laboratory. 97.1% of participants reported that they require a checklist during the transfer of specimen. CONCLUSION: Use of checklist can reduce mishaps and communication failures which is an initial link for reporting.