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)

Knowledge, Attitude and practice survey on the perspective of oral lesions and dental health in geriatric patients residing in old age homes.

AIM: The aim of the study is to analyse the knowledge about oral lesions, the symptoms of such lesions and their attitude towards the treatment of these problems faced by institutionalised geriatric individuals. METHODS AND MATERIALS: This questionnaire-based survey was conducted among 103 institutionalised elders residing at various institutions. The questionnaire consisted of questions that addressed the medical and dental issues faced by the institutionalised elders and assessed their knowledge and attitude towards dental health. All the received responses were tabulated and the results were represented graphically. RESULTS: The results of the study showed that 44.66% of the elders underwent medical check-up once yearly and 72.82% of them visited the dentist. Of all 103 elders, none of them used dentures in spite of being edentulous and only 29.13% had any knowledge about oral lesions while the rest had no knowledge of the oral lesion and considered these lesions to be normal changes with increasing age. CONCLUSIONS: The findings of the present study demonstrate the need to improve access to oral healthcare and dental health education for the institutionalised elder population. In spite of the limitations of the study, we were able to record the obvious lack of dental hygiene practises, neglect and lack of motivation for proper dental care.

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.

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

BACKGROUND: 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.

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.

Cytomegalovirus and Mucoepidermoid carcinoma: A possible causal relationship? A pilot study.

BACKGROUND: Human cytomegalovirus (CMV) infection has tropism for salivary gland ductal epithelium and establishes a persistent and lifelong infection. Mucoepidermoid carcinoma (MEC) is the most common salivary gland tumor. Recent studies have established that mouse CMV-induced tumorigenesis displays histologic and molecular characteristics similar to human MEC. We wished to explore further down the lane by analyzing the expression of pp65 and the key oncogenic signaling pathway in cases of MEC and their etiological relevance in the Indian scenario as a pilot study. MATERIALS AND METHODS: Histopathologically confirmed cases of MEC (n = 4) and normal salivary gland tissue (n = 4) were subjected to immunohistochemical analysis using the markers pp65 and epidermal growth factor receptor (EGFR). RESULTS: The pp65 antigen expression was found to be negative in all the studied cases and one case of high-grade MEC showed EGFR expression. CONCLUSION: The purpose of the study was to explore the role of CMV in the development of MEC, as it might help to exploit this etiological agent as a therapeutic target. Similar to human papillomavirus (HPV), these might identify a subset of neoplasms with a varied biological behavior and alternative therapies. However, this vision is obscured by contradicting evidence in the literature. As of today, surgery remains the only best possible management for these patients unless proven otherwise.

Metastasizing Ameloblastoma - a perennial pathological enigma? Report of a case and review of literature.

The Ameloblastoma is a slow growing locally invasive odontogenic epithelial neoplasm with a high recurrence rate and a low tendency to metastasize. Metastasis in Ameloblastoma was first described by Simmons and Emura in the 1920s. Slootweg and Muller proposed the term Malignant Ameloblastoma to describe a well-differentiated ameloblastoma that metastasizes but maintains the characteristic cytologic features of the original tumour and the term Ameloblastic Carcinoma to an ameloblastoma with malignant cytological features. About 2% of ameloblastomas undergo metastasis. So far there have only been two cases of Metastasizing Ameloblastoma reported from the Indian Subcontinent. We present the case of a 22-year-old male Indian patient, who presented with a diffuse swelling in the left posterior mandible. Radiographs revealed a multilocular radiolucency in the left mandible. On histopathological examination, the lesion was diagnosed as follicular ameloblastoma. Four years later the patient presented with a swelling in the left submandibular region. Histological examination revealed metastatic ameloblastoma within the cervical lymph node.