p16 expression in urothelial carcinoma: Experience from a tertiary care center in coastal South India.

Background: Urothelial carcinomas (UC) account for 6 and 2% of all cancers in men and women, respectively. Human papillomavirus (HPV) is one of the causative agents in cancers of the uterine cervix and head and neck. The role of HPV is also being studied in cancers of the urinary bladder, penis, and prostate. As p16-INK4a is a surrogate marker for high-risk HPVE7 oncoprotein, this study aims to highlight the utility of p16 immunohistochemistry (IHC) in the evaluation of HPV-associated UC. Materials and Methods: A retrospective study was conducted on UC of the bladder received in the Pathology department between January 2013 and December 2018. Bladder biopsies from non-neoplastic lesions served as controls. IHC was done for the detection of the p16 antigen. The p16 staining was recorded as positive, when there was strong staining in >50% of tumor nuclei. The p16 positive and negative tumors were compared based on age, gender, tumor size, grade, and muscle invasion. P value <0.05 was considered statistically significant. Results: The expression of p16 was analyzed in 72 UC and compared with 20 non-neoplastic cases, of which 26.4% of the cases showed p16 expression. The p16 expression was absent in the non-neoplastic lesions. While the majority (87.5%) of the low-grade tumors were negative for p16 expression, 43.8% high-grade tumors were positive. Similarly, a larger proportion of invasive carcinomas (38.8%) expressed p16 as compared to non-invasive carcinomas (13.8%). Thus, p16 expression showed a significant association with grade and stage in these malignancies (P < 0.05). Conclusion: The p16 expression was associated with high-grade and muscle-invasive UC. The p16 was absent in all non-neoplastic and precursor lesions. Thus, it can provide essential information not only about HPV association but also on the prognostic implications for the patients.

Effect of resveratrol and combination of resveratrol and donepezil on the expression of microglial cells and astrocytes in Wistar albino rats of colchicine-induced Alzheimer's disease.

Aim: The goal was to evaluate the effect of resveratrol (RS) and combination therapy of RS and donepezil (DPZ), on the numerical expression of microglial cells and astrocytes, in the frontal cortex, regions of the hippocampus in colchicine-induced Alzheimer's disease (AD) model. Methods: The study involved male albino Wistar rats of three months, age and consisted of 6 groups, with six animals each. The immunohistochemical staining with mouse monoclonal anti-human CD 68 and mouse monoclonal anti-GFAP was performed to assess the number of microglial cells and astrocytes, respectively. Results: AD group showed an increase in the number of microglia, and the numbers declined in the treatment groups, RS 10, RS 20, RS10/10 and DPZ + RS (p < 0.001). Astrocyte count was increased in the treatment groups in contrast to the AD group (p < 0.05). The DPZ + RS combination group revealed substantial elevation in the number of astrocytes and decreased microglial number among all the groups (p < 0.001). Conclusion: RS administration has diminished the microglial number and elevated the number of astrocytes. The elevated reactive astrocytes have decreased the microglial population. However, the limitation of our study is utilizing the colchicine for the induction of neurodegeneration. Using the transgenic models of AD may give a better insight into the pathogenesis and effect of RS. Another limitation of this study is the administration of RS and DPZ through different routes. The prospects of this research include studying the probiotic nature of RS and the effect of RS in other neurodegenerative disorders.

Web in the Neck - An Interesting Case Report.

Lateral neck masses are common in children, ranging from simple benign diseases to pathologies with malignant potential. Plexiform neurofibromas are extremely rare peripheral nerve sheath tumours involving multiple nerve sheath fascicles. They are typically seen in the paediatric population, with the majority affecting the craniofacial area and neck. Due to the close clinical and histological resemblance with other benign neck lesions such as lymphadenitis and branchial cysts, these cases can often go misdiagnosed. We describe a lesion in a young girl who presented with a progressive lateral neck swelling and how it was managed.

Diagnostic conundrum in an ambiguous posterior mandibular osseous lesion: Case report and radiological review.

INTRODUCTION AND IMPORTANCE: Non-odontogenic osseous lesions of the mandible are relatively uncommon entities compared to odontogenic lesions. Although the posterior mandible is not a usual site, it is not rare either for these osseous lesions, making the diagnosis ambiguous, and if misdiagnosed leading to different treatment protocols. CASE PRESENTATION: A 43-year-old female presented with a hard tissue lesion of the posterior mandible, misdiagnosed as a sialolith of the submandibular salivary gland in two other centers due to overlapping of symptoms, anatomic complexity, and inadequate investigations. The lesion was later diagnosed to be an osteoma of the posterior mandible with added investigations, and surgically excised. Histopathology confirmed the diagnosis. CLINICAL DISCUSSION: A variety of hard tissue lesions are known to occur in the posterior mandible like Submandibular sialolith, Osteomas, Calcified Submandibular lymph nodes, Phlebolith, and Tonsillolith. However, due to the region's structural complexity, localization of a hard tissue lesion may not always be forthright, even with radiographs. Moreover, in cases with conflicting symptoms, as was in this case there are more chances of misdiagnosis. The reasons for such diagnostic challenges are deliberated with radiological review of posterior mandibular osseous lesions. Recommendations are also suggested for proper investigations, thereby management of these posterior mandibular osseous lesions. CONCLUSION: Misdiagnosis of these posterior mandibular lesions may lead to the patient undergoing unnecessary surgical procedures as different lesions require different management. Differential diagnosis and adequate protocol for investigations are required.

Study of pre-operative neutrophil-lymphocyte ratio in urothelial carcinoma.

OBJECTIVES: Neutrophil-lymphocyte ratio (NLR), as an indicator of heightened systemic inflammatory response, predicts increased disease burden and poor oncological outcomes in urothelial carcinoma (UC). The study was undertaken with an aim to evaluate the association of NLR with clinicopathological variables and survival outcomes. METHODS: A total of 80 patients of UC were enrolled in the current retrospective study. Pre-operative NLR (within one month prior to the procedure), patient age, sex, tumour grade, pathological stage, recurrence free survival (RFS), progression free survival (PFS) and cancer specific survival (CSS) were recorded. We chose a cut-off value of 2.7 for NLR and patients were divide into two groups (NLR <2.7 and ≥2.7). RESULTS: NLR ≥2.7 was significantly associated with advanced tumour stage (p=0.001), but not with tumour grade (p=0.116). Progression (p=0.032) and death rates (p=0.026) were high in patients with NLR ≥2.7. Mean RFS (p=0.03), PFS (p=0.04) and CSS (p=0.04) were reduced in patients with NLR ≥2.7. On univariate analysis, NLR ≥2.7 predicted worse RFS (HR=2.928, p=0.007), PFS (HR=3.180, p=0.006) and CSS (HR=3.109, p=0.016). However, it was not an independent predictor of outcomes on multivariate analysis. CONCLUSIONS: Tumour stage and grade are the only independent predictors of RFS, PFS and CSS. High NLR at a cut-off value of ≥2.7 is associated with advanced pathological stage, but does not have an independent predictive value for RFS, PFS and CSS.

Immunohistochemical expression of androgen receptors in urothelial carcinoma of urinary bladder. Is it significant? Experience from coastal India.

BACKGROUND: Bladder carcinoma (BC) ranks second among the genitourinary cancers worldwide. Influence of androgens and expression of androgen receptors in neoplasms are recent findings which were implicated in the development of BC. We aimed to study androgen receptor (AR) expression in bladder urothelial neoplasms and correlate its expression with grade and stage of the tumor. METHODS: Immunohistochemistry (IHC) was done on samples collected in a tertiary care hospital over one year consisting of 71 urothelial BC and 20 non-neoplastic urothelial conditions. Two pathologists graded the IHC and nuclear staining was considered as positive expression. RESULTS: AR was expressed in 23.9% (17/71) of bladder urothelial neoplasms. AR was expressed in 25.7% and 22.3% of high and low-grade tumors and 25% and 22.3% of non muscle-invasive and muscle-invasive BC. AR expression had no significant correlation with gender, age (> 50 years), muscle invasion or grade. AR expression was significantly absent in non-neoplastic conditions (p = 0.018). CONCLUSIONS: AR has varied expression in BC and it is relatively lower in this study population.

Semantic segmentation of low magnification effusion cytology images: A semi-supervised approach.

Cytopathologists examine microscopic images obtained at various magnifications to identify malignancy in effusions. They locate the malignant cell clusters at a low magnification and then zoom in to investigate cell-level features at a high magnification. This study predicts the malignancy at low magnification levels such as 4X and 10X in effusion cytology images to reduce scanning time. However, the most challenging problem is annotating the low magnification images, particularly the 4X images. This paper extends two semi-supervised learning (SSL) models, MixMatch and FixMatch, for semantic segmentation. The original FixMatch and MixMatch algorithms are designed for classification tasks. While performing image augmentation, the generated pseudo labels are spatially altered. We introduce reverse augmentation to compensate for the effect of the spatial alterations. The extended models are trained using labelled 10X and unlabelled 4X images. The average F-score of benign and malignant pixels on the predictions of 4X images is improved approximately by 9% for both Extended MixMatch and Extended FixMatch respectively compared with the baseline model. In the Extended MixMatch, 62% sub-regions of low magnification images are eliminated from scanning at a higher magnification, thereby saving scanning time.

Wolf in sheep's clothing: Primary orbital Ewing sarcoma in a child masquerading as internal angular dermoid.

Ewing sarcoma is one of the rare, highly malignant neuroectodermal tumors frequently involving bones. Primary orbital Ewing sarcoma is extremely uncommon. We report a rare case of a 5-year-old boy who presented to us with a painless, slow-growing mass above the medial canthus of the left eye, the clinical appearance of which was masquerading as an internal angular dermoid. The child was subsequently diagnosed to have primary orbital Ewing sarcoma. Primary orbital Ewing sarcoma is a rare tumor with poor prognosis, poses diagnostic challenges, and demands a high index of clinical suspicion.

Immunohistochemical comparison of p63 and p40 in head and neck Spindle Cell Carcinoma.

BACKGROUND: Spindle Cell Carcinoma (SpCC) is a rare type of squamous cell carcinoma (SCC) with prominent malignant spindle cell component. This unique biphasic feature on histopathological examination makes its diagnosis problematic. Detection of p63 antigens in SpCC can be helpful however its expression in variousb proliferating soft tissue lesions demands for better marker. METHODS: In this study, histopathologically diagnosed SpCC of head and neck region were considered as cases, and 22 soft tissue sarcomas, reactive lesions and spindle cell lesions of the body were taken as controls. Immunohistochemistry (IHC) was done using Anti-p63 and p40 clone and the results were compared. CK was done for negative cases to prove their epithelial origin. P. value < 0.05 considered statistically significant. RESULTS: Among 22 cases of SpCC, 19 cases showed positive immunoreactivity to p63, and 18 cases for p40. IHC of controls showed no immunoreactivity in any of the sarcomas, reactive lesions or spindle cell lesions. The sensitivity of p63 is 86% while that of p40 is 82%. Specificity of both the markers was 100% CONCLUSION: Though p63 is a slightly (4%) more sensitive marker than p40, percentage of cell positivity for p40 is higher compared to p63. Both of these markers are 100% specific for SpCC.

Deep Learning Model based Ki-67 Index estimation with Automatically Labelled Data.

Ki-67 labelling index is a biomarker which is used across the world to predict the aggressiveness of cancer. To compute the Ki-67 index, pathologists normally count the tumour nuclei from the slide images manually; hence it is timeconsuming and is subject to inter pathologist variability. With the development of image processing and machine learning, many methods have been introduced for automatic Ki-67 estimation. But most of them require manual annotations and are restricted to one type of cancer. In this work, we propose a pooled Otsu's method to generate labels and train a semantic segmentation deep neural network (DNN). The output is postprocessed to find the Ki-67 index. Evaluation of two different types of cancer (bladder and breast cancer) results in a mean absolute error of 3.52%. The performance of the DNN trained with automatic labels is better than DNN trained with ground truth by an absolute value of 1.25%.

Diagnostic dilemma of a thumb swelling: An unpropitious turn of events.

Benign and malignant bone tumors arise in small bones of the hands and feet. Nevertheless, secondary deposits at these sites are extremely rare. We report a peculiar case of an adult man who presented with thumb swelling, which was later discovered to be a metastasis from renal cell carcinoma. Such cases have a sinister prognosis with a survival rate of 6-12 months from the time of diagnosis. We intend to discuss the diagnostic dilemma and treatment of acrometastases.

Clear cell carcinoma of breast lipid-rich variant.

Clear cell breast carcinomas are one of the rare types of invasive carcinoma of the breast. Among them, lipid-rich variant is still rare comprising <1%. The tumor derives its name from the intracytoplasmic neutral lipid that gives the cytoplasm a vacuolated and foamy appearance. These tumors are usually hormone receptors negative and Her-2 Neu positive. Here, we report a case of lipid-rich breast carcinoma which showed hormone receptor positivity and Her-2 Neu negative.

Use of the term atypical cells in the reporting of ascitic fluid cytology: A caveat.

BACKGROUND: Abdominal paracentesis is a routine diagnostic procedure for assessment of patients with recent onset or worsening of ascites. OBJECTIVES: The objective of the study is to (1) review clinically confirmed cases of malignancy with negative, atypical, and suspicious cytology reports and provide reasoning for discrepancies and (2) recalculate sensitivity, specificity, and predictive values after review. MATERIALS AND METHODS: Papanicolaou smears of ascitic fluid paracentesis samples received over one calendar year were reviewed retrospectively by an expert in cytopathology blinded to the final clinical and/or histopathological diagnoses. Cases with discrepancies after review were noted. Sensitivity, specificity, and predictive values were calculated before and after review of slides. Data were analyzed using SPSS version 16. RESULTS: Malignant etiology was identified in 49/115 cases (42.6%) with female genital tract being the most common site of malignancy (22, 44.8%). The remaining 66 (57.4%) had a benign etiology with hepatic cirrhosis in 42 cases (63.6%). A review revealed discrepancies in five cases, three of which were earlier called negative for malignant cells (one case each of ovarian adenocarcinoma, cecal adenocarcinoma, and cholangiocarcinoma). Two cases of ovarian adenocarcinoma that were reported as atypical/reactive mesothelial hyperplasia showed malignant cells upon review. Sensitivity and specificity after review were 69.4% and 100%, respectively, with 100% positive predictive value. CONCLUSION: Being a minimally invasive procedure, abdominal paracentesis continues to be an important diagnostic tool in guiding patient management. A proper morphological assessment with adequate clinical information and correlation with other investigations can be used to arrive at a definitive diagnosis in most cases. The term "atypical" can be misleading and is often used for want of clinical information and is best avoided.

Clinicopathological spectrum of solitary Plasmacytoma: a single center experience from coastal India.

BACKGROUND: Plasma cell disorders are a rare group of hematological malignancies that accounts for 10% of all hematological neoplasms. Solitary plasmacytomas are rarer entities accounting for less than 5% of all the plasma cell dyscrasias. They encompass three subtypes - Solitary Plasmacytoma of Bone (SPB) and Solitary Extramedullary Plasmacytoma (SEP) and multiple solitary plasmacytomas (MSP). In this study, we discuss the clinical, histopathological and immunohistochemical characteristics of solitary plasmacytomas. METHODS: A 13 year retrospective analysis of solitary plasmacytomas was performed from a single tertiary care center. Bone marrow evaluation was done concurrently at the time of diagnosis to rule out the presence of multiple myeloma. RESULTS: A total of 29 cases fulfilled the diagnostic criteria for SP during the study period. SPB accounted for 55.2%, SEP for 44.4% and MSP for 3.4% of the cases. The most common sites involved were the paranasal sinuses and vertebrae. Other infrequent sites included lymph node, tonsil and lungs. The mean age of presentation of SPB was a decade later than SEP. A male preponderance was observed in both subtypes. CONCLUSION: Solitary plasmacytoma is a rare entity, the diagnosis of which requires a systematic approach. There is limited data available in the literature on the clinico-pathological characteristics of SP from India.

Utility of fine-needle aspiration cytology in the diagnosis of HIV lymphadenopathy.

BACKGROUND: India, being a developing country, harbors the third largest human immunodeficiency virus (HIV)-infected population in the world, and HIV-associated lymphadenopathy is commonly encountered. HIV lymphadenopathy is more commonly generalized and pathology ranges from reactive lymphoid hyperplasia to infections like tuberculosis to neoplasms such as lymphoma and Kaposi sarcoma. The study intended to assess the utility of fine-needle aspiration (FNA) cytology in HIV lymphadenopathy. MATERIALS AND METHODS: A retrospective FNA slide review of HIV-infected cases with lymphadenopathy received over a period of 2 years in the cytopathology department was performed. The clinicopathological characteristics, absolute lymphocyte count (ALC), and CD4 counts were analyzed. RESULTS: Seventy-nine lymph node aspirates were received from HIV patients over 2 years. The mean age at presentation was 39 years with a male:female ratio of 2.4:1. Cervical lymph nodes (62%) were more commonly affected. Tuberculous lymphadenitis was the commonest lesion (41.8%), followed by reactive lymphadenitis (24%), nonspecific granulomatous lymphadenitis (14%), suppurative lymphadenitis (8%), cryptococcal lymphadenitis (2%), lymphoma (9%), and metastasis (1%). CONCLUSION: Lymph node FNA in HIV/AIDS is not only useful in identifying those cases that require further evaluation, but also aids in categorizing various etiologies such as opportunistic infections, non-neoplastic, and neoplastic lesions. FNA is a less expensive, expeditious minimally invasive method for an early diagnosis that abets in deciding the treatment strategy, thus curtailing the associated morbidity and mortality.

Image-guided Fine Needle Aspiration Cytology of Intrathoracic Lesions.

BACKGROUND: Percutaneous, image-guided transthoracic fine needle aspiration cytology (TTFNAC) is a rapid, yet accurate, and well-established diagnostic method used in the cytological evaluation of intrathoracic lesions. The study was done to determine the utility of image-guided TTFNAC in diagnosis of intrathoracic lesions. SUBJECTS AND METHODS: A retrospective analysis of all cases who underwent image-guided TTFNAC of a suspected intrathoracic lesion, in a tertiary care hospital was done over a period of 3 years. RESULTS: During the study period, 124 cases of image-guided FNAC of intrathoracic lesions were obtained. The mean age at presentation was 60.5 years with M:F: 3.6:1. Neoplastic lesions (71.5%) outnumbered the nonneoplastic lesions (28.5%). The most common tumor was adenocarcinoma (25%) followed by squamous cell carcinoma (SCC, 11%), and small cell carcinoma (5%). There was one case each of anaplastic carcinoma, plasmacytoma, bronchoalveolar carcinoma, and non-Hodgkin lymphoma (NHL). Most of the lesions were found on the right side and upper lobe. Among the mediastinal lesions, we found two cases of thymoma and one case each of NHL)/primitive neuroectodermal tumor (PNET), NHL, and small cell carcinoma metastasis to lymph node followed by ten cases of inflammatory lesions and seven cases of tuberculosis (TB). CONCLUSION: Image-guided TTFNAC of intrathoracic lesions is a safe method when done by well-trained medical personnel with lesser rate of complications. An early accurate diagnosis of malignancy can be made based on the cytological features; however, further subtyping of the malignancy may sometimes be difficult due to overlapping cytological features. TTFNAC can be a diagnostic tool for identifying nonneoplastic lesion such as TB. Hence, image-guided FNAC aids in early diagnosis and management of patients with intrathoracic lesions.

Neoplastic Meningitis: A Study from a Tertiary Care Hospital from Coastal India.

INTRODUCTION: Neoplastic involvement of cerebrospinal fluid (CSF) secondary to known or unknown primaries elsewhere is a poor prognostic factor and is equivalent to stage IV disease. AIM: The aim of the study is to analyse the cytological features of neoplastic meningitis in a tertiary care center. MATERIALS AND METHODS: A retrospective study of 400 consecutive CSF samples was done in the cytology laboratory of our hospital. The fluid obtained by spinal tap was sent for microbiological, biochemical and cytological evaluation. Smears that showed the presence of malignant cells were included in this study. RESULTS: Out of 400 cases, 36 (9%) showed neoplastic meningitis. Of which, 13 cases (36%) revealed leukemic infiltration, 2 (6%) lymphomatous infiltration and 21 (58%) carcinomatous meningitis. The leukemia cases included seven cases of acute lymphoblastic leukemia and six cases of acute myeloid leukemia. Among the carcinomatous meningitis cases, eight were metastasis from carcinoma breast, six from lung carcinoma and one each from malignancies of gallbladder, stomach and retinoblastoma. Four cases were metastatic adenocarcinoma from unknown primary. Pleocytosis was a significant finding seen in 58% cases (n = 21). Elevated protein and hypoglychorrhachia was noted in 68% cases (n = 18). CONCLUSION: A combined diagnostic approach including biochemical, microbiological and pathological evaluation was useful in eliminating infectious meningitis and confirming neoplastic meningitis in these cases. Cytology should be performed on cerebrospinal specimens from all patients with known or suspected malignancy with meningismus. Detection of malignant cells on cytological examination of CSF is the diagnostic gold standard for neoplastic meningitis.

Is MPV a Predictive Marker for Neonatal Sepsis? A Pilot Study.

Neonatal sepsis (NS) continues to be a diagnostic challenge and a prime cause of mortality. Forage for a lucid, cost-effective yet highly sensitive and specific marker in diagnosing this entity is an incessant task. This study aimed to evaluate the predictive value of mean platelet volume (MPV) in diagnosing NS. Neonates diagnosed with sepsis from January 2016 to March 2016 were included in the study. The subjects were stratified into the following: (i) culture-proven sepsis (group I); (ii) culture-negative clinical sepsis (group II); and (iii) control group (group III). Several hematologic markers such as hemoglobin, total leukocyte count, platelet count, MPV, plateletcrit, platelet distribution width, immature-to-mature neutrophil ratio, toxic change, serum urea, bilirubin, and C-reactive protein were analyzed. The results were compared among the groups, and their efficacy in diagnosing NS was appraised. The study involved 210 neonates, of which, groups I, II, and III constituted 64, 75, and 71 cases, respectively. The mean MPV among groups I, II, and III was 9.56, 8.86, and 8.58 fL, respectively (P<0.05). Strikingly higher values of platelet count, immature-to-mature neutrophil ratio, MPV, plateletcrit, and C-reactive protein were found in group I in contrast to those in groups II and III (P<0.05). The baseline MPV of patients with culture-proven sepsis was comparatively higher than controls and was found to be statistically significant. Hence, MPV can be a simple, economical, and specific predictor of NS.