Study of morphology with assessment of expression of proliferation marker Ki67 antigen and P53 protein in lesions of gall bladder.

OBJECTIVE: To study the spectrum and distribution of histopathological changes and evaluate immunohistochemistry markers p53 protein and Ki67 antigen in various lesions of gall bladder. MATERIALS AND METHODS: A total of 804 consecutive gall bladder specimens were evaluated. Forty cases were selected for immunohistochemical analysis to evaluate expression of p53 and ki67 proliferation index, including 20 carcinoma gall bladder cases and 20 cases of inflammatory pathology associated with metaplasia, atypia, hyperplasia, dysplasia, and adenoma. p53 immunostaining was categorized as wild type and mutant type. ki67 of >20% was considered high expression. RESULTS: The majority of the gall bladder lesions were inflammatory in origin, most common being chronic cholecystitis. In the group of 20 gall bladder carcinoma cases, 65% were p53 mutant and the remaining 35% cases had a p53 wild-type immunophenotype. 55% cases showed high expression for ki67 labeling. However, significant correlation ( P < 0.05) was seen with lympho-vascular invasion. Among non-malignant lesions, normal/wild-type p53 expression was seen with increasing intensity and positivity in lesions with atypia and intra-epithelial neoplasms. Ki67 index also showed the same trend in all cases. CONCLUSIONS: p53 and ki-67 expression increases in inflammation, and further increment occurs in premalignant and malignant lesions of the gall bladder epithelium and can be used as a marker of aggression of histopathological lesions. The results emphasize the potential of Ki-67 and p53 as biomarkers of carcinogenesis in gall bladder carcinoma.

A retrospective clinicopathological analysis of rhino-orbital mucormycosis in second wave of COVID-19.

Background: During the present surge of COVID-19 positive cases, concurrent multifold increase in the incidence of mucormycosis cases has resulted into significant morbidity and mortality. We retrospectively evaluated the clinicopathological features along with microbiological examination findings in histologically diagnosed cases of rhino-orbital mucormycosis. Material and Methods: All the H and E and special stained slides of included mucormycosis cases were retrieved from the records and were evaluated with microbiological findings including screening KOH mount examination and culture results. Results: Out of 16 cases with available details, 10 cases had the previous history of diabetes mellitus. The most frequent single site of involvement was maxillary sinus (7/25) followed by nasal cavity, orbit, ethmoid and sphenoid sinuses. While comparing the histological diagnosis with KOH mount findings and culture results, 15 cases revealed consistent results. Conclusion: A high clinical suspicion, monitoring, early diagnosis, and timely management can improve the morbidity and mortality of this life-threatening complication.

Evaluation of variable p16 immunostaining patterns, Ki-67 indices and HPV status in cervical SILs and squamous cell carcinomas: An institutional experience.

Objective: The present study evaluates the immunoexpression of p16 and Ki-67 in cervical squamous intraepithelial lesion (SIL) and carcinomas and correlates their expression with clinicopathological features and HPV-DNA status. Material and Methods: A total 36 included cases of SIL and squamous cell carcinoma (SCC) were subjected to p16 and Ki-67 immunostaining. p16 staining was evaluated depending on grading, distribution, localization pattern, intensity and IHC score. Ki-67 expression was graded based on percentage of positive cells. Results: Incidence of HSIL and SCC cases was found to be significantly increased with parity > 5. p16 grade III diffuse nucleocytoplasmic immunostaining was observed in 62.5% LSIL, 80% HSIL and 87% SCC cases. Significant association of p16 staining intensity, IHC score and Ki-67 indices was noted with increasing grades of SILs and carcinomas. Conclusion: Our experience indicates that a combination of p16 and Ki-67 immunostaining may be useful to determine the severity of dysplastic change.

Immunoexpression of PTEN, HER2/neu, and Ki-67 in endoscopic gastric carcinoma biopsies; their correlation with histological subtypes and one-year survival.

BACKGROUND: Gastric carcinoma is a major cause of cancer-related morbidity and mortality worldwide. Gastric neoplasms arise from genetic and epigenetic changes in various genes. Present study evaluates the immunoexpression of PTEN, HER2/neu, and Ki-67 in endoscopic gastric carcinoma biopsies and correlates the expression of these proteins with clinicopathological features. MATERIAL AND METHODS: Adequate endoscopic biopsies of 27 cases of gastric carcinoma were evaluated for World Health Organization (WHO) and Lauren's classification subtypes along with HER2/neu, PTEN, and Ki-67 immunoexpression. HER2/neu immunostaining was scored as proposed in the Trastuzumab for gastric cancer (ToGA) trial while PTEN staining and downregulation were assessed using an immunoreactive score. The cut-off for Ki-67 expression was taken as 90th percentile of the values in adjacent non-neoplastic tissue. All statistical analysis was done at 5% level of significance with SPSS v22 statistical software. RESULTS: Tubular adenocarcinoma was the commonest WHO histological subtype and 56% of cases were of intestinal type as per Lauren's classification. 55.6% of cases showed a complete loss of PTEN expression in neoplastic tissue. 17 of the 19 cases with adjacent non-neoplastic tissue showed PTEN downregulation in neoplastic tissue. 81.5% of cases had a high Ki-67 index and HER2/neu overexpression was noted in 36% of cases. All the four cases who died had high Ki-67 proliferation indices; 3 patients had loss of PTEN expression and HER2/neu overexpression. CONCLUSION: We conclude that these immunomarkers can play important role in the behavior of gastric carcinomas and can be targeted for new therapies.

Flow cytometry in reactive lymph node aspirates: Evaluation of distribution of populations with immunophenotypic outliers.

BACKGROUND: Diagnostic accuracy of fine needle aspiration cytology for lymphomas has markedly improved when used in conjunction with flow cytometry (FCM), especially to differentiate reactive lymph node aspirates from low-grade lymphomas, a challenge for cytopathologists. Lymphoma diagnosis by FCM depends on detection of immunophenotypic B- and T-cell outliers, also present in benign lymph nodes albeit in subsets of cells. These can mimic heterogenous populations obtained from lymphomas partially involving lymph nodes. METHODS: In the present study, FCM of fine needle aspiration/scrape material from 30 cases of benign lymph nodes was evaluated for the distribution of lymphoid populations and the presence and percentages of immunophenotypic outliers. RESULTS: The majority of the samples were cervical nodes with all cases showing T-cell predominance. CD5/CD19 co-expression was seen in 23.94 ± 9.51% (20 cases) and CD10/CD19 co-expression in 3.2%-20.2% (8 cases) of B-cell events. None of these cases showed light chain restriction. Loss of CD7 (2%-28.4%) and CD5 (12.7%) expression was seen in 17 cases and 1 case, respectively, of T-cell events. CONCLUSION: Immunophenotypic outliers suggestive of lymphomas could be seen in non-lymphomatous lymph node aspirates in a subset of the cell population. However, they could be defined as benign due to absence of light chain restriction and normal CD4:CD8 ratios in all but one case, which was confirmed to be benign after clinical correlation and excisional biopsy. Thus, multiparameter FCM along with clinicoradiological correlation can prevent overdiagnosis of lymphomas.

Nasopharyngeal melanoma: An unusual entity.

Nasopharyngeal melanoma is a rare condition with an estimated incidence of about 0.3/10 lac per year and has a slight female predominance. It can arise from stem melanocytes and mature melanocytes of the submucosa that have acquired genetic alterations, may be due to tobacco, trauma and oxidative stress. It resembles other common polypoidal lesions; therefore, histopathological examination with immunohistochemistry plays a pivotal role in confirming the diagnosis. Lack of specific clinical features often leads to a delay in diagnosis.

Primary epithelial splenic cyst: A rare encounter.

Splenic cysts are rare with an overall incidence of 0.07% in a large autopsy series and 0.5% among all the splenectomies done. The parasitic splenic cysts, usually caused by Echinococcus granulosus, account for 60% of all primary splenic cysts. The primary epithelial cysts account for 10% of all splenic cysts. We report a case of 30-year-old female presenting with left upper abdominal pain and heaviness. Computed tomography revealed a multiloculated cyst in spleen. Hydatid serology was negative. Total splenectomy was done. Histopathological evaluation was done and a diagnosis of primary epithelial splenic cyst was given.

Role of CD71 in acute leukemia- An immunophenotypic marker for erythroid lineage or proliferation?

BACKGROUND: CD71 or Transferrin receptor is expressed on the surface of erythroid lineage cells. CD71 expression has been found to be significantly increased in rapidly proliferating cells. METHODS: This cross-sectional study included 37 bone marrow samples of acute leukemia cases diagnosed between October 2016 to April 2018. The samples were analysed on BD FACS Canto II. We evaluated the expression of CD71 on leukemic blasts and compared median fluorescent intensities (MFI) of blasts in different types of acute leukemias. RESULTS: The 37 cases comprised of 21 Acute Myeloid Leukemia (AML), 13 B-Acute Lymphoblastic Leukemia (B-ALL), 2 T- Acute Lymphoblastic Leukemia (T-ALL) and 1 mixed phenotypic acute leukemia (MPAL), T/Myeloid. CD 71 expression was noted in 70.3% (n= 26/37) of acute leukemia cases. CD71 expression was most commonly observed in AML (n= 15/21;71.4%), followed by B-ALL (n= 9/13;69.2%) and T-ALL (n= 1/2;50%). Single case of MPAL revealed blasts positive for CD71. MFI of leukemic blasts of single CD71 positive T-ALL was found to be highest, followed by AML, MPAL (T/Myeloid) and least in B ALL. Of the AML cases, the blasts of AML-M6, acute promyelocytic leukemia and AML-M1 showed higher CD71 expression in terms of MFI. CONCLUSIONS: Surface CD71 expression is not only found in erythroid lineage cells, but also in proliferating cells. CD71 MFI is highest in T lymphoblasts followed by leukemic erythroblasts, myeloblasts and least in B lymphoblasts.

Cytopathology of Pulmonary Lesions: A Tertiary Care Center Experience.

BACKGROUND: Cytological evaluation of specimens from respiratory tract is the initial investigation in patients suspected to have pulmonary diseases. The various cytological specimens submitted for analysis include bronchoalveolar lavage, bronchial wash, brush smears, transbronchial needle aspiration, guided fine needle aspiration cytology (FNAC) smears and pleural fluid. Present study was undertaken to study the spectrum of lesions diagnosed by cytomorphological analysis of various cytological specimens. MATERIALS AND METHODS: Centrifuged and direct smears from received samples were stained with MGG and PAP stain. Special stains (ZN and PAS) were used wherever required. Cytohistological correlation was done wherever biopsy was available. RESULTS: This study included 671 samples from 583 patients suspected clinicoradiologically of having a respiratory pathology. A male preponderance (65.87%) was noted with 73.59% of patients in age group 40-80 years. Cytological diagnoses were classified as non-diagnostic (14.90%), negative for malignancy (59.76%), specific benign lesions (5.22%), positive for atypical cells (3.87%) and malignant (16.25%). Tuberculosis (TB) was found in 17 cases. Adenocarcinoma (both primary and metastatic) was the commonest malignancy of the 99 cases positive for malignancy. Incidence of primary lung adenocarcinoma and squamous cell carcinoma were found to be equal. CONCLUSION: Lung carcinoma is presently the leading cause of cancer deaths while TB is still a common cause of death in developing countries. Cytopathology has a definite role in diagnosis of malignant lesions and also contributes in diagnosis of unsuspected chronic infections. Cytological diagnosis is fairly accurate if the specimen obtained is adequate and representative.

Diagnostic Utility of Epithelial Membrane Antigen (EMA) and Calretinin (CAL) in Effusion Cytology.

INTRODUCTION: Fluid cytology and subsequent utilization of immunocytology on cell block is being used commonly for increasing the diagnostic accuracy in effusion cytology. AIM: To do cytological analysis of fluids and evaluate the role of Epithelial Membrane Antigen (EMA) and Calretinin (CAL) to differentiate between reactive and malignant cell in pleural and peritoneal fluids. MATERIALS AND METHODS: This was a prospective study carried out in Department of Pathology, Shri Guru Ram Rai Institue of Medical and Health Science, for a period of 18 months from January 2014 to June 2015. A total of 253 samples of pleural and peritoneal fluid were studied by May-Gruenwald-Giemsa (MGG) stain and Papanicolaou (Pap) staining. In 73 cases which were suspicious for malignancy, cell blocks were prepared and IHC was done using two immunomarkers- EMA and CAL. RESULTS: A total of 253 cases of effusion cytology were studied out of which 73 were found positive for malignant cells. Maximum cases of malignant cells or atypical cells were seen in peritoneal fluid. A total of 34 cases were histologically positive for malignancy. All these cases showed strong membranous and cytoplasmic positivity for EMA. Most of cases also showed 2%-5% positivity for CAL. There were 38 cases categorized as atypical or reactive mesothelial cell hyperplasia. These cases showed nuclear and cytoplasmic staining for calretinin and none of these were positive for EMA, although 5 cases showed score 2-3, which was considered as negative. CONCLUSION: The distinction between reactive and malignant mesothelial cells in cytological specimens can be problematic. A combination of CAL and EMA may help in accurate diagnosis.