Evaluation of Effusion Fluid (Pleural and Peritoneal) with Special Reference To Cell Block Preparation and Immunohistochemistry for Differentiating Between Reactive Mesothelial and Malignant Effusion

Background: The serous cavities are lined by a single layer offlat mesothelial cells called the serosa. Normally these cavitiesare collapsed and contain only a small amount of fluid, enoughto lubricate the adjacent surfaces. Cytological examination ofserous fluid is of paramount importance. It reveals informationabout inflammatory conditions of serous membrane, infectionby bacteria, fungi, virus and presence of malignant cells.Differentiation of population of reactive mesothelial cells fromthose of malignant cells remains a diagnostic challenge inconventional cytological smear. To overcome this challenge,cell block technique along with immunocytochemistry gives abetter histoarchitectural pattern and support immensely forcategorising the effusion to be reactive or malignant.Aims and Objectives: To evaluate utility of cell blocktechnique in effusion fluid (pleural and peritoneal) using limitedimmunohistochemistry markers for differentiating betweenreactive mesothelial and malignant mesothelial cells.Materials and Methods: The present study was carried out inDepartment of Pathology at M.K.C.G MCH, Berhampur,Odisha over a time period from July 2016- July 2018. A total of90 cases were evaluated. The fluids were stained with routinecytological stains. Cases on evaluation of cytomorphology ifsuspicious for malignancy, cell block was prepared. Cell blockwere stained both for routine hematoxylin and eosin andimmunohistochemistry with EMA (Epithelial marker antigen) forepithelial cells and Calretinin for mesothelial cells.Results: A total of 90 cases were evaluated cytologically. 40cases showed benign features and 24 cases showedmalignant features on cytomorphology alone. 26 cases weresuspicious for malignancy which on cell block preparation andimmunocytochemistry were differentiated as benign (10 cases)or malignant (16 cases). EMA showed 97.5 % sensitivity and98% specificity. Calretinin showed 100 % sensitivity and 97.5%specificity.Conclusion: The use of cytopathology of pleural andperitoneal effusion is helpful for early diagnosis and treatment.The technique is cheap, easy to perform and produces speedydiagnosis. In the identification of malignant cells in effusion andits differentiation from cells showing reactive and degenerativechanges there were diagnostic difficulties in some of the cases.Immunocytochemistry is an important diagnostic tool ineffusion cytology.

Utility of MOC-31 monoclonal antibody in differentiating metastatic adenocarcinoma cells and reactive mesothelial cells in effusion cytology

In effusion cytology, a clear distinction between reactive mesothelial cells and metastatic adenocarcinoma cells is sometimes challenging mainly due to similarities in the cytomorphological features. In such cases for definitive diagnosis, paraffin-embedded cell block examination and immunohistochemistry are helpful in making this distinction. MOC-31 is one of the proposed immunomarker for adenocarcinoma cells. We undertook to evaluate the role of MOC-31 as a marker for identifying adenocarcinoma cells in effusion specimen. A total of 185 paraffin-embedded cell blocks of effusion samples were identified, of these 111 cases were of metastatic adenocarcinoma. MOC-31 was positive in 101 of the 111 cases of metastatic adenocarcinoma. Minimal focal cytoplasmic staining was also seen in 7 of the 74 cases of reactive mesothelial cells, but these were taken negative as they did not show membrane positivity. The sensitivity and specificity of MOC-31 for metastatic adenocarcinoma cells were 92.5%, and 100% respectively, positive and negative predictive value (NPV) was 100% and 91.14%, respectively. MOC-31 can be used as a reliable marker in effusions for distinguishing metastatic adenocarcinoma from reactive mesothelial cases.

Usefulness of E-Cadherin Expression in Malignant Effusion / 대한세포병리학회지

The usefulness of E-cadherin immunostaining as a marker of malignancy in the body fluids was investigated in the present study. Thirty-three histologically proven cases of cell blocks from the pleural, peritoneal, and pericardial fluids were studied by immunocytochemistry for E-cadherin antibody using LSAB method. These cases were cytologically diagnosed as adenocarcinoma (25 cases) and atypical cells (8 cases). Tumor cells showed strong positive membranous staining for E-cadherin antibody in 21 out of 25 cases (84%) of adenocarcinoma. E-cadherin staining was not found in 6 of 8 cases of suspicious maligancy. The sensitivity and specificity were 84% and 75%, respectively. Reactive mesothelial cells and inflammatory cells scattered were all negative. In conclusion, E-cadherin is an useful adjunctive marker to distinguish reactive mesothelial cells from the carcinoma cells in the body fluids.

Azathioprine Therapy in Henoch-Schonlein Purpura Nephritis Accompanied by Nephrotic syndrome

In 1989, the Bethesda System (TBS) was introduced as an attempt to standardize cervical/vaginal reporting systems. TBS nomenclature was created for reporting cytologic diagnoses to replace the currently used Cervical Intraepithelial Neoplasia (CIN) and Papanicolaou Class System, which are deemed less reproducible. The name for preinvasive squamous lesions was changed to squamous intraepithelial lesion (SIL), subdivided into low-grade and high-grade types. TBS recommends a specific format for cytologic report, starting with explicit statement on the adequacy of the specimen, followed by general categorization and descriptive diagnosis. Pathologic and epidemiologic studies performed over last 10 years have provided evidence that human papillomavirus (HPV) plays a significant role in the development of cervical neoplasia. TBS corresponds not only to currently held views of the behavior of preinvasive lesions and their HPV distribution, but also to the current guidelines for clinical management.

The Significance of AgNOR Count in Body Fluid: Differential between reactive mesothelial cells & malignant cells / 대한세포병리학회지

To distinguish reactive mesothelial cells from malignant cells in body fluid, we applied silver staining of nucleolar organizer regions(AgNORs) to ethanol fixed cytologic preparations. Fifty aspirated samples of benign(22 cases) and malignant(26 cases) body fluids were studied using the one step silver staining method. Two cytologically atypical samples were also included in the study. In malignant cases the mean AgNOR count was 3.56+/-0.81, while in benign cases the mean AgNOR count was 2.02+/-0.33. The difference of AgNOR counts between these two groups were statistically significant(p<0.001). The mean of atypical cases was 2.91. Both were diagnosed as malignant in follow-up cytology. In malignant effusions, there is statistically significant difference in AgNOR counts between cells forming complex papillae or clusters and singly scattered cells(p<0.05), 3.29+/-0.95 and 3.83+/-0.55, respectively. We concluded that AgNOR count appears to be useful as a diagnostic tool especially when the cytologic differentiation is difficult.

Immunocytochemical Characteristics of the Short-term cultured Mesothelial Cells / 대한세포병리학회지

Reactive human mesothelial cells were examined by immunocytochemical stain with intermediate filaments(cytokeratin [CK1, CK7, CK8, CK18, CD19/, vimentin, desmin, actin), epithelial membrane antigen, carcinoembryonic antigen(CEA), MHC class II antigen(HLA-DR), LeuM-1(CD15), alpha1-antitrypsin(ACT), alpha1-antichymotrypsin (ACHT), CD68(KP-1) and FcgammaRIII(CD16). The mesothelial cells were isolated from patients with liver cirrhosis and pleural effusion, and short-term cultured in RPMI 1640 media containing 10% heat inactivated fetal calf serum and 1% identical supernatant fluid of the patients' transudates. The results obtained are as follows. 1. The cultured-reactive mesothelial cells were positive for the protein of cytoskeleton such as cytokeratin and vimentin, but negative for desmin and actin. The resting mesothelial cells showed positive reactions for cytokeratin, but negative for vimentin, desmin and actin. 2. The primary antibodies to the cytokeratin were strongly reactive for CK1, CK8 and CK18 but negative for CK7 and CK19 in both reactive and resting mesothelial cells. 3. Resting mesothelial cells showed negative reactions for CEA, but strong positive reactions in cultured-reactive mesothelial cells. 4. The markers for the monocytes\histiocytes (CD11b, CD14, CD16, CD68, lysozyme and alpha1-antitrypsin and alpha1-antichymotrypsin) were nonreactive in resting mesothelial cells, but lysozyme and alpha1-antitrypsin were weakly reactive in reactive and proliferative mesothelial cells. 5. MHC Class II molecule(HLA-DR antigen) was negative in both resting and reactive mesothelial cells. These results suggest that the short-term cultured, reactive mesothelial cells show a newly aberrant expression of the vimentin and carcino-embryonic antigen. The reason of the aberrant expression of the intermediate filament and oncofetal antigen in reactive and proliferative mesothelial cells should be further evaluated.