Role of Maspin, CK17 and Ki-67 Immunophenotyping in Diagnosing of Pancreatic Ductal Adenocarcinoma in Endoscopic Ultrasound-Guided Fine Needle Aspiration Cytology.

BACKGROUND AND STUDY AIM: One of the problems in diagnosing pancreatic ductal adenocarcinoma (PDAC) is differentiation between PDAC cells and benign pancreatic tissue cells in cytologic samples. This study aimed to evaluate the usefulness of Maspin, CK17 and Ki-67 immunocytochemistry (ICC) in differentiation between these two groups of cells. MATERIALS AND METHODS: This retrospective study was carried on 80 cases of PDAC and 25 cell blocks of benign pancreatic tissue cells as a control group for evaluation of Maspin, CK17 and Ki-67 ICC. PDAC cases were sampled by endoscopic ultrasound guided fine needle aspiration cytology (EUS-FNAC), while cell blocks of control group were aspirated from benign pancreatic tissues that were obtained from the pancreatic surgically resected specimens. Immunostaining patterns, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of each antibody as well as possible antibody combined panels of these markers in differentiation between the two groups were evaluated. RESULTS: Positive immunoreactivity for Maspin, CK17 and Ki-67 were 92.5%, 80% and 72.5% in PDAC cases, respectively. In contrast to PDAC cases, all the cell blocks of benign pancreatic tissue cells were negative for these markers. Regarding different panels, combined use of Maspin, CK17 and Ki-67 together as a triple test (at least one of them is positive) achieved the highest sensitivity of 98.8%, specificity of 100%, PPV of 100%, NPV of 96.2% and accuracy of 99% in the differentiation between PDAC and benign pancreatic tissue. CONCLUSION: Employing this short panel [Maspin, CK17 and Ki-67] is helpful for better differentiation between PDAC and benign pancreatic tissue.

Risk Assessment of Salivary Gland Cytological Categories of the Milan System: A Retrospective Cytomorphological and Immunocytochemical Institutional Study.

OBJECTIVE: The Milan System for Reporting Salivary Gland Cytology (MSRSGC) has been recently published to help communication between cytopathologists and clinicians. The aim was to assess our institutional experience with salivary gland fine needle aspiration cytology (FNAC) and the potential applicability of the MSRSGC for the estimation of the risk of neoplasm (RON) and risk of malignancy (ROM) for each category. MATERIAL AND METHOD: Salivary gland FNAC procedures performed at NCI, Cairo University in a three-year period from 2016 to 2018 and had a corresponding histopathological diagnosis were included in the current study. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy were estimated. Histopathological final diagnosis was the gold standard. Cytological diagnoses were re-stratified according to MSRSGC with estimation of RON and ROM for each category. RESULTS: A total of 118 cases were included in the current work. Sensitivity, specificity, PPV, NPV and accuracy were 84.6%, 88.2%, 78.6%, 91.8% and 87%, respectively. Cytological diagnoses were re-classified as non-diagnostic (2.5%), non-neoplastic (14.4%), atypia of undetermined significance (AUS) (6.8%), benign neoplasm (40.7%), salivary gland neoplasm of uncertain malignant potential (SUMP) (7.6%), suspicious for malignancy (8.5%), and malignancy (19.5%). The RON and ROM for each category were as follows: non-diagnostic (100%, 33.3%), non-neoplastic (17.6%, 11.8%), AUS (50%, 37.5%), benign neoplasm (97.9%, 2.1%), SUMP (88.9%, 44.4%), suspicious (90%, 60%), and malignancy (100% for each). CONCLUSION: The Milan System for Reporting Salivary Gland Cytology is a helpful classification system. The calculated ROM for each category of the studied cases was slightly above the published MSRSGC rates but still supported the recommended management for the patient.

Inter-observer reproducibility of the Royal College system for reporting thyroid cytology: Experience of the Egyptian National Cancer Institute.

PURPOSE: To assess the inter-observer agreement between 3 cytopathologists of thyroid FNAC using Royal College of Pathology reporting system. PATIENTS AND METHODS: The study is a retrospective one conducted on 204 cases retrieved from the archives of the Cytology Unit, Pathology Department, National Cancer Institute, Cairo University during the time period from January 2016 to December 2016. Cases were diagnosed separately by 3 cytopathologists using the Royal College of Pathology classification system (RCPath), where Thy1, nondiagnostic; Thy2, nonneoplastic; Thy3a, atypical, Thy3f, follicular lesion; Thy4, suspicious of malignancy; and Thy5, malignant. Kappa statistics were used where combination of the agreement between the 3 observers simultaneously was done. RESULTS: There was a good overall agreement between the three observers regarding all categories (kappa statistics was 0.679). Perfect agreement was reported for Thy5 category (κ = 0.874), good agreement was observed for Thy1 and Thy2 (κ = 0.784 and 0.719, respectively). For Thy3a, Thy 3f and Thy 4, a moderate agreement was reported (κ = 0.407, 0.446 and 0.453 respectively). Combination of surgical categories (Thy3f, Thy4, and Thy5) achieved a good agreement (κ = 0.701) as well as for non-surgical categories (Thy1, Thy2, and Thy3a) (κ = 0.712). CONCLUSION: RCPath reporting system for thyroid FNAC is clinically applicable and can be used for differentiation between benign cases needing observation and follow up on one hand, and malignant cases requiring surgical intervention on the other. The least inter-observer agreement (moderate agreement) was detected for Thy3a, Thy3f and Thy4 categories, necessitating multidisciplinary team meeting before any clinical decision.

Can GATA3 Immunocytochemistry be Utilized as a Reliable Diagnostic Marker for Metastatic Breast Carcinoma in Cytological Materials? A Comparative Study with Mammaglobin and GCDFP-15 Expression.

OBJECTIVE: Cytomorphologic differentiation of metastatic breast carcinoma from non breast metastases in cytological materials can be difficult. Current breast immunocytochemical markers have low sensitivities. Transcription factor GATA 3 is a promising marker for detecting breast differentiation in cytological materials. The aim of the study was to assess the diagnostic value of GATA 3 as a breast differentiation marker in metastatic cytological materials and to compare it with expression of mammaglobin and gross cystic disease fluid protein-15 (GCDFP-15). MATERIAL AND METHOD: We retrospectively retrieved 133 cases of metastatic breast carcinoma from the archive the of Cytology Unit between December 2013 and June 2015. They included 77 fine needle aspiration and 56 serous effusion samples. Forty-five cytological materials from non mammary metastatic tumors were used as a control. Immunostaining was performed on cell blocks for the presence of GATA 3, mammaglobin and GCDFP-15. RESULTS: GATA 3 nuclear staining was detected in 82.7% of metastatic breast carcinomas, and 11.1% of metastatic non mammary adenocarcinomas (p < 0.001). GATA 3 sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 82.7%, 88.9%, 95.7%, 63.5% and 84.3%, respectively. Mammaglobin and GCDFP-15 staining of metastatic breast carcinoma cases was positive in 70.7% and 47.1%, respectively. GATA 3 staining was significantly higher compared with mammaglobin and GCDFP-15 (p < 0.001). CONCLUSION: GATA 3 is more sensitive marker than mammaglobin and GCDFP-15 for diagnosing metastatic breast carcinoma in cytological cell block materials. Adding mammaglobin to GATA 3 resulted in improvement in its sensitivity. GATA 3 was occasionally positive in some metastatic non mammary carcinoma that may cause misdiagnosis.

Expression of cyclooxygenase 2 and vascular endothelial growth factor in gastric carcinoma: Relationship with clinicopathological parameters.

BACKGROUND: Gastric cancer is one of the most common cancers and the second most common cause of cancer-related death worldwide. Identification of specific prognostic indicators might allow a better prognostic stratification and more effective therapy. AIM: To assess the expression and relationship between COX-2 and VEGF protein in gastric adenocarcinoma and whether these markers are useful in predicting clinicopathological prognostic parameters. MATERIALS AND METHODS: The study included 83 formalin-fixed paraffin embedded tissue samples of excised gastric adenocarcinoma and 20 non tumorous tissue controls. The slides were subjected to COX-2 and VEGF immunohistochemical staining using a streptavidin-biotinperoxidase according to the manufacturer's protocol. The results were assessed independently by two pathologists. The relationships among COX-2 and VEGF expression and clinicopathological parameters were statistically analyzed. RESULTS: COX-2 and VEGF expressions were obviously higher in carcinoma tissues compared to normal mucosae (p<0.001). The expression rate of COX-2 was 54.2% and of VEGF was 68.7%. COX-2 positive tumors were significantly correlated with Lauren classification, tumor depth and Helicobacter pylori infection (p<0.001, p=0.008, p=0.035). VEGF was significantly associated with lymph node metastasis and tumor depth (p<0.001). There was a positive association between VEGF and COX-2 expression in gastric adenocarcinoma (Kappa value=0.55). CONCLUSION: In gastric adenocarcinoma, COX-2 expression might serve as a powerful indicator for intestinal type carcinoma, locally advanced disease and H. pylori infection, while VEGF was related to loco-regional progression. COX-2 might be involved in the development of angiogenesis in gastric carcinoma through VEGF upregulation.

Reliability of fine needle aspiration cytology (FNAC) as a diagnostic tool in cases of cervical lymphadenopathy.

PURPOSE: The aim of this work is to evaluate the reliability and diagnostic accuracy of fine needle aspiration cytology (FNAC) of cervical lymph nodes with an emphasis on discordant cases between the cytology and the histopathology. PATIENTS AND METHODS: This retrospective study was conducted on 157 selected patients with cervical lymphadenopathy that had undergone FNAC. Cervical nodal enlargement was the first clinical manifestation of the patients in all cases. Hypocelluar slides were excluded from the current study. The cytopathological diagnoses were compared with the histopathological results of the same excised nodes. For all discordant cases, special attention was focused on the cytomorphological features. Diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy, and discordance rate were calculated. RESULTS: The cytological diagnoses were found to be benign in 48 cases (30.6%) and malignant in 109 cases (69.4%). The overall diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of FNAC of cervical lymph nodes were 90.9%, 67.2%, 82.6%, and 81.3%, respectively. The overall diagnostic accuracy was 82.2% (129/157), while the overall discordance rate was 17.8% (28/157). The diagnostic accuracy of reactive lymphoid hyperplasia, chronic necrotizing lymphadenitis, chronic granulomatous lymphadenitis, metastatic carcinoma, Hodgkin lymphoma, and Non Hodgkin lymphoma was 85%, 83.3%, 70%, 100%, 77.8%, and 75%, respectively. CONCLUSION: The overall diagnostic accuracy of FNAC of cervical lymph nodes was 82.2% while the overall discordance rate was 17.8%. The evaluation of FNA in patients with no previously diagnosed malignancy should be interpreted by an experienced cytopathologist in the context of clinical, radiological, and laboratory finding and if any of these findings is suspicious, further investigation is justified to overcome the limitations and pitfalls of the cytomorphological features when applied alone.

Diagnostic value of p53 and ki67 immunostaining for distinguishing benign from malignant serous effusions.

BACKGROUND: The differentiation of benign mesothelial cells from malignant tumor cells, primary, or metastatic, in serous effusions based on cytomorphologic features alone can be problematic. PURPOSE: This study was conducted to evaluate the utility of p53 and ki67 immunocytochemical markers in differentiating benign from malignant tumor cells in serous effusions. PATIENTS AND METHODS: Archival Papanicolaou-stained smears of 91 pleural and peritoneal effusions were retrieved from Cytology Unit, Pathology Department, NCI, Cairo University between 2008 and 2010. Forty-one cases were positive for malignant cells and 50 cases were benign based on cytomorphologic features. Cases having doubt were excluded from the study. The slides were destained and subjected to immunocytochemical staining for p53 and ki67. Histologic sections of colonic carcinoma and tonsillar tissue were used as positive control for p53 and ki67, respectively. Smears having >5% positively stained nuclei for p53 were taken as positive and labeling index ⩾10% of ki67 was considered positive. Frequencies of the individual immunocytochemical stains; p53 and ki67, in benign and malignant effusion as well as the combination of both stains were calculated. RESULTS: p53 immunostaining showed nuclear positivity in 31 out of 41 malignant effusions (75.6%) and in 3 out of 50 benign effusions (6%), p<0.005. p53 had 75.6% sensitivity, 94% specificity, 91.2% PPV, and 82.5% NPV. ki67 immunostaining was positive in 30 out of 41 malignant effusions (73.2%) and in 17 out of 50 benign effusions (34%), p<0.05. ki67 had 73.2% sensitivity, 66% specificity, 63.8% PPV, and 75% NPV. Cases were then analyzed for combined immunoprofile of p53 and ki67. Among the 24 cases that coexpressed both antigens, 22 cases (91.7%) were malignant. Thirty two out of 34 cases (94.1%) that showed negative results for both antigens were benign. For the cases that showed p53 immunostaining only, 9 out of 10 cases (90%) were malignant. Fifteen out of 23 cases (65.2%) that showed ki67 immunostaining were benign. CONCLUSION: Benign and malignant effusions showed significantly different staining pattern for p53 and ki67. When used individually, p53 immunostaining can truly diagnose 75.6% and 94% of the malignant and benign cases, respectively. ki67 immunostaining can correctly identify 73.2% and 66% of the malignant and benign cases, respectively. When used in combination, 91.7% of p53 and ki67 positive cases were malignant while 94% of p53 and ki67 negative cases were benign. Hence they could be used when the cytomorphology fails to provide a definitive diagnosis.

The Role of P63 Immunocytochemistry for Myoepithelial Cells in the Diagnosis of Atypical and Suspicious Cases in Breast Fine Needle Aspiration Cytology (FNAC).

PURPOSE: Evaluation of the usefulness of p63 immunocytochemical marker for myoepithelial cells in the diagnosis of atypical and suspicious lesions in breast cytology. PATIENTS AND METHODS: This is a retrospective study on 122 selected patients presented at Cytology Unit, Pathology Department, NCI, Cairo University, in three years interval from 2007 to 2009, with breast lumps who underwent preoperative FNAC and diagnosed cytologically as atypical or suspicious breast lesion for biopsy then they were followed by excisional biopsy for histopathologic assessment that was considered as the golden standard diagnosis against which FNAC diagnoses were compared. Paucicellular cytologic slides as well as cases with no corresponding final histopathological diagnosis were excluded. The destained cytologic slides were subjected to p63 immunocytochemical staining. Only the nuclear immunoreactivity for p63 was considered specific, cytoplasmic and membranous staining was considered nonspecific. The stained slides with p63 marker were quantified according to the percentages of positive epithelial cell clusters and positive single bare nuclei in the BACKGROUND. The immunocytochemical results were compared with histopathologic diagnoses. RESULTS: Of the 122 studied breast aspirates, 84 cases with atypical findings and 38 cases with suspicious findings were included. The two categories yielded malignant diagnoses in 53 cases (63.1%) and 31 cases (81.6%), respectively. Invasive duct carcinoma was the most common malignant diagnosis in both categories. The most common benign diagnosis in the atypical group was fibrocystic changes (48.4%), while atypical ductal hyperplasia was the most common non-malignant diagnosis in the suspicious group (42.8%). P63 consistently stained the nuclei of myoepithelial cells, either overlying clusters and/or single bare nuclei. Of the histologically confirmed malignant cases 69% and 91.7% showed no p63 nuclear staining in cell clusters or bare nuclei, respectively; while cases showed staining pattern similar to that of benign lesions. On the other hand, 84.2% and 57.9% of the benign cases showed staining in more than 75% of the clusters and bare nuclei, respectively. The staining pattern of p63 was significantly different between malignant and benign lesions (p-value <0.005). The p63 sensitivity, specificity, positive, and negative predictive value were 90.5%, 84.2%, 92.7%, and 80%, respectively. Scattered p63 positive ductal cells ( <10% of duct cells) were detected in 6% of all malignant cases. CONCLUSION: The p63 was a reliable nuclear marker of myoepithelial cells in breast cytology. Benign and malignant breast lesions showed significantly different staining pattern for p63 on inconclusive breast cytology. The diagnostic sensitivity, specificity, positive and negative predictive value of p63 marker were 90.5%, 84.2%, 92.7%, and 80% respectively. The p63 immunostaining may be used as a diagnostic adjunct to the routine fine needle aspiration cytology in cases of breast lesions with atypical and suspicious results. KEY WORDS: P63 immunocytochemistry - Breast FNAC - Inconclusive diagnoses.

Assessment of the reliability of immunocytochemical detection of estrogen and progesterone receptors status on the cytological aspirates of breast carcinoma.

PURPOSE: Evaluation of the reliability of immunocytochemical staining for estrogen and progesterone receptor status on previously papanicolaou-stained fine needle aspiration smears of breast carcinoma cases. PATIENTS AND METHODS: This is a retrospective study conducted on destained smears of fine needle aspirates (FNA) obtained from 90 breast carcinoma cases. These cases underwent subsequent tumor resection and immunohistochemical detection of estrogen and progesterone receptors allowing a comparison between the immunocytochemical and immunohistochemical results (Gold Standard). Hypocelluar slides were excluded from the current study. Only the nuclear staining was considered specific. The results were scored on the basis of the percentage of the positive nuclei among the total epithelial malignant cells after examination of the entire slide. Smears were interpreted as positive if 10% of the examined cells demonstrated nuclear staining. These results were then compared with the immunohistochemical results. RESULTS: For estrogen receptor immunocytochemistry, the overall cyto-histologic accuracy was 91.1% (82÷90) while the discordance rate was 8.9% (8÷90). The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value were 93%, 84.2%, 95.7%, and 76.2% respectively. For progesterone receptor immunocytochemistry, the overall cyto-histologic accuracy was 88.9% (80÷90) while the discordance rate was 11.1% (10÷90). The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value were 87.1%, 95%, 98.4%, and 67.9% respectively. CONCLUSION: Immunocytochemistry is considered as an efficient tool in evaluating estrogen and progesterone receptor status in breast carcinoma. The application of estrogen and progesterone receptor immunocytochemistry on previously Papanicolaou-stained slides provides an overall accuracy of 91.1% for estrogen receptor and 88.9% for progesterone receptor when compared with the immunohistochemical results. KEY WORDS: Estrogen receptor- Progesterone receptor- Breast carcinoma- FNA- Immunocytochemistry.