Husain A. Saleh

MicroRNA profiling of diagnostic needle aspirates from patients with pancreatic cancer

Background:A major challenge to the development of biomarkers for pancreatic cancer (PC) is the small amount of tissue obtained at the time of diagnosis. Single-gene analyses may not reliably predict biology of PC because of its complex molecular makeup. MicroRNA (miRNA) profiling may provide a more informative molecular interrogation of tumours. The primary objective of this study was to determine the feasibility of performing miRNA arrays and quantitative real-time PCR (qRT–PCR) from archival formalin-fixed paraffin-embedded (FFPE) cell blocks obtained from fine-needle aspirates (FNAs) that is the commonest diagnostic procedure for suspected PC.Methods:MicroRNA expression profiling was performed on FFPE from FNA of suspicious pancreatic masses. Subjects included those who had a pathological diagnosis of pancreatic adenocarcinoma and others with a non-malignant pancreatic histology. Exiqon assay was used to quantify miRNA levels and qRT–PCR was used to validate abnormal expression of selected miRNAs.Results:A total of 29 and 15 subjects had pancreatic adenocarcinoma and no evidence of cancer, respectively. The RNA yields per patient varied from 25 to 100 ng. Profiling demonstrated deregulation of over 228 miRNAs in pancreatic adenocarcinoma of which the top 7 were further validated by qRT–PCR. The expression of let-7c, let-7 f, and miR-200c were significantly reduced in most patients whereas the expression of miR-486-5p and miR-451 were significantly elevated in all pancreas cancer patients. MicroRNAs let-7d and miR-423-5p was either downregulated or upregulated with a significant inter-individual variation in their expression.Conclusion:This study demonstrated the feasibility of using archival FFPE cell blocks from FNAs to establish RNA-based molecular signatures unique to pancreatic adenocarcinoma with potential applications in clinical trials for risk stratification, patient selection, and target validation.

Correlation of bcl-2 Oncoprotein Immunohistochemical Expression with Proliferation Index and Histopathologic Parameters in Colorectal Neoplasia

Thebcl-2 oncogene plays an important role in carcinogenesis by inhibiting cell death (apoptosis). It was initially discovered in follicular B cell lymphoma with t(14, 18), and subsequently found in other malignant and premalignant lesions. Alteration of the normal controls of cell proliferation is also a significant factor in the multistep process of tumorigenesis. The proliferative activity of a given lesion is commonly evaluated by MIB1, a monoclonal antibody to Ki67 proliferation antigen. Immunohistochemical (IHC) staining expression of bcl-2 and Ki67 was retrospectively investigated in a series of 52 colorectal carcinomas and 56 adenomas according to the avidin-biotin-complex method. The aim of the study was twofold: 1) to investigate any correlation between MIB1 and bcl-2 immunostaining expression in colonic adenomas and carcinomas, 2) to identify any relationship between either marker and several histopathologic parameters including tumor size, pathologic stage, lymph node metastasis, angiolymphatic invasion, tumor grade and differentiation in colon carcinomas. Bcl-2 was consistently higher in adenomas than in carcinomas. There were 44/56 (78.6%) adenomas, and 27/52 (51.9%) carcinomas positive for bcl-2 (p=0.004). The mean Ki67 labeling index (LI) was 30.05±7.6 and 38.12±11.01 in adenomas and carcinomas, respectively (p=0.0001). Expression of bcl-2 in carcinoma was significantly associated with a lower mean Ki67 LI and with favorable histopathologic parameters. We conclude that bcl-2 oncoprotein expression is probably an early step in the process of colon carcinogenesis, and its expression may be associated with a favorable clinical course. Furthermore, an inverse relationship exists between bcl-2 and Ki67 in colonic neoplasia. Evaluation of bcl-2 and Ki67 IHC expression in colonic carcinoma should be performed prospectively to determine if their expression is of value in predicting the clinical course in these patients.

Differentiating reactive mesothelial cells from metastatic adenocarcinoma in serous effusions: The utility of immunocytochemical panel in the differential diagnosis†

Differentiating reactive mesothelial cells (RMs) from metastatic adenocarcinoma cells (MAC) in serous fluids based on cytomorphologic features alone can be very challenging. Various immunocytochemical (ICC) markers have been used to maximize the diagnostic accuracy, however, cytopathologists still encounter difficulties in effusion cytologic diagnosis. The aim of this study was to evaluate previous and recent ICC stains to identify the most sensitive and specific markers and the best panel for differentiating RM from MAC.

Comparison of Thin-Prep and cell block preparation for the evaluation of Thyroid epithelial lesions on fine needle aspiration biopsy

Background The objective of this study was to compare the utility of Thin-Prep (TP) cytologic preparation with that of Cell Block (CB) preparation in the diagnosis of thyroid lesions, mainly follicular epithelial lesions, by fine needle aspiration biopsy (FNAB). Feasibility of using the TP slides for immunocytochemical stains is also discussed. Methods A total of 126 consecutive cases of thyroid FNAB with TP slides and 128 consecutive cases of thyroid FNAB with CB slides were reviewed blindly by two cytopathologists. The presence of colloid, follicular cells, macrophages and lymphocytes/plasma cells were recorded and scored 0–4 on each case based on TP or CB slide review. The cytologic diagnoses were grouped as follows: cyst, colloid nodule, colloid nodule with cystic change, chronic thyroiditis, atypical/neoplastic and non-diagnostic. Results The TP slides had higher diagnostic rate than CB slides. The diagnostic yield was 68% of the TP slides whereas only 24% of the CB slides were diagnostic. Also, only 4 atypical/neoplastic lesions were diagnosed on the TP slides and the corresponding direct smears, while 5 cases of atypical/neoplastic lesions were diagnosed on the smears but could not be diagnosed on the corresponding CB slides. Additionally, the TP slides revealed cytologic features that were not observed on the direct traditional smears of the same case. Conclusion In thyroid FNAB cases, TP slide preparation is superior to CB slide preparation and is more likely to have greater cellularity for diagnosis and detect atypical/neoplastic thyroid lesions, particularly those of follicular cell origin. Furthermore, TP slides appear to detect helpful diagnostic cytologic features and should be considered complementary to, rather than replacing, direct smears.

Hepatic angiosarcoma: Aspiration biopsy cytology and immunocytochemical contribution

In the absence of previously documented sarcoma, the initial diagnosis of angiosarcoma (AS) on fine‐needle aspiration (FNA) biopsy of the liver is difficult. However, awareness of its occurrence and the assistance of immunocytochemical stains may aid in arriving at the correct diagnosis. In this paper, a 59‐yr‐old smoker and alcoholic woman presented after a syncopal episode and was found to have a palpable right abdominal mass. An abdominal CT scan confirmed multiple centrally necrotic liver masses, from which an FNA biopsy was obtained. The smears were bloody with groups of relatively dishesive and singly dispersed spindle cells in a somewhat necrotic background. The nuclei were elongated to ovoid‐round with small nucleoli. The cytoplasm was ample and ill‐defined. The cells were reactive for factor VIII‐related antigen and CD31 but negative for cytokeratin immunocytochemical stains, and a diagnosis of “suspicious for angiosarcoma” was entertained. The patient declined further studies or treatment but presented 4 mo later with light‐headedness and hypoglycemia. Laparoscopic tissue biopsies of the liver/abdominal masses were obtained and revealed AS. Rarely, liver masses may represent AS. Pathologists should be aware of their cytomorphology and characteristic immunostaining to avoid their misinterpretation. Diagn. Cytopathol. 1998;18:208–211.

Utility of a Liquid-Based, Monolayer Preparation in the Evaluation of Thyroid Lesions by Fine Needle Aspiration Biopsy Comparison with the Conventional Smear Method

OBJECTIVE To retrospectively compare the diagnostic accuracy and cytomorphologic features of thyroid lesions on ThinPrep (TP) (Cytyc Corporation, Boxborough, Massachusetts, U.S.A.) monolayer preparations with those of the conventional smear (CS) method on fine needle aspiration biopsy (FNAB). STUDY DESIGN Slides of 145 TP and 145 CS consecutive cases of thyroid FNAB were retrospectively reviewed for the following features: amount and architecture of follicular cells, nuclear and cytoplasmic details, amount and quality of colloid, background blood, cyst fluid and macrophages, and lymphocytes and plasma cells. These were semiquantitatively scored 0-4 for each parameter. RESULTS The TP slides more often had higher cellularity with flat clusters, while CS slides more often had 3-dimentional clusters. The CS slides displayed better morphology and more preserved follicular cells with intact cytoplasm and crisper nuclei, while TP slides revealed shrunken cells with fragmented cytoplasm and dark, often-naked nuclei. The amount of colloid was generally more abundant on the CS slides, while it appeared as small, dense droplets (thick colloid) or as folded tissue paper-like material (thin, watery colloid) on the TP slides. The CS more often had a bloody background obscuring the cells, while the TP slides had a clear background. Simple thyroid cysts were more often detected on TP than CS slides by the presence of cyst fluid and macrophages. The 2 methods had almost similar diagnostic rates for chronic thyroiditis (11% TP vs. 12% CS) and atypical/neoplastic lesions (3.4% each). The 2 methods had similar diagnostic correlation for colloid nodules (49% TP vs. 45.5% CS), but the nondiagnostic rate was lower in TP (24%) than CS slides (31%). CONCLUSION Although there are cytomorphologic differences between the TP and CS methods, including better cellular preservation and details on CS, the TP method shows a lower nondiagnostic rate, similar diagnostic rate for chronic thyroiditis and atypical/neoplastic lesions, and slightly better diagnostic rate for colloid nodules. The 2 methods complement each other, and we strongly recommend that they both be performed on all thyroid FNAB cases.

A unique case of urinary bladder simple melanosis: a case report and review of the literature

Melanosis refers to abnormal or excessive deposition of melanin pigment in the cells and/or tissue, which can be seen in any organ but commonly in skin and oral mucosa. Melanosis of the urinary bladder is an extremely rare benign condition and only a handful of cases been reported in the English literature before. In this article, we report a new case of urinary melanosis, describe the differential diagnostic features from pseudomelanosis and offer clues for correct diagnosis. We also provide comprehensive review of the literature on the subject.

Discriminating hepatocellular carcinoma from metastatic carcinoma on fine-needle aspiration biopsy of the liver: the utility of immunocytochemical panel.

The cytomorphologic features of hepatocellular carcinoma (HCC) in fine‐needle aspiration (FNA) biopsy are well described. However, correctly diagnosing HCC on cytologic features alone and differentiating it from metastatic adenocarcinoma (MAC) remains a challenge. Studies have recommended the use of various immunocytochemical (ICC) stains to aid in the diagnosis and distinction of these tumors with variable success rates.

P53 protein immunohistochemical expression in colonic adenomas with and without associated carcinoma

Objectives:P53 protein immunohistochemical (IHC) expression was investigated in a series of colonic adenomas and carcinomas to determine the p53 immunohistochemical expression of adenomas in general compared with carcinomas, the difference in staining pattern between adenomas with associated carcinoma and those without associated carcinoma, and the difference in p53 staining in the usual adenomas (low-grade dysplasia) compared with those harboring high-grade dysplasia.Methods:The study involved a series of 20 adenomas without concurrent carcinoma (group 1 adenoma), 29 adenomas with concurrent carcinoma (group 2 adenoma), and 20 carcinomas. Sections of the paraffin-embedded tissues were stained with DO-7 p53 monoclonal antibody after microwave antigen-retrieval method. Cases with nuclear staining in ≥ 20% of the tumor cells were considered positive.Results:Analysis of results showed that 65% of carcinomas and 37% of all adenomas were reactive with p53 IHC staining (p= 0.03). With respect to the adenomas, 30% of group 1 and 41% of group 2 adenomas were reactive for p53 protein (p= 0.42).Conclusions:Our data demonstrate a statistically significant higher p53 expression rate in colonic carcinomas than in adenomas, and that adenomas with concurrent carcinomas are more frequently p53 positive than those without concurrent carcinoma, but this was not statistically significant. Also, p53 expression is more frequent and intense in adenomas with high-grade dysplasia (10/20, 50%) than in ordinary adenomas with low-grade dysplasia (8/29, 28%), which suggests a strong correlation between the degree of dysplasia in colonic neoplasia and p53 expression pattern.

Chondroblastic Osteosarcoma of the Cuboid: A Literature Review and Report of a Rare Case

UNLABELLED Chondroblastic osteosarcoma accounts for about 25% of all cases of osteosarcoma, which is the most common primary malignancy of the skeleton. Currently, only a few cases of chondroblastic osteosarcoma have been reported to involve the bones of the foot. In this report, we describe the previously unreported occurrence of chondroblastic osteosarcoma involving the left tarsal cuboid in a 15-year-old male. Histologically, the tumor showed an unusually extensive osteoclastic giant cell reaction that initially led to an erroneous diagnosis of giant cell tumor of bone. Because treatment and prognosis are very different for chondroblastic osteosarcoma as compared to giant cell tumor of bone, it is important to make the distinction between these 2 different bone tumors. In this report, we wish to draw attention to occurrence of chondroblastic osteosarcoma in the small bones of the foot, and to the possibility of an extensive giant cell reaction making accurate diagnosis difficult. We also provide a comprehensive review of the literature related to chondroblastic osteosarcoma. LEVEL OF CLINICAL EVIDENCE 4.