Ibrahim M. Zardawi

High Notch1 protein expression is an early event in breast cancer development and is associated with the HER‐2 molecular subtype

Zardawi S J, Zardawi I, McNeil C M, Millar E K A, McLeod D, Morey A L, Crea P, Murphy N C, Pinese M, Lopez‐Knowles E, Oakes S R, Ormandy C J, Qiu M R, Hamilton A, Spillane A, Soon Lee C, Sutherland R L, Musgrove E A & O’Toole S A
(2010) Histopathology56, 286–296

Renal Fine Needle Aspiration Cytology

OBJECTIVE To audit and evaluate the pitfalls in renal fine needle aspiration (FNA) cytology. STUDY DESIGN A retrospective analysis of 180 renal FNAs from 163 patients, encountered at Canberra Hospital, Australian Capital Territory, between June 1989 and July 1997 was undertaken. The FNA procedures had been performed by radiologists under computed tomography (CT) or ultrasound (US) guidance. The study correlated the FNA results with biopsy findings and clinical outcome. RESULTS The initial cytologic diagnoses included 84 (47%) benign, 6 (3%) atypical, 7 (4%) suspicious, 70 (39%) malignant and 13 (7%) inadequate. Six of the 13 cytologically inadequate group, on further investigation, had malignant histology. The benign cytologic categories contained 79 benign conditions and 5 cases with a malignant outcome. The atypical cytologic group contained 5 benign and 1 malignant case. All nine cytologically suspicious cases had malignant histology. The cytologically malignant group contained 62 malignant, 7 benign and 1 patient lost to follow-up. The sensitivity was 92.5%, specificity was 91.9%, positive predictive value was 89.9%, negative predictive value was 94.0%, and efficacy of the test was 92.2%. CONCLUSION Renal FNA can provide an accurate diagnosis in most instances; however, aspiration cytology of the kidney has limitations and pitfalls. Low grade renal cell carcinoma has to be differentiated from oncocytoma, angiomyolipoma, renal infarct and reactive conditions. Renal FNA has a high negative predictive value, which is useful in reassuring patients with radiologically and cytologically benign lesions. Negative FNA does not exclude malignancy in the presence of a radiologic suspicion.

Immunohistological evaluation of MHC class I and II antigen expression on nevi and melanoma: relation to biology of melanoma

&NA; MHC antigen expression on 20 nevi, and 35 primary and 95 metastatic melanomas was studied by immunoperoxidase techniques using monoclonal antibodies to identify the antigens on frozen tissue sections. DR antigens were not detected on nevi but were detected on 71% of primary melanomas and 56% of metastases, suggesting that this antigen may be a useful marker of malignant transformation of nevi. Expression of class II antigen could not be related to other prognostic histological features of primary melanoma such as tumour thickness, but comparison of the common phenotypes of primary and metastatic melanoma suggested that expression of DR antigens alone in the absence of DP, DQ and ABC antigens may be an indicator of metastatic potential. Class I (s HLA‐A,B,C) antigens were also expressed infrequently on nevi but were detected on 43% of primary melanomas and 34% of metastases. HLA‐A,B,C expression was inversely related to thickness of the primary melanoma. This as well as the lower expression of class I antigens on metastases, may indicate that growth and spread of melanoma may be inhibited by MHC (class I) dependent cytotoxic T cell responses. Expression of class I MHC antigens was unrelated to class II antigens. Expression of DR was more common than DP or DQ, but the latter with one exception, were not expressed in the absence of DR antigens. Significant differences were not found in MHC antigen expression on metastases in lymph nodes compared to those in subcutaneous sites, but further studies are needed to determine whether such differences may exist between metastases in other visceral sites.

Fine Needle Aspiration Cytology vs. Core Biopsy in a Rural Setting

OBJECTIVE To compare, contrast and analyze the value and limitations of fine needle aspiration (FNA) cytology and core biopsy (CB) in a rural setting. STUDY DESIGN Retrospective analysis of 100 FNA cytology and 100 CB results of mass lesions from 193 patients matched for age, sex and body organs, and referred for FNA or CB in rural New South Wales, Australia, between September 1990 and May 1996. RESULTS FNA cytology and CB results from 193 patients were analyzed, based on anatomic location and cytologic criteria. Sites included lung, retroperitoneum, liver, breast, kidney, pancreas and ovary. The FNA group contained 6 inadequate, 14 benign, 3 atypical, 6 suspicious and 71 malignant cases, whereas the CB group had 1 inadequate, 24 benign and 75 malignant conditions. The inadequate samples in both groups were due to technical difficulty in obtaining representative material. The indeterminate (atypical and suspicious) group, which was the main pitfall of FNA, contained 4 low grade carcinomas, 3 low grade non-Hodgkins lymphomas and 2 fibrocystic breast changes. The benign FNA group comprised 8 cysts, 5 inflammatory/reactive conditions and 1 benign tumor/hamartoma, whereas the benign CB group contained 11 cysts, 9 inflammatory/reactive conditions and 4 benign tumors. CONCLUSION FNA was comparable to CB at most anatomic sites. CB occasionally offered additional information. This slight advantage was due to the availability of tissue from the first and often the only pass for assessment of architecture and performance of ancillary tests, which obviated the need for further sampling. On-site assessment of the core imprints at the time of the procedure by the highly skilled and experienced interventional cytopathologist was responsible for limiting the number of attempts to one core in most of the instances, therefore minimizing complications. Pathologists are encouraged to become more familiar with the criteria of aspiration cytology, which has proven its validity in the new cost-conscious environment. Despite the recent surge in the popularity of core biopsy, FNA cytology, when practiced in a multidisciplinary setting, with involvement of pathologists, radiologists and clinicians, is an extremely accurate test with very high sensitivity, which approaches that of surgical pathology, and specificity very similar to that of frozen section. FNA has a positive predictive value for a malignant diagnosis of almost 100%. FNA is a well-tolerated, relatively noninvasive test with a very low risk of complications.

Fine Needle Aspiration Cytology in a Rural Setting

OBJECTIVE To analyze the value and limitations of fine needle aspiration (FNA) cytology in a rural setting. STUDY DESIGN Prospective analysis of 1,196 FNA cytology results of superficial and deep masses from 1,088 patients in rural New South Wales, Australia, between September 1990 and May 1996. The FNA procedures were performed by palpation and image guidance using various-gauge needles and core biopsies as appropriate. RESULTS FNA cytology results were analyzed, based on body organs and cytomorphologic findings. Breast, 450 (41%); thyroid, 152 (14%); superficial lymph nodes, 150 (14%); lung, 98 (9%); and liver, 55 (5%), made up the majority of the cases. The following general cytologic categories were used: nonrepresentative (inadequate), 39 (3.58%); benign, 662 (60.85%); atypical, 45 (4.13%); suspicious, 30 (2.76%); and malignant, 312 (28.68%). Clinical and histologic follow-up (core biopsies in 100 patients and histology of the atypical, suspicious and malignant cytologic categories in 387 patients) showed over 96% sensitivity for a diagnosis of malignancy, with about a 4% false negative rate and 99.04% predictive value of a malignant FNA diagnosis. The false positive rate in the cytologically malignant group of 312 patients was 0.96%. The breast, thyroid and lymph node fine needle aspirations were mostly benign. The great majority of deep organ fine needle aspirations were malignant. Atypical and suspicious FNA cytology, seen in both superficial and deep sites, was due to either technical difficulty in obtaining material or problems of interpretation (genuine cytologic overlap or inexperience). The radiologically suspicious cases with negative cytology were either reaspirated or subjected to surgical biopsy. CONCLUSION FNA cytology, when practiced in a multidisciplinary setting with direct involvement of pathologists, radiologists and clinicians, is an extremely accurate, well-tolerated, relatively noninvasive and low-risk test that obviates the need for surgical intervention in most benign conditions and disseminated malignancies. Therefore, by taking an active role with on-site assessment of the FNA material and discussion with radiologic colleagues, the cytopathologist could offer an FNA service comparable to surgical pathology in sensitivity and very similar to frozen section in specificity.

The True Nature of Atypical Breast Cytology

Background: Atypical breast cytology is a poorly understood heterogeneous category with limited clinical utility but significant implications for patient management. Objective: To provide an insight into the true nature of atypical breast cytology in screening-detected (asymptomatic) and symptomatic settings, and find strategies for reducing the use of this diagnostic category. Materials and Methods: A total of 6,415 breast cytology samples were processed between January 2004 and December 2008. An atypical cytological diagnosis was rendered in 256 (4%) of the cases. A blind microscopic review of the atypical cases was conducted and results were correlated with subsequent histological and/or clinical outcomes. Results: Follow-up information by histology was available in 85.5%, by repeat fine-needle aspiration (FNA) in 3.5% and by imaging or clinical follow-up in 10.2% of the cases. Two patients (0.8%) were lost to follow-up. Of the 254 cases with follow-up, 62.6% were benign and 37.4% were malignant. The benign to malignant ratios were 1:1 and 2:1 in the screening and symptomatic groups, respectively. The atypical category in the screening population mostly yielded fat necrosis, complex sclerosing lesions and low- to intermediate-grade carcinoma on follow-up. The main outcomes in the symptomatic group were papilloma, fibroadenoma, ductal carcinoma in situ and lobular carcinoma. Preanalytical (suboptimal samples) factors were encountered in 34.8% and interpretative factors in 65.2% of the cases. Uncertainty about cellular morphology was attributed to such a diagnosis in 38 (14.8%) of the cases, architectural complexity in 137 (53.5%) and morphology and architecture in 70 (27.3%); 4.3% of cases were considered nondiagnostic. Conclusion: The atypical category is a necessary diagnosis but of limited use from a patient management perspective. Some preanalytical factors such as poor sample quality can be minimized by the involvement of cytopathologists in the FNA procedure. The use of the atypical category is partly dependent on the experience and confidence of the reporting pathologist. Assigning a case to this category is also likely to be unduly influenced by clinical or radiological findings. Our study indicates that the use of the atypical category can be reduced by up to 40% by appreciating these contributing factors. The practical utilization of the atypical category in breast cytology remains subjective and further study is required to identify useful objective criteria.

Investigating a cluster of vulvar cancer in young women: a cross-sectional study of genital human papillomavirus prevalence

BackgroundVulvar cancer is a relatively rare malignancy, which occurs most often in postmenopausal women. We have previously identified a geographic cluster of vulvar cancer in young Indigenous women living in remote communities in the Arnhem Land region of Australia. In this population, we investigated the prevalence of oncogenic human papillomavirus (HPV) infection in anogenital samples (vulvar/vaginal/perianal area and cervix) and compared the overall, type-specific and multiple infection prevalence between sites.MethodsA cross-sectional survey of 551 Indigenous women aged 18–60 years was undertaken in 9 Arnhem Land communities. Women were consented for HPV detection and genotyping collected by a combined vulvar/vaginal/perianal (VVP) sweep swab and a separate PreservCyt endocervical sample collected during Pap cytology screening. HPV DNA testing was undertaken using PCR with broad spectrum L1 consensus PGMY09/11 primers with genotyping of positive samples by Roche Linear Array. The primary outcomes were the prevalence of cervical and VVP high-risk (HR) HPV.ResultsThe prevalence of VVP HR-HPV was 39%, which was significantly higher than the cervical HR-HPV prevalence (26%, p<0.0001). HPV-16 was the most common genotype detected in both sites (VVP 11%, cervical 6%). HPV-16 infection peaked in women aged <20 years; however, there was a marked decline in cervical HPV-16 prevalence with age (p=0.007), whereas following an initial decline, the prevalence of VVP HPV-16 remained constant in subsequent age-groups (p=0.835).ConclusionsIn this population experiencing a cluster of vulvar cancer, the prevalence of cervical oncogenic HPV infection was similar to that reported by studies of other Australian women; however there was a significantly higher prevalence of vulvar/vaginal/perianal infection to cervical. The large discrepancy in HPV prevalence between anogenital sites in this population may represent more persistent infection at the vulva. This needs further investigation, including the presence of possible environmental and/or genetic factors that may impair host immunity.

Clinical Value of Repeat Pap Smear at the Time of Colposcopy

OBJECTIVE To determine the clinical value of a repeat (second) Pap smear at the time of colposcopy in the management of patients with recent cytologic abnormalities. STUDY DESIGN A study of paired Pap smears and their corresponding cervical biopsies during a two-year period, commencing in June 1996, was undertaken. Pap smears and cervical biopsies from 614 patients were evaluated in the Department of Pathology, Royal Darwin Hospital, Northern Territory, Australia. To maintain uniformity, the cytologic and histologic findings were assessed according to the Bethesda System. RESULTS The original (first) Pap smears included 288 high grade and 326 low grade lesions. The second smears showed 200 high grade, 221 low grade, 167 normal and 26 unsatisfactory cases. Punch biopsies revealed 242 high grade, 300 low grade and 72 inflammatory/reactive lesions. The changes noted in the second Pap smears and in the punch biopsies in the group originally diagnosed as having high grade disease were generally less advanced. The second Pap smears and corresponding cervical punch biopsies showed more advanced changes in the group originally diagnosed as having low grade disease. Removal of part of the abnormal epithelium during the first Pap smear and the desire of the colposcopist not to damage the surface epithelium prior to performing a cervical biopsy may account for some of these findings. Sampling errors and morphological misinterpretation may explain some of the findings. CONCLUSION In the second smears, new cases of high grade abnormality were discovered mainly in patients with low grade changes on the first smears. Therefore, a second Pap smear at the time of colposcopy is justifiable in the group with low grade changes on the first smear.

DNA Ploidy in Thin Melanoma

&NA; DNA ploidy in benign nevi (BN), thin non‐metaslasizing melanomas (TNM) and thin metastasizing melanomas (TMM) was investigated using an image analyser to determine whether characteristics such as nuclear area (NA) and nuclear integrated optical density (IOD) could be used to distinguish between these lesions. NA measurements showed significant differences between samples of nevus cells and melanoma cells and nuclear IOD differences were significant between TNM and TMM samples. Differences in NA and nuclear IOD were detected across the three groups (BN, TNM and TMM) but the large variability within samples and within groups indicate further studies would be necessary to determine the usefulness of these results in terms of the rate of correct group classification of a single sample for diagnostic purposes.

The microscopic complexities of C3 in breast cytology.

Background: Fine-needle aspiration (FNA) of difficult breast lesions often results in an atypical (C3) report. The assortment of outcomes generated by C3 reports varies widely, and this has given rise to different clinical management pathways. Objective: To identify and objectively assess microscopic features associated with atypical/C3 breast FNA cases. Materials and Methods: A total of 230 atypical breast FNAs were subjected to a blind microscopic rescreen using a range of robust qualitative and quantitative cytological criteria including cellularity, architectural qualities, cytomorphology and background features. A logistic regression with a receiver operating characteristic (ROC) curve and the resultant forward stepwise analysis were conducted to assess the results. This statistical testing was measured against malignant, benign proliferative and benign non-proliferative outcomes. Results: The malignant and benign proliferative outcomes showed a mixture of opposing protective and predictive individual cytological criteria. The stepwise analysis produced models demonstrating the best combination of individual cytological criteria for malignancy, proliferative and benign non-proliferative entities. In the malignancy model, discohesion, nuclear crowding within sheets, diminished numbers of bare bipolar nuclei and myoepithelial cells, the presence of tubules or necrosis and the absence of a cystic background were important features. The benign proliferative model suggested the same criteria but with the opposite implication and with the addition of several others, such as the presence of apocrine metaplasia, retained polarity and a speckled or coarse chromatin pattern. Age was a significant factor in malignant and proliferative outcomes. The benign non-proliferative stepwise analysis produced a model with fewer criteria (complex sheets, bare bipolar nuclei and a cystic background) limiting clinical application. Conclusion: Atypical/C3 breast cytology remains a legitimate reporting category. However, it is associated with a number of different histological outcomes. The incidence of the C3 category can be significantly reduced by controlling extrinsic factors and understanding the associated microscopic features.