Anal Cytology: Institutional Statistics, Correlation With Histology, and Development of Multidisciplinary Screening Program With Review of the Current Literature.

CONTEXT.­: The incidence of anal cancer in the United States is on the rise in high-risk populations. The anal Papanicolaou test (APT) is advocated as a screening tool, in addition to digital rectal examination and high-resolution anoscopy. OBJECTIVE.­: To review our experience and the current literature to create, in cooperation with clinicians, a standardized screening and treatment algorithm given our large volume of APTs. DATA SOURCES.­: All APTs collected between January 2013 and June 2015 were reviewed and correlated with follow-up/concurrent biopsy diagnoses, and clinical and social history. In total, 1417 APTs were performed on 1185 patients and APT results were as follows: 17.4% (247 of 1417) unsatisfactory; 27.9% (395 of 1417) negative; 19.5% (276 of 1417) atypical squamous cells of undetermined significance (ASC-US); 24.1% (342 of 1417) low-grade squamous intraepithelial lesion (LSIL); 3.6% (51 of 1417) atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (HSIL) (ASC-H); and 7.5% (106 of 1417) HSIL. In total 376 cases (26.5%) had concurrent/follow-up biopsy. Review of all unsatisfactory cases with squamous intraepithelial lesion (SIL) on biopsy showed LSIL in 19.2% (5 of 26). Anal Papanicolaou test with cytologic abnormality (ASC-US+) had an 83.8% (315 of 376) rate of biopsy-proven disease, and sensitivity was higher (92%) for high-grade anal intraepithelial neoplasia or worse (AIN2+). Overall detection of AIN2+ using ASC-US+ showed specificity of 26%, negative predictive value of 92%, and positive predictive value of 26%. CONCLUSIONS.­: Anal cytology has a high abnormal rate (54.7%) and sensitivity but poor correlation with histologic grade. High unsatisfactory rate indicates need for improvement in sampling with 68.4% of cases having SIL on biopsy. Multidisciplinary effort led to improvements in sampling, cytologic interpretation, and development of a standardized management algorithm.

Sarcomatoid urothelial carcinoma of the bladder: a contemporary clinicopathologic analysis of 37 cases.

INTRODUCTION: Sarcomatoid urothelial carcinoma is a dedifferentiated biphasic tumor that exhibits morphological and/or immunohistochemical evidence of epithelial and mesenchymal differentiation. In this series, we analyzed the clinicopathologic features of this rare variant of urothelial carcinoma. MATERIALS AND METHODS: A search was made through our surgical pathology files and consultation files of the senior author for cases of sarcomatoid urothelial carcinoma of the bladder from 2005-2014. All the slides were retrieved and re-reviewed, and clinical data was also obtained including follow up. RESULTS: Thirty-seven cases of sarcomatoid urothelial carcinoma of the bladder were identified. Mean patient age was 71 years (range: 51 to 88 years). Twenty-six of 37 (70%) patients were male and 11/37 (30%) patients were female. Twenty-five cases were from cystectomy/cystoprostatectomy specimens, 8 cases from transurethral resection of bladder tumor specimens and 4 cases were from biopsy specimens. The mean tumor size was 5 cm (range: 1.4 cm to 13.0 cm). Four of 37 (10%) cases had focal heterologous components; 1 case with both chondroid and osteoid, 2 cases with chondroid and 1 case rhabdoid elements. Twenty-one of 37 (56%) patients died within a year of presentation. CONCLUSIONS: Sarcomatoid urothelial carcinoma of the bladder is more prevalent in males, with the mean age of 71 years in our series. Smoking is an important risk factor. Sarcomatoid urothelial carcinoma is an aggressive variant of urothelial carcinoma which commonly presents at an advanced stage, and over 50% of patients in our series died of disease within 1 year of presentation.

GATA3 expression in sarcomatoid urothelial carcinoma of the bladder.

The current available data on GATA-binding protein 3 (GATA3) expression in sarcomatoid urothelial carcinoma are limited, especially in the non-tissue microarray-based setting. In this study, we analyzed the expression of GATA3 in sarcomatoid urothelial carcinoma of the bladder in cystectomy/cystoprostatectomy specimens. A search was made through our surgical pathology and consultation files for cystectomy/cystoprostatectomy specimens with a diagnosis of sarcomatoid urothelial carcinoma. Only cases with available tissue blocks were selected. Immunohistochemical staining for GATA3 was performed, and staining in adjacent/overlying conventional urothelial carcinoma and/or benign urothelium was also documented. Twenty-two cases were obtained. Of 22 cases, 16 (73%) of sarcomatoid urothelial carcinoma were positive for GATA3. In the 7 (27%) of 22 cases that were negative for GATA3, it was observed that these cases were predominantly composed either of pleomorphic undifferentiated sarcomatoid areas or foci composed of extensive heterologous elements (chondroid, osteoid, or rhabdoid). GATA3 staining was positive in the adjacent/overlying conventional urothelial carcinoma and/or benign urothelium in all cases. This is one of the largest studies to date analyzing the expression of GATA3 in sarcomatoid urothelial carcinoma in cystectomy/cystoprostatectomy specimens. GATA3 is expressed in most cases of sarcomatoid urothelial carcinoma. Negative expression may, however, be observed in cases composed predominantly of pleomorphic undifferentiated sarcomatoid areas or extensive heterologous elements. We recommend including GATA3 in the panel of immunohistochemical stains for sarcomatoid carcinomas of unknown origin, especially if a bladder primary is being considered in the differential diagnosis.

Arginase-1: a highly specific marker separating pancreatic adenocarcinoma from hepatocellular carcinoma.

BACKGROUND: Arginase-1 and HepPar-1 are effective immunohistochemical (IHC) markers for hepatocellular carcinoma (HCC). In this study, we explored the possible efficacy of these stains in diagnosing pancreatic adenocarcinoma (PAD). STUDY DESIGN: Arginase-1 and HepPar-1 IHC was performed on formalin-fixed, paraffin-embedded fine needle aspiration (FNA) cell blocks (CB) of PAD (n = 46), tissue microarray (TMA) of PAD (n = 33), FNA CB of HCC (n = 44) and TMA of HCC (n = 85). Negative controls without carcinoma were also applied (pancreas CB, n = 7; pancreas TMA, n = 3). RESULTS: PAD CB demonstrated arginase-1 positivity in 0 of 46 cases and HepPar-1 positivity in 7 of 46 cases (15%). PAD TMA demonstrated arginase-1 positivity in 0 of 33 cases and HepPar-1 positivity in 4 of 33 cases (12%). HCC CB demonstrated arginase-1 positivity in 37 of 44 cases (84%) and HepPar-1 positivity in 32 of 44 cases (72%). HCC TMA demonstrated arginase-1 positivity in 75 of 85 cases (88%) and HepPar-1 positivity in 80 of 85 cases (94%). CONCLUSION: Both arginase-1 and HepPar-1 are effective IHC markers of hepatocellular differentiation. Arginase-1 demonstrates superior sensitivity and specificity compared with HepPar-1 in the diagnosis of HCC. However, both arginase-1 and HepPar-1 have a low sensitivity and a very high specificity for PAD.

Automated and manual human papilloma virus in situ hybridization and p16 immunohistochemistry: comparison in metastatic oropharyngeal carcinoma.

OBJECTIVES: We compared the efficacy of automated and manual human papilloma virus (HPV) in situ hybridization (ISH) and p16 immunohistochemical staining (IHC) in fine needle aspiration (FNA) of metastatic oropharyngeal carcinoma. STUDY DESIGN: A total of 41 FNA cell blocks (CB) were evaluated. HPV ISH was interpreted as positive if a minimum of one tumor cell showed punctate dot-like nuclear positivity. p16 was interpreted as positive if ≥70% of tumor cells showed brown nuclear and cytoplasmic staining. RESULTS: Thirty of 41 CB (73%) were positive by automated HPV ISH, 25 of 41 CB (60%) with manual HPV ISH. Eighteen of 41 CB (43%) were positive for p16 IHC. Twelve of 41 CB (29%) with automated HPV ISH and 2 of 41 CB (4%) with the manual method were positive at 10× magnification. Three of 41 CB (7%) with automated HPV ISH and 14 of 41 CB (34%) with the manual method were positive at 20× magnification. Fifteen of 41 CB (36%) with automated HPV ISH and 9 of 41 CB (21%) with the manual method were positive at 40-60× magnification. CONCLUSION: Automated HPV ISH plays a more significant role in determining the HPV status in CB. However, the failure to use high magnification in the evaluation can give false-negative results.

Thyroid transcription factor 1 and Napsin A double stain: utilizing different vendor antibodies for diagnosing lung adenocarcinoma.

OBJECTIVE: A combined thyroid transcription factor 1 (TTF-1) and Napsin A double stain has been shown to be useful in the diagnosis of adenocarcinoma (ADC). This study compares differences in double staining patterns among vendor antibodies (Leica, Dako, and Biocare). STUDY DESIGN: The cohorts included 35 FNA cell blocks of lung ADC and 24 cell blocks of lung squamous cell carcinoma (SqCCA). Double-staining immunohistochemistry was performed with TTF-1 as a brown nuclear stain and Napsin A as a red cytoplasmic stain, using three sets of double stains. Additionally, FISH expression was performed on SqCCAs with aberrant TTF-1 expression. RESULTS: The sensitivity for the double stains ranged from 40 to 74%, while the specificity ranged from 88 to 96%. Two Leica TTF-1-positive SqCCAs also showed low-level amplification by FISH assay not seen in the TTF-1-negative control SqCCAs. CONCLUSION: The use of Dako TTF-1 antibody paired with Leica Napsin A antibody as a double stain yielded the best results for diagnosing ADC; additionally, the Leica Napsin A-only staining results had the highest positive predictive value at 97%. Both Dako and Biocare antibodies expressed less staining of SqCCAs than Leica staining.

Combined double CK5/P63 stain: useful adjunct test for diagnosing pulmonary squamous cell carcinoma.

Increasing demand for accurate differentiation of pulmonary squamous cell carcinoma (SQCC) from other subtypes can be challenging for pathologists. This is more so in fine-needle aspirations (FNA) since the sample is small and SQCC may show degenerative changes and necrosis that distort the cellular features. Immunohistochemistry (IHC) is a valuable adjunct, and CK5/6 and P63 immunoreactivity is found to be basically restricted to SQCC. In our study, we evaluated the efficiency of CK5/P63 double staining in the diagnosis of pulmonary SQCC in cell blocks (CB) of lung FNA. We used a cohort including 24 CB of lung SQCC and 34 CB of lung adenocarcinomas (ADC). IHC was performed for CK5/P63 double stain. Seventeen of 24 (70%) lung SQCC were positive for the double stain CK5/P63. Two (8%) were positive for CK5 alone and two (8%) were positive for P63 alone. Thus, a total 19 of 24(79%) SQCC of the lung were positive for CK5 and P63 each. In ADC, no immunoreactivity was detected for CK5 alone or combined CK5/P63. Three of 34(8%) ADC were positive for P63. This first study of double staining of CK5/P63 in FNA CB shows a sensitivity of 70% and specificity of 100% for SQCC of the lung. When each marker staining alone is included, the sensitivity for CK5 and P63 increases to 79% each. This double stain can help in the diagnosis of pulmonary SQCC with an accuracy of 88% and a positive predictive value of 100%.

Controversial role of Epstein-Barr virus in multiple sclerosis.

The role of Epstein-Barr virus (EBV) in the pathogenesis of multiple sclerosis (MS) is still elusive. In 2007, Serafini et al demonstrated the direct role of EBV in brain lesions of MS patients. They found positive immunohistochemistry (IHC) staining for latency membrane protein 1 (LMP1), and EBV-encoded RNA (EBER) by in-situ hybridization (ISH) within postmortem brains of MS patients. The goal of this study was to attempt to demonstrate LMP1 by IHC and EBER by ISH in brains of patients with MS, to either support or refute their findings. Seventeen MS (16 brain biopsies and 1 autopsy brain) and 12 autopsy brains with no pathologic abnormalities, as normal controls, were studied. To control for the possibility that inflammation owing to other etiologies could result in EBV-positive cell accumulation, 11 brain biopsies of encephalitis and 4 brain biopsies of progressive multifocal leukoencephalopathy were also studied. Known positive (Hodgkins and non-Hodgkins lymphoma) and negative (with antibody primary replaced by buffer) controls were used. All positive and negative controls showed appropriate staining. However, there were no positive LMP1 or EBER results in any of the groups studied. The negative results of IHC and ISH in our study sharply contrast to those previously mentioned by Serafini et al, 2007 and suggest that EBV is not directly related to MS as an etiology.

DOG1 utility in diagnosing gastrointestinal stromal tumors on fine-needle aspiration.

BACKGROUND: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the gastrointestinal (GI) tract, and the majority contain KIT or PDGFRA activating mutations. Fine-needle aspiration biopsy (FNAB) is a valuable technique in the diagnosis of GIST and may allow for preoperative therapy with tyrosine kinase inhibitors (TKI). Because of the morphologic diversity of these tumors, routine diagnosis of GIST often relies on C-Kit immunohistochemical staining in addition to morphologic findings. However, up to 15% of GISTs are C-Kit negative. Antibodies with increased sensitivity and specificity for detection of C-Kit-negative GIST cases may be of value, especially because some of these cases may also benefit from TKI therapy. METHODS: Immunohistochemical staining for DOG-1, C-Kit (CD117) and protein kinase C theta (PKCθ) was performed on FNA cell-block preparations representing 30 GISTs, 17 leiomyosarcomas, 16 melanomas, 16 schwannomas, 11 adenoid cystic carcinomas, and 8 leiomyomas. RESULTS: DOG-1 was found to have 100% sensitivity and 100% specificity in diagnosis of GIST. C-Kit demonstrated 70% sensitivity and 76% specificity, and PKCθ showed 40% sensitivity and 86% specificity. When only spindle-cell neoplasms were considered (adenoid cystic carcinomas excluded), the specificity of C-Kit increased to 89%. Of interest, all C-Kit-negative cases showed DOG-1 positivity. CONCLUSIONS: DOG-1 was the most sensitive and specific of the 3 markers for the diagnosis of GIST in cell-block preparations and may be of particular use in the diagnosis of C-Kit-negative GIST.

TTF-1 and Napsin A double stain: a useful marker for diagnosing lung adenocarcinoma on fine-needle aspiration cell blocks.

BACKGROUND: Immunohistochemistry (IHC) for thyroid transcription factor-1 (TTF-1) is used to confirm the diagnosis of lung adenocarcinoma. Napsin A also has shown positivity in lung adenocarcinoma. A combined double stain for TTF-1 and napsin A has been proposed to achieve higher sensitivity and specificity. In this study, the authors evaluated the utility of this double stain in the diagnosis of lung adenocarcinoma in cell blocks of fine-needle aspirates (FNA). METHODS: The authors used a cohort comprising 35 FNA cell blocks of lung adenocarcinoma and 24 FNA cell blocks of lung squamous cell carcinoma (SqCCA). IHC was performed; expressions of TTF-1 as brown nuclear stain and of napsin A as red cytoplasmic stain were identified. RESULTS: Twenty-six of 35 (74%) lung adenocarcinomas were positive for double staining with TTF-1/napsin A. Of 35 lung adenocarcinomas, only 2 (5%) were positive for TTF-1 alone and 3 (8%) were positive for napsin A alone. For the double stain TTF-1/napsin A, 3 of 24 (12%) lung SqCCAs were positive for both. Six of 24 (25%) cases were positive for TTF-1 alone, and none were positive for napsin A alone. For lung adenocarcinoma, TTF-1/napsin A has a sensitivity of 74%, specificity of 87%, accuracy of 79%, and a positive predictive value of 89%. CONCLUSIONS: The double IHC stain, TTF-1/napsin A, for the identification of pulmonary adenocarcinoma in FNA cell block materials was diagnostically useful. The use of napsin A alone demonstrated a greater degree of accuracy and appeared diagnostically useful as a single IHC stain.