Characterizing specimen quality of cell block samples in an era of personalized diagnostics: analysis of 221 lymph node fine-needle aspirations.

INTRODUCTION: Cell block (CB) preparations of fine-needle aspirates (FNAs) are utilized for patient management, which requires retention of representative material on slides. Personalized medicine demands quality CB specimens. There is no standard protocol for CB preparation, often resulting in suboptimal slides. The utility of using two CB slides in lymph node (LN) FNA cases was investigated. MATERIALS AND METHODS: We cut 10 serial sections from each CB, slides 1 and 5 are stained and considered the first and second cuts, respectively; the remaining slides are reserved for ancillary studies. Hematoxylin and eosin-stained CBs of 221 consecutive LN FNA cases were reviewed; qualitative and quantitative assessment of diagnostic value was made on sections 1 and 5. RESULTS: Of the 221 cases, 46.1% (102) had comparable diagnostic cellularity (equally representative) on both slides, whereas 26.7% (59) and 27.1% (60) had more representative material on the first and second cuts, respectively (P = 0.52). Differences between the representativeness of first and second CB cuts of intrathoracic lymph nodes were minor (n = 192, P = 0.065). Differences between the first and the second slide representativeness of superficial (n = 22, P = 0.98) and intra-abdominal lymph nodes (n = 7, P = 0.38) are limited because of small sample sizes. CONCLUSION: One CB cut can be suboptimal for diagnosis. In our study, inclusion of a second slide increases equal representativeness from 46.1% to 73.2%. These limited observations recognize the need for additional investigations regarding the collection and preparation of CBs.

Pleural mass forming extramedullary hematopoiesis masquerading as a malignant neoplasm.

Extramedullary hematopoiesis (EMH) represents the presence of immature hematopoietic elements and their differentiation into mature blood components outside of the medullary bone and may be seen in a variety of circumstances in the postnatal period, but is most strongly associated with disorders of the hematopoietic system. Postnatally, EMH is typically identified at sites of fetal hematopoiesis, the spleen, and liver, but occasional reports have identified it in nearly every tissue of the body. We report a case of EMH presenting as pleural mass, initially suspected to represent a neoplastic process in a patient with multiple comorbidities, including history of carcinoma, but without co-existing hematologic disorder. On-site evaluation of the fine-needle aspiration specimen was initially suspicious for a malignant neoplasm, but further evaluation revealed the lesion to be a mass forming focus of non-hepatosplenic EMH. In the era of increasing utilization of imaging, mass forming EMH is increasingly detected. When unsuspected, EMH may present a diagnostic challenge for the pathologist and may be confused for a neoplastic process.

Higher Expression of miR-182 in Cytology Specimens of High-Grade Urothelial Cell Carcinoma: A Potential Diagnostic Marker.

OBJECTIVE: MicroRNAs (miRs) are short noncoding RNA molecules that posttranscriptionally modulate protein expression. There are distinct miR alterations characterizing urothelial cell carcinoma (UCC) of the urinary bladder. STUDY DESIGN: In this study, we investigate the possibility of using miR as a noninvasive marker in the screening of UCC. The total RNA was extracted from 75 cytology specimens including bladder or renal washings and voided urines. Cases comprise UCC (21 high grade and 6 low grade), 25 normal controls and 23 cases with a history of UCC but negative at the time of testing (negative with a positive history). The expressions of miR-96, miR-182, miR-183, miR-200c, miR-21, miR-141 and miR-30b were determined using quantitative TaqMan real-time PCR. RESULTS AND CONCLUSION: This study shows that the level of miR-182 is higher in cytology specimens from high-grade UCC patients as compared to normal controls. Measuring miR-182 may provide a potential alternative or adjunct approach for screening high-grade UCC.

Correlation of fine-needle aspiration with fluorodeoxyglucose positron emission tomography and ultrasonography imaging in head and neck lymph nodes.

INTRODUCTION: Fluorodeoxyglucose positron emission tomography (FDG-PET) is a well-established tracer technique, particularly useful for oncologic conditions. However, in the head and neck region, this modality may have limitations due to the complex anatomy and high frequency of normal variants in FDG-PET uptake resulting in diagnostic errors. Hence, FDG-PET results often require additional diagnostic investigations including ultrasonography (US), and guided fine-needle aspirations (FNAs) in order to improve diagnosis and patient care. MATERIALS AND METHODS: A total of 87 cases of head and neck lymph node FNA were accessioned in the cytopathology laboratory between 2010 and 2013, 75 cases with corresponding standard uptake values (SUVs) were selected to form the current study cohort. Patient demographics, primary tumor site, size of the biopsied lymph node, PET SUV, and US findings, as well as FNA diagnoses and surgical follow-ups were reviewed. SUVs were grouped into 5 ranges: A (2.2-2.9), B (3.0-4.9), C (5.0-6.9), D (7.0-9.9), and E (10.0 +). RESULTS: The SUV cutoff of 5.0 was seen in the groups C, D, and E, which increased the probability of malignancy in the FNA specimens from 67% to 100%. The average size of the lymph nodes was 18.57 mm (range 6-41 mm); it was 13.4, 14.2, 19.7, 18.4, and 20.2 for groups A, B, C, D, and E, respectively. The lymph node measurements represented similarities in groups A and B (13.4, 14.2), and groups C, D, and E (19.7, 18.4, 20.2). Albeit a few exceptions, an increase in the lymph node size 14.2 to 19.7 (ie, from groups A and B to groups C, D, and E) correlated with SUV and US abnormalities. Abnormal US findings were pronounced (100%) in group E, and ranged between 67% and 73% of the cases in other groups. CONCLUSIONS: This study documents the usefulness of SUV range in FDG-PET results and not an absolute "cutoff" number for diagnosing suspicious head and neck lymph nodes due to an overlap in the SUVs between malignant and benign lesions. False positive and negative results may occur in hyperplastic and necrotic lymph nodes. An increase in the lymph node size generally compares well with SUVs and US abnormalities; use of US-guided FNAs in these cases can be helpful to achieve definite diagnosis.

Perspective on cells: evolution of cytopathology and the University of Pennsylvania, Philadelphia PA.

This document briefly captures the development of cytopathology at the Hospital of the University of Pennsylvania (HUP) in Philadelphia PA; the first medical school and the teaching hospital in the Country. Literature suggests that cells from malignancy have been described since early 1830s. While earlier accounts are not available, in the year 1895, 9th edition of the book written by Professor of Pathology, Dr. Tyson of the University of Pennsylvania describes urothelial cell morphology. It is also noted that both gynecologic as well as non-gynecologic cytopathology is routinely being practiced at this Institution since 1949. Following the administrative consolidation of the department of pathology and laboratory medicine, a separate section of cytopathology within the department was established. Growing academic and clinical enterprises of HUP have resulted in establishment of Ruth and Raymond Perelman Center for Advanced Medicine (PeCAM) in the year 2010. Currently, the section has seven full time cytopathologists. It has patented a cart for onsite cytopathology interpretation, offers state of the art cytopathology services including onsite interpretation and reporting, point of care specimen triaging, molecular cytopathology testing as well as hosts telecytopathology systems within the Health System and training program.

TeleCyP (Telecytopathology): real-time fine-needle aspiration interpretation.

OBJECTIVE: To assess the application of TeleCyP for real-time fine-needle aspiration interpretation (RFI) necessary for case management and specimen triage. STUDY DESIGN: Twenty-two endobronchial ultrasound (EBUS)-guided mediastinal and pulmonary cases were included in the learning phase to determine the time and efficiency of TeleCyP. Slides were scanned by a cytopathology fellow in real time, and high-speed transmitted images over a secure network were interpreted by a cytopathologist while maintaining audio communication. In the validation phase, an additional 38 pancreas cases from endoscopic ultrasound (EUS) were evaluated recapitulating the RFI scenario from the learning phase. The cytopathologist was blinded to the results of the diagnosis in both phases. RESULTS: The time to provide assessment of specimen adequacy and a preliminary diagnosis was 53 s in the learning phase and 49 s in the validation phase. There was 100% correlation between RFI and TeleCyP assessment for specimen adequacy. TeleCyP particularly posed challenges in providing definitive interpretation on EUS-fine-needle aspiration of some of the pancreatic solid masses (11%, 4/36). CONCLUSION: TeleCyP can serve as a powerful alternative, time-efficient strategy to provide RFI, and for specimen triaging which is critical for personalized medicine and patient management.

Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman Disease): A case report and review of 49 cases with fine needle aspiration cytology.

Rosai-Dorfman disease (RDD), a rare, benign, self-limiting histiocytic proliferative disorder, can be encountered in both nodal and extranodal locations, and fine needle aspiration (FNA), a simple, accurate and economic tool, has been widely used for the diagnosis of superficial and deep-seated lesions. Familiarity with the cytomorphologic features of RDD is important as prognosis and treatment are quite different from other benign or malignant diseases for which it may clinically masquerade. Although large numbers of RDD cases have been reported, review of the literature has revealed 49 reported cases of RDD diagnosed by FNA. Here, we report a case of RDD with nasal and sinus involvement. The patient was seen at our institution, carrying a diagnosis of inflammatory pseudotumor rendered by an outside institution, based on material obtained by nasal and sinus surgical biopsies. Cervical lymph node FNA performed at our institution revealed typical features of RDD. The case, as well as a brief review of the literature and 49 RDD cases with FNA cytology, will be discussed.

Progression from on-site to point-of-care fine needle aspiration service: Opportunities and challenges.

BACKGROUND: Standard-of-care requires the availability of an efficient, economical and accurate on-site fine needle aspiration (FNA) service. Presence of a trained individual during the procedure ensures an improved patient care. Appropriate selection of the equipment, interaction with the clinicians and compliance with the various regulations during the procedure is essential. This is often done by an on-site FNA service. Organization and implementation of such a system in a large academic center is challenging. METHOD: we reviewed the ambulatory care needs in the new Perelman Center for Advanced Medicine (PeCAM). Multiple (9) FNA sites have been established keeping in view the patient's convenience, clinic demands, various regulatory requirements and laboratory staff. Each location has dedicated FNA station with microscopes and supplies. In addition, state- ofthe -art technologies including a mobile FNA cart (Penn-A- Cart), remote specimen evaluation (TeleCyP) have been incorporated. RESULTS: The new set up is extremely efficient and much valued by the patients and the clinicians. It has improved patient care. CONCLUSION: With necessary investments and resources a point-of-care FNA service has been created which has improved patient care. This, albeit with certain modifications may serve as a model for FNA service.

University of Pennsylvania aspiration cart (Penn-A-Cart): an innovative journey in fine needle aspiration service.

On-site evaluation of fine needle aspiration specimens is now considered the standard of care. The procedure is performed at the bedside and at various other locations within the hospital using a mobile cart fitted with essentials, including a microscope, stains and supplies. Earlier, an open specimen cart was used with various supplies and a binocular microscope. The mobile cart during the past 25 years has been variously modified. A second-generation cart containing 2 folding pods, a double-headed microscope, supplies and stains had been used for the past 20 years; it did not meet the current regulatory standards and needs, and little attention was paid to meet the Joint Commission on the Accreditation of Healthcare Organizations, Health Insurance Portability and Accountability Act and infection control standards. Also, the repeated lifting of the microscope resulted in much wear and tear. We have designed Penn-A-Cart, which has a pneumatic lift for the microscope, a high-definition camera with a TV screen and access to the Internet. It has a sharps container and storage space for supplies, stains and slides. This cart meets the various regulatory agency standards. It is user friendly and valuable for multiviewing. The cart is valuable for remote access, telecytopathology and improved patient care.

Premalignant and malignant cells in sputum from lung cancer patients.

BACKGROUND: The objective of this study was to assess the frequency of premalignant and malignant cells in sputum from patients with lung cancer and to measure the dependence of these cells on cancer stage, histologic type, tumor size, and tumor location. METHODS: This analysis included 444 patients with lung cancer. First, all patients were asked to produce sputum spontaneously; then, they underwent sputum induction. Slide preparations of the sputa were screened for the presence of abnormal cells. RESULTS: Of all patients with lung cancer who had produced adequate specimens, 74.6% had sputum that was positive for premalignant or worse cells, whereas 48.7% had sputum that was positive for malignant cells alone. Surprisingly, the presence of premalignant or worse cells in sputum depended only moderately on disease stage (82.9% of stage IV cancers vs 65.9% of stage I cancers), tumor size (78.6% of tumors >2 cm vs 64.7% of tumors

Value of P63 and CK5/6 in distinguishing squamous cell carcinoma from adenocarcinoma in lung fine-needle aspiration specimens.

The current FDA-approved standard of care for nonsmall cell lung cancer is Carboplastin/Taxol/Avastin based upon an impressive survival benefit; however, patients with squamous carcinoma (SQCC) cannot receive Avastin because of a 30% mortality rate due to fatal hemoptysis. In this study we evaluated the role of cytomorphology and immunohistochemistry in differentiating SQCC from adenocarcinoma (ADC) in lung FNA specimens. The case cohort included 53 FNA cases of nonsmall cell lung carcinoma with surgical pathology follow-up. All FNA specimens were reviewed independently by a panel of cytopathologists to differentiate between SQCC and ADC. The cell block material was available in 23 cases (11 ADC and 12 SQCC) to perform immunohistochemical stains for TTF-1, CK7, CK20, P63, and CK5/6. On surgical resection, 35/53 (66%) cases were diagnosed as ADC and 18/53 (34%) as SQCC. The number of cases classified correctly on the basis of cytomorphology was 66% for ADC and 53% for SQCC (combined accuracy 60%). By immunohistochemical staining, 14/23 (61%) cases expressed TTF-1. Nine cases were TTF-1 negative; eight of the TTF-1 negative cases (89%) were SQCC. Twenty-three cases expressed CK7 (87%); one ADC case (4%) showed focal CK20 positivity. Both P63 and CK5/6 expression was seen in 9/12 (75%) SQCC cases; none of the ADC cases showed this dual expression. Cytomorphology alone may not be able to stratify all cases of nonsmall cell lung carcinoma into ADC and SQCC in FNA specimens. The immune-panel of TTF-1, CK7, CK20, P63, and CK5/6 is useful in differentiating SQCC from ADC.

Utility of Thyroglobulin measurement in fine-needle aspiration biopsy specimens of lymph nodes in the diagnosis of recurrent thyroid carcinoma.

INTRODUCTION: The most common site for the metastasis of papillary carcinoma of the thyroid (PTC) is regional lymph nodes. Ultrasound (US) imaging may identify abnormal appearing lymph nodes, suspicious for PTC recurrence. Although fine needle aspiration biopsy (FNAB) of abnormal lymph nodes is often diagnostic of recurrence, small or cystic lymph nodes may be non-diagnostic due to lack of tumor cells. The measurement of thyroglobulin (TG) levels in FNAB specimens from lymph nodes suspicious for recurrent PTC can serve as an adjunct to the cytologic diagnosis. MATERIALS AND METHODS: 115 abnormal appearing lymph nodes were aspirated under ultrasound guidance in 89 patients with history of thyroid carcinoma. In addition to obtaining material for cytologic interpretation, an additional aspirate was obtained by FNAB and rinsed in 1 ml of normal saline for TG level measurements. RESULTS: The cytologic diagnoses included: 35 (30%) reactive lymph node, no tumor seen (NTS), 39 (34%) PTC, 23 (20%) inadequate for evaluation due to lack of lymphoid or epithelial cells (NDX) 15 (13%) atypical/suspicious for PTC, and 3 (3%) other (e.g. paraganglioma, poorly differentiated carcinoma and carcinoma not otherwise specified). TG levels were markedly elevated (median 312 ng/ml; normal < 10 ng/ml) in 28 (72%) cases of PTC lymph node recurrence identified on cytology. TG measurements were also elevated in 5 lymph nodes classified as NTS and 4 NDX on cytology which resulted in 5 and 3 carcinoma diagnoses respectively on histological follow-up. Of the 9 atypical/suspicious cases with elevated TG levels all resulted in carcinoma diagnoses on follow-up. CONCLUSION: The measurement of TG in FNAB specimens from lymph node in patients with history of PTC is useful in detecting recurrent disease, especially in cases when the specimen is known to be or likely to be inadequate for cytologic evaluation.

Entamoeba gingivalis pulmonary abscess - diagnosed by fine needle aspiration.

Entamoeba gingivalis (E. gingivalis ) is a parasitic protozoa of the oral cavity, most often found in gingival tissues around the teeth associated with poor oral hygiene. Here, we report a case of E. gingivalis in a pulmonary CT guided fine needle aspiration material, from a 60-year-old man with newly found lung mass. On site Diff-Quik smear examination revealed the presence of marked acute inflammation, colonies of actinomyces, and a number of 'large macrophages-like organisms'. Upon examination of the additional material, organisms morphologically consistent with E. gingivalis were identified. Pulmonary mass resolved after six weeks of treatment with antibiotics (Clindamycin followed by Penicillin). Proper recognition and distinction between E. gingivalis and other species of Entamoeba is important for the management of patients.

Case report and review of lupus erythematosus cells in cytology fluids.

Systemic lupus erythematosus (SLE) is a chronic inflammatory disease typically diagnosed by a combination of physical findings and clinical laboratory testing. Several decades ago, the diagnosis of lupus included the lupus erythematosus (LE) cell assay. Autoimmune serology techniques such as anti-nuclear antibody staining have replaced the LE cell assay. However, as presented in this report and review of the literature, the in vivo finding of LE cells by cytopathology remains an important finding consistent with the diagnosis of SLE.

Concordance between thyroid nodule sizes measured by ultrasound and gross pathology examination: effect on patient management.

Ultrasound examination (US) is an essential tool in the evaluation of thyroid nodules. The size determined by US is used to distinguish between clinical vs. nonclinical thyroid nodules i.e. greater than or equal to or less than 1 cm. In this study, we evaluated the concordance between the sizes of thyroid nodules measured by US and by gross examination after thyroidectomy. This study included 664 nodules that underwent fine-needle aspiration (FNA) and subsequent excision in 621 patients; 580 had single and 41 patients had multiple (39 with 2, and 2 with 3 nodules) nodules. Both US and gross pathology measurements were taken in three dimensions. The nodule sizes as measured by US were stratified into five groups: A: or=5.1 cm. FNA diagnoses were categorized into: Benign (n = 59), Neoplastic / Indeterminate (n = 342), Suspicious (n = 123), Malignant (n = 106), and nondiagnostic (n = 34). Upon excision 278 (42%) nodules were classified as malignant and 386 (58%) as benign. In group A the concordance between US and excisional size was 78.5%, group B 56%, group C 34.5%, group D 40% and group E 52.5%. Only 14 (14/664 2%) nodules measured

Endoscopic ultrasound guided fine needle aspiration of non-pancreatic lesions: an institutional experience.

BACKGROUND: Endoscopic ultrasound guided fine needle aspiration biopsy (EUS-FNA) has proven to be an effective diagnostic modality for the detection and staging of pancreatic malignancies. In recent years EUS-FNA has also been used to diagnose lesions of non-pancreatic sites such as structures in close proximity to the gut wall within the mediastinum, abdomen, pelvis and retro-peritoneum. AIMS: To evaluate experience with EUS-FNA of non-pancreatic sites at a large university medical centre. METHODS: The study cohort included 234 patients who underwent EUS-FNA of 246 lesions in non-pancreatic sites (122 peri-pancreatic and coeliac lymph nodes; 9 peri-pancreatic masses; other sites: mediastinum 12, gastric 25, liver 27, oesophagus 17, duodenum/colon/rectum 15, retro-peritoneum 8, lung 7, miscellaneous 4). RESULTS: The cytology diagnoses were classified as non-neoplastic/reactive in 82 (33%), atypical/suspicious for malignancy in 25 (10%), malignant in 86 (35%) and non-diagnostic in 53 (22%) cases. Surgical pathology follow-up was available in 75 (31%) cases. Excluding the non-diagnostic cases there were 7 false negative and 3 false positive cases. The sensitivity, specificity and positive predictive value of EUS-FNA in the diagnosis of lesions of non-pancreatic sites was 92%, 98% and 97%, respectively. CONCLUSIONS: EUS-FNA can be effectively used as a diagnostic modality in the diagnosis of lesions from non-pancreatic sites.

Is fine-needle aspiration (FNA) of multiple thyroid nodules justified?

BACKGROUND: The clinical management of patients with solitary thyroid nodule is well established; however, the evaluation of patients with multiple thyroid nodules is controversial. The aim of this study was to assess if there is a correlation between the risk of malignancy and number of thyroid nodules. DESIGN: The study cohort included 2,884 patients (2,410 females and 474 males) with 3,274 thyroid nodules that underwent FNA under ultrasound guidance between November 1997 and April 2004. Multiple thyroid nodules were aspirated in 360 patients; 2 in 332 (291 females, 41 males), 3 nodules in 26 (23 females, 3 males), and 4 nodules in 2 patients (1 female, 1 male). Subsequent information regarding the histological diagnosis was obtained in the cases that underwent surgical excision. RESULTS: The average age for patients with single or multiple nodules was the same (51 yr). The FNA specimens were diagnosed as benign (n = 1,663, 51%), neoplasm (n = 880, 27%), suspicious for neoplasm or papillary carcinoma (n = 234, 7%), malignant (n = 187, 6%), and non-diagnostic (n = 310, 9%). Surgical excision was performed in 1,135 patients: 993 patients with single nodules and 142 patients with multiple nodules. The surgical pathology diagnosis was benign in 656 nodules (52%) and malignant in 596 nodules (48%). In the malignant category 153 tumors were papillary microcarcinoma (<1 cm). Excluding these cases, the malignancy rate was 39% in patients with single nodules, 41% with two nodules, and 21% with three nodules. CONCLUSIONS: The cancer risk is similar for patients with one or two nodules over 1 cm and decreases with three or more thyroid nodules.