Prospective, Multi-Institutional, Real-Time Next-Generation Sequencing of Pancreatic Cyst Fluid Reveals Diverse Genomic Alterations That Improve the Clinical Management of Pancreatic Cysts.

BACKGROUND & AIMS: Next-generation sequencing (NGS) of pancreatic cyst fluid is a useful adjunct in the assessment of patients with pancreatic cyst. However, previous studies have been retrospective or single institutional experiences. The aim of this study was to prospectively evaluate NGS on a multi-institutional cohort of patients with pancreatic cyst in real time. METHODS: The performance of a 22-gene NGS panel (PancreaSeq) was first retrospectively confirmed and then within a 2-year timeframe, PancreaSeq testing was prospectively used to evaluate endoscopic ultrasound-guided fine-needle aspiration pancreatic cyst fluid from 31 institutions. PancreaSeq results were correlated with endoscopic ultrasound findings, ancillary studies, current pancreatic cyst guidelines, follow-up, and expanded testing (Oncomine) of postoperative specimens. RESULTS: Among 1933 PCs prospectively tested, 1887 (98%) specimens from 1832 patients were satisfactory for PancreaSeq testing. Follow-up was available for 1216 (66%) patients (median, 23 months). Based on 251 (21%) patients with surgical pathology, mitogen-activated protein kinase/GNAS mutations had 90% sensitivity and 100% specificity for a mucinous cyst (positive predictive value [PPV], 100%; negative predictive value [NPV], 77%). On exclusion of low-level variants, the combination of mitogen-activated protein kinase/GNAS and TP53/SMAD4/CTNNB1/mammalian target of rapamycin alterations had 88% sensitivity and 98% specificity for advanced neoplasia (PPV, 97%; NPV, 93%). Inclusion of cytopathologic evaluation to PancreaSeq testing improved the sensitivity to 93% and maintained a high specificity of 95% (PPV, 92%; NPV, 95%). In comparison, other modalities and current pancreatic cyst guidelines, such as the American Gastroenterology Association and International Association of Pancreatology/Fukuoka guidelines, show inferior diagnostic performance. The sensitivities and specificities of VHL and MEN1/loss of heterozygosity alterations were 71% and 100% for serous cystadenomas (PPV, 100%; NPV, 98%), and 68% and 98% for pancreatic neuroendocrine tumors (PPV, 85%; NPV, 95%), respectively. On follow-up, serous cystadenomas with TP53/TERT mutations exhibited interval growth, whereas pancreatic neuroendocrine tumors with loss of heterozygosity of ≥3 genes tended to have distant metastasis. None of the 965 patients who did not undergo surgery developed malignancy. Postoperative Oncomine testing identified mucinous cysts with BRAF fusions and ERBB2 amplification, and advanced neoplasia with CDKN2A alterations. CONCLUSIONS: PancreaSeq was not only sensitive and specific for various pancreatic cyst types and advanced neoplasia arising from mucinous cysts, but also reveals the diversity of genomic alterations seen in pancreatic cysts and their clinical significance.

Should screening for cervical cancer go to primary human papillomavirus testing and eliminate cytology?

This review will systematically highlight the pros and cons of cervical cancer screening with HPV (human papillomavirus) testing and cytological methods (Papanicolaou (Pap) test). When comparing the screening modalities, various facets will be addressed, such as cost effectiveness, and harms and benefits across different demographics and age groups. It is important to note that due to the expansive variance in material costs, practices, and resource availability across different geographical regions, these comparisons are far from straight forward, and ultimately make it challenging to render definitive global recommendations. Thus, the intent of this review is to highlight some of the differences in difference cervical cancer screening modalities that can help one to choose an optimal screening method in their specific situation.

Epithelioid Vascular Lesions: The Differential Diagnosis and Approach in Cytology and Small Biopsies.

Vascular neoplasms are rare tumors with a multitude of clinical presentations and behavior, which make accurate identification and subclassification challenging on limited small biopsies. Within the spectrum of these lesions, the ones with epithelioid morphology, such as epithelioid hemangioendothelioma and epithelioid angiosarcoma, are particularly challenging given the morphologic overlap with nonvascular lesions and the limited cells due to hemodilution on sampling. Herein, we review the differential diagnosis of epithelioid vascular neoplasms, with a focus on the cytomorphology, differential diagnoses, and ancillary studies that pathologists should be aware of when evaluating small biopsies and aspirates, including novel translocations, and associated monoclonal immunohistochemistry antibodies, that can help in the diagnosis of some of these tumors. Awareness of these morphologic and ancillary study findings in these rare tumors will hopefully allow pathologists to recognize and render-specific diagnoses on limited samples of these challenging lesions.

Adding the "US" in FNA: Pearls for starting a pathologist-performed ultrasound-guided fine needle aspiration service (USG FNA).

Ultrasound-guided fine needle aspiration (USG-FNA) biopsies have traditionally been performed in the radiology department, with radiologists performing the procurement with or without on-site cytotechnologists or pathologists to provide adequacy or diagnostic evaluation of the specimen. However, more recently, these image-guided biopsies have been performed by endocrinologists and now cytopathologists. Starting an USG-FNA service is a big task that requires consideration of multiple factors, including training, certification, privileges, equipment, documentation, information technology (IT) issues, and the overall business plan or financial component. In this review, the issues confronted when bringing on an USG-FNA service are discussed in detail in an effort to highlight the issues and challenges that many cytopathology laboratories are facing when implementing this new service.

State-of-the-art cytology of pleural fluid, focusing on the diagnosis of mesothelioma.

Mesothelioma has always been a challenging diagnosis to render in body cavity cytology samples. This review is a timely update on pleural fluid cytology and ancillary studies that should be considered in the diagnosis of mesothelial proliferations, specifically mesotheliomas. Information about new diagnostic approaches and ancillary studies in mesothelioma was obtained from the peer-reviewed literature and the authors' experiences. Although the morphological diagnosis of mesothelioma is fraught with numerous challenges given the overlap with other diagnostic entities, there are a variety of immunohistochemical and fluorescence in situ hybridization studies available to help in determining mesothelial origin and in distinguishing malignant proliferations from the more common benign or reactive mesothelial proliferations. Although ancillary studies can be helpful, there are important pitfalls to be aware of when interpreting these cases, and this review highlights some of the challenges that require caution.

Integrating next-generation sequencing to endoscopic retrograde cholangiopancreatography (ERCP)-obtained biliary specimens improves the detection and management of patients with malignant bile duct strictures.

OBJECTIVE: Despite improvements in imaging, serum CA19-9 and pathological evaluation, differentiating between benign and malignant bile duct strictures remains a diagnostic conundrum. Recent developments in next-generation sequencing (NGS) have opened new opportunities for early detection and management of cancers but, to date, have not been rigorously applied to biliary specimens. DESIGN: We prospectively evaluated a 28-gene NGS panel (BiliSeq) using endoscopic retrograde cholangiopancreatography-obtained biliary specimens from patients with bile duct strictures. The diagnostic performance of serum CA19-9, pathological evaluation and BiliSeq was assessed on 252 patients (57 trainings and 195 validations) with 346 biliary specimens. RESULTS: The sensitivity and specificity of BiliSeq for malignant strictures was 73% and 100%, respectively. In comparison, an elevated serum CA19-9 and pathological evaluation had sensitivities of 76% and 48%, and specificities of 69% and 99%, respectively. The combination of BiliSeq and pathological evaluation increased the sensitivity to 83% and maintained a specificity of 99%. BiliSeq improved the sensitivity of pathological evaluation for malignancy from 35% to 77% for biliary brushings and from 52% to 83% for biliary biopsies. Among patients with primary sclerosing cholangitis (PSC), BiliSeq had an 83% sensitivity as compared with pathological evaluation with an 8% sensitivity. Therapeutically relevant genomic alterations were identified in 20 (8%) patients. Two patients with ERBB2-amplified cholangiocarcinoma received a trastuzumab-based regimen and had measurable clinicoradiographic response. CONCLUSIONS: The combination of BiliSeq and pathological evaluation of biliary specimens increased the detection of malignant strictures, particularly in patients with PSC. Additionally, BiliSeq identified alterations that may stratify patients for specific anticancer therapies.

Telecytology rapid on-site evaluation: Diagnostic challenges, technical issues and lessons learned.

INTRODUCTION: Rapid on-site evaluation (ROSE) has been widely used to improve diagnostic adequacy and facilitate specimen triage. Telecytology ROSE has gained popularity recently and shown high concordance with traditional ROSE. However, telecytology involves multiple personnel and technical devices that could introduce additional errors. The aim of this paper is to share errors encountered and lessons learned since employing telecytology for ROSE at our institution. METHODS: The laboratory information system was searched for all documented telecytology ROSE errors from 2017 to 2019. These errors were subclassified as technical errors, cytotechnologist-related errors and pathologist-related errors. The following details were recorded for each reported event: type of error, reason for error, ROSE diagnosis, final diagnosis and actions taken to avoid future errors. RESULTS: Telecytology ROSE errors were documented in 46 (1.3%) sessions. Ten (22%) had technical errors, 13 (28%) were owing to cytotechnologist errors and 23 (50%) were attributed to pathologist interpretation errors. The majority of the technical (90%) and cytotechnologist errors (85%) occurred within the first year of implementation of telecytology. Common ROSE misinterpretation errors included missing microorganisms, misclassifying neuroendocrine tumours as other neoplasms and overcalling malignancy on gastrointestinal endoscopic procedures. CONCLUSIONS: A variety of errors may occur during telecytology ROSE. While some errors are inevitable (eg, information technology downtime), certain telecytology errors can be reduced by increasing staff familiarity with the system, providing timely feedbacks and taking prompt corrective actions. We recommend establishing a mechanism to document and act upon recorded errors as part of a telecytology quality improvement programme.

The utility of cell blocks for international cytopathology teleconsultation by whole slide imaging.

INTRODUCTION: Telecytology for second opinion consultation has largely been limited by technical issues, such as the inability to focus well on cellular material. Nevertheless, international telecytology consultation was undertaken at our institution with partners in China and Italy. To overcome issues with scanning cytology slides, we adopted a cell-block (CB) preference for teleconsultation. METHODS: Telecytology consultation cases received over a 7.5-year period were retrospectively reviewed. Cytology glass slides were scanned without Z-stacking using different whole slide scanners. For one referring site, only haematoxylin-eosin-stained CBs were scanned, as well as immunostains requested by consultants. For another host centre, aspirate smears were also scanned in some cases. RESULTS: A total of 51 non-gynaecological cases (44 CB only) were evaluated from 48 patients. The specimens included pleural fluids (19), pancreas (14), lymph nodes (6), peritoneal fluids (2) and miscellaneous samples (10). The cytological diagnoses spectrum included 16 (31.37%) cases positive for malignancy, 7 (13.72%) positive for neoplasm, 6 (11.76%) suspicious for malignancy, 10 (19.60%) atypical, 10 (19.60%) negative for malignancy and 2 (3.92%) non-diagnostic. In 42 (82.35%) cases, immunocytochemistry was requested. Turn-around-time ranged from 1.5 to 306 hours. CONCLUSIONS: Our experience shows that international telecytology for consultation purposes involving non-gynaecological cases is feasible. A second opinion interpretation was rendered in the majority (64.7%) of cases. Utilising CB only for cytology consultations by whole slide imaging solved focus issues that typically plague evaluation of cytology aspirate smears.

Quantitative image analysis for CD8 score in lung small biopsies and cytology cell-blocks.

INTRODUCTION: Immunotherapy has shown promising results in non-small cell lung cancer (NSCLC), for which tumour-infiltrating cytotoxic (CD8+) T cells play a critical role. We investigated the utility of image analysis (IA) to quantify CD8+ T cells in a series of matched small biopsies and resections of NSCLC. METHODS: CD8 immunohistochemistry was performed on cell-blocks (CB), core needle biopsies (CNB) and corresponding resections from primary NSCLCs. Slides were digitised using an Aperio AT2 scanner (Leica) and annotated by whole slide image (WSI) or fields of view occupied by tissue spots (TS). Quantitative IA was performed with a customised Aperio algorithm (Leica). CD8 scores (number of T cells with 1-3+ staining/total area) were then compared. RESULTS: Forty-four cases with CB or CNB material and a corresponding resection were analysed. Average CD8 score was determined in CB (7.67 WSI, 77.67 TS) and/or CNB (47.35 WSI, 325.67 TS), and corresponding resections (190.35 WSI, 336.58 TS). CD8 score concordance was highest (78.6%) for CNBs using WSI annotation. Overall, small biopsies (CB or CNB) correlated with the resection in 71.4% cases using WSI and 63.3% cases using TS annotation. IA performed better for low CD8 scores. CONCLUSIONS: These findings show that CD8 density in NSCLC can be quantified by IA in small biopsies and cell blocks, achieving the best concordance using WSI scores. Discrepancies were attributed to values near the cut-off and background detection of staining. These data warrant future studies with more cases and follow-up data to further investigate the clinical utility of IA for CD8 analysis in NSCLC.

Assessing competency for remote telecytology rapid on-site evaluation using pre-recorded dynamic video streaming.

INTRODUCTION: Telecytology using real-time microscopy has gained popularity for rapid on-site evaluations (ROSE). Although proficiency testing is routinely used in cytopathology, no established means of competency assessment is currently available for telecytology. Our aim was to determine the feasibility of a dynamic (real-time) platform to assess telecytology competency. METHODS: Remote Medical Technology dynamic (real-time) video streaming platform for ROSE is used at our institution, and short video clips of telecytology cases were recorded using Camtasia Studio 8 software during different ROSE sessions. Selected MP4 videos (range 13-88 seconds, mean 33 seconds), along with clinical histories, were used to build a multiple-choice question test with one training case and 20 test cases, utilising Tutor (Philips) software to host the web-based test. The test was voluntary for cytopathologists and cytotechnologists. Answers and feedback from test takers were analysed. RESULTS: Thirteen participants-four cytopathologists and nine cytotechnologists-previously trained to use telecytology, volunteered to take the test. Individual scores ranged from 10 (50%) to 19 (95%) with a median of 16 (80%). Most feedback received involved technical difficulties. CONCLUSIONS: We present, to the best of our knowledge, the first tool to assess telecytology competency for ROSE using pre-recorded dynamic streaming videos. Despite technical challenges related to incorporating videos into a web-based test, the test was feasible and provided users with valuable feedback about their ROSE performance. Future effort will be devoted to establishing a more user-friendly test platform and establishing a benchmark for passing scores.

Characteristics of Malignant Pleural Effusion Resident CD8+ T Cells from a Heterogeneous Collection of Tumors.

While T cell-based cancer immunotherapies have shown great promise, there remains a need to understand how individual metastatic tumor environments impart local T cell dysfunction. At advanced stages, cancers that metastasize to the pleural space can result in a malignant pleural effusion (MPE) that harbors abundant tumor and immune cells, often exceeding 108 leukocytes per liter. Unlike other metastatic sites, MPEs are readily and repeatedly accessible via indwelling catheters, providing an opportunity to study the interface between tumor dynamics and immunity. In the current study, we examined CD8+ T cells within MPEs collected from patients with heterogeneous primary tumors and at various stages in treatment to determine (1) if these cells possess anti-tumor activity following removal from the MPE, (2) factors in the MPE that may contribute to their dysfunction, and (3) the phenotypic changes in T cell populations that occur following ex vivo expansion. Co-cultures of CD8+ T cells with autologous CD45- tumor containing cells demonstrated cytotoxicity (p = 0.030) and IFNγ production (p = 0.003) that inversely correlated with percent of myeloid derived suppressor cells, lactate, and lactate dehydrogenase (LDH) within the MPE. Ex vivo expansion of CD8+ T cells resulted in progressive differentiation marked by distinct populations expressing decreased CD45RA, CCR7, CD127, and increased inhibitory receptors. These findings suggest that MPEs may be a source of tumor-reactive T cells and that the cellular and acellular components suppress optimal function.

Preoperative next-generation sequencing of pancreatic cyst fluid is highly accurate in cyst classification and detection of advanced neoplasia.

OBJECTIVE: DNA-based testing of pancreatic cyst fluid (PCF) is a useful adjunct to the evaluation of pancreatic cysts (PCs). Mutations in KRAS/GNAS are highly specific for intraductal papillary mucinous neoplasms (IPMNs) and mucinous cystic neoplasms (MCNs), while TP53/PIK3CA/PTEN alterations are associated with advanced neoplasia. A prospective study was performed to evaluate preoperative PCF DNA testing. DESIGN: Over 43-months, 626 PCF specimens from 595 patients were obtained by endoscopic ultrasound (EUS)-fine needle aspiration and assessed by targeted next-generation sequencing (NGS). Molecular results were correlated with EUS findings, ancillary studies and follow-up. A separate cohort of 159 PCF specimens was also evaluated for KRAS/GNAS mutations by Sanger sequencing. RESULTS: KRAS/GNAS mutations were identified in 308 (49%) PCs, while alterations in TP53/PIK3CA/PTEN were present in 35 (6%) cases. Based on 102 (17%) patients with surgical follow-up, KRAS/GNAS mutations were detected in 56 (100%) IPMNs and 3 (30%) MCNs, and associated with 89% sensitivity and 100% specificity for a mucinous PC. In comparison, KRAS/GNAS mutations by Sanger sequencing had a 65% sensitivity and 100% specificity. By NGS, the combination of KRAS/GNAS mutations and alterations in TP53/PIK3CA/PTEN had an 89% sensitivity and 100% specificity for advanced neoplasia. Ductal dilatation, a mural nodule and malignant cytopathology had lower sensitivities (42%, 32% and 32%, respectively) and specificities (74%, 94% and 98%, respectively). CONCLUSIONS: In contrast to Sanger sequencing, preoperative NGS of PCF for KRAS/GNAS mutations is highly sensitive for IPMNs and specific for mucinous PCs. In addition, the combination of TP53/PIK3CA/PTEN alterations is a useful preoperative marker for advanced neoplasia.

Langerhans cell histiocytosis and Erdheim-Chester disease, both with cutaneous presentations, and papillary thyroid carcinoma all harboring the BRAF(V600E) mutation.

Langerhans cell histocytosis (LCH) and Erdheim-Chester disease are two rare histiocytic disorders. Their occurrence in the same patient is more infrequent, but has been described. We report a case of a 38-year-old woman who presented with a diagnosis of single system cutaneous LCH. Subsequently, she developed multiple papules on her extremities consistent with a non-LCH xanthogranuloma type lesion. BRAF(V600E) mutation immunostain, VE1 was positive in the skin lesion, which was confirmed by molecular polymerase chain reaction (PCR) studies, initiating a complete systemic workup for Erdheim-Chester disease. Systemic involvement was confirmed with bilateral sclerotic bone lesions and retroperitoneal and pelvic fibrosing disease. She was also found to have a BRAF(V600E) mutation positive papillary thyroid carcinoma. New suspicious cutaneous lesions presenting in patients with a history of LCH need to be biopsied. A BRAF(V600E) mutation in a non-LCH histiocytic lesion with a xanthogranuloma phenotype (CD163/CD68/CD14/fascin/Factor 13a) should prompt an Erdheim-Chester disease workup. This is a unique case of a woman with BRAF(V600E) mutation positive Erdheim-Chester disease and cutaneous LCH, while also being, to our knowledge, the first reported case in the English literature of it occurring in a patient with a BRAF(V600E) mutation positive papillary thyroid carcinoma.

Cytopathology of extracranial ectopic and metastatic meningiomas.

INTRODUCTION: Extracranial meningiomas may infrequently be encountered as ectopic or metastatic tumors. Their rarity and unique cytomorphology often pose significant diagnostic dilemmas. The aim of this study was to report our experience with a series of ectopic and metastatic meningiomas, characterizing their cytomorphology with histological correlation. MATERIALS AND METHODS: A retrospective analysis involving 13 patients with cytological preparations from extracranial meningiomas was performed. Cytology cases were correlated with available surgical resection specimens. Data regarding clinical findings, tumor information, cytomorphology, follow-up histological features and immunohistochemistry were recorded and analyzed. RESULTS: There were 5 cases with metastases and 8 ectopic meningiomas. Metastases occurred in the scalp/skull, lung, paraspinal soft tissue and liver. Primary ectopic meningiomas were located in the paranasal sinuses and ear, orbit and neck. Cytomorphological features characteristic of meningiomas were identified in the majority of samples including tightly cohesive clusters of spindled cells, whorls, intranuclear inclusions, nuclear grooves and psammomatous calcification. Unusual cytomorphological features identified in only a few cases included epithelioid cell predominance, abundant inflammatory cells, small-cell change, papillary structures and pseudoacinar growth. Metastatic tumors exhibited more nuclear atypia and occasionally mitoses or necrosis. Meningiomas were shown to be immunoreactive for epithelial membrane antigen, pancytokeratin and vimentin. CONCLUSION: Although rare, extracranial meningiomas can be encountered in cytologic specimens and should be included in the differential diagnosis when characteristic morphological features of meningiomas are seen. Cytopathologists should be aware that these lesions could be mistaken for other tumors, especially when confounded by atypia and unusual cytomorphological features.

Trichorhinophalangeal Syndrome Type 1 Is a Highly Sensitive and Specific Marker for Diagnosing Triple-Negative Breast Carcinomas on Cytologic Samples.

CONTEXT.­: Definitive diagnosis of metastatic triple-negative breast carcinoma (TNBC) is challenging on cytologic samples. Recent studies demonstrated that trichorhinophalangeal syndrome type 1 (TRPS1) is a highly sensitive and specific marker for diagnosing breast carcinomas, including TNBC, on surgical specimens. OBJECTIVE.­: To evaluate TRPS1 expression in TNBCs on cytologic samples and a large series of nonbreast tumors on tissue microarray sections. DESIGN.­: Immunohistochemical (IHC) analysis of TRPS1 and GATA-binding protein 3 (GATA3) was performed on 35 TNBC cases on surgical specimens, and 29 consecutive TNBC cases on cytologic specimens. IHC analysis of TRPS1 expression was also performed on 1079 nonbreast tumors on tissue microarray sections. RESULTS.­: Of the surgical specimens, 35 of 35 TNBC cases (100%) were positive for TRPS1, all with diffuse positivity, whereas 27 of 35 (77%) were positive for GATA3, with diffuse positivity in 7 cases (26%). Of the cytologic samples, 27 of 29 TNBC cases (93%) were positive for TRPS1, with diffuse positivity in 20 cases (74%), whereas 12 of 29 (41%) were positive for GATA3, with diffuse positivity in 2 cases (17%). Of the nonbreast malignant tumors, TRPS1 expression was seen in 9.4% (3 of 32) of melanomas, 10.7% (3 of 28) of small cell carcinomas of the bladder, and 9.7% (4 of 41) of ovarian serous carcinomas. CONCLUSIONS.­: Our data confirm that TRPS1 is a highly sensitive and specific marker for diagnosing TNBC cases on surgical specimens as reported in the literature. In addition, these data demonstrate that TRPS1 is a much more sensitive marker than GATA3 in detecting metastatic TNBC cases on cytologic samples. Therefore, inclusion of TRPS1 in the diagnostic IHC panel is recommended when a metastatic TNBC is suspected.

Practical applications of three-dimensional printing for process improvement in the cytopathology laboratory.

With the increased availability of three-dimensional (3D) printers, innovative teaching and training materials have been created in medical fields. For pathology, the use of 3D printing has been largely limited to anatomic representations of disease processes or the development of supplies during the coronavirus disease 2019 pandemic. Herein, an institution's 3D printing laboratory and staff with expertise in additive manufacturing illustrate how this can address design issues in cytopathology specimen collection and processing. The authors' institutional 3D printing laboratory, along with students and trainees, used computer-aided design and 3D printers to iterate on design, create prototypes, and generate final usable materials using additive manufacturing. The program Microsoft Forms was used to solicit qualitative and quantitative feedback. The 3D-printed models were created to assist with cytopreparation, rapid on-site evaluation, and storage of materials in the preanalytical phase of processing. These parts provided better organization of materials for cytology specimen collection and staining, in addition to optimizing storage of specimens with multiple sized containers to optimize patient safety. The apparatus also allowed liquids to be stabilized in transport and removed faster at the time of rapid on-site evaluation. Rectangular boxes were also created to optimally organize all components of a specimen in cytopreparation to simplify and expedite the processes of accessioning and processing, which can minimize errors. These practical applications of 3D printing in the cytopathology laboratory demonstrate the utility of the design and printing process on improving aspects of the workflow in cytopathology laboratories to maximize efficiency, organization, and patient safety.

Benchmarking cytology support for ROSE during endoscopic and bronchoscopic procedures.

INTRODUCTION: There has been an increase in endoscopic and bronchoscopic biopsies as minimally invasive methods to obtain specimens from gastrointestinal (GI) or pancreatobiliary lesions and thoracic or mediastinal lesions, respectively. As hospitals undertake more of these procedures, it is important to consider the staffing implications that this has on cytopathology laboratories with respect to support for rapid on-site evaluation (ROSE). MATERIALS AND METHODS: Volume and time data from endoscopic ultrasound and bronchoscopic procedures (including endobronchial ultrasound-guided transbronchial needle aspirations and small biopsies with touch preparation) in the GI suite, bronchoscopy suite, or operating room were reviewed for 2 months at 2 different medical centers with ROSE services provided by cytologists or fellows physically present at the procedure and cytopathologists located remotely using telecytology. Statistical analysis was performed to investigate significant trends based on the location of the biopsies and other factors. RESULTS: A total of 16 proceduralists performed 159 procedures and submitted 276 different specimens during 16 total weeks at 2 institutions. The total ROSE time for the on-site personnel to cover these procedures was 109.3 hours (bronchoscopy, 62.3 hours [57%]; GI, 29.8 hours [27%]; OR, 17.2 hours [16%]), which represents an average of 0.69 hour (41.4 minutes) per procedure or 0.40 hour (24.0 minutes) per part, with the shortest procedure times per sample recorded during bronchoscopy. When stratified by practice volume for individual proceduralists, the average time per specimen sample submitted was shorter for proceduralists with high volume practices and was most pronounced during bronchoscopy procedures. CONCLUSIONS: Endoscopic and bronchoscopic procedures account for an increasing amount of the ROSE time for the cytology team. On average, each ROSE procedure takes 0.69 hour (41.4 minutes) or approximately 0.40 hour (24.0 minutes) per specimen, with shorter time requirements for specimens obtained in bronchoscopy procedures and for operators with high volume practices for endobronchial ultrasound-guided transbronchial needle aspirations. This provides important benchmarking data to calculate staffing needs for cytology to provide ROSE support for different proceduralists.

American Society of Cytopathology Telecytology validation recommendations for rapid on-site evaluation (ROSE).

Telecytology has multiple applications, including rapid onsite evaluation (ROSE) of fine-needle aspiration (FNA) specimens. It can enhance cytopathology practice by increasing productivity, reducing costs, and providing subspecialty expertise in areas with limited access to a cytopathologist. However, there are currently no specific validation guidelines to ensure safe practice and compliance with regulations. This initiative, promoted by the American Society of Cytopathology (ASC), intends to propose recommendations for telecytology implementation. These recommendations propose that the validation process should include testing of all hardware and software, both separately and as a whole; training of all individuals who will participate in telecytology with regular competency evaluations; a structured approach using retrospective slides with defined diagnoses for validation and prospective cases for verification and quality assurance. Telecytology processes must be integrated into the laboratory's quality management system and benchmarks for discrepancy rates between preliminary and final diagnoses should be established and monitored. Special attention should be paid to minimize discrepancies that downgrade malignant cases to benign (false positive on telecytology). Currently, billing and reimbursement codes for telecytology are not yet available. Once, they are, recommendation of the appropriate usage of these codes would be a part of the recommendations. These proposed guidelines are intended to be a resource for laboratories that are considering implementing telecytology. These recommendations can help to ensure the safe and effective use of telecytology and maximize its benefits for patients.

Indeterminate pediatric thyroid fine needle aspirations: a study of 68 cases.

OBJECTIVE: The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) provides a reporting scheme for thyroid fine needle aspiration (FNA) and includes three indeterminate categories with different management strategies. This study analyzes indeterminate thyroid FNAs in children, and correlates these findings with the histological features. METHODS: A total of 179 thyroid FNA specimens were retrieved from children. Cases were categorized by TBSRTC. Only cases diagnosed as atypia (AUS)/follicular lesion of undetermined significance (FLUS), suspicious for follicular or oncocytic neoplasm (SFON), or suspicious for malignancy (SM) were selected and correlated with the nodule size and histological follow-up. RESULTS: Sixty-eight cases were identified, including 43 (63%) AUS/FLUS diagnoses, 19 (28%) SFON, and 6 (9%) SM. On follow-up, 48% were malignant, including 28% AUS/FLUS cases, 58% SFON, and 100% SM. The average size of the malignant lesions diagnosed preoperatively as AUS/FLUS was 1.5 cm (range 0.7-4.5), compared to 3.3 cm (range 1.2-6.6) in SFON and 2.8 cm (range 0.7-3.8) in SM. Malignancies included 92% papillary thyroid carcinoma (PTC), 77% of which were the follicular variant of PTC (FVPTC) and 8% follicular carcinomas. The AUS/FLUS cases were largely due to compromised specimens (49%) and the highest malignancy rate occurred in those with cytological atypia (50%). CONCLUSIONS: This study shows an incremental risk of malignancy within the indeterminate categories using TBSRTC in children. Malignant nodules with a preoperative AUS/FLUS diagnosis tended to be smaller than those with a SFON or SM diagnosis, and the vast majority of malignancies were PTC, with a high proportion being FVPTC.

Current Approach to Undifferentiated Neoplasms, With Focus on New Developments and Novel Immunohistochemical Stains.

CONTEXT.­: Workup of the poorly differentiated or undifferentiated tumor remains a significant and challenging entity in the practice of anatomic pathology. Particularly in the setting of small biopsies and limited material, these cases demand a balanced approach that considers the patient's clinical and radiologic presentation, a basic assessment of tumor morphology, a reasonably broad immunohistochemical panel, and diligent preservation of tissue for prognostic and therapeutic studies. OBJECTIVE.­: To illustrate some of the new and emerging immunohistochemical markers in the evaluation of tumors with undifferentiated or poorly differentiated morphology, with a focus on the workup in limited tissue samples to raise awareness of the issues involved with the pathologic workup in these challenging tumors. DATA SOURCES.­: A literature review of new ancillary studies that can be applied to cytologic specimens was performed. CONCLUSIONS.­: Knowledge of the patient's history and communication with the patient's clinical team is essential in formulating a differential diagnosis that can appropriately limit the differential diagnosis based on morphology, especially in small specimens. This information, in conjunction with classifying the tumor morphology (eg, epithelioid, spindled, neuroendocrine, basaloid/biphasic, mixed) gives a logical approach to choosing an initial immunohistochemical panel. Fortunately, immunohistochemistry is evolving quickly in the wake of groundbreaking molecular studies to develop new and better markers to further classify these difficult tumors beyond where we traditionally have been able to go.