The cytological features of effusions with mesothelioma in situ: A report of 9 cases.

INTRODUCTION: The diagnosis of mesothelioma in situ (MIS) is now accepted by the WHO as a pre-invasive neoplastic mesothelial proliferation and considered a diagnosis based on histologic evaluation only. Although the definition of MIS includes recurrent effusions, little is known about the cytologic features of such effusions. Since mesothelioma is usually diagnosed at an advanced stage and has a poor prognosis, early detection of a neoplastic mesothelial population in such effusions can potentially have a positive impact on the management of such a dire disease. MATERIALS AND METHODS: We reviewed a total of 18 pleural effusions from nine patients with recurrent effusions. Of these, five patients had follow-up biopsies diagnosed as MIS and the remaining four cases had negative radiology and malignant cytology proven by molecular markers (BAP1, MTAP or CDKN2A deletion) and at least 1 year follow-up with no overt mass identified by radiology. RESULTS: Initial effusions may mimic reactive mesothelial hyperplasia or exhibit atypia. As effusions recur, the cellularity and atypia increase and the mesothelial proliferation becomes morphologically indistinguishable from mesothelioma. Molecular alterations diagnostic of mesothelioma can be detected in these effusions, even in the initial-benign/reactive appearing ones. The cellularity and atypia detected in such effusions surpassed those noted on the biopsies, raising questions regarding the cause of such discrepancy. CONCLUSION: The diagnosis of MIS can be suspected based on malignant effusion cytology supported by molecular alterations. We propose that the proliferation of neoplastic mesothelial clones represent a clinically silent "liquid phase MIS stage" corresponding to in situ stage in other organs.

The cytologic diagnosis of mesothelioma: are we there yet?

INTRODUCTION: Mesothelioma is a rare but highly aggressive malignancy with poor prognosis that frequently present with recurrent effusions. Establishing the diagnosis by cytology can lead to early diagnosis and treatment and consequently improve prognosis. MATERIALS AND METHODS: This review examines the cytological diagnosis of mesothelioma in the context of its historical and morphologic evolution and provides an update of the current reporting systems. Clues to identify the mesothelial and malignant nature of the sample are detailed as well as the supporting ancillary tests. RESULTS: Cytologically, the samples are overwhelmingly cellular and malignancy is recognized by both architectural and cytological atypia. Numerous variably sized clusters and enlarged cells are easily identified, some with papillary architecture and collagen cores. Recognizing the mesothelial nature of the cells and supportive immunostains are essential to rule out the differential diagnosis of metastatic carcinomas and reactive mesothelium. Current ancillary tests such as homozygous deletion of CDKN2A, loss of BRCA1-associated protein, and methylthioadenosine phosphorylase expression can provide further support of malignancy. CONCLUSIONS: At this time with the aid of current ancillary tests and in the hands of cytopathologists with adequate experience with the interpretation of effusions, the diagnosis of mesothelioma can be established with accuracy in most cases.

The World Health Organization Reporting System for Lung Cytopathology.

The International Academy of Cytology has joined with the International Agency for Research on Cancer (IARC) to bring together a group of experts in lung cytopathology to develop a WHO Reporting System for Lung Cytopathology (WHO System). This WHO System defines five categories for reporting lung cytopathology, that is, "Insufficient"/"Inadequate"/"Non-diagnostic," "Benign," "Atypical," "Suspicious for malignancy," and "Malignant," each with a clear descriptive term for the category, a definition, a risk of malignancy and a suggested management algorithm. The key diagnostic cytopathology features of each of the lesions within each category have been established by consensus and will be presented more fully in a subsequent IARC e-book and published hard cover book.The WHO System provides the best practice application of ancillary testing, including immunocytochemistry and molecular pathology, and provides a review to guide sampling and processing techniques to optimize the handling and preparation of the cytopathology sample emphasizing the cytomorphological differential diagnosis to aid low-resourced settings. The authors recognize that local medical and pathology resources will vary, particularly in low- and middle-income countries, and have developed the WHO System to make it applicable worldwide based on cytomorphology with options for further diagnostic management of the patient.The online WHO System provides a direct link to the WHO Tumour Classification for Thoracic Tumours 5th Edition. It will raise the profile and use of cytopathology by increasing awareness of its current role and its potential role in the era of personalized medicine based on molecular pathology utilizing "small biopsies." Ultimately, the System will improve patient care and outcomes.This System aims to improve and standardize the reporting of cytopathology, facilitate communication between cytopathologists and clinicians and improve patient care. The System is based on the current role of lung cytopathology and synthesizes the existing evidence while highlighting areas requiring further research and the future potential role of lung cytopathology.

Automated analysis of computerized morphological features of cell clusters associated with malignancy on bile duct brushing whole slide images.

BACKGROUND: Bile duct brush specimens are difficult to interpret as they often present inflammatory and reactive backgrounds due to the local effects of stricture, atypical reactive changes, or previously installed stents, and often have low to intermediate cellularity. As a result, diagnosis of biliary adenocarcinomas is challenging and often results in large interobserver variability and low sensitivity OBJECTIVE: In this work, we used computational image analysis to evaluate the role of nuclear morphological and texture features of epithelial cell clusters to predict the presence of pancreatic and biliary tract adenocarcinoma on digitized brush cytology specimens. METHODS: Whole slide images from 124 patients, either diagnosed as benign or malignant based on clinicopathological correlation, were collected and randomly split into training (ST , N = 58) and testing (Sv , N = 66) sets, with the exception of cases diagnosed as atypical on cytology were included in Sv . Nuclear boundaries on cell clusters extracted from each image were segmented via a watershed algorithm. A total of 536 quantitative morphometric features pertaining to nuclear shape, size, and aggregate cluster texture were extracted from within the cell clusters. The most predictive features from patients in ST were selected via rank-sum, t-test, and minimum redundancy maximum relevance (mRMR) schemes. The selected features were then used to train three machine-learning classifiers. RESULTS: Malignant clusters tended to exhibit lower textural homogeneity within the nucleus, greater textural entropy around the nuclear membrane, and longer minor axis lengths. The sensitivity of cytology alone was 74% (without atypicals) and 46% (with atypicals). With machine diagnosis, the sensitivity improved to 68% from 46% when atypicals were included and treated as nonmalignant false negatives. The specificity of our model was 100% within the atypical category. CONCLUSION: We achieved an area under the receiver operating characteristic curve (AUC) of 0.79 on Sv , which included atypical cytological diagnosis.

Rapid On-site Evaluation Practice Variability Appraisal (ROSE PETAL) survey.

BACKGROUND: Rapid on-site evaluation (ROSE) is frequently used during diagnostic procedures in patients with or suspected to have lung cancer. There is variation in ROSE use among bronchoscopists, and discussion of ROSE results can have significant consequences for patients. This study was performed to define ROSE practice and result disclosure patterns among bronchoscopists. METHODS: This cross-sectional study was performed using an electronic survey disseminated to the members of the American Association for Bronchology and Interventional Pulmonology and the Society for Advanced Bronchoscopy. The questions centered around ROSE availability, utilization, barriers, and discussion of results with patients. RESULTS: There were 137 respondents. Most identified themselves as interventional pulmonologists (109, 80%); most respondents worked in an academic setting (71, 52%). Availability of ROSE was reported by 121 (88%) respondents. Time constraints (28%), availability of cytology (22%), and scheduling conflicts (20%) were the most reported barriers to ROSE use. Endobronchial ultrasound transbronchial needle aspiration (85%) and nonrobotic peripheral bronchoscopy (65%) were the most reported procedures that used ROSE. There was heterogeneity regarding discussion of ROSE results with the patient or their caregiver in the immediate postprocedure setting: yes - always (40, 33%), yes - sometimes (32, 26%), yes - rarely (18, 15%), or no (31, 26%). Thirty-eight respondents reported they believed ROSE was ≥90% concordant with final cytology results. CONCLUSIONS: The results confirmed the heterogeneity of practice patterns. Estimates of ROSE-final cytology concordance were lower than previously published concordance results. Notably, the discussion of ROSE results varied significantly.

The concept of mesothelioma in situ, with consideration of its potential impact on cytology diagnosis.

Diffuse malignant mesothelioma (MM) is an incurable tumour of the serosal membranes, which is often caused by exposure to asbestos and commonly diagnosed at advanced stage. Malignant mesothelioma in situ (MMIS) is now included as diagnostic category by the World Health Organization (WHO). However, our international survey of 34 pulmonary pathologists with an interest in MM diagnosis highlights inconsistency regarding how the diagnosis is being made by experts, despite published guidelines. Whilst the WHO restricts the diagnosis to surgical samples, the very concept has implication for cytological diagnosis, which is already regarded as controversial in itself by some. MMIS is currently only applicable as precursor to MM with an epithelioid component, and raises the possibility for different molecular pathways for different histological MM subtypes. The clinical implications of MMIS at this stage are uncertain, but aggressive therapies are being initiated in some instances. Based on the results of the survey we here present a critical appraisal of the concept, its clinical and conceptual implications and provide practice suggestions for diagnosis. A low threshold for ancillary testing is suggested. The designations of 'malignant mesothelioma, cannot exclude MMIS' or 'atypical mesothelial proliferation with molecular indicators of malignancy, so-called MMIS' could be used on cytology samples, adding 'no evidence of invasion in sample provided' for surgical samples. Clinical and radiological correlation are integral to diagnosis and best done at multidisciplinary meetings. Finally, collaborative studies are required to improve our understanding of MMIS.

Immunochemistry in the work-up of mesothelioma and its differential diagnosis and mimickers.

The differential diagnosis in cellular effusions with cytological atypia often includes malignant mesothelioma (MM), reactive mesothelial proliferation, and malignancies of metastatic origin, particularly carcinomas. The International Reporting System for Serous Fluid recently established guidelines for reporting MM. In conjunction with the cytomorphologic evaluation, the role of immunochemistry (IC) was emphasized as a very useful tool in the workup of serous fluids, especially with the availability of novel markers. Utilizing a panel of markers, IC allows the characterization of the cells, whether mesothelial or not, and when mesothelial origin is established, IC can frequently assist in delineating its benign or malignant nature. IC can also confirm metastatic disease, allowing the identification of the primary origin in most cases. This review summarizes the current status of IC and its role in the diagnosis of MM and its differential diagnosis in serous fluids.

Serous fluid cytopathology: Past, present, and future.

Serous fluids/effusions are common cytology samples that reflect a wide range of diseases and lend themselves to a multitude of investigations that include microscopy, chemical analysis, cell count, cultures, and analysis for biomarkers and immunomarkers. In recent years, effusions have also served as a liquid biopsy that can be interrogated by molecular tests for thoranostic and prognostic markers and selection of targeted therapy. The recently published International Reporting System for Serous Fluid Cytopathology (IRSSF) provides a standardized reporting terminology with well- defined diagnostic criteria. This editorial provides a global review of the progress in the work-up of effusions and a summary of the IRSSF book and its most significant contributions. The editorial also includes a summary of the diagnostic categories including their definition and the significant relevant information.

Adenoviral cytopathic changes in pleural fluid: A unique cytological presentation mimicking malignancy.

Adenoviruses are emerging as important viral pathogens in immunocompromised patients due to immunodeficiency diseases and recently hematopoietic stem cell and solid organ transplant recipients, impacting morbidity and even mortality. Immunocompromised children are prone to respiratory infection, due to alterations in their immune system. When confronted with diseases involving the pleural effusions, such as viral infections, the diagnostic problem becomes more complex and special effort is needed to recognize and characterize them accurately and to differentiate them from other pathologies such as malignancies. However, cytology of adenoviral infection in pleural effusions has not been reported before. We report a case of an adenovirus infection of the pleural effusion which included lymphocytosis associated with background atypical cells showing a cytopathic effect (cytoplasmic viral particles). The differential diagnosis included lymphoma and infections. Immunohistochemical stain for adenovirus was positive and confirmed by molecular studies. Usually in viral infections there are cytopathic changes due to viral particles affecting epithelial cells but this case is unique as the viral particles were identified in macrophages. We discuss the significance of such infection in comparison with other viral changes indigenous to the pleural effusions, which could also occur in such specimens.

Root cause analysis of indeterminate diagnoses in serous fluids cytopathology.

INTRODUCTION: The International System for Reporting Serous Fluid Cytopathology proposed five diagnostic categories: Nondiagnostic (ND), Negative for Malignancy (NFM), Atypia of Undetermined Significance (AUS), Suspicious for Malignancy (SFM) and Malignant (MAL) (Primary or Metastatic). The indeterminate (AUS/SFM) categories are challenging for management. The goal of this study is to reveal the root causes contributing to indeterminate diagnoses (ID). MATERIALS AND METHODS: We searched our archives between 1 January 2017 and 30 June 2019, and performed a root cause analysis (RCA) using the "5 whys" method to determine the contributing factors of ID. RESULTS: Nine hundred eleven specimens were evaluated and diagnosed: ND (9, 1%), NFM (667, 73.2%), AUS (51, 5.6%), SFM (27, 3%) and MAL (157, 17.2%). More than one factor contributed to 38/78 ID. Low volume (<50 cc), and low cellularity were identified in 31 and 51 cases, respectively. Three cases were simply deferred to concurrent biopsy. Eleven cases were called atypical, favor reactive mesothelium despite confirmatory IHC. Atypical lymphoid population was reported in seven cases. Cellblocks (CB) were low in cellularity despite volume >1000 mL in 13 cases. Two mesotheliomas were underdiagnosed as suspicious. CONCLUSIONS: Low cellularity and low volume were the most common contributing factors, highlighting the importance of adequate sample collection. Adequate volume specimens with low cellularity may benefit from a close inspection and a second CB. Some IDs can be switched to NFM or MAL with careful consideration of clinical, radiologic findings and ancillary testing, and concurrent surgical pathology correlation when available.

Rapid on-site evaluation (ROSE) for fine needle aspiration of thyroid: benefits, challenges and innovative solutions.

Rapid on-site evaluation/adequacy assessment (ROSE) is considered an essential component of thyroid fine needle aspiration (FNA) and many reported that it significantly decreases the nondiagnostic (ND) rate. The average reported ND rate without ROSE is about 20% and is improved by 12% when ROSE is implemented. However, the data also suggest that the improvement in ND rate after implementation of ROSE is directly related to the ND rate prior to ROSE and that it is mostly beneficial to aspirators with less experience. Several studies have also raised concerns regarding the impact of ROSE as it prolongs the procedure time, requires additional resources and increases the cost incurred by the additional fees. This resulted in a wide variation in the methodology applied to acquire the sample and implement ROSE across the globe with variation in the number of passes performed, stain utilized and the personnel reviewing the slides, e.g., cytotechnologists versus pathologist. This review summarized the literature reporting the impact of ROSE including its pros and cons, its accuracy and reproducibility, concordance between cytotechnologists and pathologists based on final diagnosis and highlights the different ways laboratories attempted to circumvent the challenges. In particular, the review highlights a unique approach practiced in Ito Hospital, Tokyo, Japan.

Use of cytology centrifuged supernatants improves cost and turnaround time for targeted next generation sequencing.

BACKGROUND: Molecular testing is an essential step in providing patients with advanced non-small-cell lung cancer (NSCLC), the most appropriate front-line targeted therapies. We recently implemented targeted NGS on previously discarded cytology centrifuged supernatant (CCS). METHODS: In this study, we reviewed our implementation process to evaluate its performance. Performance and turnaround time (TAT) of molecular testing on all cytology NSCLC cases submitted for targeted NGS from June 2018 to September 2019 were evaluated, which included 46 and 62 cytology cases before and after implementation of CCS, respectively. Associated cost savings using CCS was also analyzed. RESULTS: The mean TAT defined as the time of collection to time of reporting was 8.5 ± 1.8 days in CCS cohort (range 5-13) as compared with 12.2 ± 5.3 days in the (FFPE) cell block (CB) cohort (range: 6-27). The success rate of sequencing was similar for both cohorts (100% in CCS and 96% in FFPE CB). CONCLUSION: Our results demonstrate that NGS using CCS improves TAT, preserves FFPE CB for other testing, and results in cost savings of $50 per case.

Nondiagnostic fine-needle aspirates of the pancreas: A root cause analysis.

BACKGROUND: Fine-needle aspiration (FNA) of the pancreas is considered the primary and least invasive diagnostic method in the evaluation of pancreatic lesions. A nondiagnostic sample may trigger repeat FNA or a more invasive diagnostic procedure. The goal of this study was to identify the root causes of nondiagnostic samples. METHODS: We performed a retrospective review of FNAs of the pancreas categorized as nondiagnostic at our institution between 2008 and 2019. Medical records and slides were reviewed to identify the features described by imaging, rapid on-site evaluation, fluid chemistry, final cytology diagnosis, and final histology. A root cause analysis was performed using the Ishikawa (or fishbone) diagram and the 5 Whys method. RESULTS: A total of 30 cases were identified: 11 adenocarcinomas, 6 cases of pancreatitis, 4 intraductal papillary mucinous neoplasms, 3 serous cystadenomas, 3 neuroendocrine tumors, 1 mucinous cystic neoplasm, 1 retention cyst, and 1 case of Brunner gland hyperplasia. The root causes identified were: man in 8 cases, machine in 1 case, method in 17 cases, and material in 18 cases. In many cases, more than 1 root cause contributed to the problem. CONCLUSION: Material related errors contributed to the majority of nondiagnostic results and were primarily related to fibrotic cancers, chronic pancreatitis, absence of diagnostic criteria of cystic lesions, and technically challenging cases. Only 1 major interpretation error was identified. Sampling and interpretive errors contributed equally to man-related causes. For mucinous cysts, neoplastic mucin was difficult to identify in liquid-based preparations. Pathologists tended to issue a nondiagnostic categorization when epithelial cells are lacking and particularly when the nature and radiological impression of the cyst was not communicated.

Collection and Handling of Thoracic Small Biopsy and Cytology Specimens for Ancillary Studies: Guideline From the College of American Pathologists in Collaboration With the American College of Chest Physicians, Association for Molecular Pathology, American Society of Cytopathology, American Thoracic Society, Pulmonary Pathology Society, Papanicolaou Society of Cytopathology, Society of Interventional Radiology, and Society of Thoracic Radiology.

CONTEXT.­: The need for appropriate specimen use for ancillary testing has become more commonplace in the practice of pathology. This, coupled with improvements in technology, often provides less invasive methods of testing, but presents new challenges to appropriate specimen collection and handling of these small specimens, including thoracic small biopsy and cytology samples. OBJECTIVE.­: To develop a clinical practice guideline including recommendations on how to obtain, handle, and process thoracic small biopsy and cytology tissue specimens for diagnostic testing and ancillary studies. METHODS.­: The College of American Pathologists convened an expert panel to perform a systematic review of the literature and develop recommendations. Core needle biopsy, touch preparation, fine-needle aspiration, and effusion specimens with thoracic diseases including malignancy, granulomatous process/sarcoidosis, and infection (eg, tuberculosis) were deemed within scope. Ancillary studies included immunohistochemistry and immunocytochemistry, fluorescence in situ hybridization, mutational analysis, flow cytometry, cytogenetics, and microbiologic studies routinely performed in the clinical pathology laboratory. The use of rapid on-site evaluation was also covered. RESULTS.­: Sixteen guideline statements were developed to assist clinicians and pathologists in collecting and processing thoracic small biopsy and cytology tissue samples. CONCLUSIONS.­: Based on the systematic review and expert panel consensus, thoracic small specimens can be handled and processed to perform downstream testing (eg, molecular markers, immunohistochemical biomarkers), core needle and fine-needle techniques can provide appropriate cytologic and histologic specimens for ancillary studies, and rapid on-site cytologic evaluation remains helpful in appropriate triage, handling, and processing of specimens.

Tiny but mighty: use of next generation sequencing on discarded cytocentrifuged bile duct brushing specimens to increase sensitivity of cytological diagnosis.

Bile duct brushing (BDB) is used to evaluate pancreatobiliary lesions as it widely samples lesions with a low complication rate. Cytological evaluation of BDB is a specific but insensitive test. There is limited literature on the use of post-cytocentrifuged (PCC) samples, which are usually discarded, for next-generation sequencing (NGS) as an adjunct to cytological diagnosis of BDB. In this study we investigate whether molecular analysis by NGS of PCC specimens improves the sensitivity of diagnosis. PCC samples from 100 consecutive BDB specimens spanning 93 unique patients were retained. DNA was extracted and mutational analysis was performed agnostic of morphologic or clinical findings. Each BDB specimen was characterized as negative, atypical or positive based on morphological analysis by trained cytopathologists. Performance characteristics for mutational profiling and morphological analysis were calculated on the basis of clinicopathologic follow-up. There was sufficient clinicopathologic follow-up to classify 94 of 100 cases as either malignant (n = 43) or benign (n = 51). Based on morphologic analysis of cytology, these 94 cases were classified as either benign (n = 55), atypical (n = 18), or as at least suspicious or positive for malignancy (n = 21). Morphologic analysis of cytology showed a sensitivity of 49% and a specificity of 100% if atypical cases were considered negative. NGS revealed oncogenic alterations in 40/43 (93%) of malignant cases based on clinicopathologic follow-up. The most common alterations were in KRAS and TP53, observed in 77% and 49% of malignant cases respectively. No alterations were observed in the 51 benign cases classified based on clinicopathologic follow-up. Supplementing cytomorphologic analysis with molecular profiling of PCC by targeted NGS analysis increased the sensitivity to 93% and maintained specificity at 100%. This study provides evidence for the utility of NGS molecular profiling of PCC specimens to increase the sensitivity of BDB cytology samples, although studies with larger cohorts are needed to verify these findings.

Can we predict histological outcome of distinctive cohorts of patients with glandular cell abnormalities on ThinPrep Papanicolaou testing based on human papillomavirus status, age, and associated squamous abnormalities?

BACKGROUND: The cytology diagnosis of glandular cell abnormalities (GCAs) is diagnostically challenging, causing inadequate reproducibility. Histological outcome of GCA on cytology varies from benign to malignant diseases. The goal of this study is to evaluate histological outcome and identify distinctive cohorts of patients with GCA based on human papillomavirus (HPV) status, age, and associated squamous abnormality to stratify the patient into high risk for squamous/glandular lesions. METHODS: From 2012 to 2017, out of 162 088 ThinPrep Papanicolaou tests performed, 998 (0.61%) were reported as GCAs. Histologic follow-up was available in 638 cases and 429 had concurrent HPV results. RESULTS: The overall rate of high-risk human papillomavirus (hrHPV)-positivity (hrHPV+) was 33.6% (144/429 cases). Among the hrHPV+ cases, 18.1% had cervical intraepithelial neoplasia 2/3 (CIN2/3), 3.5% squamous cell carcinoma (SCC), 3.5% cervical adenocarcinoma in situ (AIS)/adenocarcinoma (ADC), and 2.8% endometrial carcinoma. Among hrHPV- cases, 1.4% had CIN2/3, 1.1% AIS/ADC, and 17.5% endometrial carcinoma. The high-grade cervical lesions (CIN2/3/AIS/ADC) were significantly higher in women with hrHPV+ and associated squamous abnormalities compared to hrHPV- and no squamous abnormality in all age groups except patients >65 years. Endometrial carcinoma was most commonly present in women >65 years especially with HPV- and no associated squamous abnormalities. CONCLUSIONS: HPV testing is useful for predicting the risk of high-grade cervical neoplasia in women with GCA especially with associated squamous abnormalities on cytology. The endometrial carcinoma is more frequent in hrHPV- older women. The combination of cytology with knowledge of associated squamous abnormality, hrHPV status, and age can significantly aid in stratifying the patient into high risk for glandular/squamous lesions which facilitates appropriate management of these patients.

Association of Bacterial Vaginosis and Human Papilloma Virus Infection With Cervical Squamous Intraepithelial Lesions.

OBJECTIVES: Human papillomavirus (HPV) is known to be associated with squamous intraepithelial lesions (SILs). However, there is limited and conflicting literature on the relationship between bacterial vaginosis (BV) and SIL. The aim of this study is to determine the prevalence of BV and evaluate the association between BV and SIL. METHODS: A retrospective study was performed on 10,546 cases between 2012 and 2017. HPV results were available in 7,081 cases. RESULTS: BV was present in 17.6% of cases. There was significant association between BV, positive HPV infection, and high-grade SIL. BV patients with negative HPV infection showed more squamous abnormalities than BV-negative HPV-negative patients. CONCLUSIONS: We found there is a significant association between BV and SIL. BV is more common among patients with HPV infection and is independently associated with squamous abnormalities in cervical smears and surgical follow-up.

Comparison of cytocentrifugation supernatant fluid and formalin-fixed paraffin-embedded tissue for targeted next-generation sequencing.

BACKGROUND: The emergence of less invasive procedures coupled with the growth of molecular testing have created a need for clinical laboratories to optimize workflows to enable tissue preservation and ancillary testing. In the preparation of formalin-fixed paraffin-embedded cell blocks (FFPE CBs), there is a cytocentrifugation step for cell pellet extraction that results in postcentrifugation supernatant fluid (SN). This SN, which in most routine workflows is discarded, has been suggested to contain adequate cellular material for molecular testing. In the current study, the authors describe the use of DNA and RNA extracted from SN for the detection of clinically relevant biomarkers by next-generation sequencing (NGS). METHODS: After cell pellet removal, cytocentrifugation SN from 30 endobronchial fine-needle aspiration rinses that were positive for malignancy on FFPE CB were collected. DNA and RNA were extracted from the SN and tested using an in-house NGS Solid Tumor Focus Assay. The NGS results were compared with findings from corresponding FFPE samples. RESULTS: Testing was successful in all 30 samples. There was 100% concordance between variants observed in the SN and corresponding FFPE specimens, which included 50 single-nucleotide variants, 9 copy number amplifications, 3 structural variants, and 2 indels. Furthermore, there was excellent correlation (correlation coefficient, 0.93) between the variant allele frequency of mutations observed in SN compared with that noted in corresponding FFPE CBs. CONCLUSIONS: Cytocentrifugation SN is a valuable source for NGS, is comparable to FFPE that preserves tissue for other ancillary testing, and can reduce the failure rate of testing that may result from insufficient material being available in the CB.

The pathological outcome of ThinPrep Pap tests diagnosed as glandular cell abnormalities alone versus combined glandular and squamous abnormalities.

BACKGROUND: Although the incidence of glandular cell abnormalities (GCA) on cervical cytology is low, the clinical and histologic findings are often significant. A combined diagnosis of squamous intraepithelial lesion (SIL) and GCA indicates concern for either two distinct lesions or single entity. The goal of this study is to evaluate the outcome of the diagnosis of GCA alone or in combination with squamous abnormality (SqA). METHODS: From January 2012-June 2017, our laboratory processed 162 088 ThinPrep Pap tests. 998 (0.61%) cases were reported as GCA. Histologic follow-up was available in 569 cases after excluding adenocarcinoma, NOS and atypical endometrial cells. HPV results were available in 429 (67.2%) cases. RESULTS: The surgical follow-up on 271 cases with GCA alone diagnosis revealed negative/benign lesions in 183 (67.5%) cases, glandular lesions in 40 (14.8%) cases; SqA in 47 (17.3%) cases; combined in 1 (0.4%) case. Surgical follow-up on 298 cases with dual interpretation revealed negative/benign lesions in 108 (36.2%) cases, SqA in 159 (53.4%) cases, GCA in 21 (7.0%) cases and only 10 (3.4%) cases were combined lesions. The mean age was 44 ±13.36 years. The overall hrHPV-positive rate was 36.2%. Endometrial carcinoma was most common abnormality in patients >65 years (71.4%) especially with hrHPV-negative results. CIN 1-3 was the most common finding in patients <30 years (50%). CONCLUSION: A cytological diagnosis of GCA has a higher risk of glandular abnormality on surgical follow-up especially in the older and hrHPV-negative group (P < .0001) while a combined diagnosis has a higher risk of a squamous lesion especially in <30 years (P < .0001). The combination of cytology, hrHPV-status and patient age can significantly aid in the stratification of the patient into high risk for glandular/squamous lesions which results in appropriate management.

Cytohistological correlation, immunohistochemistry and Murine Double Minute Clone 2 amplification of pulmonary artery intimal sarcoma: A case report with review of literature.

Pulmonary artery intimal sarcoma is a rare aggressive intraluminal tumor often misdiagnosed as acute or chronic pulmonary thromboembolism due to its clinical presentation and radiological findings. Thus early diagnosis is very crucial and may improve patient outcome. There is limited literature on diagnosis of pulmonary artery sarcoma by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). Herein, we report a case of mass-like lesion in the PA diagnosed on cytological material obtained by EBUS-TBNA with rapid on-site evaluation (ROSE). The aspirate showed pleomorphic malignant spindled cells arranged in loosely cohesive clusters. The intraluminal origin of PAIS was supported by radiographic findings. Subsequently, the patient received preoperative chemotherapy and underwent tumor resection with reconstruction. This report describes the cytomorphologic features of this rare intravascular tumor and demonstrates how limited cytological sample obtained from EBUS-TBNA with ROSE can be triaged efficiently for ancillary studies like immunohistochemistry and MDM2 amplification, thus expediting the management.