Evaluation of Lineage/Site-specific Nuclear Immunohistochemical Markers SATB2, Cyclin D1, SALL4, and BCOR in High-grade Endometrial Carcinomas.

Poorly differentiated malignant neoplasms involving the gynecologic tract routinely include a poorly differentiated endometrial carcinoma (EC) in the differential diagnosis. Some nuclear lineage/site-specific immunohistochemical markers are utilized in this diagnostic setting including SATB2, cyclin D1, SALL4, and BCOR, but their specificity and use in small samples are not clear across the spectrum of ECs. Cases of undifferentiated/dedifferentiated endometrial carcinomas (UEC/DDEC), clear cell carcinoma (CCC), uterine serous carcinoma (USC), FIGO grade 3 endometrial endometrioid carcinoma (EEC), and uterine carcinosarcoma (UCS) were identified and diagnoses confirmed. Whole-section immunohistochemical stains for SATB2, cyclin D1, SALL4, BCOR, and PAX8 were performed. A total of 113 cases were utilized: 15 CCC, 26 EEC, 19 UCS, 22 USC, and 31 UEC/DDEC. Cases were distributed across both low (49%) and high (51%) FIGO clinical stages. SATB2 was expressed by UCS (8/19, 42%), EEC (10/26, 38%), UEC/DDEC (11/30, 37%), and USC (6/22, 27%). Cyclin D1 was expressed by EEC (24/26, 92%), USC (17/22, 77%), UEC/DDEC (15/20 EEC component, 75%; 22/30 UEC, 73%), UCS (10/16 carcinoma, 63%; 11/19 sarcoma, 58%), and CCC (8/15, 53%). SALL4 was expressed most frequently by UEC/DDEC (12/30, 40%), but also USC (7/22, 32%), EEC (5/26, 19%), and UCS (4/16 carcinoma, 25%; 3/19 sarcoma, 16%). BCOR was expressed at low levels in 2 USC, 2 UEC/DDEC, and 2 UCS. PAX8 was generally positive but showed lower expression in UEC/DDEC (17/30, 57%) and in the sarcomatous portions of UCS (6/19, 32%). SATB2, cyclin D1, SALL4, and BCOR stain variable numbers of poorly-differentiated EC and must be carefully interpreted within morphologic and clinical context.

Impact of education on institutional and faculty rates of atypical squamous cells.

INTRODUCTION: Papanicolaou test quality metrics include the ASC rate, ASC:SIL ratio, and ASC HPV+ rate. What a laboratory should do when metrics show a worrisome trend is not well defined. In 2015, our laboratory noted a worrisome trend in our quality metrics and decided to implement a systemic education program in 2016; we monitored the effectiveness of our program. METHODS: An educational intervention was designed for March/April 2016. Cytotechnologist education consisted of: group meeting on March 10 to discuss metrics, lecture, and written materials on ASC-US criteria, a quiz on challenging ASC-US cases, encouragement to seek consultation, and each cytotechnologist received quarterly individual metrics. The cytopathologist education consisted of: group meeting on April 16 to discuss metrics, encouragement to bring borderline cases to consensus conference, and each faculty received quarterly individual metrics. The ASC rate, ASC:SIL ratio, and ASC HPV+ rate was collected for the institution and each individual faculty in 2016 for January to March (pre-interventions, Q1), April to June (post-interventions, Q2), and July to September (post-interventions, Q3). ASC-H was included in the calculation of ASC %, ASC:SIL, and ASC HPV+ rates. RESULTS: There was a substantial decline in the lab ASC rate and ASC:SIL ratio, and the ASC HPV+ rate increased. Individual faculty changes in ASC:SIL ratio and ASC HPV+ rate also improved. CONCLUSIONS: In our institution, an educational program has been very effective in improving Papanicolaou test metrics. It is helpful to perform re-education at all levels within the department.

ACGME Milestones 2.0: why and what's new for cytopathology?

BACKGROUND: Primary stakeholders in the Accreditation Council for Graduate Medical Education (ACGME) Milestones Project are: ACGME, Residency Programs, Residents, Fellowship Programs, Fellows, and Certification Boards. The intent of the Milestones is to describe the educational and professional developmental trajectory of a trainee from the first stages of their postgraduate education through the completion of their clinical training. The Milestones 2.0 project includes changes made based on experience with Milestones 1.0. METHODS: The ACGME solicited volunteers to participate in the development of subspecialty Milestones 2.0. The workgroup was charged with reviewing/making any additions to the four "Harmonized Milestones", developing subspecialty specific milestones for the Patient Care and Medical Knowledge competencies, and creating a supplemental guide. The Milestones were finalized following review of input from an open comment period. RESULTS: The Cytopathology Milestones 2.0 will go into effect July 2021. They include additional subcompetencies in the 4 harmonized competency areas and cytopathology-specific edits to the patient care and medical knowledge subcompetencies. Although the number of subcompetencies has increased from 18 to 21, within each subcompetency, the number of milestone trajectories has decreased. Additionally, within each subcompetency, the wording has been streamlined. A supplemental guide was created and Milestones 1.0 were compared to 2.0; however, curriculum mapping has been left to programs to develop. CONCLUSIONS: The ultimate goal of the Cytopathology Milestones 2.0 is to provide better real-time documentation of the progress of cytopathology fellows. The expected outcome is to produce highly competent cytopathologists, improving the care they provide, regardless of the program at which they trained.

Cytopathology milestones: can you get to level 5?

INTRODUCTION: ACGME Milestones describe 6 areas of proficiency, indicating readiness for practice. Each is divided into 5 levels of mastery; Level 1 (new trainees) through Levels 4 (graduation) and 5 (aspirational). Milestones reporting began Spring 2016. We used Milestones to assess graduated fellows. MATERIALS AND METHODS: We conducted phone interviews with previous fellows and collected demographic information including practice setting. We asked graduates if they fulfilled each example of mastery and recorded their answers. RESULTS: A total of 22 fellows graduated from 2010 to 2017; 15 responded (10 academic, 5 private). Milestones in which nearly all respondents performed well (Level 4+) were: PC1, MK1, SBP2, SBP4, PROF1-4, ICS1-3. Some were more challenging (PC2, MK2, SBP1/3/5, PBL1). For PC2, 2 respondents achieved Level 1 (did not perform fine-needle aspirations). For MK2, 2 respondents achieved Level 1 (did not evaluate Papanicolaou). For SBP1, 80% in private practice achieved Level 5; 50% in academics achieved Level 3. For SBP3, 80% in private practice achieved Level 4+; 100% in academics achieved maximum Level 2. For SBP5, 60% of all respondents achieved maximum Level 3; only 1 achieved Level 5. CONCLUSIONS: Many Milestones are attainable. Eleven of 18 yielded Level 4+ from most respondents. Three (PC2, MK1, MK2) yielded rare Level 1 due to scope of practice. Others (SBP1, SBP3) reflect more of an all-or-nothing phenomenon. For SBP5, most respondents achieved Level 3; only 1 achieved Level 5. Some Milestones are highly dependent on practice setting, and others remain aspirational.

Cytology and curetting diagnosis of endocervical adenocarcinoma.

INTRODUCTION: Papanicolaou testing is effective in identifying squamous intraepithelial lesions of the cervix. Endocervical adenocarcinoma (EAC) and adenocarcinoma in situ (AIS) are far less commonly identified. Endocervical curettings (ECCs) are usually obtained after colposcopic biopsy, sample the endocervical canal, and aid in the detection of endocervical glandular lesions. Here, we examine the utility of Papanicolaou testing and endocervical curetting for detecting AIS and EAC. MATERIALS AND METHODS: Cases from 2007 to 2019 with a histologically confirmed diagnosis of AIS and EAC were identified and the clinical data and diagnostic material, including the cytology and surgical specimens, obtained. RESULTS: A total of 108 cases of AIS and EAC were identified, Papanicolaou tests were performed in 97 of these cases, and ECC in 87. AIS or EAC were detected in 46.4% of Papanicolaou tests; 41.4% of ECC showed AIS or EAC. A total of 92.7% of cases were positive for high-risk human papillomavirus (HPV) and concurrent squamous intraepithelial lesion was present in 53.3% of cases. AIS or EAC were more commonly identified in cases without concurrent squamous intraepithelial lesions. CONCLUSIONS: Papanicolaou testing and endocervical curettings have a low detection rate for AIS and EAC. The majority of AIS and EAC cases test positive for high-risk HPV. Papanicolaou test and ECC show different detection rates and are complementary tools in patients with AIS and EAC. In some settings, an ECC can increase the diagnostic sensitivity and specificity of the pathologic diagnosis.

Primary salivary gland-type tumors of the tracheobronchial tree diagnosed by transbronchial fine needle aspiration: Clinical and cytomorphologic features with histopathologic correlation.

BACKGROUND: Primary salivary gland-type tumors of the tracheobronchial tree are rare; their cytologic features have been seldom reported. We aim to describe the clinical and cytomorphologic features of tracheobronchial salivary gland-type tumors diagnosed by transbronchial fine needle aspiration (TBNA) at our institution, and correlate the findings with a corresponding surgical specimen. METHODS: We searched our laboratory information system to identify patients with a primary salivary gland-type neoplasm of the tracheobronchial tree diagnosed by TBNA and with a corresponding surgical pathology specimen, over 10 years. RESULTS: The study cohort consisted of 11 patients (7F/4M; mean age 58 years, range 41-78). Presenting symptoms included hemoptysis (4), cough (3), dyspnea (1), stridor (1), and shoulder pain (1). Most had a tracheal mass (5), while 3 had mainstem bronchi masses and 3 had lung masses. Radiographically, the masses were lobulated, rounded, or polypoid in six patients. All underwent TBNA with a 21- or 22-gauge needle and endobronchial biopsy. The most frequent diagnosis was adenoid cystic carcinoma (4/11, 36%), followed by mucoepidermoid carcinoma (3/11, 27%), epithelial-myoepithelial carcinoma (2/11, 18%), oncocytoma (1/11, 9%), and hyalinizing clear cell carcinoma, salivary gland type (1/11, 9%). The surgical pathology specimens confirmed the diagnosis in all cases. CONCLUSIONS: To our knowledge, this is one of the largest cytomorphologic studies of primary salivary gland tumors of the tracheobronchial tree in the literature. Salivary gland tumors of the tracheobronchial tree are rare, and recognizing cytomorphologic changes that occur in salivary gland-type tumors is important for establishing a definitive diagnosis.

Barriers to the recognition of medullary thyroid carcinoma on FNA: Implications relevant to the new American Thyroid Association guidelines.

BACKGROUND: The 2016 American Thyroid Association guidelines recommend multiple endocrine neoplasia testing and evaluation for pheochromocytoma before thyroidectomy after a thyroid fine-needle aspiration biopsy (FNA) is positive for medullary thyroid carcinoma (MTC). In the current study, the authors examined the reasons why FNA was unable to definitively diagnose MTC preoperatively, with attention to morphologic patterns that can be misleading. METHODS: Cases of MTC diagnosed on thyroid surgical resection for which there was a prior FNA and slides available for review were included (28 cases). Clinicopathologic data were collected. Slides were reviewed for numerous features blinded to the original FNA interpretation. Morphologic features were compared between concordant cases (20 cases) ("positive for MTC" or "suspicious for MTC") and discordant cases (4 cases) (any other interpretation). Three cases of microscopic MTC (measuring <1 cm) were excluded from statistical analysis, as was 1 case of sampling error (benign thyroid tissue). RESULTS: Nine men and 19 women were diagnosed with MTC. Four patients ultimately were diagnosed with multiple endocrine neoplasia, and 1 had bilateral pheochromocytomas. At the time of surgical excision, the mean tumor size was 2.3 cm (range, 0.1-7.5 cm). Review of morphologic features demonstrated that the discrepant cases were significantly more likely to have limiting factors (air-drying artifact/excess blood), a cohesive pattern, or to lack plasmacytoid morphology. None of the discordant cases had pseudoinclusions or amyloid (finding was not statistically significant). CONCLUSIONS: The majority (86%) of thyroid FNAs from patients with MTC are concordant (positive/suspicious for MTC). Patterns of failure include sampling error and limited typical morphologic features, particularly a lack of plasmacytoid morphology and cellular dyshesion. A high level of suspicion for MTC is critical to ensure patients receive appropriate preoperative testing. Cancer Cytopathol 2018;126:397-405. © 2018 American Cancer Society.

Utility of GATA3, mammaglobin, GCDFP-15, and ER in the detection of intrathoracic metastatic breast carcinoma.

INTRODUCTION: Breast carcinoma (BC) metastatic to the intrathoracic cavity is difficult to diagnose due to low sensitivity of current immunohistochemical (IHC) stains. Mammaglobin, gross cystic disease fluid protein-15 (GCDFP-15), and estrogen receptor (ER) immunomarkers show variable results. GATA3 is a recently described marker for detecting urothelial and breast cancers. Our goal is to test the utility of GATA3 in cell blocks from thoracic cytology specimens. MATERIALS AND METHODS: We retrieved cases of BC that metastasized to the thoracic cavity from January 1, 2005 to September 30, 2013. IHC was performed on the cell blocks for the presence of GATA3, ER, GCDFP-15, and mammaglobin. Stains were scored quantitatively and qualitatively. RESULTS: Fifty cases of metastatic BC found in pleural effusions and endobronchial ultrasound-guided fine-needle aspirates were identified in 48 patients. Thirty-four cases had sufficient material for IHC (19 pleural effusions, 15 endobronchial ultrasound-guided fine-needle aspirates). GATA3 showed strong nuclear positivity in 31 of 34 cases (91.2%). ER (25 of 34, 73.5%), mammaglobin (23 of 34, 67.6%) and GCDFP-15 (11 of 34, 32.6%) were positive in fewer cases. GATA3 and ER were concordant in 26 of 34 cases (76.5%) (24 ER/GATA3-positive, 2 ER/GATA3-negative). Discordant results were found in 8 of 34 cases (23.5%). Of these, GATA3 was positive and ER was negative in 7 cases. GATA3 was negative and ER was positive in 1 case. CONCLUSIONS: GATA3 is more sensitive than ER, mammaglobin, or GCDFP-15 in detecting metastatic BC in cytologic specimens. GATA3 may be positive when ER is negative. In cytologic specimens with limited diagnostic material, GATA3 may be used as a first-line marker in a limited IHC panel to support metastatic BC.

Documentation of quality control and operator training at point-of-care testing: a College of American Pathologists Q-Probes study of 106 institutions.

CONTEXT: Operator training, quality control, and proper follow-up for out-of-range quality control (QC) events are crucial steps that must be adequately performed and documented to ensure excellent patient care and regulatory compliance. OBJECTIVE: To examine point-of-care testing (POCT) personnel training and QC documentation/compliance. DESIGN: Participants in a POCT documentation study of the College of American Pathologists Q-Probes program collected data retrospectively for glucose and urine dipstick testing regarding test operators, operator competency assessment, and QC documentation. Documentation was assessed for participant adherence to 4 quality indicators: (1) whether test operator training was up to date, (2) whether the test operator names were noted in the test records, (3) whether QC was performed, and (4) whether out-of-range QC events were followed up. Data were analyzed for associations with institutional demographic and practice variables. RESULTS: The institutional median number of POCT personnel was 648 for blood glucose and 76 for urine dipstick testing, with a median number of 105 948 glucose tests and 9113 urine tests performed. Ninety-four percent (3830 of 4074) of the test operators completed training or competency assessment within the prior 12 months, 96.8% (21 603 of 22 317) of the test records documented the operator, and 95.7% (19 632 of 20 514) of the expected QC events (per institutional regulations) were documented. Approximately 3% (659 of 20 514) of the QC events were outside the designated range (an average of 6 out-of-range QC events were identified per institution [n = 106]). Of the out-of-range QC events, 92.6% (610 of 659) had documentation of appropriate follow-up. Most laboratories (176 of 179; 98.3%) violated specimen requirements by storing POCT urine specimens for less than 24 hours. CONCLUSIONS: There was greater than 90% compliance for POCT documentation and nearly 96% of expected QC events were properly documented.

Laboratory compliance with the American Society of Clinical Oncology/College of American Pathologists human epidermal growth factor receptor 2 testing guidelines: a 3-year comparison of validation procedures.

CONTEXT: The American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) published guidelines in 2007 regarding testing accuracy, interpretation, and reporting of results for HER2 studies. A 2008 survey identified areas needing improved compliance. OBJECTIVE: To reassess laboratory response to those guidelines following a full accreditation cycle for an updated snapshot of laboratory practices regarding ASCO/CAP guidelines. DESIGN: In 2011, a survey was distributed with the HER2 immunohistochemistry (IHC) proficiency testing program identical to the 2008 survey. RESULTS: Of the 1150 surveys sent, 977 (85.0%) were returned, comparable to the original survey response in 2008 (757 of 907; 83.5%). New participants submitted 124 of 977 (12.7%) surveys. The median laboratory accession rate was 14,788 cases with 211 HER2 tests performed annually. Testing was validated with fluorescence in situ hybridization in 49.1% (443 of 902) of the laboratories; 26.3% (224 of 853) of the laboratories used another IHC assay. The median number of cases to validate fluorescence in situ hybridization (n = 40) and IHC (n = 27) was similar to those in 2008. Ninety-five percent concordance with fluorescence in situ hybridization was achieved by 76.5% (254 of 332) of laboratories for IHC(-) findings and 70.4% (233 of 331) for IHC(+) cases. Ninety-five percent concordance with another IHC assay was achieved by 71.1% (118 of 168) of the laboratories for negative findings and 69.6% (112 of 161) of the laboratories for positive cases. The proportion of laboratories interpreting HER2 IHC using ASCO/CAP guidelines (86.6% [798 of 921] in 2011; 83.8% [605 of 722] in 2008) remains similar. CONCLUSIONS: Although fixation time improvements have been made, assay validation deficiencies still exist. The results of this survey were shared within the CAP, including the Laboratory Accreditation Program and the ASCO/CAP panel revising the HER2 guidelines published in October 2013. The Laboratory Accreditation Program checklist was changed to strengthen HER2 validation practices.

Rapid on-site evaluation of endobronchial ultrasound-guided fine-needle aspirates: correlation of adequacy assessment and final diagnosis in patients with bronchogenic carcinoma.

INTRODUCTION: We review endobronchial ultrasound-guided fine-needle aspirate samples and investigate cases with discrepancies between rapid on-site evaluation (ROSE) and final diagnosis in patients with bronchogenic carcinoma. MATERIALS AND METHODS: Endobronchial ultrasound-guided fine-needle aspirates from 2009 to 2010 were studied. On-site adequacy assessments were compared with final diagnoses. Concordant diagnoses showed agreement between ROSE interpretation and final diagnosis. If the initial interpretation differed from the final diagnosis, the case was discordant. Slides from discordant aspirates were reviewed. Discordant results were categorized as sampling error or interpretive/screening error at ROSE. RESULTS: A total of 340 endobronchial ultrasound-guided procedures were performed in 335 patients (168 men, 167 women, median age 65 years). Diagnostic discrepancies between ROSE and final diagnoses occurred in 65 aspirates (11%) from 51 patients with carcinoma. Of the 65 discrepant cases, 52 (83%) were subsequently called positive for carcinoma. Rescreening of slides in 47 available cases with a final positive diagnosis showed insufficient tumor for diagnosis in 28 of 47 cases (60%). The remaining 19 of 47 cases (40%) were classified as interpretive/screening errors at ROSE. Most errors occurred in aspirates called atypical or atypical suspicious, which upon rescreening were considered diagnostic (16 aspirates, 84%). CONCLUSIONS: Initial and final diagnoses were concordant in 89% of aspirates from patients with carcinoma. All aspirates that were positive at ROSE were concordant. In discordant cases, all aspirates deemed "atypical suspicious for malignancy" and 86% of aspirates deemed "atypical cells" on ROSE had a final diagnosis of carcinoma. The majority of discordant cases with a positive final diagnosis were due to sampling (60%).

Utility and morphologic features of granulomas on rapid on-site evaluation of endobronchial ultrasonography-guided fine-needle aspiration.

INTRODUCTION: Endobronchial ultrasonography (EBUS)-guided fine-needle aspiration (FNA) is increasingly used to sample central lung lesions and mediastinal lymphadenopathy. We investigate the utility of EBUS-guided FNA and concomitant rapid on-site evaluation (ROSE) to diagnose granulomas, the morphologic characteristics of granulomas on ROSE, and how the diagnosis of granulomas changed the clinical impression. MATERIALS AND METHODS: All pathologic reports and associated clinical records of patients who had EBUS-guided FNA of the lungs or mediastinal lymph nodes that yielded granulomas were reviewed with at least a 1-year follow-up after EBUS-guided FNA. All ROSE slides were rereviewed to evaluate granulomas for quantity, necrosis, and cohesion. RESULTS: Over a 3-year period, 882 EBUS-guided FNAs were performed. One hundred and twelve patients (49% male, average age 50.8 years, range 16-83) had 161 EBUS-guided FNAs that yielded granulomas (18%). The etiologies of the granulomas were as follows: sarcoidosis (54%), infection (12%), malignancy (5%), inflammatory bowel disease-related lymphadenopathy (1%), and no specific clinical etiology (28%). Of the patients with EBUS-guided FNAs, 98 had ROSE performed (87.5%) and granulomas were seen in 70 of these patients (71%). Granulomas associated with sarcoidosis were mostly well-formed and non-necrotizing (90%). The results of the EBUS-guided FNA changed or redefined the clinical diagnosis in 79 patients (71%). CONCLUSIONS: EBUS-guided FNA with concurrent ROSE is a useful technique for the diagnosis of granulomas. The quality and quantity of granulomas detected during ROSE may suggest an etiology and help direct ancillary testing.

Endobronchial ultrasound-guided fine-needle aspiration cytology of bronchial low-grade mucoepidermoid carcinoma: Rapid on-site evaluation of cytopathologic findings.

Bronchial mucoepidermoid carcinoma (MEC) is rare, comprising about 0.2% of primary lung tumors. Endobronchial ultrasound (EBUS) guided fine-needle aspiration (FNA) cytology is an integral tool in the diagnosis and staging of malignant lung tumors. Rapid on-site evaluation (ROSE) has been proven useful as a guide for assessing the adequacy and accuracy of the FNA samples. Therefore, comprehensive knowledge of diagnostic cytomorphologic findings of MEC is critical for ROSE. We reported a 46-year-old woman with 6 weeks of cough productive of yellow sputum that did not improve on antibiotics. A chest CT demonstrated a well-circumscribed nodule in the right lower lobe bronchus that extended into the lung parenchyma. EBUS-guided FNA was performed to obtain diagnostic materials. The ROSE of cytology specimen revealed numerous tight clusters of cells with well-defined, but scant cytoplasm. These cells were relatively small and bland with high N/C ratio, resembling benign ductal cells. Neither cilia nor intranuclear inclusions were noted. Focal extracellular metachromatic mucinous materials were also noted. A preliminary diagnosis of "low-grade epithelial neoplasm, favor low grade MEC" was rendered. The definitive diagnosis was confirmed by both cytology and core biopsy. EBUS-guided FNA cytology can be a reliable method for the diagnosis of bronchial low grade MEC. The cyto-morphology of ROSE can indicate the diagnosis of low grade MEC and direct the appropriate follow-up triage of the specimen.

Immunoarchitectural patterns of germinal center antigens including LMO2 assist in the differential diagnosis of marginal zone lymphoma vs follicular lymphoma.

OBJECTIVES: To examine the immunoarchitectural patterns of the germinal center (GC)-associated markers CD10, BCL6, and LMO2 and their utility in the differential diagnosis of marginal zone lymphoma (MZL) vs follicular lymphoma (FL). METHODS: Forty-two cases of MZL involving lymph nodes and 88 cases of FL were examined. RESULTS: Interfollicular staining for GC markers was uncommon in MZL but common in FL, including BCL2-positive and BCL2-negative cases. Two atypical patterns of intrafollicular GC staining were identified that were more common in MZL than in FL. CONCLUSIONS: Staining for LMO2 in addition to CD10 and BCL6 facilitates the detection of a GC phenotype in FL. Lymph nodes involved by MZL frequently show characteristic alterations of GC immunoarchitecture. Recognizing these altered patterns assists in the distinction between MZL and FL.