Simultaneously Detection of 50 Mutations at 20 Sites in the BRAF and RAS Genes by Multiplexed Single-Nucleotide Primer Extension Assay Using Fine-Needle Aspirates of Thyroid Nodules.

Fine-needle aspiration (FNA) is commonly used for primary evaluation of thyroid nodules. Twenty to 30 percent of thyroid nodules remain indeterminate after FNA evaluation. Studies show the BRAF p.V600E to be highly specific for papillary thyroid carcinoma (PTC), while RAS mutations carry up to 88 percent positive predictive value for malignancy. We developed a two-tube multiplexed PCR assay followed by single-nucleotide primer extension assay for simultaneous detection of 50 mutations in the BRAF (p.V600E, p.K601E/Q) and RAS genes (KRAS and NRAS codons 12, 13, 19, 61 and HRAS 61) using FNA smears of thyroid nodules. Forty-two FNAs and 27 paired formalin-fixed, paraffin-embedded (FFPE) tissues were tested. All BRAF p.V600E-positive FNA smears (five) carried a final diagnosis of PTC on resection. RAS mutations were found in benign as well as malignant lesions. Ninety-two percent concordance was observed between FNA and FFPE tissues. In conclusion, our assay is sensitive and reliable for simultaneous detection of multiple BRAF/RAS mutations in FNA smears of thyroid nodules.

Efficacy and safety of EMR to completely remove Barrett's esophagus: experience in 41 patients.

BACKGROUND: EMR is typically used to remove focal abnormalities of the esophageal mucosa. However, larger areas of Barrett's esophagus (BE) can be resected through side-by-side resections. OBJECTIVE: To assess the efficacy and safety of EMR to completely remove BE. DESIGN: Retrospective, single-center study. SETTING: University of Iowa Hospitals and Clinics. PATIENTS: Between January 2006 and December 2010, 46 patients underwent EMR for complete removal of BE. Three were lost to follow-up, one died of unrelated causes before completion, and one was still undergoing EMR treatment at the conclusion of the study. The remaining 41 patients were included for analysis. The worst histologic grade was low-grade dysplasia in 4 patients, high-grade dysplasia without cancer in 26 patients, and high-grade dysplasia with superficial adenocarcinoma in 11 patients. BE was circumferential in 65.9% of cases, and the mean (± SD) length was 3.3 ± 2.3 cm. INTERVENTION: EMR was performed by using a cap (n = 4), a multiband ligator device (n = 31), or both (n = 6), with a mean (± SD) of 2.4 ± 1.2 sessions per patient. MAIN OUTCOME MEASUREMENTS: Remission rates and complications. RESULTS: Remission of high-grade dysplasia and cancer, all dysplasia, and all BE was achieved in 94.6%, 85.4%, and 78.0%, respectively. Complications included minor bleeding (31.7%), perforations (4.9%), and strictures (43.9%). All complications were managed conservatively. LIMITATIONS: Retrospective design. CONCLUSION: Complete removal of BE with EMR is effective but associated with a high complication rate, which is mainly related to stricture formation. This needs to be considered when choosing between available treatment modalities.

Human Papillomavirus (HPV)-Related Carcinoma of the Oropharynx with Squamous Cell and Adenoid Cystic Carcinoma-Like Features: Report of a Case.

We describe the heretofore unreported case of an HPV-related carcinoma of the palatine tonsil with distinct areas of squamous cell- and adenoid cystic carcinoma-like differentiation in a 54-year old patient. The morphological, immunophenotypic and molecular findings of the tumor are illustrated. We discuss the parallels between the tumor and HPV-related multiphenotypic sinonasal carcinoma (HMSC) which is well-known to exhibit adenoid cystic carcinoma-like features. A review of the literature of high-risk HPV-associated non-squamous carcinomas of the oropharynx is presented.

Catalase Modulates the Radio-Sensitization of Pancreatic Cancer Cells by Pharmacological Ascorbate.

Pancreatic cancer cells (PDACs) are more susceptible to an oxidative insult than normal cells, resulting in greater cytotoxicity upon exposure to agents that increase pro-oxidant levels. Pharmacological ascorbate (P-AscH-), i.e., large amounts given intravenously (IV), generates significant fluxes of hydrogen peroxide (H2O2), resulting in the killing of PDACs but not normal cells. Recent studies have demonstrated that P-AscH- radio-sensitizes PDAC but is a radioprotector to normal cells and tissues. Several mechanisms have been hypothesized to explain the dual roles of P-AscH- in radiation-induced toxicity including the activation of nuclear factor-erythroid 2-related factor 2 (Nrf2), RelB, as well as changes in bioenergetic profiles. We have found that P-AscH- in conjunction with radiation increases Nrf2 in both cancer cells and normal cells. Although P-AscH- with radiation decreases RelB in cancer cells vs. normal cells, the knockout of RelB does not radio-sensitize PDACs. Cellular bioenergetic profiles demonstrate that P-AscH- with radiation increases the ATP demand/production rate (glycolytic and oxidative phosphorylation) in both PDACs and normal cells. Knocking out catalase results in P-AscH- radio-sensitization in PDACs. In a phase I trial where P-AscH- was included as an adjuvant to the standard of care, short-term survivors had higher catalase levels in tumor tissue, compared to long-term survivors. These data suggest that P-AscH- radio-sensitizes PDACs through increased peroxide flux. Catalase levels could be a possible indicator for how tumors will respond to P-AscH-.

Expansion of a cell population expressing stem cell markers in parathyroid glands from patients with hyperparathyroidism.

OBJECTIVE: We sought to examine abnormal parathyroid glands for the presence of stem cells. SUMMARY BACKGROUND DATA: Cancer stem cells have been identified in cancers from a variety of tissues as a CD44/CD24 cell population. We hypothesize that stem cells (SC) may also be involved in the pathogenesis of benign clonal expansion characteristic of hyperparathyroidism (HPT). METHODS: Under institutional review board approval, parathyroid tissue was obtained from 20 patients with HPT and analyzed by fluorescence-activated cell sorting (FACS) for the CD44/CD24 cell population. Immunohistochemistry (IHC) with CD44 antibody was correlated with FACS results. RESULTS: Parathyroid tissue was obtained for FACS analysis from 25 enlarged parathyroid glands from 20 patients, 17 with primary HPT, and 3 with secondary HPT. The average percent of SC defined as CD44/CD24 population was 10.93% for enlarged parathyroid glands. IHC using CD44 antibody was performed on 27 abnormal parathyroid glands and 7 normal parathyroid gland biopsies from the same patients. Although IHC was not as sensitive as FACS, comparison of IHC and FACS results for 24 abnormal glands gave a correlation coefficient of 0.52, which was statistically significant (P = 0.01, Spearman rank). By IHC, 13 of 27 abnormal glands stained 1+ to 3+ (average, 0.93) compared with no CD44 staining in normal glands, which was statistically different (mean IHC of 0 vs. 0.93, P = 0.03, Wilcoxon). CONCLUSIONS: These novel findings demonstrate expansion of a resident cell population that expresses SC markers in abnormal parathyroid glands from patients with HPT. Our results suggest that clonal expansion of a resident SC population occurs in the pathogenesis not only of cancer, but also in benign parathyroid tumors occurring in HPT.

EUS-guided Trucut biopsies may enable the diagnosis of lymphoepithelial cysts of the pancreas. Report of two cases.

CONTEXT: Lymphoepithelial cysts are rare benign lesions of the pancreas. Endoscopic ultrasound guided fine needle aspiration (FNA) has reportedly been helpful in reaching a diagnosis of lymphoepithelial cyst. In cases where it may be difficult to obtain a diagnosis using FNA a histological sample may be obtained using EUS guided Trucut biopsy. CASE REPORTS: We describe two cases of lymphoepithelial cysts where a diagnosis of lymphoepithelial cyst was made using EUS guided Trucut biopsy. CONCLUSION: In some case, lymphoepithelial cysts may present a diagnostic dilemma. EUS with FNA and Trucut biopsy may enable the clinician to make a definitive diagnosis.

[Long-term, tumor-free survival after radiotherapy combining hepatectomy-Whipple en bloc and orthotopic liver transplantation for early-stage hilar cholangiocarcinoma].

OBJECTIVE: To report the experience in surveillance and early detection of cholangiocarcinoma (CC) and in using en bloc total hepatectomy-pancreaticoduodenectomy-orthotopic liver transplantation (OLT-Whipple) to achieve complete eradication of early-stage CC complicating primary sclerosing cholangitis (PSC). METHODS: Asymptomatic PSC patients underwent surveillance using endoscopic ultrasound and endoscopic retrograde cholangiopancreatography (ERCP) with multilevel brushings for cytological evaluation. Patients diagnosed with CC were treated with combined extra-beam radiotherapy, lesion-focused brachytherapy, and OLT-Whipple. RESULTS: Between January 1988 and February 2001, 42 of 119 PSC patients were followed according to the surveillance protocol. CC was detected in 8 patients, 6 of whom underwent OLT-Whipple. Of those 6 patients, 4 had stage I CC, and 2 had stage II CC. All 6 OLT-Whipple patients received combined external-beam and brachytherapy radiotherapy. The median time from diagnosis to OLT-Whipple was 144 days. One patient died 55 months post-transplant of an unrelated cause, without tumor recurrence. The other 5 were well without recurrence at 79, 82, 108, 128, 129 and 145 months. CONCLUSIONS: For patients with PSC, ERCP surveillance cytology and intralumenal endoscopic ultrasound examination allow for early detection of CC. Broad and lesion-focused radiotherapy combined with OLT-Whipple to remove the biliary epithelium en bloc offers promising long-term, tumor-free survival. All patients tolerated this extensive surgery well with good quality of life following surgery and recovery. These findings support consideration of the complete excision of an intact biliary tree via OLT-Whipple in patients with early-stage hilar CC complicating PSC.

Long-term, tumor-free survival after radiotherapy combining hepatectomy-Whipple en bloc and orthotopic liver transplantation for early-stage hilar cholangiocarcinoma.

This retrospective study reviews our experience in surveillance and early detection of cholangiocarcinoma (CC) and in using en bloc total hepatectomy-pancreaticoduodenectomy-orthotopic liver transplantation (OLT-Whipple) to achieve complete eradication of early-stage CC complicating primary sclerosing cholangitis (PSC). Asymptomatic PSC patients underwent surveillance using endoscopic ultrasound and endoscopic retrograde cholangiopancreatography (ERCP) with multilevel brushings for cytological evaluation. Patients diagnosed with CC were treated with combined extra-beam radiotherapy, lesion-focused brachytherapy, and OLT-Whipple. Between 1988 and 2001, 42 of 119 PSC patients were followed according to the surveillance protocol. CC was detected in 8 patients, 6 of whom underwent OLT-Whipple. Of those 6 patients, 4 had stage I CC, and 2 had stage II CC. All 6 OLT-Whipple patients received combined external-beam and brachytherapy radiotherapy. The median time from diagnosis to OLT-Whipple was 144 days. One patient died 55 months post-transplant of an unrelated cause, without tumor recurrence. The other 5 are well without recurrence at 5.7, 7.0, 8.7, 8.8, and 10.1 years. In conclusion, for patients with PSC, ERCP surveillance cytology and intralumenal endoscopic ultrasound examination allow for early detection of CC. Broad and lesion-focused radiotherapy combined with OLT-Whipple to remove the biliary epithelium en bloc offers promising long-term, tumor-free survival. All patients tolerated this extensive surgery well with good quality of life following surgery and recovery. These findings support consideration of the complete excision of an intact biliary tree via OLT-Whipple in patients with early-stage hilar CC complicating PSC.

The value of touch imprint cytology in EUS-guided Trucut biopsy.

BACKGROUND: EUS-guided Trucut biopsy (TCB) enables the acquisition of tissue cores for histologic assessment. Touch imprint cytology (TIC) can be performed at the time of a biopsy to assess the adequacy of the sample; however, limited information is available on the diagnostic value of TIC of these specimens. OBJECTIVE: To investigate the diagnostic accuracy of TIC compared with a TCB. PATIENTS AND DESIGN: Consecutive EUS-guided TCB and TIC (n = 109) were retrospectively and independently reviewed by a surgical pathologist (for the TCB) and a cytopathologist (for TIC) blinded to the final diagnoses. SETTING: University of Iowa Hospitals and Clinics, Iowa. MAIN OUTCOME MEASUREMENTS: Diagnostic accuracy of a TCB, TIC, and combined TCB + TIC. RESULTS: The diagnostic accuracy of a TCB was 92.7% (95% CI, 83.1%-97.3%), TIC was 82.6% (95% CI, 74.3%-88.6%), and TCB + TIC was 95.4% (95% CI, of 89.4%-98.3%). The diagnostic accuracy of a TCB alone was superior to TIC alone (P = .038); a TCB was diagnostic in 14 cases that were nondiagnostic by TIC. The addition of TIC allowed for the identification of 3 malignancies (2.8%) that were not identified on TCB alone. In 22 cases, TIC was considered diagnostic, but a TCB provided additional specific diagnostic information. LIMITATIONS: Retrospective study and relatively low numbers. CONCLUSIONS: TIC is a valuable tool for use in a EUS-guided TCB; TIC is independently diagnostically accurate, which allows for confidence in a rapid preliminary diagnosis, and it provides additional diagnostic value when combined with TCB.

Simultaneous detection of single-nucleotide variant, deletion/insertion, and fusion in lung and thyroid carcinoma using cytology specimen and an RNA-based next-generation sequencing assay.

BACKGROUND: Molecular testing for epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) and ROS proto-oncogene 1, receptor tyrosine kinase (ROS1) fusion is routinely performed in patients with stage IV lung adenocarcinoma to assess their eligibility for targeted therapy. Fine-needle aspiration (FNA)-derived material frequently is the only pathologic material available. The identification of genomic aberrations in thyroid nodules from FNA smears may help stratify cancer risk and spare patients from a second surgery. In the current study, the authors tested nucleic acid extracted from the cytology smears of lung and thyroid carcinomas for simultaneous detection of single-nucleotide variant, insertion/deletion, and gene fusion using an RNA-based next-generation sequencing assay. METHODS: A total of 27 cases (17 lung and 10 thyroid carcinomas, the majority of which had known variants) were tested. Areas of interest were scrapped from stained smears using a scalpel. Total nucleic acid was extracted. Gene fusion and mutational analysis was performed using the Comprehensive Thyroid and Lung FusionPlex Assay. Data were analyzed using the analysis pipeline provided by the vendor. Eleven cases with available formalin-fixed, paraffin-embedded (FFPE) tissue were tested in parallel. RESULTS: Gene fusions were detected in 6 cases; common single-nucleotide variants in EGFR, RAS, and BRAF in 14 cases; and in-frame deletions within EGFR in 3 cases. A concordance rate of 100% was observed between FNA and FFPE tissue. CONCLUSIONS: Cytology preparations can be a reliable source for the detection of both DNA and RNA aberrations. The ability to simultaneously detect multiple types of genomic variants is crucial for patients with advanced cancer and maximizes the usefulness of cytology specimens. Cancer Cytopathol 2018;126:158-69. © 2018 American Cancer Society.

Using Focused Laboratory Management and Quality Improvement Projects to Enhance Resident Training and Foster Scholarship.

Training in patient safety, quality, and management is widely recognized as an important element of graduate medical education. These concepts have been intertwined in pathology graduate medical education for many years, although training programs face challenges in creating explicit learning opportunities in these fields. Tangibly involving pathology residents in management and quality improvement projects has the potential to teach and reinforce key concepts and further fulfill Accreditation Council for Graduate Medical Education goals for pursuing projects related to patient safety and quality improvement. In this report, we present our experience at a pathology residency program (University of Iowa) in engaging pathology residents in projects related to practical issues of laboratory management, process improvement, and informatics. In this program, at least 1 management/quality improvement project, typically performed during a clinical chemistry/management rotation, was required and ideally resulted in a journal publication. The residency program also initiated a monthly management/informatics series for pathology externs, residents, and fellows that covers a wide range of topics. Since 2010, all pathology residents at the University of Iowa have completed at least 1 management/quality improvement project. Many of the projects involved aspects of laboratory test utilization, with some projects focused on other areas such as human resources, informatics, or process improvement. Since 2012, 31 peer-reviewed journal articles involving effort from 26 residents have been published. Multiple projects resulted in changes in ongoing practice, particularly within the hospital electronic health record. Focused management/quality improvement projects involving pathology residents can result in both meaningful quality improvement and scholarly output.

Double slide viewing as a cytology quality improvement initiative.

Few studies have measured the effect of pre-sign out double viewing of cytology cases as a means to decrease error. Three Agency for Healthcare Research and Quality-funded project sites performed pre-sign out double viewing of 431 pulmonary cytology cases. Two-step or more differences in diagnosis were arbitrated as interpretive errors, and the effect of double viewing was measured by comparing the frequency of cytologic-histologic correlation-detected errors in the previous 2 years with the double-viewing period. The number of interpretive errors detected by double viewing for the 3 institutions was 2.7%, 0% and 1.9%, respectively. Double viewing did not lower the frequency of cytologic-histologic correlation false-negative errors. We conclude that double viewing detects errors in up to 1 of every 37 cases and that biases in the double-viewing process limit error detection.

Differentiating Small Cell Carcinoma From Squamous Cell Carcinoma in Cytologic Specimens: A Head-to-Head Comparison of p40 and p63 Using Cell Block Immunocytochemistry.

Distinguishing small cell carcinoma (SCC) from poorly differentiated squamous cell carcinoma (SCCA) can be challenging on fine needle aspiration (FNA). p63 immunocytochemistry is frequently used to support the presence of squamous differentiation. However, issues relating to specificity have arisen, and the common pan-p63 clone (4A4) has been challenged by antibodies specific for the ΔN isoform of p63 (p40), which show superior specificity for SCCA. A total of 50 SCC and 25 SCCA FNA cell blocks were stained with antibodies for p40, p63, K903, and CK5/6. For each marker, both extent and intensity of immunoreactivity were recorded, blinded to diagnosis, and H-scores (extent×intensity) were calculated. A total of 4%, 34%, 4%, and 2% of SCCs were positive for p40, p63, K903, and CK5/6, respectively. The rate of p63 positivity was significantly higher than for the other markers. Median H-scores in p63-positive SCCs (40) and SCCAs (270) were significantly different (P<0.0001), though, and 94% of p63-positive SCCs showed an H-score <150. SCCs frequently express p63 (34%). In the SCCA versus SCC differential diagnosis, p40 and K903 are each marginally more sensitive (92%) than p63 (88%), whereas CK5/6 boasts the greatest specificity (98%). p63's poor specificity (66%) can be improved to 94%, if an H-score ≥150 is used as the cutoff for a positive result.

Correlating thyroid cytology and histopathology: Implications for molecular testing.

BACKGROUND: A gene expression classifier (GEC) has been advocated in management of some indeterminate nodules without surgery. We assessed the potential negative predictive value (NPV) of the GEC at our academic center. METHODS: Retrospectively, all cytologically indeterminate fine-needle aspirates (FNAs) diagnosed by University of Iowa cytopathologists over a 3-year period were identified. Histopathologic findings were recorded. Using published sensitivity and specificity, NPVs were calculated. RESULTS: Of 178 nodules (17, 135, and 26 in classes III, IV, and V, respectively), 71 (40%) were malignant. Prevalence of malignancy was 41%, 29%, and 96% for classes III, IV, and V, respectively. Using sensitivities and specificities for the GEC, NPVs were 91% for the cohort: 88%, 92%, and 26% for classes III, IV, and V, respectively. CONCLUSION: Molecular testing should be associated with an NPV no lower than that from clinical criteria alone. With the prevalences reported, GEC use may result in more missed cancer diagnoses. © 2016 Wiley Periodicals, Inc. Head Neck 38: 1104-1106, 2016.

Multiplatform comparison of molecular oncology tests performed on cytology specimens and formalin-fixed, paraffin-embedded tissue.

BACKGROUND: Molecular oncology testing is important for patient management, and requests for the molecular analysis of cytology specimens are increasingly being made. Formalin-fixed, paraffin-embedded (FFPE) cell blocks of such specimens have been routinely used for molecular diagnosis. However, the inability to assess cellularity before cell block preparation is a pitfall of their use. In this study, various cytologic preparations were tested with several molecular test platforms, and the results were compared with paired FFPE tissue. METHODS: Seventy-seven cytology cases, including fine-needle aspiration smears, touch preparations, and SurePath thin-layer preparations, were selected from the archives. Areas of interest were removed from the slide with a matrix capture solution. DNA extracted from the cells was evaluated by mutation analysis for BRAF, epidermal growth factor receptor (EGFR), RAS, and a 50-gene panel with various testing platforms (single-nucleotide primer extension assay, Sanger sequencing, and next-generation sequencing). Thirty-eight tumors with available FFPE tissue were tested in parallel. RESULTS: The average DNA concentration was 299 ng/µL for the cytology specimens and 171 ng/µg for the paired FFPE tissue. Point mutations and large deletions were detected in BRAF, KRAS, NRAS, HRAS, and EGFR genes. In comparison with FFPE tissue, 5- to 8-fold less input DNA was needed when cytology preparations were used. The concordance between cytology specimens and FFPE tissue was 100%. CONCLUSIONS: Cytologic preparations were found to be a reliable source for molecular oncology testing. DNA derived from cytology specimens performed well on multiple platforms, and 100% concordance was observed between cytology specimens and FFPE tissue.

A cytomorphometric analysis of pulmonary and mediastinal granulomas: differentiating histoplasmosis from sarcoidosis by fine-needle aspiration.

BACKGROUND: Histoplasmosis and sarcoidosis are commonly included in the differential diagnosis of mass lesions at lung and mediastinal sites. Once cancer is excluded on aspiration biopsy, further classification is essential for proper treatment. METHODS: A search identified patients with histoplasmosis and sarcoidosis for whom the diagnosis was made by clinicopathologic correlation. Cases were reviewed for various cytologic parameters along with patient demographic, clinical, and laboratory data. RESULTS: Fifty-eight cases of histoplasmosis and 44 cases of sarcoidosis were reviewed. Thirty-seven of 58 (64%) Histoplasma cases exhibited abundant, bland necrosis, and 76% of cases contained <2 angular and ragged granulomas per slide. Yeasts were identified in 36 of 37 (97%) specimens with necrosis and in 44 of 58 (76%) cases overall. These cases had an acute (14%) and/or chronic (67%) inflammatory component and uncommonly had a giant cell infiltrate (12%). Sarcoid granulomas were round with crisp, sharp borders: 80% of these granulomas contained >3 granulomas per slide, and 32% contained >10 granulomas per slide. All sarcoid granulomas had a chronic inflammatory background without acute inflammatory cells, and 50% contained giant cell infiltrates. CONCLUSIONS: Differentiation between histoplasmosis and sarcoidosis is possible in the majority of cases. Histoplasmosis usually exhibits few angular, ragged granulomas (<2 granulomas per slide) in a background of bland necrosis. Yeasts are identified on special stains performed in aspirate smears. Sarcoidosis typically contains many more granulomas (often >10 per slide) than histoplasmosis and has a rounded morphology with crisp and sharp borders. Typically, there is no necrosis or acute inflammation, and giant cell infiltrates are frequent.

Pathology Milestones: Assessing Clinical Competency by Committee.

All Accreditation Council for Graduate Medical Education accredited pathology residency training programs are now required to evaluate residents using the new Pathology Milestones assessment tool. Similar to implementation of the 6 Accreditation Council for Graduate Medical Education competencies a decade ago, there have been challenges in implementation of the new milestones for many residency programs. The pathology department at the University of Iowa has implemented a process that divides the labor of the task in rating residents while also maintaining consistency in the process. The process is described in detail, and some initial trends in milestone evaluation are described and discussed. Our experience indicates that thoughtful implementation of the Pathology Milestones can provide programs with valuable information that can inform curricular changes.

Bile duct brushings in a pig model: examination of intraobserver variability and variability in specimen quality obtained in sequential animals and between two different brushes.

PURPOSE: In patients with bile duct malignancy, bile duct brushing is plagued by a low yield diagnosing underlying malignancy. There are few data explaining why this is so. This porcine model was designed to examine three variables: 1) examination of inter-observer variability, 2) variability in specimen quality obtained in sequential animals, and 3) variability between two different brushes (one designed for colon with large bristles, one for duodenum with short bristles). METHODS: En bloc resection of liver, bile ducts, duodenum, and pancreas was performed on three 6-mo-old crossbred pigs at the time of commercial slaughter. In each pig, one common hepatic duct and one common bile duct brushing, all performed by the same investigator, were done. Ten identical vigorous passes were done with each brush (long bristle or short bristle) on virgin epithelium. Specimens were graded for cellularity by three cytopathologists who were blinded to the site or brush size. Interobserver variability as well as variability among sequential animals and between the two different brushes was compared. RESULTS: Interobserver variability among the three cytologists was almost nil. Cellularity obtained using the short brush alone varied from unsatisfactory to high. Cellularity obtained using the long brush alone varied from unsatisfactory to moderate. Variability of cells obtained from one pig to the next ranged from unsatisfactory to high. CONCLUSIONS: 1) While interobserver variability was very low, variability in cellularity obtained from one pig to the next, and from one brush to the next, was very high. This sampling variability may partially explain the low yield in malignant cells in human malignant biliary brushing. Multiple brushings in one patient may alleviate part of this problem. 2) There was no advantage to either brush type (large bristle or small bristle).

Fine-needle aspiration of histoplasmosis in the era of endoscopic ultrasound and endobronchial ultrasound: cytomorphologic features and correlation with clinical laboratory testing.

BACKGROUND: Histoplasmosis has a textbook cytologic description with numerous intracellular organisms that are readily apparent on routine stains. This is based on series and reports describing histoplasmosis in immunosuppressed patients with disseminated disease. With the advent of ultrasound-guided (US) fine-needle aspiration (FNA) techniques, a marked increase in the cytologic diagnosis of histoplasmosis in immunocompetent patients is noted. METHODS: A search identified all cytology cases diagnosed with Histoplasma within the past 10 years. Cases were reviewed, along with patient demographic, clinical, and laboratory data. RESULTS: A total of 40 FNA cases of histoplasmosis were identified. Patients ranged in age from 15 years to 86 years. There were 23 female patients and 17 male patients; 37 were immunocompetent and 3 were immunosuppressed. Sixteen patients were being staged for primary tumors of other sites; others presented with primary pulmonary symptoms or histoplasmosis was noted incidentally. Specimens were composed of bland acellular necrosis, most commonly with granulomas (77.5%); only rare intracellular organisms were present on routine stains, and variable extracellular organisms were noted on Grocott methenamine silver stain (GMS) stain. GMS stain on direct smears was found to be more sensitive than cell block. Laboratory studies for urine antigen, yeast, and mycelial antibody (by compliment fixation), serum antibody (by immunodiffusion), and culture were positive in 11.8%, 59.1%, 4.5%, 47.6%, and 3.4% of cases, respectively. CONCLUSIONS: In an endemic region, histoplasmosis presents more commonly in immunocompetent patients as localized fibrocaseous disease on FNA and is often identified by high-resolution imaging. FNA is increasingly used in the diagnosis because of endoscopic ultrasound and endobronchial ultrasound. GMS stain on direct smears is more sensitive than cell block. In general, laboratory tests have low sensitivity in this patient population.

Accuracy of fine-needle aspiration and imaging in the preoperative workup of salivary gland mass lesions treated surgically.

OBJECTIVES/HYPOTHESIS: Fine-needle aspiration (FNA) biopsy and imaging are commonly used in the preoperative assessment of salivary gland mass lesions. The goal of this retrospective study was to clarify the role of FNA and imaging in the workup of salivary gland masses. STUDY DESIGN: Retrospective cohort study. METHODS: A computer search identified patients with an FNA of a salivary gland lesion with subsequent excision during a 10-year study period. Chart review of study patients was performed, and information on site of lesion, age, gender, radiologic diagnosis, pain in the tumor area, and facial paralysis was recorded and analyzed. RESULTS: There were 543 patients who had an FNA and subsequent histopathology. The majority of the tumors were in the parotid gland (n = 492, 90.9%), followed by submandibular gland (n = 45, 8.3%). The incidence of malignancy across all sites was 29.7%. The mean patient age was 54.1 years, and 54.1% were female. The sensitivity and specificity of FNA were 85.7% and 99.5%, respectively. Positive predictive value (PPV) was 98.6%, and negative predictive value (NPV) was 94.3%. A total of 464 patients had available radiologic studies. For the radiological diagnoses, sensitivity was 81.8% and specificity was 67.3%, whereas PPV and NPV were 52.7% and 89.3%, respectively. CONCLUSIONS: FNA is a reliable method of preoperatively assessing both benign and malignant salivary gland lesions. Preoperative imaging has a lower sensitivity and specificity than FNA in differentiating malignant from benign tumors. Older age, pain, and facial paralysis are clinical features independently associated with malignancy.