Promoting improved utilization of laboratory testing through changes in an electronic medical record: experience at an academic medical center.

This case study over time describes five years of experience with interventions to improve laboratory test utilization at an academic medical center. The high-frequency laboratory tests showing the biggest declines in order volume post intervention were serum albumin (36%) and erythrocyte sedimentation rate (17%). Introduction of restrictions for 170 high-cost send-out tests resulted in a 23% decline in order volume. Targeted interventions reduced mis-orders involving several "look-alike" tests: 1,25-dihydroxyvitamin D, 25-hydroxyvitamin D; manganese, magnesium; beta-2-glycoprotein, beta-2-microglobulin. Lastly, targeted alerts reduced duplicate orders of germline genetic testing and orders of hepatitis B surface antigen within 2 weeks of hepatitis B vaccination.

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.

Data on the utilization of paraneoplastic syndrome autoantibody testing at an academic medical center.

Paraneoplastic syndromes are rare conditions associated with characteristic autoantibodies produced by malignancy, although similar autoantibodies and clinical presentations may occur in the absence of any neoplasm. Testing for paraneoplastic syndromes often involves panels of autoantibody assays. While autoantibody testing may reveal or confirm actionable clinical diagnoses, inappropriate utilization of testing may be low yield and further lead to false positives that may confuse the clinical picture. There is thus opportunity to improve patient care by analyzing patterns of paraneoplastic autoantibody test utilization. The data in this article provides results from detailed retrospective review of patients tested by 7 autoantibody tests or test panels offered by two large reference laboratories in the United States. The data include 1,446 tests performed on 1,338 unique patients at an academic medical center. For all results, detailed chart review revealed main category of presenting symptoms, patient location at time of testing (either inpatient or outpatient), sex, age, whether cancer was present at the time of testing or later detected, and the specific results of the testing. The data are summarized by category of testing and specific autoantibodies.

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.

Cribriform adenocarcinoma of minor salivary gland: a report of two cases with an emphasis on cytology.

Cribriform adenocarcinoma of minor salivary gland (CAMSG) is a recently characterized low grade salivary gland malignancy that most commonly presents as a mass in the base of the tongue, frequently with regional lymph node metastasis. Given its relative rarity and overlapping cytomorphology, CAMSG may be confused with polymorphous low grade adenocarcinoma (PLGA) in minor salivary gland sites and papillary thyroid carcinoma (PTC) in cervical metastasis, in both fine-needle aspiration and excisional specimens. As there are no cytology reports in the literature, we present two new cases of CAMSG and describe the aspiration cytology of the tumor taken from bench top aspirates, compare it with the histomorphology, and discuss the features that may help one avoid misdiagnosis of PTC in the setting of cervical lymph node metastasis. We found that like PTC, aspirates of CAMSG contain polymorphic fragments of epithelial cells arranged in monolayer sheets, papillary fronds and tips, and occasional cribriform configurations, and metachromatic stromal fragments, which may be misinterpreted as colloid. A background of myxoid/mucoid material also reminiscent of colloid was prominent. Differentiation from PLGA is more difficult based strictly on cytology. A review of the most current literature in relation to the molecular and immunohistochemical profiles, therapeutic options, and prognosis is also presented. It is critical for pathologists and clinicians to be aware of this tumor when presented with patients having a cervical lymph node mass in the absence of a primary tumor.

Immunohistochemistry for the novel markers glypican 3, PAX8, and p40 (ΔNp63) in squamous cell and urothelial carcinoma.

OBJECTIVES: To examine squamous cell carcinomas (SCCs) from diverse anatomic sites and invasive urothelial carcinomas (UCs) for expression of the oncofetal antigen glypican 3 (GPC3), the paired box transcription factor PAX8, and the ΔN isoform of p63 (p40). METHODS: Immunohistochemistry for GPC3, PAX8, and p40 was performed on whole sections of 107 SCCs from 11 anatomic sites and 49 UCs; evaluation included extent and intensity of staining. RESULTS: GPC3 was detected in 20% of SCCs and 12% of UCs and PAX8 in 3% of SCCs, limited to the uterine cervix, and 10% of UCs. p40 Was found in 99% of SCCs and 96% of UCs. CONCLUSIONS: GPC3 expression is frequent in SCC/UC, awareness of which should guard against an incorrect diagnosis of hepatocellular carcinoma, while PAX8, limited in distribution, may have some use in suggesting a cervical or urothelial tract origin in a metastatic squamotransitional carcinoma of unknown primary. There is no drop-off in sensitivity for the diagnoses of SCC or UC with ΔNp63-specific immunohistochemistry, and if this performance can be extended to other applications, p40 may supplant the dominant "pan-p63" antibody clone.

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.

Nuclear MTA1 overexpression is associated with aggressive prostate cancer, recurrence and metastasis in African Americans.

Metastasis-associated protein 1 (MTA1), a negative epigenetic modifier, plays a critical role in prostate cancer (PCa) progression. We hypothesized that MTA1 overexpression in primary tumor tissues can predict PCa aggressiveness and metastasis. Immunohistochemical staining of MTA1 was done on archival PCa specimens from University of Mississippi Medical Center and University of Iowa. We found that nuclear MTA1 overexpression was positively correlated with the severity of disease progression reaching its highest levels in metastatic PCa. Nuclear MTA1 overexpression was significantly associated with Gleason > 7 tumors in African Americans but not in Caucasians. It was also a predictor of recurrent disease. We concluded that MTA1 nuclear overexpression may be a prognostic indicator and a future therapeutic target for aggressive PCa in African American men. Our findings may be useful for categorizing African American patients with a higher probability of recurrent disease and metastasis from those who are likely to remain metastasis-free.

Ascending aortic graft thrombosis and diffuse embolization from early endoluminal Aspergillus infection.

We present a 43-year-old man who underwent emergent replacement of the ascending aorta for type A dissection and hemiarch reconstruction with a 28-mm prosthetic graft. Dramatic neurologic symptoms, renal failure, and bowel ischemia developed on postoperative day 5. A computed tomography scan showed a large floating thrombus in the ascending aortic graft and massive peripheral embolization throughout the body.