Highly sensitive noninvasive early lung cancer detection using DNA methylation topology in sputum-derived epithelial cells.

Objective: Sputum is a source of exfoliated respiratory epithelial cells transformed early in lung carcinogenesis. Malignant cells are hypomethylated and contain less genomic 5-methylcytosine (5mC). Validating a test that recognizes and quantifies aberrantly hypomethylated cells in sputum, we assessed its potential as a screening tool for detecting early-stage non-small cell lung cancer. Methods: Cells extracted from sputum were immunofluorescence labeled with an anti-5-methylcytosine antibody and counterstained with 4',6-diamidino-2-phenylindole (DAPI) delineating global nuclear DNA (gDNA). Via confocal scanning and 3-dimensional image analysis, fluorescence 5mC and DAPI signals were measured in segmented cell nuclei, and a 5mC/DAPI co-distribution map was generated for each imaged cell. Cells were classified as hypomethylated based on 5mC load and 5mC/DAPI co-distribution. The proportion of hypomethylated epithelial cells in the sputum determines whether a patient has lung cancer. Results: A total of 88 subjects were enrolled: 12 healthy subjects; 34 high-risk subjects with benign chronic lung disorders (10 with chronic obstructive pulmonary disease, 24 with idiopathic pulmonary fibrosis), and 43 subjects with non-small cell lung cancer (27 with stage I-II and 16 with stage III-IV). The test identified early-stage non-small cell lung cancer and distinguished it from the high-risk group with 95.8% (95% confidence interval, 78.9-99.9) sensitivity and 41.2% (95% confidence interval, 24.6-59.3) specificity applying only 5mC, 95.8% (95% confidence interval, 78.9-99.9) sensitivity and 26.5% (95% confidence interval, 12.9-44.4) specificity using solely 5mC/DAPI index, and 100% (95% confidence interval, 98.7-100) sensitivity and 26.1% (95% confidence interval, 26.2-27.8) specificity with the combined parameters. Conclusions: We tested and validated a novel, noninvasive, highly sensitive screening test for non-small cell lung cancer. With the use of sputum, our test may impact lung cancer screening, evaluation of pulmonary nodules, and cancer surveillance algorithms.

Intraductal papillary lesions of the breast: clinical and pathological correlation.

Papillary lesions of the breast range from a spectrum of benign intraductal papillomas with and without atypia to papillary carcinoma. Distinction between benign and malignant lesions on core needle biopsy (CNB) is difficult without surgical excision. We examined if clinical findings in patients with benign intraductal papillomas (IP) on CNB correlate with pathology at surgical excision. Between 1998 and 2011, 103 patients were identified with a papillary lesion on CNB. Clinical variables were studied to determine if there was clinical correlation with pathological outcomes at final surgical excision. Of the 103 patients, 59 (57%) patients had IP on initial CNB and were included in our analysis. On final pathology, 17 (29%) of these were upstaged to intraductal papilloma with atypia and six (10%) were found to have carcinoma. A clinically palpable mass was the only significant predictor of upstaging to malignancy (P<0.05). No radiographic findings were found to be significant predictors of pathological upstaging. In conclusion, surgical excision is still recommended for benign papillary lesions diagnosed on CNB because the correlation with clinical and radiological findings does not assure benign pathology.

The use of virtual microscopy for proficiency testing in gynecologic cytopathology: a feasibility study using ScanScope.

CONTEXT: ScanScope software can digitize entire cytology slides. OBJECTIVE: To test the feasibility of using virtual microscopy methods and "virtual Papanicolaou tests" for proficiency testing (PT) in gynecologic cytopathology. DESIGN: Two PT exercises were conducted using virtual microscopy. Five cytopathologists and 1 cytotechnologist interpreted images using 2 different schema as follows: (1) the College of American Pathologists graded diagnostic codes (CAP-GDCs) and (2) the Center for Medicare and Medicaid Services test scoring categories (CMS-TSCs). The number of diagnostic errors using the CAP-GDCs and the CMS-TSCs, the mean length of time spent diagnosing each case, and the impressions by the users regarding the facility of the technology and image quality were studied. RESULTS: In the first PT exercise, the participants provided incorrect diagnoses in 4 to 8 of the 10 test cases using the CAP-GDCs and in 1 to 4 of the 10 test cases using the CMS-TSCs. In the second PT exercise, the number of errors decreased to 1 to 6 using the CAP-GDCs and to 0 to 6 using the CMS-TSCs. The results did not achieve statistical significance. The mean time of 9.4 minutes spent per case in the second PT exercise was significantly shorter than the 14.4 minutes spent per case in the first PT exercise (P < .001). The ease of use of the software and the image quality were scored by all participants as 3+ or as 4+. CONCLUSIONS: This preliminary study shows that virtual microscopy and virtual Papanicolaou tests prepared using ScanScope may provide effective tools for PT. Technical issues that require further investigation are discussed.