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Background: The Papanicolaou (Pap) screen has been successful in reducing cervical cancer; but exhibits low sensitivity when detecting cervical dysplasia. Use of molecular biomarkers in Pap tests may improve diagnostic accuracy., Design: Monoclonal antibodies to Minichromosome Maintenance Protein 2 (MCM2) and DNA Topoisomerase II α (TOP2A) were selected for use in IHC based on their ability to differentiate normal from diseased cervical tissues in tissue microarrays. Enhanced Green Fluorescent Protein Western blot analysis was used to help identify binding epitopes specific to MCM2 and TOP2A antibody clones. Antibody affinity was determined by solution phase affinity measurement and immunohistochemistry was performed using high affinity MCM2 or TOP2A antibodies on serial histological sections., Results: Antibody clones to MCM2 and TOP2A clones were selected based on their ability to detect over expression in abnormal cervical epithelia. In IHC, MCM2-27C5.6 and MCM2-26H6.19 demonstrated superior staining in abnormal cervical tissue over the MCM2-CRCT2.1 antibody. A combination of MCM2 and TOP2A antibodies showed greater staining when compared to staining with any of the antibodies alone on serial histological sections. Distinct linear epitopes were elucidated for each of the MCM2 and TOP2A clones. Affinity values (Kd) for MCM2 or TOP2A antibodies had a similar range. In a research study, the MCM2 and TOP2A (BD ProEx™ C) antibody cocktail showed increased epithelia staining with increasing dysplasia. The use of BD ProEx™ C in combination with H&E staining enhanced immunohistochemical discrimination of dysplastic and non-dysplastic FFPE cervical tissue specimens., Conclusions: BD ProEx™ C containing MCM2 and TOP2A antibodies showed strong specific nuclear staining that correlated with increased dysplasia and lesion severity. Enhanced performance of the antibodies was linked to their unique topography recognition. BD ProEx™ C incorporates antibodies that enhance detection of CIN2+ cervical disease., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The best way to quantitate venous reflux is still a matter of debate. Duplex-derived valve closure time (VCTs) have been used recently because they can be measured easily. We examined the relationships between VCT and duplex-obtained quantitation of venous volume and between VCT and air plethysmography (APG). Sixty-nine legs in 45 patients with varying clinical degrees of chronic venous insufficiency were studied by duplex scan and APG. VCTs were compared with duplex-derived flow calculations and with APG-derived venous filling index and residual volume fraction. The patient's mean age was 47.5 +/- 13.9 years; the mean duration of their symptoms was 13 +/- 4 years. Twenty percent had a history of deep venous thrombosis, and 29% had undergone venous surgery. No correlation was found between VCT and flow volume or between VCT and flow at peak reflux at any of the anatomic locations studied: saphenofemoral junction, greater saphenous vein, lesser saphenous vein, superficial femoral vein, profunda femoris vein, and popliteal vein. Likewise, no correlation was found between total VCT and APG-derived venous filling index or between total flow volumes and APG-derived residual volume fraction. Total VCT and total flow volumes did, however, have a moderate correlation (r = 0.65; p = 0.0003). Duplex-derived VCTs, although extremely useful in determining the presence of reflux, do not correlate with the magnitude of reflux, and should not be used to quantitate the degree of reflux.