Anti-Pro-Brain Natriuretic Peptide polyclonal antibody

Chloride anion is critical for hypochlorous acid (HOCl) production and microbial killing in neutrophil phagosomes. However, the molecular mechanism by which this anion is transported to the organelle is poorly understood. In this report, membrane-enclosed and functionally active phagosomes were isolated from human neutrophils by using opsonized paramagnetic latex microspheres and a rapid magnetic separation method. The phagosomes recovered were highly enriched for specific protein markers associated with this organelle such as lysosomal-associated membrane protein-1, myeloperoxidase (MPO), lactoferrin, and NADPH oxidase. When FITC-dextran was included in the phagocytosis medium, the majority of the isolated phagosomes retained the fluorescent label after isolation, indicative of intact membrane structure. Flow cytometric measurement of acridine orange, a fluorescent pH indicator, in the purified phagosomes demonstrated that the organelle in its isolated state was capable of transporting protons to the phagosomal lumen via the vacuolar-type ATPase proton pump (V-ATPase). When NADPH was supplied, the isolated phagosomes constitutively oxidized dihydrorhodamine 123, indicating their ability to produce hydrogen peroxide. The preparations also showed a robust production of HOCl within the phagosomal lumen when assayed with the HOCl-specific fluorescent probe R19-S by flow cytometry. MPO-mediated iodination of the proteins covalently conjugated to the phagocytosed beads was quantitatively measured. Phagosomal uptake of iodide and protein iodination were significantly blocked by chloride channel inhibitors, including CFTRinh-172 and NPPB. Further experiments determined that the V-ATPase-driving proton flux into the isolated phagosomes required chloride cotransport, and the cAMP-activated CFTR chloride channel was a major contributor to the chloride transport. Taken together, the data suggest that the phagosomal preparation described herein retains ion transport properties, and multiple chloride channels including CFTR are responsible for chloride supply to neutrophil phagosomes. (C) 2012 Elsevier Inc. All rights reserved.

The aim of this study was to investigate the proliferation, adhesion, and migration function of chloride ion channels in HLEC B-3, a human lens epithelial cell line, and provide experimental basis for studying the mechanism of human after-cataract as well as the effective methods for controlling and treating it. MTT and Scratch assay methods were applied to detect the proliferation, adhesion, and migration of HLEC B-3 after applying 5, 10, 20, 50, 100, and 200 mu mol/L of chloride ion channel blockers (NPPB). In the cell proliferation experiment, no apparent difference was found in the first 12 and 24 h after treatment with 10, 20, 50, 100 and 200 mu mol/L NPPB or 48 h after treatment with 20, 50, 100 and 200 mu mol/L NPPB. The cell proliferations were obviously inhibited compared with the control group(without NPPB treatment), and the difference was significant (P < 0.05 or P < 0.01). Cell adhesion experimental results showed that when the cells were treated with 10, 20, 50, 100, and 200 mu mol/L NPPB for 12 or 24 h, cell adhesions were obviously inhibited. The difference was very significant (P < 0.05 or P < 0.01) compared with the control group. The activity of chloride ion channels can inhibit the proliferation, adhesion, and migration of LECs; therefore, inhibiting its activity can control the occurrence of human after-cataracts.