Regulated Cleavages at the West Nile Virus NS4A-2K-NS4B Junctions Play a Major Role in Rearranging Cytoplasmic Membranes and Golgi Trafficking of the NS4A Protein

A common feature associated with the replication of most RNA viruses is the formation of a unique membrane environment encapsulating the viral replication complex. For their part, flaviviruses are no exception, whereupon infection causes a dramatic rearrangement and induction of unique membrane structures within the cytoplasm of infected cells. These virus-induced membranes, termed paracrystalline arrays, convoluted membranes, and vesicle packets, all appear to have specific functions during replication and are derived from different organelles within the host cell. The aim of this study was to identify which protein(s) specified by the Australian strain of West Nile virus, Kunjin virus (KUNV), are responsible for the dramatic membrane alterations observed during infection. Thus, we have shown using immunolabeling of ultrathin cryosections of transfected cells that expression of the KUNV polyprotein intermediates NS4A-4B and NS213-34A, as well as that of individual NS4A proteins with and without the C-terminal transmembrane domain 2K, resulted in different degrees of rearrangement of cytoplasmic membranes. The formation of the membrane structures characteristic for virus infection required coexpression of an NS4A-NS4B cassette with the viral protease NS2B-3pro which was shown to be essential for the release of the individual NS4A and NS4B proteins. Individual expression of NS4A protein retaining the C-terminal transmembrane domain 2K resulted in the induction of membrane rearrangements most resembling virus-induced structures, while removal of the 2K domain led to a less profound membrane rearrangement but resulted in the redistribution of the NS4A protein to the Golgi apparatus. The results show that cleavage of the KUNV polyprotein NS4A-4B by the viral protease is the key initiation event in the induction of membrane rearrangement and that the NS4A protein intermediate containing the uncleaved C-terminal transmembrane domain plays an essential role in these membrane rearrangements.

Ba0.5Sr0.5Co0.8Fe0.2O3-delta ceramic hollow-fiber membranes for oxygen permeation

Setf-supported asymmetric hollow-fiber membranes of mixed oxygen-ionic and electronic conducting perovskite Ba0.5Sr0.5Co0.8Fe0.2O3-delta (BSCF) were prepared by a combined phase-inversion and sintering technique. The starting inorganic powder was synthesized by combined EDTA-citrate complexing process followed by thermal treatment at 600 degrees C. The powder was dispersed in a polymer solution and then extruded into hollow-fiber precursors through a spinneret. ne fiber precursors were sintered at elevated temperatures to form gastight membranes, which were characterized by SEM and gas permeation tests. Performance of the hollow fibers in air separation was both experimentally and theoretically studied at various conditions. The results reveal that the oxygen permeation process was controlled by the slow oxygen surface exchange kinetics under the investigated conditions. The porous inner surface of the prepared perovskite hollow-fiber membranes considerably favored the oxygen permeation. The maximum oxygen flux measured was 0.031 mol-m(-2).s(-1) at 950 degrees C with the sweep gas flow rate of 0.522 mol(.)m(-2).s(-1). To improve the oxygen flux of BSCF perovskite membranes, future work should be focused on surface modification rather than reduction of the membrane thickness. (c) 2006 American Institute of Chemical Engineers.