p97 and p47 function in membrane tethering in cooperation with FTCD during mitotic Golgi reassembly

p97ATPase‐mediated membrane fusion is required for the biogenesis of the Golgi complex. p97 and its cofactor p47 function in soluble N‐ethylmaleimide‐sensitive factor (NSF) attachment protein receptor (SNARE) priming, but the tethering complex for p97/p47‐mediated membrane fusion remains unknown. In this study, we identified formiminotransferase cyclodeaminase (FTCD) as a novel p47‐binding protein. FTCD mainly localizes to the Golgi complex and binds to either p47 or p97 via its association with their polyglutamate motifs. FTCD functions in p97/p47‐mediated Golgi reassembly at mitosis in vivo and in vitro via its binding to p47 and to p97. We also showed that FTCD, p47, and p97 form a big FTCD‐p97/p47‐FTCD tethering complex. In vivo tethering assay revealed that FTCD that was designed to localize to mitochondria caused mitochondria aggregation at mitosis by forming a complex with endogenous p97 and p47, which support a role for FTCD in tethering biological membranes in cooperation with the p97/p47 complex. Therefore, FTCD is thought to act as a tethering factor by forming the FTCD‐p97/p47‐FTCD complex in p97/p47‐mediated Golgi membrane fusion.     

Assessment of mouse strain on bacterial translocation from the gastrointestinal tract

The translocation of indigenous bacteria from the gastrointestinal tract to the mesenteric lymphnodes was compared in ten strains of mice. Indigenous Escherichia coli were cultured from the mesenteric lymphnodes of only two of the six mouse strains examined. Thus, spontaneous translocation of indigenous enteric bacteria across the intestinal barrier did not occur to any significant extent in any of the mouse strains examined. Since bacterial overgrowth in the gastrointestinal tract promotes bacterial translocation, bacterial translocation was tested in ten mouse strains including B10 series after antibiotic-decontaminated and subsequent colonization with streptomycin-resistant E. coli C25. E. coli C25 populated the ceca of the mice at levels of 10(8) to 10(9) per gram and translocated to 90-100% of the mesenteric lymphnodes with mean of 10(1.13) to 10(1.86) per mesenteric lymphnode. However, there were no significant differences between mouse strains as to the translocation incidence or the numbers of viable E. coli C25 per mesenteric lymphnode. Thus, genetic differences between mouse strains did not influence bacterial translocation from the gastrointestinal tract to the mesenteric lymphnodes.     

Inactivation of Escherichia coli K12 Using Atmospheric Gas Plasma Produced from Humidified Working Gas

The bactericidal effect of atmospheric gas plasma (AGP) on Escherichia coli K12 cells in 0.9% [w/v] sodium chloride was investigated under normal atmospheric pressure. A plasma‐generating unit was supplied with working gas (air) at 25 °C and a relative humidity (RH) of 0, 14, 32, 43, 50, 60 or 70%. Gas plasma was generated using radio frequency discharge (20 kHz) under atmospheric pressure, and blown onto the surface of an E. coli cell suspension. Seven log‐cycles of cells were completely inactivated within 15 minutes of the start of AGP treatments using working gas at 32, 43, 50 or 60% RH. AGP at 14% RH inactivated only 2 log‐cycles of cells, but no inactivation was observed when air at 0% or 70% RH was used. The inactivation curves were biphasic and the rate constants for both stages were closely related to the RH of the working gas. The rate of cell inactivation was at its maximum at an RH of 43%. The bactericidal effect of the AGP treatment was not a result of ozone generation, however, the moisture content of the working gas was a significant factor in ensuring that E. coli K12 inactivation occurred. The interpretation of these data was that chemical species generated from the water molecules in the working gas were bactericidal in their effects.