DOC2B: A Novel Syntaxin-4 Binding Protein Mediating Insulin-Regulated GLUT4 Vesicle Fusion in Adipocytes

OBJECTIVE— Insulin stimulates glucose uptake in skeletal muscle and adipose tissues primarily by stimulating the translocation of vesicles containing a facilitative glucose transporter, GLUT4, from intracellular compartments to the plasma membrane. The formation of stable soluble N-ethyl-maleimide–sensitive fusion protein [NSF] attachment protein receptor (SNARE) complexes between vesicle-associated membrane protein-2 (VAMP-2) and syntaxin-4 initiates GLUT4 vesicle docking and fusion processes. Additional factors such as munc18c and tomosyn were reported to be negative regulators of the SNARE complex assembly involved in GLUT4 vesicle fusion. However, despite numerous investigations, the positive regulators have not been adequately clarified

Distillation of hydrogen isotopes for polarized HD target

We have developed a cryogenic distillation system to purify Hydrogen-Deuteride (HD) gas for a polarized HD target in LEPS experiments at SPring-8. A small amount of ortho-H$_2$ ($\sim$0.01%) in the HD gas plays an important role in efficiently polarizing the HD target. Since there are 1$\sim$5% impurities of H$_2$ and D$_2$ in commercially available HD gases, it is inevitable that the HD gas is purified up to $\sim$99.99%. The distillation system has a cryogenic pot (17$\sim$21 K) containing many small stainless steel cells called Heli-pack. Commercial HD gas with an amount of 5.2 mol is fed into the pot. We carried out three distillation runs by changing temperatures (17.5 K and 20.5 K) and gas extraction speeds (1.3 ml/min and 5.2 ml/min). The extracted gas was analyzed by using a gas analyzer system combining a quadrupole mass spectrometer with a gas chromatograph. The HD gas of 1 mol with a purity better than 99.99% has been successfully obtained. The effective NTS (Number of Theoretical Stages), which is an indicator of the distillator performances, is obtained as 37.2$\pm$0.6. This value is in reasonable agreement with a designed value of 37.9. The HD target is expected to be efficiently polarized under a well-controlled condition by doping an optimal amount of ortho-H$_2$ to the purified HD gas.Comment: 7 pages, 8 figures, 2 tables, updated 2011-12-1

Naturally occurring mutations in the PA gene are key contributors to increased virulence of pandemic H1N1/09 influenza virus in mice

We examined the molecular basis of virulence of pandemic H1N1/09 influenza viruses by reverse genetics based on two H1N1/09 virus isolates (A/California/04/2009 [CA04] and A/swine/Shandong/731/2009 [SD731]) with contrasting pathogenicities in mice. We found that four amino acid mutations (P224S in the PA protein [PA-P224S], PB2-T588I, NA-V106I, and NS1-I123V) contributed to the lethal phenotype of SD731. In particular, the PA-P224S mutation when combined with PA-A70V in CA04 drastically reduced the virus's 50% mouse lethal dose (LD50), by almost 1,000-fold