Yong Man Yu

Formation of size-controlled nano carrier systems by self-assembly

Nano carrier systems were prepared by forming self-assembled liposomes having a size distribution in the nano range through use of an ultrasonic homogenizer. Phosphatidylcholine and cholesterol were utilized as an amphiphilic compound and a shape stabilizer, respectively. The size of prepared samples was decreased (up to 150 nm) by elevating ratio of lecithin and extending homogenization time (2 ∼ 6 min). After secondary coating with alginic acid (0.1, 0.3 and 0.5%, W/V), size was remarkably changed in the range of ±30 nm and zeta-potential was altered (only chitosan coating (molecular weight: 30 000 Da, 0.2%, W/V): 10.3 mV, Alginic acid coating (0.5%, W/V) after the chitosan coating: −21.8 mV). The low molecular weight chitosan (0.1%, W/V)-coated nano-liposomes had a lower absolute value of zeta-potential than the high molecular weight chitosan (0.1%, W/V)-coated nano-liposomes. The encapsulation efficiency was measured by gas chromatography. The efficiency was decreased slightly by elevating chitosan concentration (0.1 ∼ 0.5%, W/V).

Actin Cytoskeleton and Golgi Involvement in Barley stripe mosaic virus Movement and Cell Wall Localization of Triple Gene Block Proteins

Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treatments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.