Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cul-tured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn’t stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.

The Riptortus (stinkbug) has a specialized symbiotic organ, M4 midgut, to harboring symbiont Burkholderia. M4 midgut is located in abdomen and surrounded with insect hemolymph. Recently our group demonstrated that symbiotic Burkholderia showed different physiology after adapting in M4 gut compare with in vitro cultured Burkholderia. And population of symbiotic Burkholderia in the M4 midgut is regulated by special organ. However, the molecular mechanism to prevent spreading and migrating symbiont bacteria to other host tissues from symbiotic organ is not clear. Therefore, we assumed that symbiont Burkholderia are susceptible to host humoral immunity after established infection in M4 midgut to prevent spreading and migrating into the other host tissues through Riptortus hemolymph. To prove this assuming, we tested the susceptibility and survival rate of symbiont Burkholderia in hemolymph of Riptortus in vitro and in vivo. We also examined the susceptibility of symbiont Burkholderia using purified antimicrobial peptides (AMP), pyrrhocoricin-like, thanatin-like and defensin-like AMPs. Finally, we tested inducing ability for AMPs by systemic infection of symbiotic Burkholderia. Gene expression of purified AMPs was not different after systemic infection of both symbiont and in vitro cultured Burkholderia. Surprisingly, in vitro cultured Burkholderia resisted on bacteria injected hemolymph and purified AMPs but symbiont Burkholderia were highly susceptible in bacteria injected hemolymph and purified AMP. These results suggest that symbiont Burkholderia can't survive in the hemolymph after escaping symbiotic organ. Moreover, humoral immunity of host Riptortus is important to prevent spreading and migrating symbiont Burkholderia into the other host tissue or organ from symbiotic organ.

Background : ROS produced by oxidative stress damaged endothelial cells, and cause a variety of vascular complications. In diabetic hyperglycemia state, ROS increase. The polyol pathway occur in diabetic complications, the excess glucose is absorbed into the polyol pathway when aldose reductase increased, NADPH changes it to sorbitol. Glutathione (GSH) removes ROS. GSH level is reduced by glutathione reductase, using NADPH as an electron donor. Activation of the polyol pathway decrease NADPH, and GSH also reduced. As a result, ROS is increased. In diabetic hyperglycemia state, Glycolysis increases. Effects of increased glycolysis, protein kinase C (PKC) is increased. NAD(P)H oxidase, stimulated by PKC-dependent pathway, increases ROS in the cell. In this study, we measured the ROS scavenging activity of 5 natural products (Lycii fructus, Astragalus membranaceus, Cassia Tora, Polygonatum odoratum, Rubus Coreanus), to confirm the efficacy as diabetic antioxidants. Methods and Results : We extracted 5 natural product by distilled water and ethanol. DPPH radical scavenging activity was significantly higher in Lycii fructus, Rubus coreanus. ABTS radical scavenging activity was better Rubus coreanus, Lycii fructus, Cassia Tora. In addition to, Rubus coreanus, Cassia Tora, Lycii fructus was comparatively higher reducing power activity than other natural products. And total phenolic and flavonoid contents were much higher in Rubus coreanus compared with other extracts. Conclusion : These results suggest that Lycii fructus, Rubus coreanus can be applied as diabetic antioxidant that prevent vascular complications caused by ROS.