The cytotoxicity and apoptotic effect of phytoceramide IV (N-palmitoyl-phytospingosine) were investigated by preparaing the ceramide containing liposomes using cholesteryl hemisuccinate (CHEMS) and methoxypolyethylene glycol (2000) cholesteryl succinate (PEGCHS). The ceramide-containing liposomes prepared using ultrasonication contained CHEMS and the phytoceramide IV at a molar ratio of 1:1. Gel chromatography and high performance liquid chromatography equipped with an evaporative light-scattering detection (HPLC-ELSD) were used to analyze the liposomes. The results show that the ceramide entrapment efficiency is over 90% and the molar ratio of phytoceramide IV to CHEMS is 1:1.4. The ratio of ceramide to CHEMS as well as the ultrasonication duration affects the liposome properties. The phytoceramide Ⅳ encapsulated in the liposomes reduces the cellular activity of the murine mastocytoma cell line P815 in a dose-dependent manner and the reduction of cellular activity is due to cell apoptosis. 4. 6-diamidino- 2-phenylindole dihydrochloride (DAPI) staining further supports this result. The encapsulation of highly hydrophobic ceramides into liposomal formulations could become a potential candidate for enhanced apoptosis.
A series of polyhydroxyalkanoate(PHA)copolymers consisting of short-chain-length(SCL)and medium-chain-length(MCL)3-hydroxyalkanoate(3HA)monomers were synthesized in the recombinant Ralstonia eutropha PHB - 4 harboring a low-substrate-specificity PHA synthase PhaC2Ps from Pseudomonas stutzeri 1317. These polyesters,whose monomer compositions varied widely in chain length,were purified and characterized by acetone fractionation,nuclear magnetic resonance(NMR),gel-permeation chromatography(GPC),and differential scanning calorimetry(DSC).This was the first time that the physical properties of PHA copolymers polymerized by PhaC2Ps were characterized.The results indicated that the variation in MCL 3HA contents did not have an obvious influence on the molecular weights of these PHA copolymers but was effective in changing their physical properties. The variation in the thermal property of PHA copolymers with 3-hydroxyoctanoate(3HO)content was also inves- tigated in this study.