The bioemulsifier of Acinetobacter radioresistens KA53, referred to as alasan, is a high-molecular-weight complex of polysaccharide and protein. The emulsifying activity of the purified polysaccharide (apo-alasan) is very low. Three of the alasan proteins were purified by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had apparent molecular masses of 16, 31, and 45 kDa. Emulsification assays using the isolated alasan proteins demonstrated that the active components of the alasan complex are the proteins. The 45-kDa protein had the highest specific emulsifying activity, 11% higher than the intact alasan complex. The 16-and 31-kDa proteins gave relatively low emulsifying activities, but they were significantly higher than that of apo-alasan. The addition of the purified 16-and 31-kDa proteins to the 45-kDa protein resulted in a 1.8-fold increase in the specific emulsifying activity and increased stability of the oil-in-water emulsion. Fast-performance liquid chromatography analysis indicated that the 45-kDa protein forms a dimer in nondenaturing conditions and interacts with the 16-and 31-kDa proteins to form a high-molecular-mass complex. The 45-kDa protein and the three-protein complex had substrate specificities for emulsification and a range of pH activities similar to that of alasan. The fact that the purified proteins are active emulsifiers should simplify structurefunction studies and advance our understanding of their biological roles.

Laboratory aquarium experiments demonstrated thatVibrio strain AK-1 caused rapid and extensive bleaching of the coral Oculina patagonica at 29°C, slower and less-complete bleaching at 23°C, and no bleaching at 16°C. At 29°C, the application of approximately 100 Vibrio strain AK-1 cells directly onto the coral caused 50 and 83% bleaching after 10 and 20 days, respectively. At 16°C, there was no bleaching, even with an initial inoculum of 1.2 × 108 bacteria. To begin to understand the effect of seawater temperature on bleaching ofO. patagonica by Vibrio strain AK-1, adhesion of the bacteria to the coral as a function of temperature was studied. Inoculation of 107 Vibrio strain AK-1 organisms into flasks containing 20 ml of seawater at 25°C and a fragment ofO. patagonica resulted in net levels of bacterial adhesion to the coral of 45, 78, and 84% after 2, 6, and 8 h, respectively. The adhesion was inhibited 65% by 0.001%d-galactose and 94% by 0.001% methyl-β-d-galactopyranoside (β-M-Gal). After the incubation of Vibrio strain AK-1 with the coral for 6 h, 42% of the input bacteria were released from the coral with 0.01% β-M-Gal, compared to less than 0.2% when β-M-Gal was present during the adhesion step. Adhesion did not occur whenVibrio strain AK-1 was grown at 16°C, regardless of whether the corals were maintained at 16 or 25°C, whereas bacteria grown at 25°C adhered to corals maintained at 16 or 25°C. Bacteria grown at 25°C adhered avidly to Sepharose beads containing covalently bound β-d-galactopyranoside but failed to bind if grown at 16°C. These data suggest that elevated seawater temperatures may cause coral bleaching by allowing for the expression of adhesin genes of Vibrio strain AK-1.

Background Neuroblastoma (nbl) is one of the most common solid cancers in children. Prognosis in advanced nbl is still poor despite aggressive multimodality therapy. Furthermore, survivors experience severe long-term multi-organ sequelae. Hence, the identification of new therapeutic strategies is of utmost importance. Cannabinoids and their derivatives have been used for years in folk medicine and later in the field of palliative care. Recently, they were found to show pharmacologic activity in cancer, including cytostatic, apoptotic, and antiangiogenic effects.Methods We investigated, in vitro and in vivo, the anti-nbl effect of the most active compounds in Cannabis, Δ9-tetrahydrocannabinol (thc) and cannabidiol (cbd). We set out to experimentally determine the effects of those compounds on viability, invasiveness, cell cycle distribution, and programmed cell death in human nbl SK-N-SH cells.Results Both compounds have antitumourigenic activity in vitro and impeded the growth of tumour xenografts in vivo. Of the two cannabinoids tested, cbd was the more active. Treatment with cbd reduced the viability and invasiveness of treated tumour cells in vitro and induced apoptosis (as demonstrated by morphology changes, sub-G1 cell accumulation, and annexin V assay). Moreover, cbd elicited an increase in activated caspase 3 in treated cells and tumour xenografts.Conclusions Our results demonstrate the antitumourigenic action of cbd on nbl cells. Because cbd is a non psychoactive cannabinoid that appears to be devoid of side effects, our results support its exploitation as an effective anticancer drug in the management of nbl.