Molecular rotors as conditionally fluorescent labels for rapid detection of biomolecular interactions.

We demonstrate the use of fluorescent molecular rotors as probes for detecting biomolecular interactions, specifically peptide-protein interactions. Molecular rotors undergo twisted intramolecular charge transfer upon irradiation, relax via the nonradiative torsional relaxation pathway, and have been typically used as viscosity probes. Their utility as a tool for detecting specific biomolecular interactions has not been explored. Using the well characterized p53-Mdm2 interaction as a model system, we designed a 9-(2-carboxy-2-cyanovinyl) julolidine-based p53 peptide reporter, JP1-R, which fluoresces conditionally only upon Mdm2 binding. The reporter was used in a rapid, homogeneous assay to screen a fragment library for antagonists of the p53-Mdm2 interaction, and several inhibitors were identified. Subsequent validation of these hits using established secondary assays suggests increased sensitivity afforded by JP1-R. The fluorescence of molecular rotors contingent upon target binding makes them a versatile tool for detecting specific biomolecular interactions.

Antihyperlipidemic and antioxidant effects of extracts from Pleurotus citrinopileatus.

Pleurotus citrinopileatus is a popular edible mushroom which is physiologically active in both humans and animals. In the study we investigate the effects of this mushroom on hyperlipidemic hamster rats. Four dietary forms of the mushroom were created as follows. The powdered dry fruiting body, hot-water extract, and two kinds of elutes were obtained, from ethyl acetate extract and methanol extract, respectively, in different mixed proportion solvents over silica gel column chromatography (referred to as EAE and MOE, respectively). They were tested at different dosages as a supplement to a high-fat diet in hyperlipidemic rats. Serum triglycerides and total cholesterol levels were significantly lower in groups supplemented with the highest dosages of EAE and MOE (0.5 g/kg, body weight daily) as compared with the control groups that received no mushroom additive. High-density lipoprotein levels in these same two experimental groups were also significantly higher than those in the negative control group. The tested rats that were fed with EAE had the highest serum glutathione peroxidase and superoxide dismutase activity, and those with the MOE and EAE had the highest DPPH free radical scavenging activities and ferric-reducing abilities, tested in vitro. The major constituents of MOE and EAE were identified as ergosterol and nicotinic acid, respectively. P. citrinopileatus extracts may have a significant antihyperlipidemia effect. Furthermore, antioxidant activities and antihyperlipidemic effects of MOE and EAE seemed to display similar tendencies.

Antihyperglycemic effect of polysaccharide from fermented broth of Pleurotus citrinopileatus.

Pleurotus citrinopileatus is an edible mushroom, which has recently become very popular, with a consequent increase in industrial production. Water-soluble polysaccharides (WSPS), extracted from edible mushrooms, have been found to have antitumor and immunoenhancing effects. In this study, we investigate the effects of WSPS extracted from submerged fermented medium of P. citrinopileatus on hyperglycemia and damaged pancreatic cells in rats with streptozotocin (STZ)-induced diabetes. The diabetic rats fed with water-soluble polysaccharide of P. citrinopileatus (SPPC) lost less body weight than those fed SPPC-free regular diet. Serum total cholesterol and triglyceride levels in the diabetic rats fed with SPPC at a dose of 0.4 g/kg bw daily was lower than in the groups fed with SPPC at doses of 0.04 and 0.12 g/kg bw. The fasting blood glucose levels of diabetic rats fed with SPPC were 44% lower than the negative controls. The degree of damage to the islets of Langerhans of the rats fed with the highest dosage of SPPC was significantly lower than those fed with SPPC at doses of 0.04 and 0.12 g/kg bw. The results showed that STZ-induced diabetic rats fed with SPPC might help alleviate the elevation of the level of that in fasting blood glucose.