Smooth muscle relaxant evaluation of Jatropha Curcas Linn (Euphorbiaceae) and isolation of triterpenes

Jatropha curcas is a herbal preparation used in the tropics for the treatment of threatened abortion and related problems associated with pregnancy. The Stem bark of Jatropha curcas is used ethno medicinally in Nigeria especially in the eastern part of the country for the treatment of infertility and spontaneous abortion (miscarriage). The present study was undertaken in order to validate the folkloric claim, using scientific experimental procedures and bioassay guided fractionation. The crude powdered sample was subjected to phytochemical screening testing for the presence of alkaloids, tannins, saponins and carbohydrates. Chromatographic analysis (TLC and VLC) were carried out using various solvent systems. The effect of methanolic extracts on rat uterine contractions was studied in vitro, in 40ml organ baths containing physiological salt solution of De Jalon maintained at 370C, aerated with 95% O2 and 5% CO2 with an isometric transducer connected an UgoBasile recorder under a resting tension of 750mg. The result of the phytochemical screening revealed the presence of glycosides, tannins, saponins and alkaloids. The extract abolished significantly the spontaneous contraction of the uterus and reduced acetylcholine induced uterine contractions at a dose of 50mg/ml. The tocolytic effects indicate the presence of active principle(s) which would explain the ethno medicinal use of the stem bark of Jatropha curcas to treat spontaneous abortion.     

Screening one bead one compound libraries against serum using a flow cytometer: Determination of the minimum antibody concentration required for ligand discovery

One bead one compound (OBOC) libraries can be screened against serum samples to identify ligands to antibodies in this mixture. In this protocol, hit beads are identified by staining with a fluorescent labeled secondary antibody. When screens are conducted against two different sets of serum, antibodies, and ligands to them, can be discovered that distinguish the two populations. The application of DNA-encoding technology to OBOC libraries has allowed the use of 10?µm beads for library preparation and screening, which pass through a standard flow cytometer, allowing the fluorescent hit beads to be separated from beads displaying non-ligands easily. An important issue in using this approach for the discovery of antibody biomarkers is its analytical sensitivity. In other words, how abundant must an IgG be to allow it to be pulled out of serum in an unbiased screen using a flow cytometer? We report here a model study in which monoclonal antibodies with known ligands of varying affinities are doped into serum. We find that for antibody ligands typical of what one isolates from an unbiased combinatorial library, the target antibody must be present at 10–50?nM. True antigens, which bind with significantly higher affinity, can detect much less abundant serum antibodies.