Abstract: The present invention is directed to Protein-fragment Complementation Assays (PCAs) and assay compositions based on fluorescent proteins. The invention provides methods for fragmenting fluorescent proteins and generating mutant fragments with desired spectral characteristics for PCA. The invention encompasses assays and compositions based on fluorescent proteins from the species Aequorea, Anemonia and Anthozoa. In particular, the invention is directed to fragments of mutant fluorescent proteins having improved spectral properties over the wild-type proteins. The invention encompasses fragments of mutant versions of A. Victoria green fluorescent protein (GFP), in particular yellow fluorescent proteins (EYFP and super-EYFP), ‘Venus’, cyan, ‘citrine’, blue, cyan-green, and photoactivatable variants of GFP The invention also encompasses red fluorescent PCAs based on Discosoma red fluorescent protein (RFP PCA) and a kindling fluorescent protein PCA (KFP1 PCA) derived from Anemonia sulcata.

Abstract: The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library and therefore finds numerous applications in the study of protein-protein interactions. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed into E. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We applied these selection processes to a library-versus-library sample of 2.

Abstract: The invention provides a method of screening a candidate drug, a compound library or a biological extract to identify activators or inhibitors of G-protein-coupled receptors or G-protein-coupled pathways, comprising: (A) using a fluorescent protein fragment complementation assay to construct an assay for one or more steps in a G-protein-coupled pathway; (B) testing the effects of the candidate drugs, compound library, or biological extract on the receptor or pathway of interest; and (C) using the results of the screening to identify specific agents that activate or inhibit the receptor or pathway of interest. The invention also provides a method for identifying a drug lead that modulates the activity of a G-protein-coupled pathway using a fluorescent protein fragment complementation assay. The method of the invention is used to identify agonists, antagonists, activators or inhibitors of G-protein coupled receptors or G-protein-coupled pathways.

Abstract: The present invention provides protein single-color and multi-color protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter such as monomeric enzymes and fluorescent proteins, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. The development of such assays provides for a broad, flexible and biologically relevant platform for drug discovery.

Abstract: The present invention provides a pharmaceutical formulation suitable for filling softgel capsules comprising: (a) a therapeutically effective amount of a non-steroidal anti-inflammatory drug selected from the group consisting of selected from the group consisting of (1) the propionic acid derivatives; (2) the acetic acid derivatives; (3) the fenamic acid derivatives; (4) the biphenylcarboxylic acid derivatives; and (5) the oxicams; as well as Cox 2 inhibitors and mixtures thereof; and (b) a solvent system comprising 40% to 60% by weight a polyoxyethylene ether of the formula: wherein n=1 to 6; 15% to 35% by weight of glycerin and 15% to 35% by weight water. In the case of compositions containing an NSAID having carboxyl function an alkaline hydroxide may be added to neutralize said hydroxide function.

Abstract: Bacillus licheniformis O.W.U. 138B and 88B and other bacterial strains capable of degrading feathers isolated from wild birds, are disclosed. The strains and the keratinases produced by these strains are useful for degrading waste feathers produced by commercial poultry processing; producing animal feed, fertilizer, or natural gas from poultry waste; and cleaning of certain fabrics.

Abstract: Nutriceutical compositions useful as a dietary supplement which have antioxidant/free radical scavengers and also having a cytoprotective effect are disclosed. The compositions contain a cytoprotective and antioxidant/free radical scavenging amount of at least one of D-histidine, L-histidine, a racemic mixture thereof, a non-racemic mixture thereof, and nutriceutically acceptable salts thereof in combination with phytonutrients having antioxidant properties such as vitamin C, vitamin E, vitamin A and .beta.-carotene. The compositions can be prepared in capsule form, tablets, sustained release tablets, suspensions and oral rehydration solutions.