Abstract: The present invention relates to a pknB kinase and a pstP phosphatase as well as their use for identifying antibacterial substances.

Abstract: Provided is a method of identifying a target cell using a virus. The method includes infecting the target cell with the virus by contacting the virus to the target cell and culturing the target cell to propagate the virus; adding a chromogenic substrate to the resultant cell culture to induce enzyme reaction converting the chromogenic substrate to a chromogenic product; and measuring an optical signal emitted from the chromogenic product, wherein the virus contains in its genome a gene encoding an enzyme capable of converting the chromogenic substrate to the chromogenic product and a gene encoding a ligand allowing the virus to specifically bind with a receptor of the target cell to infect the target cell with the virus.

Abstract: Methods, compositions, kits, and a system are disclosed for detecting one or more analytes in a sample. A mixture comprising the (i) sample, (ii) a first binding reagent comprising a cleavage-inducing moiety and a first binding agent specific for an analyte, and (ii) one or more electrophoretic probes each having a second binding agent is subjected to conditions under which binding of respective binding agents occurs. The interaction between the binding agents and the analyte brings the cleavage-inducing moiety within a proximity effective for cleaving a cleavable linkage tethering an electrophoretic tag to the second binding agent, thereby releasing the tag for electrophoretic separation. Separation of different tags occurs by virtue of their distinct electrophoretic mobilities. After separation, a signal amplification moiety on each tag is activated to generate a signal to indicate the presence of a particular analyte in the sample.

Abstract: Renal injury and inflammation is diagnosed by detecting an elevation in GSK3b level or activity. Inflammation of bodily tissues such as renal tissue is inhibited by administration of GSK3b inhibitory compositions.

Abstract: We disclose methods of sorting or separating mixtures of living cells (e.g., eukaryotic, prokaryotic, mammalian, pathogenic, bacterial, viral, etc.). We perform our methods by activating cell-selective photophoric labels, which photosensitize and chemically reduce a photosensitive metal compound to form metal grains, particles or crystals. The metal adheres to the cells and forms the basis for sorting or separating different cell types. Photophoric labels may include chemiluminescent agents such as peroxidase enzymes activated with peroxidase substrates capable of luminescence. Photosensitive metal compounds may be present in a light-sensitive matrix or emulsion containing photosensitizable metal compounds, which form metal grains, particles or crystals upon exposure to a developer solution. Developer solutions are formulated to substantially allow living cells to remain viable after exposure to the developing solution.

Abstract: In a nonhuman animal model that produces a secretory protein, which is obtained by transplanting secretory protein-expression vector-transfected cells into a nonhuman animal, an amount of the secretory protein is measured and, based on the amount of the secretory protein, transcriptional activity, number of the transplanted cells, and tumor volume is measured. Further, screening of a compound that affects a transcriptional activity, number of transplanted cells, or tumor volume is performed by using the nonhuman animal model to which a compound has been administered.

Abstract: This invention relates to screening assay methods that can be applied to broad members of the protein kinase gene family and are useful in the detection and evaluation of kinase inhibitors.

Abstract: The present invention relates to protein modulators, especially modulators of kinesin bioactivities. The present invention provides methods for screening protein modulators, e.g., kinesin modulators that bind to a region of a kinesin and/or that modulate kinesin bioactivities. The present invention also provide methods and compositions for treatment of disorders mediated by abnormal cellular proliferation activities.

Abstract: The present invention provides a DNA coding for a protein defined in the following (A) or (B) is obtained from Brevibacterium flavum chromosomal DNA library by cloning a DNA fragment that complicates serB deficiency of Escherichia coli as a open reading frame in the DNA fragment. (A) A protein which comprises an amino acid sequence of SEQ ID: 2 in Sequence Listing; or (B) A protein which comprises an amino acid sequence including substitution, deletion, insertion, addition or inversion of one or several amino acids in the amino acid sequence of SEQ ID NO: 2 in Sequence Listing, and which has phosphoserine phosphatase activity.

Abstract: The present invention is related to a method for the identification and/or the quantification of a target compound obtained from a sample, preferably a biological sample, comprising the steps of putting into contact the target compound with a capture molecule in order to allow a specific binding between the target compound with a capture molecule, the capture molecule being fixed upon a surface of a solid support according to an array comprising a density of at least 20 discrete regions per cm2, each of the discrete regions being fixed with one species of capture molecules, performing a reaction leading to a precipitate formed at the location of the binding, determining the possible presence of precipitate(s) in discrete region(s), and correlating the presence of the precipitate(s) at the discrete region(s) with the identification and/or a quantification of the target compound.

Abstract: The present invention is directed to methods and compositions for improving pulmonary surfactant catabolism. More specifically, the specification describes methods and compositions for making and using a lysosomal phospholipase A2 in methods for the diagnosis, and treatment of disorders of phospholipid catabolism such as pulmonary alveolar proteinosis.

Abstract: Follicular thyroid adenoma (FTA) is distinguished from follicular thyroid carcinoma (FTC) by comparing amount of an expression product of at least one gene selected from the group consisting of DDIT3, ARG2, ITM1, C1orf24, TARSH, and ACO1 in a test follicular thyroid specimen to a normal control thyroid specimen. The test follicular thyroid specimen is identified as FTA if the amount of expression product of TARSH is equal to or greater in the test follicular thyroid specimen than in the normal control thyroid specimen. The test follicular thyroid specimen is identified as FTC if the amount of expression product of DDIT3, ARG2, ITM1, C1orf24, or ACO1 is greater in the test follicular thyroid specimen than in the normal control thyroid specimen.

Abstract: Compositions for an alkaline phosphatase and methods for over-expression and purification of thermolabile Antarctic phosphatase (TAP) are provided. Uses for TAP include dephosphorylation of nucleic acids, sugars, peptides and proteins. TAP as described herein has advantages over phosphatases from other sources with respect to thermolability at 65° C. and efficiency of dephosphorylation activity at approximately neutral pH.

Abstract: A method for detecting transferase activity of a sample includes contacting the sample with a substrate and at least one of a phosphate group donor and a phosphate group acceptor. The substrate includes a reporter compound and amino acids. A peptidase is added that cleaves a non-phosphorylated substrate at a first rate and a phosphorylated substrate and a second rate. The output of the reporter compound is detected. In a preferred embodiment, the transferase activity detected is a kinase activity. In another preferred embodiment, the transferase activity detected is a phosphatase activity. Also provided is a method of screening for alterations in a transferase reaction. Kits and peptide substrate are also provided for carrying out at least one of the methods of the invention.

Abstract: An assay is disclosed for measuring activity of enzymes, such as kinases, phosphatases, and proteases. Measurements of enzymatic activity are accomplished in a homogenous assay format utilizing a fluorescence quenching technique employing paramagnetic metal ions.

Abstract: The present invention provides ligand-detection reagents, ligand analogs and methods for determining the presence of a ligand in a sample. The ligand-detection reagent comprises a ligand-binding antibody and a ligand analog to form an antibody-ligand analog complex wherein the ligand analog is covalently bonded to a reporter molecule. This complex may additionally comprise a labeling protein non-covalently bonded to the antibody to form a ternary complex wherein the labeling protein comprises a monovalent antibody fragment or a non-antibody protein that is covalently bonded to a label moiety. The reporter molecule is either quenched by the ligand-binding antibody or by the label moiety of the labeling protein, depending on the reporter molecule and the ligand-binding antibody, wherein the amount of quenching is directly related to the amount of ligand present in the sample.

Abstract: The present invention relates to novel chemical compounds, methods for their discovery, and their therapeutic use. In particular, the present invention provides benzodiazepine derivatives and methods of using benzodiazepine derivatives as therapeutic agents to treat a number of conditions associated with the faulty regulation of the processes of programmed cell death, autoimmunity, inflammation, and hyperproliferation, and the like.

Abstract: The present invention concerns the development of a cell-based assay system having improved sensitivity to HCV NS3 protease activity when compared to known assays, which is useful for screening test compounds capable of modulating (particularly inhibiting) HCV NS3 protease activity. This system provides a first construct comprising a transactivator domain joined downstream of the NS3–5 domains of HCV under the control of a non-cytopathic viral promoter system. A second construct is also provided that comprises a reporter gene under the control of an operator sensitive to the binding of the transactivator. The NS3–5 domains encodes the NS3 polyprotein which comprises: the NS3 protease, followed by the NS4A co-factor, the NS4B and NS5A proteins (including any derivative, variant or fragment thereof), terminated by the NS5B protein (including any derivative, variant or fragment thereof) sufficient to constitute a NS5A/5B cleavage site.

Abstract: The infection of a mammalian host by a microorganism can be prevented or treated through the alteration of the C. albicans homologue of the high affinity phosphodiesterase, PDE2, gene and/or the adenylate cyclase-associated protein gene. These methods may be used in the identification, prevention or treatment of microbial infection of mammalian hosts such as immunocompromised or immunosuppressed humans, for example, those having AIDS or undergoing transplantation or anti-cancer therapy.

Abstract: Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enzymes.

Abstract: Methods, compositions and kits are disclosed for determining one or more target polypeptides in a sample where the target polypeptides have undergone a post-translational modification. A mixture comprising the sample and a first reagent comprising a cleavage-inducing moiety and a first binding agent for a binding site on a target polypeptide is subjected to conditions under which binding of respective binding moieties occurs. The binding site is the result of post-translational modification activity involving the target polypeptide. The method may be employed to determine the target polypeptide itself. In another embodiment the presence and/or amount of the target polypeptide is related to the presence and/or amount and/or activity of an agent such as an enzyme involved in the post-translational modification of the target polypeptide.

Abstract: The present invention relates to a method for quickly determining cAMP content or an adenylate cyclase activity in a biological sample containing non-cyclic adenine nucleotides without the use of radioactive agents. Particularly, the present invention provides a method of determining cAMP content or an adenylate cyclase activity in a biological sample containing non-cyclic adenine nucleotides selected from the group consisting of cAMP produced by endogenous adenylate cyclase, and AMP, ATP, ADP and a mixture thereof, which comprises (1) combining a biological sample with effective amounts of apyrase, adenosine deaminase and alkaline phosphatase to enzymatically remove non-cyclic adenine nucleotides other than cAMP, and glucose-6-phosphate in the sample; (2) enzymatically converting cAMP into AMP; (3) determining an amount of AMP without the use of radioactive agents, and a kit to carry out the method.

Abstract: This invention has as its object a method for detecting catalytic activity of a sample, characterized in that it comprises: the incubation of a substrate (S) with the sample that may have the catalytic activity that it is desired to detect, the addition of a reagent (X) that can react either with a chemical group of unconsumed substrate (S) or with a chemical group of product (P) that is formed after an incubation period with the sample, the addition of a developer (R) that can react with reagent (X), and the detection of the transformation of developer (R).

Abstract: The invention provides a purified or recombinant phytase enzyme (SEQ ID NO:2) initially derived from Escherichia coli B. The enzyme has a molecular weight of about 47.1 kilodaltons and has phytase activity (SEQ ID NO:2). The enzyme can be produced from native or recombinant host cells and can be used to aid in the digestion of phytate where desired. In particular, the phytase of the present invention can be used in foodstuffs to improve the feeding value of phytate rich ingredients.

Abstract: HPTPbeta is useful as a target in screening agents effective for the treatment of angiogenesis mediated disorders.

Abstract: The invention provides monoclonal antibodies that selectively bind to ectodermally- and endodermally-derived stem cells and methods for the diagnosis of a neoplasm in a subject by contacting a tissue sample from the subject with the antibodies. Also disclosed are methods for isolating such stem cells from a heterogeneous cell population by contacting the population with antibodies which selectively bind to stem cells.

Abstract: Benzodiazepine compounds, and methods for using those compounds are provided. Some of the benzodiazepine compounds include 1,4-benzodiazepine-2-one and 1,4-benzodiazepine-2,5-dione compounds of the following structures: wherein R1, R2, R3 and R4 are as defined. The invention also includes enantiomers, pharmaceutically acceptable salts, prodrugs or derivatives of the benzodiazepine compounds. Any one or more of these benzodiazepine compounds can be used to treat a variety of dysregulatory disorders related to cellular death. Such disorders include autoimmune disorders, inflammatory conditions, hyperproliferative conditions, viral infections, and atherosclerosis. In addition, the above compounds can be used to prepare medicaments to treat the above-described dysregulatory disorders. The benzodiazepines can also be used in drug screening assays and other diagnostic methods.

Abstract: The present invention provides filter-based assays for measuring binding of compounds to phosphodiesterases (PDEs). The assay permits stoichiometric binding of compounds to PDEs, thereby providing a highly sensitive measure of a PDE binding and inhibition.

Abstract: The present invention provides high throughput screening methods for characterizing compounds that modulate the biological activity of a biochemically functional sarcomere. Compounds characterized by such methods are useful for treating congestive heart failure.

Abstract: The invention discloses methods, compositions and kits for stabilizing a solubilized phenyl phosphate, preferably paranitrophenyl phosphate (PNPP), using charcoal. Also disclosed are methods, compositions and kits for recycling solubilized phenyl phosphate, preferably PNPP, that has an absorbance of less than 0.1 when measured at 405 nm due to non-enzymatic hydrolysis.

Abstract: Proteins are identified from human breast tumor cell lines (MDA-MB-231, T-47D and T-47D/Met) that interact specifically with the substrate-trapping mutant form of Density Enhanced Phosphatase-1 (DEP-1). These proteins include the functional component p120 catenin (p120ctn), the adaptor protein Gab 1, and the HGF/SF receptor Met. The invention relates to isolated complexes comprising DEP-1 polypeptides in specific association with Met, Gab 1, or p120ctn, identified herein as DEP-1 substrate polypeptides. Screening assays for agents that alter DEP-1 interaction with DEP-1 substrate polypeptides are also disclosed, as are methods for altering biological signals in cells that are transduced via DEP-1 pathways.

Abstract: A method for determining the level of kinase activity or phosphatase activity in a sample without the use of antibodies or radioactive labels is disclosed. The method employs a fluorescently-labeled phosphorylatable reporter peptide that is capable of being cleaved by a protease only when the peptide is in an unphosphorylated state. A change in fluorescence characteristics is an indication that the peptide is cleaved and, therefore, in an unphosphorylated state. Thus, the level of protease cleavage, as measured by the fluorescence change, provides a direct measure of phosphatase activity whereas the level of kinase activity is inversely proportion to the level of protease cleavage. The method is particularly well suited to high throughput screening, for example, for screening compounds which modulate kinase or phosphatase activity.

Abstract: Described herein are methods which identify candidate agents as binding to a protein or as a modulator of the binding characteristics or biological activity of a protein. Generally, the methods involve the use of ADP or phosphate. The assays can be used in a high throughput system to obviate the cumbersome steps of using gels or radioactive materials.

Abstract: A method is disclosed whereby the concentration of a blood substitute, such as cross-linked hemoglobin, in a serum or plasma specimen is rapidly and accurately identified and quantified. The method further takes the measured concentration of the blood substitute and uses it to correct for its effect, if any, on a measured analyte concentration, e.g., serum/plasma total protein. Further, the method allows for the determination of the concentration of true hemoglobin in the presence of blood substitutes. The method is carried out in respect of samples contained in a primary or secondary labelled tube, or a pipette tip used to dispense serum or plasma in a blood analyzer.

Abstract: A method for detecting transferase activity of a sample includes contacting the sample with a substrate and at least one of a phosphate group donor and a phosphate group acceptor. The substrate includes a reporter compound and amino acids. A peptidase is added that cleaves a non-phosphorylated substrate at a first rate and a phosphorylated substrate and a second rate. The output of the reporter compound is detected. In a preferred embodiment, the transferase activity detected is a kinase activity. In another preferred embodiment, the transferase activity detected is a phosphatase activity. Also provided is a method of screening for alterations in a transferase reaction. Kits and peptide substrate are also provided for carrying out at least one of the methods of the invention.

Abstract: The invention provides isolated nucleic acids molecules, designated MID 4460 nucleic acid molecules, which encode novel tyrosine phosphatase family members. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing MID 4460 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a MID 4460 gene has been introduced or disrupted. The invention still further provides isolated MID 4460 proteins, fusion proteins, antigenic peptides and anti-MID 4460 antibodies. Diagnostic and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: The present invention provides an assay for the identification of agents which can modulate TOR-mediated phosphorylation of substrate proteins. The assays of the invention utilize substrate proteins whose amino acid sequence contains the Ser/Thr motif recognized by TOR. Naturally occurring TOR which may be used in the methods of the invention include TOR isolated from a variety of species, particularly mammalian tissues.

Abstract: Two human MATER proteins as well as their use for fertility disorders, therapy and diagnosis are described.

Abstract: The invention provides fluorescence-based assays for determining the phosphorylating or the dephosphorylating activity of an enzyme acting on a substrate molecule. The substrate comprises at least one moiety that is capable of being phosphorylated or dephosphorylated by an enzyme to yield a product, the substrate being labelled with a fluorescent dye, and which is capable of exhibiting change in fluorescence intensity and fluorescence lifetime upon phosphorylation of the dye-labelled substrate or upon dephosphorylation of the dye-labelled phosphorylated substrate. In preferred embodiments, the substrate is labelled with a fluorescent dye selected from the acridone and the quinacridone classes of dyes. Also provided is a method is provided for screening for a test agent whose effect on the phosphorylating or dephosphorylating activity of an enzyme is to be determined.

Abstract: The invention provides human phosphodiesterases (HPDE) and polynucleotides which identify and encode HPDE. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with aberrant expression of HPDE.

Abstract: The present invention provides a method of inhibiting a member of a family of Protein Tyrosine Phosphatases (PTPases, PTPs) such as PTP1B, TC-PTP, CD45, SHP-1, SHP-2, PTP?, PTP?, PTP?, PTP?, PTP?, PTP?, PTP?, PTPD1, PTPD2, PTPH1, PTP-MEG1, PTP-LAR, and HePTP by exposing said Ptpase member by administration to a host or otherwise to at least one compound with certain structural, physical and spatial characteristics that allow for the interaction of said compound with specific residues of the active site of PTP1B and/or TC-PTP. These compounds are indicated in the management or treatment of a broad range of diseases such as autoimmune diseases, acute and chronic inflammation, osteoporosis, various forms of cancer and malignant diseases, and type I diabetes and type II diabetes, as well as in the isolation of PTPases and in elucidation or further elucidation of their biological function.

Abstract: A novel receptor-type protein tyrosine phosphatase (RPTP) protein or glycoprotein and the DNA coding therefor is expressed in a wide variety of mammalian tissues. Included in this family of proteins are human RPTP?, human RPTP? and human RPTP?. The RPTP protein or glycoprotein may be produced by recombinant means. Antibodies to the proteins, methods for measuring the quantity of the proteins, methods for screening compounds, such as drugs, which can bind to the proteins and inhibit or stimulate their activity, are provided.

Abstract: The invention is directed to methods and reagents for identifying test substances useful for the prevention or treatment of diseases involving an oxidative stress. The methods involve screening assays, including high throughput screening techniques, in which the test substances are tested for their ability to promote resistance to oxidative stress by inhibiting the activity of GADD34L, and thereby inhibiting the dephosphorylation of eIF2?, while not causing stress.

Abstract: A chemiluminescence method characterized in that when a 1,2-dioxetane derivative of the formula 1: wherein each of R1, R2, R3, R4 and R5 which are independent of one another, is a hydrogen atom, an alkyl group or an aryl group, or each pair of R2 and R3, and R4 and R5, which are independent of each other, may form together a cyclic alkyl group, R6 is a hydroxyl group, an alkoxyl group, an aralkyloxy group, a group represented by —OSi(R8R9R10) (provided that each of R8, R9 and R10 which are independent of one another, is an alkyl group or an aryl group), a phosphate group or a group represented by S(C?O)R11 (provided that R11 is an alkyl group or an aryl group), and R7 is a hydrogen atom, a halogen atom, an alkyl group or an alkoxyl group, is let generate chemiluminescence by means of an activator selected from the group consisting of a base, an acid, a salt, a fluorine compound, an enzyme, a catalyst and an amine compound, a cationic surfactant and a fluorescent material are made to coexist.

Abstract: The present invention provides a method of detecting thermonuclease-positive staphylococci that does not require inactivation of DNase-positive/TNase-negative bacteria with heat. The method includes (a) providing a culture medium selective for growing staphylococci; (b) inoculating the culture medium with a sample; (c) incubating the inoculated culture medium under conditions effective to promote the growth of staphylococci; (d) providing an indicator system that produces a differentiable, detectable signal in the presence of thermonuclease-positive staphylococci; (e) contacting the indicator system with the inoculated, incubated culture medium, thereby forming a detection assembly; (f) incubating the detection assembly under conditions effective for generating the differentiable, detectable signal; and (g) detecting the detectable signal. The present invention also provides a culture medium for the selective identification of Staphylococcus aureus.

Abstract: The present invention relates to nucleotide sequences of coryneform bacteria, coding for proteins involved in the bio-synthesis of L-serine and to methods for the isolation thereof. The invention further relates to an improved method for the production of L-serine. In addition, the present invention relates to the use of L-serine in the food, animal feed and/or pharmaceutical industries or in human medicine.

Abstract: A process for the production of a modified phytase with a desired property improved over the property of the corresponding unmodified phytase is disclosed, as well as modified phytases, polynucleotides encoding modified phytases, and animal feed including modified phytases.

Abstract: Monocolonal antibodies having a higher reactivity with tartrate-resistant acid phosphatase 5b (TRACP 5b) than tartrate-resistant acid phosphatase 5a (TRACP 5a) and having a higher specificity to TRACP 5b can be obtained by cell fusion using as antigens TRACP 5b purified from human osteoclasts. By using the monoclonal antibody, TRACP 5b in a sample can be detected specifically with a high sensitivity.

Abstract: A method for identifying phosphoprotein, which comprises: subjecting a sample protein to two-dimensional electrophoresis; dephosphorylating the sample protein with phosphatase; once again performing the two-dimensional electrophoresis under the same conditions; and detecting the spot that migrates to the alkaline side of the isoelectric focusing as the phosphoprotein is provided as a convenient method for identifying phosphoprotein with high accuracy.

Abstract: The invention concerns GENSET polynucleotides and polypeptides. Such GENSET products may be used as reagents in forensic analyses, as chromosome markers, as tissue/cell/organelle-specific markers, in the production of expression vectors. In addition, they may be used in screening and diagnosis assays for abnormal GENSET expression and/or biological activity and for screening compounds that may be used in the treatment of GENSET-related disorders.