Abstract: The invention relates to methods identify or characterize compounds that can be used to treat specified clinical disorders such as hyperglycemia and type 2 diabetes. Compounds that can be used in these methods include 4?-fluoro-17?-ethynylandrost-5-ene-3?, 7?,17?-triol, 4?-fluoro-17?-ethynylandrost-5-ene-3?,7?,17?-triol, 4?-fluoro-17?-ethynylandrost-5-ene-3?,7?,17?-triol and 4?-fluoro-17?-ethynylandrost-5-ene-3?,17?-triol-7-one.

Abstract: A relatively specific trophoblastic-associated biomarker, HSD3B1, is disclosed. A method for screening gestational trophoblastic disease using HSD3B1 is also disclosed.

Abstract: A method for assessing muscle damage in a biological sample obtained from a subject is disclosed. The method involves obtaining a biological sample from a subject being assessed for muscle damage, and evaluating the sample for the presence or absence of a myofilament protein modification product. The method can also be used to assess the extent and/or type of muscle damage in a subject by studying the profile of myofilament protein modification products detected in the sample taken from the subject. The invention further provides a method for screening for an agent which modulates the level of a myofilament protein modification product present in a biological sample or for a calcium sensitizing agent. The invention is applicable to cardiac muscle and skeletal muscle.

Abstract: The present invention relates to a method for lowering activity with respect to maltose in glucose measurement comprising a step of reacting modified pyrroloquinoline quinone dependent glucose dehydrogenase subjected to amino acid sequence modification, wherein pyrroloquinoline quinone dependent glucose dehydrogenase is reacted in the presence of at least one type of substance selected from the group comprising succinic acid, malonic acid, glutaric acid, malic acid, phthalic acid, 2-ketoglutaric acid, 3,3-dimethylglutaric acid, pimeric acid, suberic acid, adipic acid, maleic acid, potassium chloride, ammonium chloride, diammonium hydrogen citrate, L-lysine, taurine, calcium glycerate, amino-n-butyric acid, sodium glycolate, sodium ?-ketoglutarate, fumaric acid, glycine, glutamic acid, serine and citric acid.

Abstract: An assay for assessing the permeability of mitochondrial membranes is provided. The assay is based on enzymatic reactions that produce a detectable signal in the presence of intra-mitochondrial compounds. In some embodiments, the intra-mitochondrial compounds may be reduced and/or oxidized nicotinamide adenine dinucleotides (i.e., NADH and NAD+, respectively). The assay may optionally be provided in the form of a kit, and may further be useful in the discovery and interrogation of compounds that may serve to inhibit pathologic increase in mitochondrial membrane permeability, which is associated with numerous and varied mammalian disease.

Abstract: The present invention relates to a technique for measuring a glucose level by utilizing a reaction system containing an enzyme and an electron carrier. In accordance with the glucose level measuring method of the present invention, glucose dehydrogenase with cytochrome C attached thereto or glucose dehydrogenase derived from a microorganism belonging to a burkholderia genus is used as the enzyme, and a Ru compound is used as the electron carrier. The present invention further provides a glucose sensor in which glucose dehydrogenase with cytochrome C attached thereto or glucose dehydrogenase derived from derived from a microorganism belonging to a burkholderia genus is used as the enzyme, and a Ru compound is used as the electron carrier.

Abstract: The invention relates to a method and reagent system for detecting an analyte in a sample by means of an enzymatic reaction, involving the use of an enzyme-coenzyme complex as a stoichiometric reaction partner for the analyte present in the sample.

Abstract: A method of a pretreatment of a sample for quantitating cholesterol characterized by, before measuring cholesterol contained in specific lipoproteins, treating the sample containing lipoproteins with an enzyme, the substrate of which is free cholesterol, optionally together with a reaction accelerator; a method for quantitating cholesterol in specific lipoproteins by using the above method; and a kit for quantitating cholesterol in specific lipoproteins to be used in the above quantification method. By using this quantification method, cholesterol in a specific fraction can be conveniently, accurately and efficiently quantitated fundamentally without resort to polyanion, etc. Thus, this method is appropriately usable in various automatic analyzers.

Abstract: The present invention relates to a method of producing an optically active pyridineethanol derivative. More particularly, it relates to a method of producing an optically active polycyclic pyridineethanol derivative by causing an enzyme or enzyme source to act on polycyclic acetylpyridine derivatives. The present invention also relates to a novel enzyme which can be used in the production method mentioned above, a DNA coding for said enzyme, a recombinant vector having said DNA, and a transformant having said recombinant vector. The invention further relates to a method of producing an optically active polycyclic pyridineethanol derivative by causing the above novel enzyme or the above transformant to act on optically inactive polycyclic pyridineethanol derivatives.

Abstract: A method for the identification of a composition useful in the treatment of an overweight or obese person to reduce the person's body mass index is provided which comprises obtaining an extract of an ethnobotanical plant, and evaluating the activity of the extract in an assay. A composition is described for treating patient conditions made up of C. grandis standardized extract (AdipoCleave™) with cucurbitacins B and D as active ingredients, Cephalandrol, Cephalandrine A and B, and like compounds and cucurbitacins. The composition is effective to maintain normal blood sugar and normal levels of non-enzymatic protein glycosilation.

Abstract: Stabilized aqueous solution of a coenzyme for hydrogen-transferring enzymes characterized in that the solution contains NAD, NADP or a derivative thereof in an oxidized or reduced form and one or several organic compounds or salts thereof having a pKa value between 1.5 and 6.0 and/or a nitrogen compound of the general formula in which the residues R1, R2 and R3 are the same or different and denote hydrogen, or a saturated or unsaturated alkyl or aryl group as well as the use of the solution to determine dehydrogenases in particular lactate dehydrogenase or substrates thereof.

Abstract: Mediator stabilized reagent compositions and methods for their use in electrochemical analyte determination assays are provided. The subject reagent compositions include an enzyme, a redox mediator and a mediator-stabilizing buffer. Optionally, the reagent compositions may further include one or more of a wetting agent, detergent, enzyme cofactor and combinations thereof. Also provided are electrochemical test strips that include the subject reagent compositions, systems and kits that include the same as well as methods for using the same in analyte detection assays. The subject invention finds use in a variety of different applications, including glucose concentration determination applications.

Abstract: Disclosed are a method and a corresponding kit for determining the cytotoxicity of a test agent. The method includes the steps of adding a pre-determined amount of the test agent to a vessel containing living cells in culture medium. The cells are then incubated for a pre-determined amount of time. To the cells and culture medium is then added a reagent mixture that is non-toxic to the living cells. In the preferred embodiment, the reagent mixture contains a solvent, a dye, an electron transfer agent, a substrate for a cytoplasmic enzyme having a half-life greater than two hours and NAD+. The contents of the vessel are then measured for production of the reduced state of the dye, the production of the reduced state of the dye being caused by a cytoplasmic enzyme specific for the substrate in the reagent mixture. In the preferred embodiment, the production of the reduced state of the dye is caused by lactate dehydrogenase released from dead and dying cells within the vessel.

Abstract: The invention concerns novel enzymes having an arogenate dehydrogenase activity, in particular arogenate dehydrogenase enzymes of plants, and the genes encoding said enzymes. The inventive arogenate dehydrogenase enzymes catalyze the last stage of the metabolic pathway of tyrosine biosynthesis, and constitute, as such, potential targets of herbicides. Hence the invention also concerns a method for identifying herbicide compounds targeting said enzymes, said herbicide compounds preventing tyrosine biosynthesis by being fixed on said enzymes. The invention further concerns transgenic plants tolerant to herbicide compounds targeting an enzyme involved in the tyrosine biosynthesis pathway, in particular an enzyme involved in the transformation of L-tyrosine prephenate, in particular an arogenate dehydrogenase enzyme.

Abstract: The invention relates to polypeptides from phytopathogenic fungi with the biological activity of an inosine monophosphate dehydrogenase, to nucleic acids encoding them, to the use of the polypeptides and nucleic acids for identifying modulators of the polypeptides, to methods for identifying such modulators, and to the use of these modulators as fungicides.

Abstract: Methods for rapidly identifying drug candidates that can bind to an enzyme at both a common ligand site and a specificity ligand site, resulting in high affinity binding. The bi-ligand drug candidates are screened from a focused combinatorial library where the specific points of variation on a core structure are optimized. The optimal points of variation are identified by which atoms of a ligand bound to the common ligand site are identified to be proximal to the specificity ligand site. As a result, the atoms proximal to the specificity ligand site can then be used as a point for variation to generate a focused combinatorial library of high affinity drug candidates that can bind to both the common ligand site and the specificity ligand site. Different candidates in the library can then have high affinity for many related enzymes sharing a similar common ligand site.

Abstract: Disclosed is a modified glucose dehydrogenase having pyrroloquinoline quinone as a coenzyme, wherein one or more amino acid residues in a region of amino acid 349-377 of water-soluble PQQGDH derived from Acinetobacter calcoaceticus is replaced with other amino acid residues and has an inhibition constant (Ksi) of 200 mM or more. The modified water-soluble PQQGDH of the invention can be utilized for measuring glucose levels in the presence of high concentrations of glucose because of the low substrate inhibition by glucose.

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: A probe, kit comprising the probe, and methods for using the probe are provided where the probe comprises: wherein DM is a detectable marker; and L is a straight or branched chain moiety providing between 1 and 20 atom separation between DM and the ring atom to which DM is attached.

Abstract: Methods for detecting a malaria parasite are described that include: releasing the malaria parasite by allowing an erythrocyte in a specimen to undergo hemolysis; staining the released malaria parasite with a fluorescent dye to prepare a measurement sample; detecting optical information from the measurement sample; and distinguishing the malaria parasite based on the optical information obtained. Apparatuses and reagent kits are also described.

Abstract: This invention concerns a method for enzymatic preparation of 6-O-?-D-glucopyranosyl-D-sorbitol (1,6-GPS) from isomaltulose. It is specified that the isomaltulose is put into an aqueous reaction solution containing a unit having sorbitol dehydrogenase (SDH) activity, incubated, and then 1,6-GPS is recovered from the reaction solution.

Abstract: This invention provides (R)-2-octanol dehydrogenase that catalyzes oxidation-reduction reaction using NAD+ (NADH) as a coenzyme and the genes that encodes them. The enzymes of this invention can be obtained from microorganisms such as the genera Pichia, Candida, and Ogataea, and so on. It is possible to produce alcohols, in particular, alcohols such as (S)-4-halo-3-hydroxybutyric acid esters and (R)-propoxybenzene derivatives by reducing ketones with this (R)-2-octanol dehydrogenase. Moreover, the (R)-2-octanol dehydrogenase of this invention is excellent in activity and stereoselectivity.

Abstract: The present invention provides a screening method by which a prediabetic state can be more accurately determined and many test samples can be treated without physical burden on patients, and a diagnostic reagent used for it. By measuring a D-mannose concentration, preferably a D-mannose concentration and a glucose concentration, in a body fluid collected from a subject, and comparing them with their respective standard values, a patient in a prediabetic state can be detected. Further, the measurement of the mannose concentration is conducted by allowing an enzyme having an oxidizing activity on mannose by dehydrogenation to react on the mannose in a sample in the presence of an electron acceptor, and quantitatively determining the formed reductant of the electron acceptor.

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: Isolated PGE synthase, provided from encoding nucleic acid. Methods of production and use. Assays for modulators, especially inhibitors, of PGE synthase activity.

Abstract: A reagent for measuring an alanine aminotransferase activity comprising L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, and reduced nicotinamide adenine dinucleotide, characterized by further comprising a substance having an activity of inhibiting lactate dehydrogenase activity is disclosed. Further, a method for measuring an alanine aminotransferase activity, characterized by bringing a sample to be analyzed, which may contain alanine aminotransferase, into contact with L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, reduced nicotinamide adenine dinucleotide, and a substance having an activity of inhibiting a lactate dehydrogenase activity is disclosed. According to the reagent and method, an increase in the reagent blank reaction, i.e., an increase in the initial absorbance, can be suppressed.

Abstract: The cDNA sequence of the human 9-cis-retinol dehydrogenase enzyme is disclosed.

Abstract: The present invention discloses a method of evaluating the immunotoxicity of a hydrophobic test substance, comprising (1) culturing immunocompetent cells in a culture system containing a hydrophobic test substance and a surfactant, (2) obtaining a survival rate of the immunocompetent cells after the culturing (1), and (3) evaluating the immunotoxicity of the hydrophobic test substance from the survival rate obtained in (2).

Abstract: The object of the invention is to provide a handy type visually determinable ATP measuring reagent which can be easily handled and has good measuring sensitivity. The invention is an ATP measuring reagent which rendered possible visual determination by amplifying the reaction to form glucose 6-phosphate from glucose through the joint use of ATP contained in a sample and acetyl phosphate contained in the reagent with glucokinase and acetate kinase contained in the reagent, and further leading it to a visualizable color reaction by the combination of glucose-6-phosphate dehydrogenase and diaphorase.

Abstract: This invention relates to a dry analytical element using a water-soluble indicator which improves measuring accuracy, which comprises a water-impermeable support, a water-permeable layer and a spreading layer having a function to spread a liquid uniformly, wherein a water-soluble indicator for colorimetry is incorporated into the s preading layer.

Abstract: Modified water-soluble glucose dehydrogenase having pyrrolo-quinoline quinone as a coenzyme are provided wherein at least one amino acid residue is replaced by another amino acid residue in a specific region. Modified water-soluble PQQGDHs of the present invention have improved thermal stability.

Abstract: Graphitic nanotubes, which include tubular fullerenes (commonly called “buckytubes”) and fibrils, which are functionalized by chemical substitution, are used as solid supports in electrogenerated chemiluminescence assays. The graphitic nanotubes are chemically modified with functional group biomolecules prior to use in an assay. Association of electrochemiluminescent ruthenium complexes with the functional group biomolecule-modified nanotubes permits detection of molecules including nucleic acids, antigens, enzymes, and enzyme substrates by multiple formats.

Abstract: Disclosed are methods for suppressing the induction of inducible nitric oxide synthase in a cell comprising contacting said cell with an effective amount of at least one induction suppressor of inducible nitric oxide synthase or a cytokine are disclosed. The induction suppressor can be an inhibitor of mevalonate synthesis, an inhibitor of the farnesylation of Ras, an antioxidant, an enhancer of intracellular cAMP, an enhancer of protein kinase A (PKA), an inhibitor of NF-k? activation, an inhibitor of Ras/Raf/MAP kinase pathway, an inhibitor of mevalonate pyrophosphate decarboxylase or an inhibitor of farnesyl pyrophosphate.

Abstract: Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. A method to detect proteins which contain nitrosothiols involves several steps. Nitrosylated cysteines are converted to tagged cysteines. Tagged proteins can then be detected, for example, by immunoblotting and/or can be purified by affinity chromatography. The method is applicable to the detection of S-nitrosylated proteins in cell lysates following in vitro S-nitrosylation, as well as to the detection of endogenous S-nitrosothiols in selected protein substrates.

Abstract: The present invention relates, in general, to stem cells, and in particular, to a method of isolating stem cells and to reagents suitable for use in such a method. The invention further relates to stem cell populations isolatable in accordance with the present method.

Abstract: The present invention relates to a method of performing concentration determinations of aqueous solutions of polyaspartic acid and/or salts thereof.

Abstract: There is provided a method for a selective assay of component, particularly cholesterol, in very low-density lipoprotein (VLDL) which is one of serum lipoproteins. In the assay, an enzymatic reaction of lipoprotein lipase (LPL) or cholesterol esterase (CE) which well reacts with high-density lipoprotein (HDL) and VLDL is carried out at least in the presence of calixarene or a salt thereof. It is also carried out in the presence of one or more substance(s) selected from albumin and basic amino acids in addition to calixarene or a salt thereof.

Abstract: A biphasic process for rapid screening of organic reactions comprising monitoring relative rates of parallel organic reactions. The screening process is suitable to determine the efficacy of different reactants, process conditions, and process enhancers such as catalysts or promoters. The biphasic process also allows multiple samples to be analyzed/monitored simultaneously. In addition because enzymes are used to monitor the reaction product in this invention, when that product is chiral and an enantio-discriminating enzyme is used to monitor the product, in addition to the relative rates, enantioselectivities of a set of parallel organic reactions can also be determined.

Abstract: A method of producing coryneform bacteria having improved amino acid or nucleic acid productivity comprising the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence, or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and selecting a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows the construction of a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield by recombination or mutation.

Abstract: Stabilized tetrazolium dye-phenazine reagent compositions and methods for their use in the measurement of an analyte in a sample are provided. The subject reagent compositions include: (1) a tetrazolium dye component, e.g., a water soluble tetrazolium salt; (2) a phenazine component; and (3) an effective amount of one or more tetrazolium dye-phenazine stabilizing reagents, e.g., an inorganic Group IIIA compound and/or a flavin. In many embodiments, the subject reagent compositions include additional members of an analyte oxidizing signal producing system, such as: an analyte oxidizing enzyme, e.g., an analyte dehydrogenase or an analyte oxidase; and an enzyme cofactor. Also provided are test strips that include the subject reagent compositions, as well as systems and kits incorporating the subject test strips. The subject reagent compositions, test strips, systems and kits find use in the detection of a wide variety of analytes in a sample, such as a physiological sample, e.g.

Abstract: The present invention relates to a test piece having a simple structure, by which HDL cholesterol can be measured easily using a small amount of specimen. A reagent layer (2) is formed on a support (1), and an enzyme reagent for measuring cholesterol, a first surfactant that makes a solubility of a high-density lipoprotein (HDL) higher than that of a lipoprotein other than the HDL, and a second surfactant that inhibits the lipoprotein other than the HDL from dissolving are contained in the reagent layer (2).

Abstract: The present invention relates to a method and a means of diagnosing Candida infection. In particular, the present invention relates to a method of diagnosing Candida infection comprising the steps of obtaining a biological sample from a subject at risk of, or suspected to be suffering from, Candida infection, contacting the sample with a mannose depleted antigen composition comprising a soluble cytoplasmic antigen preparation comprising antigens of molecular weight 55 kDa, 30 kDa and 20 kDa and detecting the antigen/antibody reaction.

Abstract: An objective of the present invention is to provide a D-phenyllactic acid dehydrogenase. Another objective of the present invention is to provide a gene which encodes the D-phenyllactic acid dehydrogenase. A D-phenyllactic acid dehydrogenase according to the present invention is a protein comprising an amino acid sequence of the amino acid sequence of SEQ ID NO: 2 or a modified amino acid sequence of the amino acid sequence of SEQ ID NO: 2 that has one or more modifications selected from a substitution, a deletion, an addition and an insertion and has D-phenyllactic acid dehydrogenase activity. A D-phenyllactic acid dehydrogenase gene according to the present invention comprises a nucleotide sequence which encodes the D-phenyllactic acid dehydrogenase, for example the nucleotide sequence of SEQ ID NO: 1.

Abstract: The present invention provides methods for analyzing proteomes, as cells or lysates. The analysis is based on the use of probes that have specificity to the active form of proteins, particularly enzymes and receptors. The probes can be identified in different ways. In accordance with the present invention, a method is provided for generating and screening compound libraries that are used for the identification of lead molecules, and for the parallel identification of their biological targets. By appending specific functionalities and/or groups to one or more binding moieties, the reactive functionalities gain binding affinity and specificity for particular proteins and classes of proteins. Such libraries of candidate compounds, referred to herein as activity-based probes, or ABPs, are used to screen for one or more desired biological activities or target proteins.

Abstract: The present invention provides novel glutathione-dependent formaldehyde dehydrogenase that makes possible quantitative measurement of formaldehyde by cycling reaction, and a determination method of formaldehyde and biological components, such as creatinine, creatine, and homocysteine, which produces formaldehyde as a reaction intermediate. In addition, the present invention provides a reagent kit for the above-mentioned determination method. The present invention provides a novel determination method of a homocysteine using transferase utilizing homocysteine and other compound as a pair of substrates. Particularly, the present invention provides a determination method of homocysteine which includes bringing betaine-homocysteine methyltransferase and dimethylglycine oxidase into contact with a sample and measuring produced hydrogen peroxide or formaldehyde. Moreover, the present invention provides novel dimethylglycine oxidase stable to thiol compound, which is suitably used for the measurement.

Abstract: Electrochemiluminescent enzymes, their preparation and use as biosensors are disclosed. Specifically, two appendages are covalently attached to a desired dehydrogenase enzyme; (1) a nicotinamide adenine cofactor or analog thereof, and (2) a luminescent ruthenium complex. For example, glucose concentrations is the following way. A doubly-modified glucose dehydrogenase could oxidize glucose with concomitant reduction of the attached NAD+ to NADH. Because NADH, but not NAD+, is able to interact with surface ruthenium to promote ECL, only enzyme molecules that have reacted with glucose will emit light from their ruthenium label in an ECL instrument. The relative close proximity of NADH and ruthenium on the enzyme surface enhances light emission as compared to the same concentrations in free solution. When NADH reduces ruthenium, it returns to become NAD+, permitting multiple cycles of ECL light emission from a single enzyme molecule. Such biosensors can be used in solution or bound to a solid surface.

Abstract: Here we describe the molecular identification of a cDNA encoding a novel serine protease we have termed protease C-E. The deduced amino acid sequence, and it alignment with other well-characterized serine proteases indicates that it is a member of the S1 serine protease family. We have found that the protease C-E mRNA is expressed in pancreas, placenta, prostate, small intestine, stomach, spleen, fibroblasts and epidermis, as well as in certain regions of the brain i.e., cerebellum, cerebral cortex, pituitary and hippocampus. Enzymatically active protease C-E, as produced using the methodologies described herein, is amenable to further biochemical analyses for the identification of physiological substrates and specific modulators.

Abstract: This invention relates to an isolated nucleic acid fragment encoding a tetrahydrofolate metabolism enzyme. The invention also relates to the construction of a chimeric gene encoding all or a portion of the tetrahydrofolate metabolism enzyme, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the tetrahydrofolate metabolism enzyme in a transformed host cell.

Abstract: The present invention relates to polynucleotides that encode proteins having OpcA enzymatic activity. These polynucleotides can be used for increasing lysine biosynthesis in Coryneform glutamicum.

Abstract: The invention provides methods for identifying agonists and antagonists of FabK polypeptides.