Abstract: The invention relates to methods for identifying compounds which affect cellular stress. In particular, the method relates to identifying compounds which inhibit protein advanced glycation end product formation, where the compounds are carbonyl scavengers which inhibit the formation. The assay involves combing the substance of interest with histone H1 and ADP-ribose, and then measuring fluorescence and protein cross linking. Various inhibitors of protein AGE glycation have been identified, using this assay.

Abstract: A method of determining an analyte in a sample utilizing a redox reaction is provided, by which a highly reliable determined value can be obtained. In this method, a tetrazolium compound is added to a sample in the presence of a surfactant prior to a redox reaction so as to eliminate an effect of hemoglobin and a hemoglobin degradation product as reducing substances in the sample. Thereafter, a reducing or oxidizing substance derived from an analyte is produced and the amount thereof is determined by the redox reaction. Then, the amount of the analyte in the sample is determined based on the amount of the reducing or oxidizing substance thus determined. According to this method, cloudiness due to the coexistence with the hemoglobin can be prevented by the surfactant and thus an increase in the absorbance due to the cloudiness can be inhibited as shown in FIG. 1. As the surfactant, polyoxyethylene ether and the like can be used.

Abstract: The hypervariable region (E2HV) of the putative hepatitis C virus (HCV) glycoprotein E2/NS1, between about amino acid 384 to about amino acid 414, is a rapidly evolving region of HCV, and is likely to be under positive immune selection. A newly discovered motif within this hypervariable region is immunogenic and conserved with respect to the character of the amino acids. In many isolates, this motif falls between amino acids 401 to 406 or 407. The discovery of this motif allows for additional materials and methods to treat and diagnose HCV.

Abstract: The invention provides methods, compositions, apparatus, and a computer data retrieval system for conducting proteomics.

Abstract: An increased risk of a fetal chromosomal abnormality, for example, fetal Down syndrome can be detected by separating or discriminating &agr;-fetoproteins present in the body fluid of a pregnant woman, and measuring the proportion of one or more of the &agr;-fetoproteins which have a specific sugar chain structure, relative to the total &agr;-fetoproteins.

Abstract: A method for diagnosis of pre-eclampsia is disclosed, which comprises measuring the hormone inhibin A in a biological sample such as maternal serum. The method allows non-invasive, early diagnosis and can be used to predict the onset of secondary symptoms.

Abstract: With rotating and translating a rotation-translation drum installed in a vacuum chamber of a mass spectrometry interface to constitute a mass spectro-meter, a liquid sample incorporating a dissolved substance to be analyzed in its mass is emitted for the drum from a sample supplying nozzle of the mass spectrometry interface, and the dissolved substance is isolated as a spiral filament on the drum. Then, a laser beam is irradiated onto the filament from a laser source via a laser beam inlet situated at the vacuum chamber and thereby, the dissolved substance is ionized without exposing to the air.

Abstract: A method is provided for joining a microchip device to a capillary tube. The microchip device has a capillary channel opening onto an edge surface of the device. A short hole is drilled into the edge surface, aligned with the capillary channel. The drilling is done with a flat bottom, preferably by a two-step drilling process. Then, the end of the capillary can be inserted into the hole so that its end is substantially flush with the flat bottom of the hole, thereby eliminating dead volume. Testing has shown that this connection provides very little band broadening of samples transported through the capillary channel into the capillary tube. The tip of the capillary tube can be tapered, so that it is suitable for use as an electrospray source for a mass spectrometer.

Abstract: The present invention describes a variety of substituted and unsubstituted carbazolylvinyl dyes and their use for detecting and quantifying poly(amino acids), including peptides, polypeptides and proteins. The labeled proteins or peptides are highly colored, but are also detected by their strong fluorescence enhancement. Poly(amino acids) are detected in solution, in electrophoretic gels, and on solid supports, including blots and dipsticks. The present method of staining is highly sensitive, extremely facile, and relatively non-selective and can be accomplished without the use of organic solvent additives.

Abstract: The invention relates to an assay method for assessing a transferrin variant or combination of transferrin variants for the diagnosis and monitoring of alcoholism, and to kits for performing the assay. In particular, the invention provides a method of producing an algorithm for determining the content of a transferrin variant or combination of transferrin variants, preferably a CDT variant or combination of CDT variants, in a sample of body fluid, said method comprising: (a) obtaining at least two solutions each having known contents of asialo- (A1, A2, A3, etc.) and disialo-transferrins (D1, D2, D3, etc.); (b) determining the content in each of said solutions of a transferrin variant or combination of transferrin variants fractions substantially free from tri- and higher sialylated transferring; (c) determining the total transferrin variant content (T1, T2, T2, etc.

Abstract: The full amino acid and DNA sequences of placental protein 13 (PP13) are disclosed. Also described are various PP13 derived peptide fragments, and a recombinant method for the production of PP13. PP13 may be used in a screening and a diagnostic method for pregnancy-related complications.

Abstract: There are provided methods for assaying biological specimens for one or more of leptin, prorenin or renin in order to provide predictive information about the likelihood of a woman carrying a Downs Syndrome affected fetus.

Abstract: A method and kit of diagnosing endometriosis in a female patient suspected of having endometriosis is disclosed. The method includes obtaining a sample from the patient. The sample is analyzed to detect the presence of ENDO -I glycoprotein or its mRNA in the sample compared to non-endometriosis controls who do not express ENDO-I. The protein is characterized by (i) a molecular weight of 40,000 to 55,000 as determined by two-dimensional SDS-PAGE polyacrylamide gel electrophoresis; (ii) having an isoelectric point of 4.0 to 5.5; and (iii) being synthesized and secreted specifically by stromal cells of endometriotic tissue origin; and (iv) in humans having a cDNA as set forth in SEQ ID No:1. The present invention further discloses a cDNA for human ENDO-I (SEQ ID No:1) and antibody directed against ENDO-I.

Abstract: Disclosed is a method for determining alcohol intake or alcohol induced liver damage in a subject by quantifying the content of sialic acid in apolipoprotein J (Apo J). In particular the present invention involves the steps of (a) providing a sample containing Apo J from a subject; (b) purifying the Apo J from the sample; (c) determining sialylation index, i.e., moles of sialic acid peer mole of Apo J in the sample; and (d) evaluating whether the sialylation index is an indication of alcohol intake or alcohol related liver damage recovery from alcohol addiction or alcohol relapse in the subject.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: The present invention relates to a rapid method for the detection of ischemic states and to a kit for use in such a method. Provided for is a rapid method of testing for the existence of and quantifying ischemia based upon method of detecting and quantifying the existence of an alteration of the serum protein albumin which occurs following an ischemic event; methods for detecting and quantifying this alteration include evaluating and quantifying the cobalt binding capacity of circulating blood, analysis and measurement of the ability of serum albumin to bind exogenous cobalt, detection and measurement of the presence of copper in a purified albumin sample and use of an immunological assay sepcific to the alterated form of serum albumin which occurs following an ischemic event.

Abstract: There are provided methods for assaying biological specimens for one or more of leptin, prorenin or renin in order to provide predictive information about the likelihood of a woman carrying a Downs Syndrome affected fetus.

Abstract: The present invention relates to methods and compositions for lipid matrix-assisted chemical ligation and synthesis of membrane polypeptides that are incorporated in a lipid matrix. The invention is exemplified in production of a prefolded membrane polypeptide embedded within a lipid matrix via stepwise chemoselective chemical ligation of unprotected peptide segments, where at least one peptide segment is embedded in a lipid matrix. Any chemoselective reaction chemistry amenable for ligation of unprotected peptide segments can be employed. Suitable lipid matrices include liposomes, micelles, cell membrane patches and optically isotropic cubic lipidic phase matrices. Prefolded synthetic and semi-synthetic membrane polypeptides synthesized according to the methods and compositions of the invention also permit site-specific incorporation of one or more detectable moieties, such as a chromophore, which can be conveniently introduced during synthesis.

Abstract: The present invention relates to a method of identifying a molecule of a molecule-substrate complex, wherein the molecule is covalently attached directly to a substrate or indirectly by means of a linking moiety, comprising: (a) bombarding the molecule-substrate complex with energized particles to cleave the molecule from the molecule-substrate complex; and (b) determining the molecular weight of the cleaved molecule by means of mass spectrometry. The inventive method may further comprise irradiating the cleaved molecule with photons.

Abstract: A prenatal screening method for Down's syndrome involves assaying for hyperglycosylated gonadotropin in biological test samples such as urine, plasma or serum obtained from pregnant women. Hyperglycosylated gonadotropin comprises a variant population of chorionic gonadotropin, chorionic gonadotropin-free &bgr;-subunit, &bgr;-core fragment, and/or free &agr;-subunit exhibiting differences in the carbohydrate content from what is observed in samples obtained from pregnant women carrying normal fetuses. Qualitative or quantitative observation of differences in the carbohydrate content of the hyperglycosylated gonadotropin population from corresponding control samples containing a normal gonadotropin population, or direct observation of the variant species seen in Down's syndrome, indicates that the woman's fetus has Down's syndrome. Typical screens involve carbohydrate analyses, immunoassays, or combinations of these methods.

Abstract: The present invention provides an improved method for using the measurement of total urinary 17 KS-S, referred to herein as the urinary Anabolic/Catabolic Index (ACI), as part of a biomarker for biological age.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: Low molecular weight peptide derivatives which are able to act as inhibitors of the interaction between laminin and nidogen (laminin/nidogen interaction), a process for their preparation, pharmaceutical compositions prepared therefrom and their use for preparing pharmaceuticals and for identifying inhibitors of the laminin/nidogen interaction.

Abstract: Compositions and methods for determining the presence or concentration of an analyte in a sample by exposing the sample to an indicator molecule comprising a fluorescent lanthanide metal chelate complex. The presence or concentration of the analyte in the sample is determined by observing and/or measuring the change in intensity of fluorescence emitted by the lanthanide metal chelate complex upon binding of the analyte to one or more recognition elements in the complex. The fluorescent indicator molecules can be used in various types of fluorescent sensing devices and are useful in various fields, including energy, medicine and agriculture.

Abstract: The present invention relates to a method of identifying a molecule of a molecule-substrate complex, wherein the molecule is covalently attached directly to a substrate or indirectly by means of a linking moiety, comprising: (a) bombarding the molecule-substrate complex with energized particles to cleave the molecule from the molecule-substrate complex; and (b) determining the molecular weight of the cleaved molecule by means of mass spectrometry. The inventive method may further comprise irradiating the cleaved molecule with photons.

Abstract: The invention relates to the staining of poly(amino acids), including peptides, polypeptides and proteins in gels and on solid supports, using neutral or anionic complexes of transition metals.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: An isolated and purified glycoprotein designated PUP-1 and functional analogs thereof are disclosed and characterized by being a progesterone induced and estradiol inhibited secretory glycoprotein from stromal cells of endometrial origin; having an N-terminal amino acid sequence as set forth in SEQ ID No:1; having a molecular weight of 70,000 daltons and an isoelectric point of 5.7; and synthesized by endometrium at the time of fertilization, early embryogenesis, and implantation. The present invention further provides a method of determining endometrial receptivity, monitoring placental physiology during gestation, and monitoring the effects of protocols to induce ovarian hyperstimulation or ovulation induction on uterine receptivity by monitoring the cyclic presence of PUP-1 in bodily fluid and tissue samples.

Abstract: The present invention sterol-modified hedgehog polypeptides and functional fragments thereof. Methods of identifying compositions which affect hedgehog activity based on inhibition of cholesterol modification of hedgehog protein are described. In one aspect of the invention, the method provides a means for affecting cholesterol biosynthesis or transport in a cell comprising contacting a cell with an effective amount of a compound that affects hedgehog, thereby affecting cholesterol biosynthesis or transport. The effect may be inhibition or stimulation of cholesterol biosynthesis or transport.

Abstract: Sensitive methods for identifying compounds having biological activity comprising combining living cells with two fluorescent membrane permeable ionic dyes having the same charge sign, the first of which has an emission spectrum which overlaps the excitation spectrum of the second fluorescent membrane penetrative dye. The fluorescence is then induced by illuminating the dyes at a wavelength corresponding to the excitation spectrum of the first fluorescent dye and emission is then registered at a wavelength corresponding to the emission spectrum of the second fluorescent dye (FRET). The change in the FRET is indicative of a modulation of cell membrane potential by the biologically active compounds.

Abstract: A disposable high density optically readable polydeoxynucleotide array with integral fluorescence excitation and fluorescence emission channels is described. The compact array size allows integration into several types of interventional devices such as catheters, guidewires, needles, trocars and may be used intraoperatively. Highly sensitive monitoring of the metabolic and disease pathways of cells in vivo under varying chemical, genetic and environmental conditions is afforded.

Abstract: A method of screening an agent for potential use in the treatment of Alzheimer's disease, comprises reacting, in the presence of the agent, tau protein with a suitable sulphated carbohydrate under appropriate conditions to form filaments, and monitoring for the presence of filaments. Tau protein and sulphated carbohydrate, e.g. sulphated glycosaminoglycan, will react under appropriate conditions to form filaments, either paired helical filaments or straight filaments. If filament formation is affected when the reaction is carried out in the presence of an agent being screened, this is possibly due to an interfering, inhibiting or blocking effect of the agent. An agent which inhibits assembly of PHFs in vitro may also have an inhibiting effect in vivo and thus have potential therapeutic value in delaying the dementing effects of Alzheimer's disease. The invention can thus provide a screen to identify agents worthy of further investigation for use in the treatment of Alzheimer's disease.

Abstract: Secretin and secretin compositions are used for the treatment of autism and other neurological, behavioral and immunological disorders. The method includes administering an effective amount of secretin, such as Secretin-Ferring, to a patient. In one example, 2 clinical units (CU) of Secretin-Ferring was dissolved in a 7.5 ml solution of sodium chloride and was intravenously injected over 1 minute. In another example, secretin was administered transdermally by applying dimethyl sulfoxide (DMSO) to the patients skin and rubbing about 15 CU of Secretin-Ferring into the DMSO. Other methods and compositions for administering the effective amount of secretin include other transdermal carrier substances, such as gels, lotions, or patches; oral carriers, such as tablets, capsules, or lozenges; inhalation through the nose or mouth (e.g., as an aerosol); suppository forms of secretin and secretin compositions; and using acoustic waves to cause the secretin to penetrate the skin.

Abstract: There are provided methods for assaying biological specimens for one or more of leptin, prorenin or renin in order to provide predictive information about the likelihood of a woman carrying a Downs Syndrome affected fetus.

Abstract: Compositions and methods are provided for prophylactic or therapeutic treatment of balance impairments involving neuronal damage, loss, or degeneration, preferably of vestibular ganglion neurons, in an animal by administration of an effective amount of a trkB or trkC agonist, particularly a neurotrophin, more preferably NT-4/5.

Abstract: The invention is directed to methods to assess connective tissue, especially bone, metabolism in disease or to monitor therapy, which method comprises assessing the levels of native free collagen-derived crosslinks in biological fluids, especially urine. The method can be enhanced by concomitantly determining the levels of an indicator of bone formation in biological fluids of the same individual and assessing the differences between the degradation marker and the formation indicator. Antibodies which are specifically immunoreactive with forms of crosslinks which occur free in biological fluids are also disclosed.

Abstract: The present invention involves a process for characterizing the glycosylation of glyco-proteins and an in vitro process for determining the bioavailability of glyco-proteins, based on the "hypothetical charge number" (or N) and usable for both endogenic and exogenic glyco-proteins.

Abstract: A method for the determination of an individual's alcohol consumption, characterized in comprising the steps: i) determining the level of sialic acid in a body fluid sample from the individual; ii) comparing the level found in step (i) with the level for the normal population, an increased level being an indication of an increased alcohol consumption relative to the consumption (per individual) of the normal population.

Abstract: The present invention provides a measuring method most suitable for measuring the concentrations of specific constituents, especially urine protein level and sugar level. After a protein-contained liquid sample is opacified by heating or while the sample is being heated, a light is projected to the liquid sample. The concentration of protein is determined from the intensity of light transmitted through the sample or scattered from the sample. In a urinalysis, an angle of rotation of the sample is measured before the sample is opacified, and in addition, intensity of the transmitted light or scattered light of the opacified urine is measured, whereby the urine sugar and protein levels are obtained.

Abstract: Methods of calculating the platelet count of an individual, by measuring the amount of a released platelet granule protein of interest in a sample of whole blood or of platelet-rich plasma from the individual, are described. The platelet granule protein of interest is either thrombospondin or .beta.-thromboglobulin. The amount of released platelet granule protein of interest in the whole blood sample or platelet-rich plasma sample is measured using an enzyme-linked immunosorbent assay; radioimmunoassay; sandwich assay; a quantitative immunochromatographic assay; or non-solid phase nephelometry. The platelet count is directly related to the amount of released platelet granule protein of interest in the sample, and can be determined from the amount of platelet granule protein of interest that is released from a known number of platelets.

Abstract: The present invention is directed to a single test system and method for determining the integrated glycemic condition of a subject by measuring the concentration of glucose and the level of protein-bound glucose in a subject's body fluid, such as whole blood. The glucose concentration is indicative of the subject's immediate glycemic condition, whereas the protein-bound glucose concentration is indicative of either intermediate or long-term glycemic condition. Optionally, other analytes indicative of glycemic condition, such as ketone bodies or fatty acid derivatives, can also be measured. The present invention also provides a method of diagnosing diabetes. The invention additionally provides a method for analyzing the concentration of fructosamine in less than or equal to five minutes without the use of a reaction accelerator.

Abstract: Dimeric inhibin concentration can be used in the determination of the risk of a women carrying an unborn child affected by chromosomal abnormalities, e.g. Downs Syndrome. A method for antenatal screening for chromosomal abnormalities is carried out by measuring body fluid taken from a pregnant woman for the level of at least one marker or precursor or metabolite of the marker together with the gestational age of the woman are compared with reference values taken from women carrying fetuses with chromosomal abnormalities and/or women carrying normal fetuses, using dimeric inhibin as a marker. A second marker is the free beta subunit of human chorion gonadotropin. An apparatus for carrying out the method described comprises a computer with a means for comparing the values described.

Abstract: A method of determining intracellular levels of water-soluble and fat-soluble antioxidants. Peripheral blood mononuclear cell lysates are prepared from an individual and analyzed by HPLC, either directly or after extraction with organic solvents to extract fat-soluble antioxidants. The levels of antioxidants are compared to normal levels as an indication of the overall oxidative health of the individual.

Abstract: A method for continuous separation and analysis of glycated and non-glycated proteins in a blood sample by HPLC using a phenylboronic acid resin and a polyol as eluant.

Abstract: A novel relationship between pancreatico-biliary secretion and autistic syndrome is disclosed. This relationship enables a novel therapy for the treatment of the symptoms of autistic syndromes, comprising the administration of a therapeutically effective, preferably intravenous, dose of secretin to an individual with autistic syndrome. The relationship further enables a differential diagnosis for autistic syndrome, comprising an analysis of an individual's blood and/or intestinal tissue for the presence of secretin and comparison of the level of secretin to known norms.

Abstract: A method for correlating a peptide fragment mass spectrum with amino acid sequences derived from a database is provided. A peptide is analyzed by a tandem mass spectrometer to yield a peptide fragment mass spectrum. A protein sequence database or a nucleotide sequence database is used to predict one or more fragment spectra for comparison with the experimentally-derived fragment spectrum. In one embodiment, sub-sequences of the sequences found on the database which define a peptide having a mass substantially equal to the mass of the peptide analyzed by the tandem mass spectrometer are identified as candidate sequences. For each candidate sequence, a plurality of fragments of the sequence are identified and the masses and m/z ratios of the fragments are predicted and used to form a predicted mass spectrum.

Abstract: A method for antenatal screening for chromosomal abnormalities in which maternal blood from a pregnant woman is measured for levels of free beta hCG and at least a second serum marker and/or precursors and metabolites of these markers and the measured levels of these markers together with the gestational age of the pregnant woman are compared to reference values at various gestational ages of the levels for free beta hCG and the second serum marker in (a) pregnant women carrying foetuses having the abnormalitie(s) subject to the screen and (b) pregnant women carrying normal foetuses, the comparison being indicative of the risk of the pregnant woman carrying a foetus with an abnormality subject to the screen characterised in that the second serum marker is pregnancy associated plasma protein A (PAPPA) and the screen is carried out by the end of the thirteenth (13th) completed week of pregnancy.

Abstract: A sample presentation device, with a surface-bound complex including at least one molecule which chemically modifies a biomolecule, is prepared and exposed to a biomolecule. The molecular weights of the chemically modified biomolecule is then determined by mass spectrometry.

Abstract: Elevated levels of Inhibin-A in maternal serum or plasma have been shown to indicate the presence of Down's Syndrome. The assay method comprises the use of a monoclonal antibody specific for at least part of the inhibin-A .beta. sub-unit (.beta.A), and another monoclonal antibody specific for at least part of the Inhibin-A .alpha. sub-unit. The .beta.A antibody is used to capture Inhibin-A from the test sample, and the .alpha. sub-unit antibody is used as the detection antibody and is linked to a detectable marker. The method is carried out in the first or second trimester as a screening test to select patients for subsequent diagnostic testing.

Abstract: The invention provides a kit for use in assaying protein, which comprises:an alkaline copper solution containing a tartrate; anda solution of Folin reagent.An alkaline copper solution containing tartrate has surprisingly been found to remain stable for several months. This allows a kit to be provided for use in a method of protein assay, in which the alkaline copper solution is provided ready-made up for immediate use in the method.