Abstract: The invention relates to a method of determining the presence or absence of a target microbe in a liquid water sample, the method consisting of a) combining a liquid water sample and a powdered medium having at least one nutrient indicator, the nutrient indicator being selected from the group consisting of: orthonitrophenyl-B-D-glucuronide; B-napthalamide-B-D-glucuronide; &agr;-napthol-B-D-glucuronide; and methylumbelliferyl-B-D-glucuronide, to form a mixture; b) incubating the mixture for a time sufficient to produce a detectable color signal indicative of the presence or absence of a target microbe; and c) observing the signal to determine the presence or absence of the target microbe in said sample.

Abstract: The present invention generally provides a method of measuring the biological activity of Coagulation Factor VIIa (Factor VIIa). Specifically, the present invention provides a method for directly measuring the activity of Factor VIIa in a plasma sample. More specifically, the present invention provides a method for utilizing Factor VIIa activity as a bio-marker to monitor Factor VIIa inhibition to screen compounds which are able to inhibit the biological activity of Factor VIIa.

Abstract: A colloidal system for detection of a variety of analytes involves techniques which permit reconstitution of a desiccated substance such as for surface enhanced Raman spectroscopic analysis and multiple sensors at once, each having different spectra through the use of markers or the like. Competitive assay techniques and a variety of substances are explained to permit a practical an versatile system which can also be used for immunological assays and can include antibodies tagged to provide spectroscopic indicia.

Abstract: A solid growth plating medium in which Shigella organisms will grow and form colonies in the medium, and substantially, other microorganisms are inhibited or their colonies are differentiated from Shigella organisms. In one embodiment of the invention, colonies produced by Shigella appear with the color of the plating medium, usually a clear off-white color, that can be readily observed. In another embodiment, the fact that Shigella boydii and Shigella sonnei produce the enzyme alpha-galactosidase, but most Shigella dysenteriae and Shigella flexneri strains do not, is utilized with a chromogenic substrate to produce colonies of these microorganisms of a distinguishing color.

Abstract: The invention provides chemiluminescent assays that incorporate a film including at least one chemiluminescent precursor immobilized therewith which produces a triggerable chemiluminescent compound, the film being free of compounds which generate singlet oxygen and being adapted for use with a sensitizer-labeled agent or agent probative of the analyte.

Abstract: The present invention provides methods and apparatus for purifying metabolites of interest and conducting metabolic analyses. The methods generally involve determining metabolic flux values for a plurality of target analytes by monitoring the relative isotope abundance of a stable isotope in a substrate labeled with the stable isotope and/or one or more target metabolites formed through metabolism of the labeled substrate. Certain methods utilize multiple electrophoretic methods to separate the target analytes from other components within the sample being analyzed. The methods can be used in a variety of applications including screens to identify metabolites that are correlated with certain diseases and diagnostic screens for identifying individuals having, or susceptible to, a disease.

Abstract: A method for monitoring the progress of fat loss in a patient during a weight loss program which comprises, contacting a body fluid sample from said patient with a solid test strip to provide a color indication of the presence in said body fluid of &bgr;-hydroxybutyrate, optionally together with acetoacetate and/or acetone.

Abstract: One aspect of the present invention is a reagent test strip that includes: a) a top support layer including a sample aperture; b) a membrane that includes a reagent system for indicating the concentration of a target; c) a spreading layer; d) a bottom support layer including a measuring port in substantial alignment or approximate alignment with the sample aperture. Preferably, the membrane is affixed to the top support layer. Preferably, the membrane is positioned between the top support layer and the bottom support layer. Preferably, the spreading layer is positioned above the top support layer and in substantial or approximate alignment with the sample aperture. Preferably, a fluid applied to the spreading layer, passes through the sample aperture and contacts the membrane.

Abstract: A method for diagnosis and treatment of arteriosclerotic lesions is provided wherein the method is characterized by introducing a chemical compound to the patient, the compound being a complex of a photosensitive portion, and a radioactive portion. Cells which exhibit an affinity for the porphyrin element indicate sites of plaque buildup. The radioactive portion within the compound allows tomographic scanning as well as simultaneous radiation treatment. The complexed compound can be introduced to the patient a desired number of times to provide the necessary radiation treatment and ongoing monitoring of plaque removal. Further observation or treatment may be conducted through a fluorescence guided endoscopic procedure.

Abstract: A method of detecting cardiac ischemia by detecting elevated levels of serum free fatty acids in serum unbound to serum albumin (FFAU) compared to an average FFAU level in individuals without cardiac ischemia, wherein the detection method uses a free fatty acid binding protein derivatized with a fluorescent moiety, is disclosed.

Abstract: A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Abstract: Disclosed are a spectrofluorimetrically detectable luminescent composition and processes for enhancing the luminescence of one or more lanthanide-containing macrocycles. The luminescent composition comprises a micelle-producing amount of at least one surfactant, at least one energy transfer acceptor lanthanide element macrocycle compound having an emission spectrum peak in the range from 500 to 950 nanometers, and a luminescence-enhancing amount of at least one energy transfer donor compound of yttrium or a 3-valent lanthanide element having atomic number 59-71, provided that the lanthanide element of said macrocycle compound and the lanthanide element of said energy transfer donor compound are not identical. The addition of gadolinium(III) in the presence of other solutes to both the prototype and the di-functionalized europium, samarium, and terbium macrocyclic complexes, which were taught in our U.S. Pat. Nos. 5,373,093 and 5,696,240, enhances their luminescence.

Abstract: A cancer screening method is provided, wherein the method is characterized by introducing a chemical compound to a patient, the compound being a complex of a fluorescent marker and a radioactive marker. Cells are then collected preferably through non-invasive or minimally invasive means. If fluorescence is observed in the exfoliated cells, tomographic scanning is conducted to further locate and/or confirm suspected malignant areas or metastatic areas. Further observation or treatment may be conducted either through fluorescence guided endoscopy, photo-dynamic therapy, and/or radiation treatment.

Abstract: The present invention relates to non-radioactive enzymatic methods for detecting Sphingosine-1-Phosphate (S1P) in biological fluids. The present invention further relates to a method of detecting the presence of cancer in a patient by the use of these and other methods of detecting S1P in biological samples from a patient.

Abstract: The present invention relates to a method for the selected chemical activation of photo-activatable cross-linker molecules around ligand binding pockets and fluorescent groups in macromolecules, especially biological macromolecules, by using fluorescent ligands of the macromolecule and by selecting photo-activatable cross-linker molecules having specific activation energies so that a radiationless energy transfer (Förster transfer) from the fluorescent ligands to the cross-linker molecules, which become activated thereby, takes place.

Abstract: The instant invention provides methods for identifying agents that modulate an enzymatic activity of a carbohydrate sulfotransferase. The methods generally involve contacting the sulfotransferase, in a reaction solution, with a sulfate donor, a test agent, and a polymeric sulfate acceptor that is readily separated from the reaction solution. Determination of an effect of the test agent on the sulfotransferase is by detecting the amount of sulfate in the polymeric sulfate acceptor that has been separated from the reaction solution. The invention further provides kits for use in carrying out the subject methods.

Abstract: The invention relates to methods for diagnosing Alzheimer's disease from a blood specimen. The method involves incubating a blood sample with a fluorescent probe that interacts with either protein kinase C (PKC) or protein kinase A (PKA), and measuring the fluorescence intensity in different conditions. Variation in the emission spectra data relative to those of the spectra obtained with healthy subjects is a discriminatory factor vis-à-vis Alzheimer's disease. The probes may recognize the catalytic site or the regulator domain of PKC or PKA, and may be coupled to a synthetic organic compound such as bis-indolmaleimide. In one aspect, the probes are lipidic, including probes prepared from phospholipid constituents of a cell membrane. In another aspect, the probes are proteinic or peptide probes of a membrane or cytoskeletal nature. Useful probes include derivatives of substrates for PKC or PKA.

Abstract: Methods, compositions and articles of manufacture for assaying a sample for a GHB source are provided. A sample suspected of containing a GHB source is contacted with a first oxidoreductase selective for GHB and an oxidized cofactor. In the presence of GHB in the sample, the first oxidoreductase oxidizes GHB to succinic semialdehyde and reduces the cofactor. The reduced cofactor thus produced can be detected directly, or a hydride abstractor can be used that abstracts a hydride from the reduced cofactor and produces a detectable change. The hydride abstractor can be a second oxidoreductase that oxidizes the reduced cofactor and produces a detectable change in a chromogen or dye. Preferably a visual change is produced, allowing performance of the assay outside of a laboratory setting. Fusion proteins comprising the first oxidoreductase, polynucleotides encoding such proteins, host cells expressing such proteins, and vectors comprising such polynucleotides are also provided.

Abstract: Methods for monitoring and/or controlling biofouling in membrane separation systems are provided. The present invention utilizes measurable amounts of fluorogenic agent(s) added to a feed stream to monitor and/or control the level of microbial growth during membrane separation and, thus, the purification of such feed stream during membrane separation.

Abstract: An apparatus for use in carrying out a microbiological assay on samples in a petri plate has a support for holding the petri plate, a camera, and a plurality of diffuse-white-light emitting diodes. The camera optically detects microbe-growth inhibition zones arising about the diffusion disks and is disposed above the support. The diodes are disposed in a circular array above the support and below the camera for illuminating the nutrient medium and the drug diffusion disks. An electrical power and control circuit energizes the diodes with power from the computer upon an activation of the camera.

Abstract: A method of analyzing the concentration of soluble analyte in a sample involves performing a competition assay using a predetermined amount of formed bodies to which are attached at least one analyte, varying known concentrations of an unlabeled ligand that binds to analyte, and a known concentration of ligand labeled with a detectable marker. After analysis in a flow cytometer, the test sample and a plurality of control samples generate data on curves formed by plotting signal vs. concentration of labeled ligand in controls (first), and vs. concentration of total labeled and unlabeled ligand in test samples (second). The concentration of unlabeled ligand that bound to soluble analytes in test samples is determined by evaluating the difference between the concentration that corresponds with the intersection of these two curves and the constant labeled ligand concentration in the test samples.

Abstract: The invention relates to a process and a device for the light-controlled synthesis of biopolymers on surfaces. In this process, patterns of individual sequences (20) are produced by the imaging of an arrangement (1) of electrically individually controllable light diodes (2) onto the surfaces.

Abstract: The present invention discloses a process of extraction, purification and characterization of a non-polar fluorescent dye from a marine echinoderm Holothuria scabra, compositions containing the dye and various applications of the dye, said dye is a natural, cell permeant, nontoxic and environmentally friendly non-polar fluorescent dye.

Abstract: A method for measuring chemiluminescence by which measurement of chemiluminescence can be carried out efficiently and accurately. In this method, an enzyme reaction of a chemiluminescent substrate in the presence of the enzyme carrying out the enzyme reaction and a chemiluminescence enhancer is conducted further in the presence of a chemiluminescence intensity-adjusting agent; and then the resulting chemiluminescence from the reaction product is measured.

Abstract: The present invention provides a method and a kit for detecting or quantitatively determining homocysteine rapidly and simply and with high sensitivity by oxidizing the residual homocysteine cosubstrate, the produced homocysteine-converting enzyme product or an enzyme reaction product thereof in the presence of an SH reagent to produce hydrogen peroxide and determining the produced hydrogen peroxide by color development using an oxidative color-developing agent. By using the method and kit of the present invention, homocysteine in biological samples, in particular, in body fluids such as blood and urine can be detected and quantitatively determined rapidly and simply and with high sensitivity.

Abstract: The invention relates to devices and methods for carrying out quantitative fluorescence immunoassays using evanescent field excitation. Light from at least one light source is directed onto the boundary between two media which have differing refractive indices. The light source emits practically monochromatic light with a wavelength suitable for exciting a marking substance. The light is directed onto a boundary surface disposed between an optically transparent base plate, the refractive index of which is greater than that of the material above the boundary surface, and a receiving region for the sample. The receiving region is covered with a covering plate on the side disposed opposite the base plate, there being arranged between the base plate and covering plate at least one functional layer. A detector for detecting the fluorescent light is disposed on the same side of the base plate as the light source.

Abstract: The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood, derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay.

Abstract: Methods and compositions for detecting and localizing light originating from a mammal are disclosed. Also disclosed are methods for targeting light emission to selected regions, as well as for tracking entities within the mammal. In addition, animal models for disease states are disclosed, as are methods for localizing and tracking the progression of disease or a pathogen within the animal, and for screening putative therapeutic compounds effective to inhibit the disease or pathogen.

Abstract: Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting specially overlapping target substances. The dyes comprise 4,7-dichlorofluoresceins, and particularly 2′,4′,5′,7′-tetrachloro-4,7-dichloro-5-(and 6-) carboxyfluoresceins. Methods and kits for using the dyes in DNA analysis are provided.

Abstract: The excitation of label molecules usable in chemical and biochemical analysis by electrical pulses at electrodes covered with a thin insulating film, and the use of such electrodes in chemical, clinical and biochemical analysis. The electrodes include a conducting base material that has been coated with an organic or inorganic insulating film or multiple layers of such films, so that either one or several label compounds can be excited to an excited state which is deexcited by emission of ultraviolet, visible or infrared light, in aqueous solution providing the basis for reproducible analytical applications in bioaffinity assays such as immunoassays and DNA-probing assays.

Abstract: Novel heterocyclic compounds which generate chemiluminescence on reaction with a phosphatase enzyme are provided as well as a process for their preparation and intermediates useful therein. The compounds comprise a nitrogen, oxygen or sulfur-containing heterocyclic ring system bearing an exocyclic carbon—carbon double bond. The double bond is further substituted at the distal carbon with a phosphate group and an oxygen or sulfur atom-containing group. Novel compositions further comprising a cationic aromatic compound (CAC) in addition to the heterocyclic phosphate compound are provided. The addition of the CAC in the composition greatly increases the production of chemiluminescence and provides improved detection sensitivity. Compositions further comprising an anionic surfactant and a non-ionic surfactant provide additional improvements in detection sensitivity.

Abstract: The present invention is directed to methods for making and using informative nucleic acid arrays (e.g., DNA including cDNA, RNA, PNA) for research and other applications in various disciplines or areas of interest. Examples of such disciplines include, without limitation, dermatology, pharmacology, toxicology, oncology, gynecology, urology, gastroenterology, as well as studies of sentinel gene discovery, signature gene discovery, mechanism of action, drug screening, drug metabolism, etc. The informative nucleic acid arrays of the present invention may contain only the gene sequences that are of interest in a particular area of interest or application, and may exclude other gene sequences.

Abstract: The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay.

Abstract: Cytochrome P-450 assay methods and kits for the methods are provided employing a cytochrome P-450 enzyme, substrates characterized by having an oxidizable methylene group oxidized to an aldehyde and a fluorescent hydrazine. A fluorescent hydrazine is added to the reaction mixture and the resulting hydrazone analyzed by capillary electrophoresis. The method finds use in evaluating compounds for enzyme modulating activity.

Abstract: A rapid method for detecting spoilage of a food sample, particularly a fish sample, by detecting and enumerating sulfide-producing bacteria (SPB). A growth medium containing iron and sulfur is combined with the food sample forming an incubation mixture which is incubated for a period of time. A plurality of fluorescence measurements are taken during an incubation period of about 4 hours to 17 hours at 30° C. SPB are determined to be present in the sample if the fluorescence measurement initially increases and then decreases to form a fluorescence maximum (peak). The time to detection of the fluorescence peak can be used with a correlation schedule to enumerate the SPB in the food sample. A visual test can also be used to identify color changes in the incubation mixture to provide a semi-quantitative enumeration of SPB effective in about 4 hours to 17 hours at 30° C.

Abstract: The present invention relates to procedures for the detection or for the determination of solid phase-associated factors, which are multiply associated with the same solid phase. According to the invention, the sample is brought into contact with a transmitter particle, on which at least one ligand having binding affinity for a solid phase-associated factor and a transmitter are immobilized, and a receiver particle, on which at least one ligand having binding affinity for said solid phase-associated factor and a receiver is immobilized, and then the signal is determined which results when transmitter and receiver are brought sufficiently close to one another. In particular, the invention relates to the detection of cell surface receptors which can be used for the typing of cells or for the determination of cell activation states. It is thus possible to replace the hitherto widely customary flow cytometry by a more simple procedure.

Abstract: A multiplexed enzyme assay method and substrate set for performing the method are disclosed. The method includes performing a plurality of enzyme reactions in the presence of a plurality of enzymes substrates, under conditions effective to convert an enzyme substrate to a corresponding product, where the product of each substrate and the substrate have different separation characteristics from each other and from the other substrates and their corresponding products. After performing the reactions, which may be carried out in separate or combined reactions, the substrates and products in said reactions are separated in a single separation medium. For each separated product and substrate, a separation characteristic effective to identify that product and substrate and a signal related to the amount of the product and substrate is detected. From this, one can determine the amount of substrate converted to the corresponding product in each of the reactions.

Abstract: The present invention provides high throughput screening Systems for identifying antifungal compounds. The method can be performed in plurality simultaneously with fluorescence or absorbance readouts.

Abstract: This invention relates to the detection of patients at risk for developing integrin antagonist/agonist mediated disease states. This invention relates to assays useful for the detection in a patient bodily fluid sample of drug-dependent antibodies which bind to integrins, or intergrin-associated proteins or complexes thereof in the presence of an integrin antagonist/agonist. This invention also relates to assays useful for the detection in a patient bodily fluid sample of drug-dependent antibodies (DDABS) that bind to integrins, including the platelet glycoprotein IIb/IIIa (GPIIb/IIIa), in the presence of a integrin agonist and/or antagonist. This invention also relates to procedures for identifying integrin antagonists/agonists that are less prone to elicit integrin antagonist/agonist mediated disease states. This invention also relates to procedures which increase the recovery of integrin-directed antibodies in body fluids, resulting in an increased sensitivity and specificity of DDAB detection assays.

Abstract: A method for enhancing the conversion of a phenol substrate to a product by a peroxidase enzyme comprises the steps of reacting a conjugate comprising a detectably labeled phenol-containing molecule with a peroxidase enzyme in the presence of an enhancing reagent, the enhancing reagent comprising an organic compound having the structure wherein each R is independently selected from the group consisting of: hydrogen and a C1-12 substituent; where V, W, Y and Z are each independently selected from the group consisting of: H, halogen, the C1-12 substituent, NR2, OR and SR; and where X is selected from the group consisting of: H, Br, Cl, F, the C1-12 substituent, NR2, OR and SR; or synergistic mixtures of an inorganic salt and the organic compound. The C1-12 substituent is linear, branched or cyclic. The C1-12 substituent is alkyl, alkenyl, alkynyl, heteroatom substituted alkyl, heteroatom substituted alkenyl, heteroatom substituted alkynyl, aryl, arylalkyl, arylalkenyl or arylalkynyl.

Abstract: This invention relates to a method for establishing and using a decision marker by which positive samples can be discriminated from negative samples. The method employs the analysis of multiple samples from confirmed positive and negative samples. A fluorescence channel is selected so that the desired sensitivity and specificity are achieved. A microparticle having this fluorescence channel then is made and is used in conjunction with a fluorescence marker which is specific for the population of interest.

Abstract: Oxidative stress in living organisms is determined in a photochemiluminescence measuring system after separation of low-molecular weight antioxidants from protein-containing test samples that were withdrawn from these organisms using an assay kit that contains a gel chromatographic column, a photosensitizer solution, a carbonate buffer solution, and a phosphate buffer solution.

Abstract: A method for the detection of a disease comprising the steps of: (i) obtaining a biological sample from a human subject; (ii) applying the biological sample to a sialidase substrate which has been immobilized on a solid support medium; and (iii) detecting a change in the immobilized sialidase substrate is provided. The substrate is preferably 5-bromo-4-chloro-3-indoyl &agr;-D-N-acetyl neuraminic acid or a salt thereof.

Abstract: A method of measuring the enzymatic activity of a luciferase includes contacting a luminogenic protein, such as a luciferase, with a protected luminophore to form a composition; and detecting light produced from the composition. The protected luminophore provides increased stability and improved signal-to-background ratios relative to the corresponding unmodified coelenterazine.

Abstract: The present invention relates to novel methods for sequencing nucleic acid molecules. More specifically, methods are provided for sequencing nucleic acid molecules present in bacterial lysates without the need to purify nucleic acid templates from highly soluble cellular components.

Abstract: A method for analyzing a liquid test sample by electrochemiluminescence. The method includes at least one specific biochemical binding reaction that leads to the formation of a complex which contains a chemiluminescence marker and the binding of the complex to a magnetic microparticle. Detection is carried out in a measuring cell having a working electrode in order to determine the concentration of the marked microparticle. The detection cycle includes a purification step, a conditioning step, a recovery step and a measuring step. A specified potential profile is applied to the working electrode during these steps. Between the conditioning step and the recovery step, an additional potential pulse with an oxidizing and/or a reducing potential is inserted into the voltage shape of the detection cycle in order to improve the deposit of the microparticle.

Abstract: Methods for enhancing contrast during imaging to assess cell and nuclear morphology of a sample. A contrast agent such as acetic acid, toluidine blue, hypertonic saline, hypotonic saline, Lugol's iodine, an absorbing dye, a liposome, or a contrast agent linked to a marker are applied to a sample. The sample is analyzed with an imaging device to create image data, and the sample is diagnosed using the image data.

Abstract: A device and method for detecting bacteria in food substances and the like utilizing a three layer composite consisting of a transparent base, an indicator exhibiting color change when exposed to changes in pH, and a gas permeable cover placeable in proximity to the substance. The method utilizes the generation of CO2 gas as a byproduct of bacterial growth which produces carbonic acid lowering the pH of the substance in the region of the composite resulting in an observable color change as in indication of the presence of bacteria.

Abstract: The present invention discloses the process of extraction, purification and characterization of a fluorescent dye from a marine echinoderm Holothuria scabra, compositions containing the dye and various applications of the dye.

Abstract: Optical detection techniques for the assessment of the physiological state, health and/or viability of biological materials are provided. Biological materials which may be examined using such techniques include cells, tissues, organs and subcellular components. The inventive techniques may be employed in high throughput screening of potential diagnostic and/or therapeutic agents.