Abstract: A method of testing sperm quality including obtaining a sample of sperm to be tested; detecting and measuring the testis-specific HspA2 chaperone protein (or, the chaperone protein homologues to HspA2) in human and animal sperm; and determining a sperm quality parameter based upon the chaperone protein, wherein an increased amount of the chaperone protein species indicates a higher sperm quality. The chaperone protein is detected and measured either by binding one or more antibodies specific to the sperm chaperone protein to the sperm and measuring the antibody content or measuring ATP bound to the sperm chaperone protein. In the case of the latter method, the chaperone protein may be detected and measured by measuring ATP bound to the sperm chaperone protein, and such measuring is by chaperone protein-bound and CK-B generated ATP measurement, or by bioluminescence of the chaperone protein bound-ATP.

Abstract: An automated in situ heat induced antigen recovery and staining method and apparatus for treating a plurality of microscope slides. The process of heat induced antigen recovery and the process of staining the biological sample on the microscope slide are conducted in the same apparatus, wherein the microscope slides do not need to by physically removed from one apparatus to another. Each treatment step occurs within the same reaction compartment. The reaction conditions of each reaction compartment for treating a slide can preferably be controlled independently, including the individualized application of reagents to each slide and the individualized treatment of each slide. The reagents are preferably held in a reagent dispensing strip similar to a “blister pack”.

Abstract: The present invention provides a means to identify the alleles present in a DNA-containing sample by providing subsets of loci for amplification by multiplex PCR. The loci include the thirteen CODIS short tandem repeat (STR) loci and amelogenin. The loci within each subset are grouped so that, upon PCR amplification, the amplicons produced within a given subset do not overlap. Differential labeling of subsets makes it possible to further group the subsets into compound multiplexes for co-amplification in a single reaction vessel, and analysis in a single electrophoretic channel.

Abstract: Electrokinetic devices having a computer for correcting for electrokinetic effects are provided. Methods of correcting for electrokinetic effects by establishing the velocity of reactants and products in a reaction in electrokinetic microfluidic devices are also provided. These microfluidic devices can have substrates with channels, depressions, and/or wells for moving, mixing and monitoring precise amounts of analyte fluids.

Abstract: This invention provides a method of treating an animal with pyrophosphate gout or osteoarthritis comprising administering an effective amount of a calcium antagonist, thereby reducing calcium pyrophosphate crystal deposition in the animal.

Abstract: The invention relates to a process for in vitro diagnosis of an infection by human cytomegaloviruses. The process consists of contacting cells, possibly carrying the infection, with a monoclonal antibody reacting with a polypeptide of molecular weight 68,000, induced by human cytomegalovirus and which possesses a protein-kinase activity. The detection of the reaction is preferably carried out by immunofluorescence.

Abstract: Compositions and methods for prevention and treatment of uncontrolled formation of intravascular fibrin clots are provided wherein fibrinolytic or anticoagulant drugs are biocompatibly coupled to red blood cell carriers.

Abstract: A method for quantitatively and/or qualitatively assaying an analyte in a sample, wherein the analyte is a receptor binding compound, has low detection limits equivalent to those of radioreceptor assays. The method comprises the steps of a) contacting the sample with material comprising a receptor for the analyte in order for receptor-analyte binding to occur and b) further contacting the sample with a detectable ligand for the receptor in order for receptor-ligand binding to occur, followed by c) separating the resulting receptor bound and free fractions, d) subjecting the receptor bound fraction to dissociating conditions releasing the ligand from the receptor and e) assaying for the dissociated ligand in a manner known per se for the detection of the detectable ligand.

Abstract: The present invention features calcilytic compounds. “calcilytic compounds” refer to compounds able to inhibit calcium receptor activity. Also described are the use of calcilytic compounds to inhibit calcium receptor activity and/or achieve a beneficial effect in a patient; and techniques which can be used to obtain additional calcilytic compounds.

Abstract: A process and a kit are provided for detecting differences in two or more samples of protein, including proteins bearing post-translational modifications and peptides. Proteins are prepared, for example, from each of a different group of cell samples or body fluid samples to be compared. Each protein extract is labeled with a different one of a luminescent dye from a matched set of dyes. The matched dyes have generally the same ionic and pH characteristics but emit light at different wavelengths to exhibit a different color upon luminescence detection. The labeled protein extracts are mixed together and separated together by electrophoresis or a chromatographic method. The separation is observed to detect proteins unique to one sample or present in a greater ratio in one sample than in the other. Those unique or excess proteins will fluoresce the color of one of the dyes used. Proteins common to each sample migrate together and fluoresce the same.

Abstract: A general stochastic method for synthesizing random oligomers can be used to synthesize compounds to screen for desired properties. The use of identification tags on the oligomers facilitates identification of oligomers with desired properties.

Abstract: A method and apparatus for the manipulation of colloidal particulates and biomolecules at the interface between an insulating electrode such as silicon oxide and an electrolyte solution. Light-controlled electrokinetic assembly of particles near surfaces relies on the combination of three functional elements: the AC electric field-induced assembly of planar aggregates; the patterning of the electrolyte/silicon oxide/silicon interface to exert spatial control over the assembly process; and the real-time control of the assembly process via external illumination. The present invention provides a set of fundamental operations enabling interactive control over the creation and placement of planar arrays of several types of particles and biomolecules and the manipulation of array shape and size. The present invention enables sample preparation and handling for diagnostic assays and biochemical analysis in an array format, and the functional integration of these operations.

Abstract: Disclosed are materials and methods for detecting biomolecules in samples employing transponders associated with the bead(s) used as the solid phase in the assay, and information pertinent to the assay is encoded on the transponders memory elements. A dedicated read/write device is used remotely to encode or remotely to read the information. The invention can be used in direct or competitive ELISA-type assays, or in multiplex assays for the simultaneous assay of several analytes.

Abstract: Magnetic particles for separating biological mixtures consist of nacreous luster pigments with magnetic properties coated with a biological polymer.

Abstract: Methods and compositions are provided for detecting changes in membrane potential in membranes biological systems. In one aspect, the method comprises; a) providing a living cell with a first reagent comprising a charged hydrophobic molecule which is typically a fluorescence resonance energy transfer (FRET) acceptor or donor, or is a quencher and is capable of redistributing within the membrane of a biological membrane in response to changes in the potential across the membrane; b) providing the cell with a second reagent that can label the first face or the second face of a biological membrane within the cell; c) detecting light emission from the first reagent or the second reagent. One aspect of this method involves monitoring membrane potential changes in subcellular organelle membranes in a living cells. Another aspect of the invention is the use of certain embodiments of the method for the screening of test chemicals for activity to modulate the activity of a target ion channel.

Abstract: This invention provides modified phycobilisomes and phycobilisome complexes that are supramolecular complexes with diverse spectral properties, and which may optionally be immobilized on a manufactured solid support. The invention provides a versatile set of highly sensitive signal-generating systems and conjugates that may be used as highly detectable tracers and labels, or in biotransducers comprising phycobilisomes or phycobilisome complexes, and also provides methods for performing specific binding assays using signal-generating systems comprising phycobilisomes as detectable labels. The embodiments of the invention provide the art with an extremely sensitive, nonisotopic detection means for assaying analytes and for sensing molecular events and environmental conditions.

Abstract: This invention provides dyes, particularly cyanine and related dyes, with acyl fluoride activating groups, the dyes having the general formula wherein: each dotted line represents carbon atoms necessary to form a fused substituted or unsubstituted aromatic ring; n is an integer selected from the group consisting of 1, 2 and 3; X and Y are selected from the group consisting of S, O, N, CH2 and C(CH3)2; at least one of said R1 and R2 comprises a sulfonic acid or sulfonate group attached to the aromatic ring; and R3 and R4 are independently selected from the group consisting of alkylcarboxylate, activated alkylcarboxylate and an inert group; wherein at least one of said R3 and R4 groups is alcylcarboxylate or activated alkylcarboxylate with the carboxyl group converted to an acyl fluoride. The inert group is a group that is inert towards acyl fluoride and has a sterical structure which allows aminoacylation of the acyl fluoride group.

Abstract: A method of identifying embryonic or fetal red blood cells in a sample containiing maternal blood cells and embryonic or fetal red blood cells or both, the method comprising determining which cell or cells contain or express an adult liver component. A method of isolating embryonic or fetal red blood cells from a sample containing maternal blood cells and embryonic or fetal red blood cells or both, the method comprising isolating the cells which contain or express an adult liver component. A method of determining a fetal abnormality the method comprising identifying or isolating embryonic or fetal cells according to the above methods and analysing said embryonic or early fetal cells for said abnormality. Use of a means for determining whether a cell contains or expresses an adult liver component for identifying or isolating an embryonic or fetal red blood cell.

Abstract: Combinations, called matrices with memories, of matrix materials that are encoded with an optically readable code are provided. The matrix materials are those that are used in as supports in solid phase chemical and biochemical syntheses, immunoassays and hybridization reactions. The matrix materials may additionally include fluophors or other luminescent moieties to produce luminescing matrices with memories. The memories include electronic and optical storage media and also include optical memories, such as bar codes and other machine-readable codes. By virtue of this combination, molecules and biological particles, such as phage and viral particles and cells, that are in proximity or in physical contact with the matrix combination can be labeled by programming the memory with identifying information and can be identified by retrieving the stored information. Combinations of matrix materials, memories, and linked molecules and biological materials are also provided.

Abstract: The invention describes quality control devices for assays that measure analytes in cells and tissue samples, and methods of use thereof. In particular, the quality control device comprises a matrix affixed with synthetic controls in different concentrations, or different synthetic controls. The quality control device can be adhered to a microscope slide and processed simultaneously with a tissue sample.

Abstract: A device for detecting the presence of an antigen including (1) a cell having antibodies which are expressed on the surface of the cell and are specific for the antigen to be detected, where binding of the antigen to the antibodies results in an increase in calcium concentration in the cytosol of the cell, the cell further having a emitter molecule which, in response to the increased calcium concentration in the cytosol, emits a photon; (2) a liquid medium for receiving the antigen and in which the cell is immersed; and (3) an optical detector arranged for receiving the photon emitted from the cell.

Abstract: A method for detecting at least one substance (a ligand) present in a compound library using at least one additional substance (a receptor) that binds to the ligand comprises adding to the compound library such a receptor that has substantially higher molecular weight than the ligand to be identified, and performing of such a spectroscopic measurement technique with the mixture, without isolating the receptor-ligand complex, that can detect those dipolar resonance phenomena which occur upon a binding of a receptor to a ligand.

Abstract: Boron complexes of certain bis-heterocyclic compounds are provided. The complexes resemble monomethine cyanines and are useful for imparting fluorescent properties to materials by covalent and noncovalent association. The compounds have the following general formula: wherein the dotted lines Z1 and Z2 represent the atoms necessary to complete a structure selected from the group consisting of one ring, two fused rings, and three fused rings, each said ring having five or six atoms, and each said ring comprising carbon atoms and, optionally, no more than two atoms selected from oxygen, nitrogen and sulfur, and R1 through R5 represent various atoms or groups and M is Cl or F.

Abstract: This invention is thus related to a biospecific multiparameter assay employing single molecule detection. In particular, this invention is related to a new method for determination of several different biomolecules simultaneously in the same reaction solution. The method uses a fluorescent label for labelling biospecific primary probes and a combination of other labels, bound to secondary probes. Complexes are formed comprising primary probes, analyte molecules and secondary probes in the biospecific reaction. These single molecule complexes are counted selectively while discriminating the signals from other fluorescent molecules using confocal fluorometry or two-photon excitation fluorometry including an auto- and cross-correlator for the signals obtained from said fluorescent labels.

Abstract: The invention deals with the use of a reactive compound in immunological analyses. The compound is ouabain or its analog to which another compound, labeled by radioiodine or by a fluorogen, is coupled. The invention covers also the method to measure ouabain or its analog in plasma to diagnose cardiovascular and endocrine diseases and other harmful conditions.

Abstract: A new class of optical indicators which are capable of memorizing and preserving the spatial localization of intracellular analytes in a time resolved manner is described. The compounds comprise a chromophore carrying a photolabile group capable of undergoing an irreversible and detectable chemical transformation upon irradiation by light. The chromophore is linked to a binding site capable of binding an analyte, wherein binding of the analyte to the binding site alters an optical property of the chromophore, thus altering the ability of the photolabile group to undergo the chemical transformation. Methods and kits for memorizing the spatial localization of the analytes are also described.

Abstract: Identification of defective ER and subclassifying ER+ breast cancers on the basis of the presence of defective ER is described as a tool potentially useful, as previously shown by the results of a pilot study, for predicting which among the ER+ tumors will respond and which will fail to repsond to hormonal modes of therapy. Improvements are introduced in the specimen sampling and ligand introduction steps of the immunohistochemical procedure which was developed for sub-classifying estrogen receptor-positive tumors of human breast cancers and other cancers of the estrogen target organs on the basis of the presence of defective estrogen receptors. Additionally, a new monoclonal antibody reagent which has potential use as a replacement for the polyclonal anti-ER antibody reagents is also described. The modified steps include the use of tumor imprints instead of cryosections; use of ligand coated slides instead of the original ligand layering step.

Abstract: The invention relates to a method for carrying out heterogeneous immunoassays, in particular to a method for separating a coated solid phase from a liquid phase by means of precipitation and subsequent centrifugation, with a detectable activity remaining in the liquid phase.

Abstract: The invention concerns a method for the determination of an analyte in a sample liquid wherein the sample liquid is incubated with (a) a first receptor which binds to a first epitope that is only present once on the analyte and is immobilized on a particulate carrier material and (b) a second receptor which binds to a second epitope on the analyte and has at least two binding sites for the second epitope, the first epitope being different from the second epitope and the analyte is determined by means of the binding to both receptors.

Abstract: A method of detecting dioxin-like compounds consisting essentially of the group of compounds: polychlorinated dibenzodioxins, polychlorinated dibenzofurans, polychlorinated biphenyls, and structural analogues thereof which exhibit biological activity characteristic of polychlorinated dibenzodioxins, polychlorinated dibenzofurans, polychlorinated biphenyls. The assay includes: 1) an inactive Ah receptor in a form capable of binding to the dioxin-like compounds and being transformed to an active form that forms a complex with ARNT and binds a dioxin responsive element, and 2) a quantity of ARNT sufficient to optimize Ah receptor transformation. A test sample is contacted with the assay under conditions effective to bind the dioxin-like compounds to the Ah receptor and allow transformation of the Ah receptor to an active form that forms a complex with ARNT. The presence of the complex containing the transformed Ah receptor and the ARNT is detected.

Abstract: A suspension of inert particles is prepared in an aqueous solution, to which an antibody or an antigen and a carrier-bound antigen or antibody, respectively, are added in any desired order. After centrifuging, the positive, weakly positive, or negative reaction can easily be recognized on the basis of a simple pattern.

Abstract: Activating groups based on N-hydroxynaphthalimide, are disclosed herein. The activating groups can mediate the coupling of labeling moieties, such as biotin or cyanine dyes, to a variety of components, including chain terminators, nucleoside triphosphates, and oligonucleotides, which are used in nucleotide sequencing. From these activating groups, activated esters of the labeling moieties can be prepared. The activated esters react with a component, for example a derivatized nucleotide chain terminator, to give a labeled component. In additions, methods of the present invention provide for labeling a nucleoside triphosphate in organic media. The activating groups and methods of the present invention allow the activation and coupling reactions to occur at a much higher yield, compared with the prior art.

Abstract: PSP94 occurs in the serum mainly as a complex with carrier protein. Determination of bound PSP94 provides an indicator of prognosis in prostate cancer and assists in diagnosis of prostate cancer in patients with borderline elevations of PSA.

Abstract: Methods and compositions are provided for determining the potential of a membrane. In one aspect, the method comprises:(a) introducing a first reagent comprising a hydrophobic fluorescent ion capable of redistributing from a first face of the membrane to a second face of the membrane in response to changes in the potential of the membrane, as described by the Nernst equation,(b) introducing a second reagent which labels the first face or the second face of the membrane, which second reagent comprises a chromophore capable of undergoing energy transfer by either (i) donating excited state energy to the fluorescent ion, or (ii) accepting excited state energy from the fluorescent ion,(c) exposing the membrane to radiation;(d) measuring energy transfer between the fluorescent ion and the second reagent, and(e) relating the energy transfer to the membrane potential.Energy transfer is typically measured by fluorescence resonance energy transfer.

Abstract: A device for detecting the presence of an antigen including (1) a cell having antibodies which are expressed on the surface of the cell and are specific for the antigen to be detected, where binding of the antigen to the antibodies results in an increase in calcium concentration in the cytosol of the cell, the cell further having a emitter molecule which, in response to the increased calcium concentration in the cytosol, emits a photon; (2) a liquid medium for receiving the antigen and in which the cell is immersed; and (3) an optical detector arranged for receiving the photon emitted from the cell.

Abstract: The present invention is a method for identifying a patient at risk to developing fibrinogen receptor antagonist-induced thrombocytopenia which comprises incubating patient plasma with a GPIIb/IIIa receptor:GPIIb/IIIa receptor antagonist complex to form a GPIIb/IIIa receptor:GPIIb/IIIa receptor antagonist:plasma antibody complex, incubating the GPIIb/IIIa receptor:GPIIb/IIIa receptor antagonist:plasma antibody complex with a secondary anti-human detectable antibody to form a GPIIb/IIIa receptor:GPIIb/IIIa receptor antagonist:plasma antibody:secondary anti-human detectable antibody complex, and detecting the presence of the secondary anti-human detectable antibody in the GPIIb/IIIa receptor:GPIIb/IIIa receptor antagonist:plasma antibody:secondary anti-human detectable antibody complex.

Abstract: Automated assays for detecting and measuring ion channel and cell surface receptor activity and for identifying compounds that modulate or potentiate such activity are provided. Among the assays are fluorescent indicator-based assays that use cells containing effective levels of a fluorescent indicator that is responsive to changes in ion concentration. Activation of the ion channels or receptors is initiated in the automated measurement apparatus by injection into one or more predetermined cell-containing wells of a reagent which is a known or possible activator or inhibitor of the ion channels or receptors of interest. The resulting activity of the ion channels or receptors, which causes changes in ion concentration in the cytoplasm, is determined by measurement of fluorescence intensity changes of the indicator in response to an excitation wavelength.

Abstract: A method is described for determining the sequence of nucleic acids. The method employs small solid phase particles having transponders, with a primary layer of an oligonucleotide of known sequence attached to the outer surface of the particle. A read/write scanner device is used to encode and decode data on the transponder. The stored data includes the sequence of the oligonucleotide immobilized on the transponder. The sequence of sample nucleic acids is determined by detecting annealing to an oligonucleotide bound to a particle, followed by decoding the transponder to determine the sequence of the oligonucleotide.

Abstract: This invention provides a method for preserving cells which comprises the steps of (a) suspending cells in a physiologically-acceptable, isotonic medium; and (b) fixing the cells so suspended at a temperature of less than about 10.degree. C. under sufficiently hypertonic conditions so as to disperse the cells in a single, unagglutinated state, thereby preserving cells. This invention also provides a method for detecting cells separated from a sample which have been preserved according to the aforementioned method. This invention also provides a method for visualizing cells. Also provided is a method for detecting a metabolic process in cells present in a sample. This invention also provides a method for detecting the presence of rare cells in a sample which specifically possess on their surfaces a moiety recognized by a known ligand comprising preserving cells separated from the sample according to the aforementioned method for preserving cells.

Abstract: Upon activation, platelet .alpha.-granules' soluble contents are secreted and membrane-bound contents are translocated to the plasma membrane. Membrane-bound proteins include the .beta.-amyloid precursor protein (APP) from which the .beta.-amyloid (A.beta.) deposits found surrounding the cerebrovasculature of patients with Alzheimer's Disease (AD) may originate. It has been discovered that activated platelets from AD patients exhibit less APP processing, retain more of the protein on their surface and secrete less as soluble fragments, than do controls. There is little APP or CD62 on the surface of resting platelets, but upon activation, while control platelets exhibited more of both proteins on their surface, advanced AD patients exhibited CD62 in similar amounts to controls, but retained significantly more surface APP. AD platelets secreted similar amounts of most soluble .alpha.-granule contents as controls, but less APP fragments.

Abstract: A method for detecting macromolecular species present on or in particles wherein particles are collected onto a surface, and the macromolecular species are allowed to diffuse from the particles. The diffused macromolecular species in close proximity to the particles are immobilized nonspecifically, wherein an immobilized macromolecular species is sufficiently close to a particle so as to be indicative of being diffused from the particle. The presence of the macromolecular species is detected while maintaining the close proximity of the immobilized macromolecular species to the particles from which the macromolecular species diffused.

Abstract: Compounds and libraries are labeled with a galactosyl epitope and then screened in accordance with an assay involving cells having a characteristic of interest. Conveniently, the screening may embody target cells, where the compounds are brought in contact with the cells. Each of the compounds carries with it the information of its identity or method of synthesis. After washing away non-specifically bound compounds, blood may be applied to the cells, whereby antibody binding to the galactosyl epitope initiates the complement cascade. Plaques are identified and the compound associated with the plaque identified. The formation of the plaque demonstrates that the compound has specific affinity for the target cell, binding of the compound to the cell does not interfere with binding of the antibody, and that the complex is capable of cytotoxic activity by means of the complement cascade.

Abstract: This invention relates to peptide ligand discovery and is particularly directed to a method for the discovery of agonists for membrane bound receptors. The inventive detection system involves the use of a "tethered" ligand for probing receptor binding. The general detection system includes a membrane, a membrane bound receptor, and a chimeric ligand presenting molecule. This chimeric protein forms the tethered ligand and in turn includes a membrane domain, a linker domain, a ligand domain, and a cleavable terminal domain. The "ligands" of the system are exposed by the addition of a specific peptidase that cleaves at the designated sequence. The sequence of the ligand that produces signal as a result of the interaction between the agonist and receptor can be then be isolated using sib selection.

Abstract: Compounds and libraries are labeled with a galactosyl epitope and then screened in accordance with an assay involving cells having a characteristic of interest. Conveniently, the screening may embody target cells, where the compounds are brought in contact with the cells. Each of the compounds carries with it the information of its identity or method of synthesis. After washing away non-specifically bound compounds, blood may be applied to the cells, whereby antibody binding to the galactosyl epitope initiates the complement cascade. Plaques are identified and the compound associated with the plaque identified. The formation of the plaque demonstrates that the compound has specific affinity for the target cell, binding of the compound to the cell does not interfere with binding of the antibody, and that the complex is capable of cytotoxic activity by means of the complement cascade.

Abstract: A method that assays activated antithrombin III (AT III) is useful for monitoring the effect in vivo of an agent that activates AT III. It involves the steps of contacting AT III with a solution sample containing the agent, wherein the agent is a sulfated glycosaminoglycan or a sulfated oligosaccharide, followed by contacting the sample with i) a first monoclonal antibody that preferentially binds AT III activated by the agent, as opposed to binding native AT III, and ii) a second monoclonal antibody that preferentially binds denatured AT III as opposed to either AT III activated by the agent or native AT III.

Abstract: Method for treatment of a disease in a patient characterized by accumulation of .beta.-amyloid. The method includes identifying a patient potentially suffering from such a disease and contacting a neuron of the patient with a therapeutically effective amount of a tachykinin agonist. Methods for screening for compounds useful for treating such a disease are also disclosed.

Abstract: A suspension of inert particles is prepared in an aqueous solution, to which an antibody or an antigen and a carrier-bound antigen or antibody, respectively, are needed in any desired order. After centrifuging, the positive, weakly positive, or negative reaction can easily be recognized on the basis of a simple pattern.

Abstract: Disclosed is a method of diagnosing sepsis in a human infant. The method includes detecting an increase in the expression of leukocyte cell surface antigens in a blood sample from an infant at risk for developing sepsis.

Abstract: In order to diagnose the coeliac disease antiendomysial antibodies are detected through immunofluorescence techniques in culture media in which bioptic fragments of the human enteral mucosa are kept alive. The cultures of the enteral mucosa take place by dipping the bioptic fragment into nutrient liquids with or without the presence of low concentrations of peptic-tryptic digest of gliadin. The peptide fragment of gliadin containing a definite aminoacid sequence capable of production of antiendomysial antibodies to a greater extent than the PT digest, even in very little concentration, is also identified.

Abstract: HIV-1 peptides having at least one point mutation between position 593 and 611 of the HIV-1 gp160 amino acid sequence. The point mutation either is at position 604 or 610, or both positions. Immunoassays which utilize these peptides are provided, as well as, diagnostic test kits which contain these peptides.