КОМПОЗИЦИИ ДЛЯ РЕАКЦИИ ОБРАТНОЙ ТРАНСКРИПЦИИ С ГОРЯЧИМ СТАРТОМ ИЛИ ДЛЯ ПОЛИМЕРАЗНОЙ ЦЕПНОЙ РЕАКЦИИ С ОБРАТНОЙ ТРАНСКРИПЦИЕЙ С ГОРЯЧИМ СТАРТОМ

... 1. Композиция для реакции обратной транскрипции с горячим стартом, которая содержит ион Mg, четыре типа dNTP, обратную транскриптазу, пирофосфат (PPi), и фосфатазу (ППазу).2. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, в котором композиция, содержит пирофосфат (PPi) в концентрации 0,1-5 мМ, а пирофосфотазу в количестве 0,005-0,25 Ед.3. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая один или несколько праймеров для обратной транскрипции.4. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, где композиция заморожена или высушена.5. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая нуклеотидную матрицу.6. Композиция для реакции обратной транскрипции с горячим стартом по п. 5, в котором нуклеотидная матрица является РНК.7. Композиция для реакции обратной транскрипции с горячим стартом по п. 1, дополнительно содержащая краситель, который не реагирует ...

ТЕСТ-НАБОР ДЛЯ ОПРЕДЕЛЕНИЯ РАКА

... 1. Набор для испытания активностей арилсульфатазы или ее изоферментов, включающий 4-метилумбеллиферон сульфат и 4-метилумбеллиферон. ! 2. Набор по п.1, который применяется для определения рака или других болезней и соответствующего начального скрининга, мониторинга терапевтического эффекта и контроля рецидива. ! 3. Набор по п.1, где рак представляет собой рак легкого, рак желудка, рак яичника, рак поджелудочной железы, рак груди, рак печени, рак пищевода, рак почки, рак мочевого пузыря, рак лимфатической системы, рак ободочной и прямой кишки, лейкемию или злокачественную лимфому. ! 4. Набор по п.1, где биологические образцы, которые будут применяться, являются цельной кровью человека, сывороткой, мочой, слюной, мокротой, лимфатической жидкостью или другой тканевой жидкостью. ! 5. Набор по п.4, где биологические образцы, которые будут применяться, являются человеческой сывороткой. ! 6. Набор по п.4, где биологические образцы, которые будут применяться, являются человеческой мочой. ! 7. Набор ...

VERFAHREN ZUR FAERBUNG VON NICHTRADIOAKTIVEN MARKIERTEN NUKLEINSAEUREN UND ZUSAMMENSETZUNG ZUM GEBRAUCH IN DIESEM VERFAHREN.

Reagenz und Verfahren zur Bestimmung von Kupplungsverbindungen

Homogeneous fluorescent assay for kinase, phosphatase or phosphodiesterase, useful in screening for pharmaceuticals and plant-protection agents, uses polycationic polymer as quencher

Homogeneous assay for quantitative measurement of kinase, phosphatase or phosphodiesterase reactions by reacting the enzyme with a fluorescent, (de)phosphorylatable substrate (A) in presence of polycationic polymer (B) containing quencher groups. The change in phosphorylation is determined from a change in fluorescence.

FLUOROGENE PHOSPHORSAEUREESTER, VERFAHREN ZU DEREN HERSTELLUNG SOWIE VERFAHREN UND MITTEL ZUM NACHWEIS UND ZUR FLUORIMETRISCHEN BESTIMMUNG VON PHOSPHATASEN

FLUORESZENZPOLARIZATIONSBESTIMMUNGEN UNTER VERWENDUNG VON POLYIONEN

Chemiliumineszierendes Substrat von Hydrolasen wie Phosphatasen

TESTVERFAHREN ZUR MESSUNG DER PHOSPHORYLIERUNG

VERFAHREN ZUR HEMMUNG VON ADENYLSUCCINAT-SYNTHETASE-AKTIVITÄT IN METHYLTHIOADENOSIN-PHOSPHORYLASE-UNZUREICHENDEN ZELLEN

Methods

Method of testing

CHROMOGENIC AND FLUOROGENIC ESTERS

DETECTION AND IMAGING IN BIOCHEMICAL ASSAYS USING PHOSPHOR SCREENS

Monitoring bacterial contamination of a wound involving assay of adenosine triphosphate

The invention provides a method of monitoring the bacterial contamination of a wound comprising monitoring the adenosine triphosphate (ATP) concentration of wound fluid removed from the wound. Preferably, an enzyme-coupled reaction such as the luciferin-luciferase reaction is used for the monitoring. Diagnostic kits and wound dressings adapted for use in the method are also provided.

Phosphatese detection using 4-acetamidophenyl phosphate substrate

Phosphatase activity is detected electrochemically with 4-acetamidophenyl phosphate (paracetamol phosphate) as substrate.

VHZ for diagnosis and treatment of cancer

We provide VHZ for use in a method of treatment, prophylaxis or alleviation of a cancer, such as breast cancer, in an individual. We provide an anti- VHZ agent for the treatment, prophylaxis or alleviation of cancer. We further provide a kit for detecting breast cancer in an individual or susceptibility of the individual to breast cancer comprising means for detection of VHZ expression in the individual or a sample taken from him or her as well as a method of detecting a cancer cell, the method comprising detecting modulation of expression, amount or activity of VHZ in the cell.

Coumarin snd quinolinone derivatives

Compounds of the formula wherein Y is M is hydrogen, or an alkali metal or ammonium ion, X is -O- or an amino group, R is H, Cl, Br, CN or carbamoyl, R1 is H or -SO3H, and A is one of several particular organic radicals, are especially useful in the photometric or fluorimetric determination of phosphatase or sulphatase activity in a liquid sample.

Screening Assay

Methods for measuring protein kinase and phosphatase activity

Animal feed enzyme extraction

Paracetamol phosphate for immunoassays

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

Chemical assay technique of the phosphatasic enzymes allowing to determine the diseases with retrovirus: paludism with répitition hepatitis ictère AIDS and primary education cancer of the liver and to differentiate their seropositivity

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

NEW KCNQ POLYPEPTIDE AND THEIR USE WITH THE DIAGNOSIS OF MENTAL DISORDERS

PROCEDURE FOR THE IDENTIFICATION OF AT PHOSPHOINOSITID OBTAINING SIGNALLING CONNECTIONS TAKEN PART AS WELL AS USE OF IT WITH THE PRODUCTION OF DRUGS

PROCEDURE AND KIT FOR THE ANALYSIS OF A GOAL OF NUCLEINSÄUREFRAGMENTES

TEST PROCEDURE FOR THE MEASUREMENT OF THE PHOSPHORYLATION

PROCEDURE FOR THE REGULATION OF ALKALINE PHOSPHATASE UNDER REMOVAL OF HEMOGLOBIN DISTURBANCES

FLUORESCENCE SUBSTRATES TO THE PROOF OF ORGANOPHOSPHATASE ENZYMAKTIVITÄT

PROCEDURE FOR MEASURING GROUPS CHEMICAL TO BIOLOGICAL MOLECULES BOUND ONES,

Screening for compounds, e.g. proteins or peptides, that increase the activity of the protein Rac1 comprises contacting cultured endothelial cells with a test compound and measuring endothelial permeability

Screening for compounds such as proteins, peptides, peptidomimetics, antibodies, or small organic molecules, that increase the activity of the protein Rac1 by binding to the extracellular portion of this protein comprises: (a) contacting a confluent layer of cultured endothelial cells grown on porous membranes with at least one of the test compounds alone or together with a stimulant of vascular permeability; and (b) measuring the endothelial permeability with a suitable agent, which can be colorimetrically detected. Independent claims are: (1) a method for screening for compounds such as proteins, peptides, peptidomimetics, antibodies, or small organic molecules that prevent the activation of RhoA and consequentially the change in the cytoskeletal structure of endothelial cells comprising: (a) contacting a confluent layer of cultured endothelial cells with thrombin in the presence of at least one of test compounds; (b) lysing the endothelial cells with a lysation buffer; and (c) measuring ...

ENZYMATIC ANALYSIS USING A SUBSTRATE THAT NIERDERSCHLAG RESULTS IN A FLUORESCENT ONE

USE OF DERIVATISIERTER ALKALINE PHOSPHATASE AS STANDARD.

1-NAPHTHOLPHTHALEINMONOPHOSPHATE, PROCEDURE FOR THEIR PRODUCTION AS WELL AS THEIR USE FOR the REGULATION the ALKALINE PHOSPHATASE.

SUBSTRATE BEGINNING OF MIXTURES IN a 2-AMINO2-METHYL-1-PROPANIL CONTAINING BUFFER FUER ALKALIN PHOSPHATASE SAMPLE.

RECOMBINED ALKALINE PHOSPHATASE FROM CALF INTESTINE

PARACETAMOLPHOSPHAT FOR IMMUNOASSAYS

PRODUCTION AND USE OF FLUORESCENT BENZOTHIAZOLDERIVATEN

FUNCTIONAL PROOF OF LIPOPROTEIN WITH HIGH DENSITY

CONNECTIONS, COMPOSITIONS AND METHODS TO THE PRODUCTION OF CHEMILUMINESZENZ WITH PHOSPHATASE ONES ENZYMES

PROCEDURE FOR THE REGULATION OF NUCLEIC ACIDS

ANTI- MIKROBI AGENTS AND SCREENING PROCEDURES FOR IT

EUROPIUM AND TERBIUM CHELATORE FOR TIME-SOLVED FLUOROMETRI TEST

PROCEDURE AND TEST KIT FOR THE DETERMINATION OF THE 5 ' - NUKLEOTIDASE ACTIVITY

CHEMILUMINESZENZ SUBSTRATES OF HYDROLYTIC ENZYMES SUCH AS PHOSPHATASEN

PROCEDURE FOR THE IDENTIFICATION OF INDUCTORS AND RESTRICTORS OF PROGRAMMED CELL DEATH

FLUORESZENZPOLARIZATIONSBESTIMMUNGEN USING POLYIONEN

Genetically engineered enzymes and their conjugates for diagnostic assays

APPARATUS AND METHOD TO READ BIOLOGICAL INDICATOR

A method for detecting biological activity in a biological indicator comprises the steps of providing a known quantity of spores containing a fixed number of copies of an enzyme and a liquid growth medium comprising substrates of the enzyme. The enzyme substrates have a first emission spectrum, and are configured to be converted by the enzyme to substrate derivatives having a second emission spectrum. The spores are exposed to the liquid growth medium, and the liquid growth medium is measured for the second emission spectrum, and either no change in the second emission spectrum or a linear increase in the second emission spectrum, is detected as a function of time. -42 3536513vl 914-" 1000 INCUBATE MONITOR FOR CHANGE IN FLUORESCENCE is YES A LINEAR NO INCREASE IN FLUORESCENCE? MONITOR FLUID FOR ASSUME STERILIZATION CHANGE TO EXPONENTIAL CYCLE EFFECTIVE INCREASE IN AND MEDICAL DEVICE FLUORESCENCE READY FOR USE CONFIRM STERILIZATION CYCLE INEFFECTIVE AND MEDICAL DEVICE NOT READY FOR USE MEDICAL ...

Biodegradable biochemical sensor for determining the presence and/or the level of pesticides or endocrine disruptors: method and composition

The present invention is directed to biodegradable biochemical sensor method to perform in a sample multiplex detection and/or quantification of pesticides and/or endocrine disruptors and to provide and logical integrated response to the user. This biochemical sensor is a vesicle encapsulating biochemical networks using enzymes capable of generating, inhibiting or activating specific measurable signal in presence of said target analytes. The biochemical network is able to provide an integrated logical final response to the user. The present invention also relates to a composition or kit comprising said biochemical sensor vesicle.

SUBSTRATE COMPOSITION FOR ALKALINE PHOSPHATASE AND METHOD FOR ASSAY USING SAME

2-AMINO-2-METHYL-1-PROPANOL BUFFER FOR ALKALINE PHOSPHATASE ASSAYS

Method and reagent kit for determining activity of 5'-nucleotidase

Method for modulating g-protein coupled receptors

LIPID PHOSPHATASE ASSAYS IN DISEASE AND DRUG DISCOVERY

Assay

The present invention discloses an assay device (1) for detecting active enzyme in a sample. Said assay device (1) comprises the following components:- (a) a placement region (10) onto which the sample can be placed; (b) a matrix (20) operably connected to said placement region (10) such that the sample when present (such as placed) on said placement region (10) can migrate along said matrix (20); (c) at least one distinct capture location (30) on said matrix (20), wherein each distinct capture location (30) is distanced away from the placement region (10), and wherein the sample can migrate across said distinct capture location (30); (d) capture means (40) being present at or defining each distinct capture location (30), wherein said capture means (40) are capable of binding to said enzyme such that at least a portion of said sample of said enzyme is retained at at least one distinct capture location (30); and (e) selective indication means (50), or at least a component thereof, to provide ...

Methods for detection of rare subpopulations of cells and highly purified compositions of cells

Methods are provided for detection of a target cell type within a cell population, and compositions are provided comprising cells and an indicator that indicates the number of cells of the target cell type in the cell population. Examples are provided in which these methods are used to detect human embryonic stem cells within a differentiated cell population with exquisite sensitivity. Differentiated cells produced from embryonic stem cells can be characterized by these methods before transplantation into a recipient, thereby providing further assurance of safety.

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

The present invention relates generally to the field of detection of microorganisms, in particular detection of bacteria, to methods for measuring enzyme activity, such as DNA polymerase activity, and particularly relates to such methods performed on microbial crude lysates, useful for determining microbial enzyme activities which can be linked to amplification signal generators such as real-time Polymerase Chain Reaction (PCR) techniques, thereby enabling determination of microbial pathogens in samples such as unpurified blood and other body fluids. This invention also relates to reagents for use in such methods, and to test kits comprising such reagents for carrying out the methods.

METHOD FOR QUANTITATIVELY ASSAYING THE PRESENCE FO DIARRHETIC SHELLFISH POISONING TOXINS IN MARINE SAMPLES.

METHODS FOR SCREENING FOR A COMPOUND USEFUL IN THE TREATMENT OR PREVENTION OF LYMPHOCYTIC DISORDERS, FOR INHIBITING LYMPHOCYTE ACTIVITY AND PREVENTING OR TREATING LYMPHOCYTIC DISORDERS

REVERSIBLE OXIDATION OF PROTEIN TYROSINE PHOSPHATASES

Nits as modifiers of the p53 pathway and methods of use

Promls as modifiers of the p53 pathway and methods of use

Kcnmas as modifiers of the p53 pathway and methods of use

B3galts as modifiers of the p53 pathway and methods of use

LOW MOLECULAR WEIGHT PROTEIN TYROSINE PHOSPHATASE (LMW-PTP) AS A DIAGNOSTIC AND THERAPEUTIC TARGET

New markers for severe progression of idiopathic scoliosis and uses thereof to stratify scoliotic patients and predict the risk of developing scoliosis

Methods of stratifying a subject having or at risk for developing adolescent idiopathic scoliosis (AIS) into diagnostically or clinically useful subclasses are provided. The stratification is based on the subject's expression and/or activity and/or PIPK1 expression and/or activity. Also provided are methods of predicting the risk of developing a scoliosis also based on the subject's expression and/or activity and/or PIPK1 expression and/or activity; and methods of increasing GiPCR signaling in cells of a subject in need thereof comprising administering to the subject's cells an effective amount of an inhibitor of PIPK1 tyrosine phosphorylation; an activator of PIPK1Y tyrosine dephosphorylation; and/or an inhibitor of PIPK1 expression and/or activity.

Liver test

The present invention relates generally to the field of diagnostics, and the prediction of mid- to long- term clinical outcomes for subjects with liver disease. In particular, the present invention provides a new liver test involving the use of a serum panel model and the development of an algorithm to reliably 5 predict liver related mid- to long-term clinical outcomes in chronic HCV infection, and in particular, in relation to liver related death, hepatocellular carcinoma (HCC) and decompensation.

Phosphatase inhibitor sample collection system

Assays for detecting modulators of cytoskeletal function

Method of measuring bone resorption rate

Fluorescence-based high throughput screening assays for protein kinases and phosphatases

Phosphate analysis

ASSAY FOR PHOSPHATASE-TARGETING TOXINS

The invention provides an assay method for determining phosphatase targeting toxins which inhibit protein phosphatases comprising contacting a solid support having an immobilized ligand immobilized thereon with: (i) a sample suspected of being contaminated with toxin and (ii) a non-immobilized ligand, wherein said immobilized ligand is capable of generating directly or indirectly detectable signal when uncomplexed, when complexed by said toxin, when complexed by a complex of said toxin and said non-immobilized ligand or when complexed by said non-immobilized ligand or said non-immobilized ligand is capable of generating a directly or indirectly detectable signal when uncomplexed or when complexed, separating a bound fraction from a non-bound fraction; and directly or indirectly determining the non-immobilized ligand bound to the immobilized ligand (the bound fraction) or non-complexed in aqueous solution (the non-bound fraction).

EMISSION RATIOMETRIC INDICATORS OF PHOSPHORYLATION

A chimeric phosphorylation indicator is provided. A chimeric phosphorylation indicator can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. A chimeric phosphorylation indicator also can contain a phosphorylatable polypeptide and a fluorescent protein, wherein the phosphorylatable polypeptide is contained within the sequence of the fluorescent protein, or wherein the fluorescent protein is contained within the sequence of the phosphorylatable polypeptide. Also provided are polynucleotides encoding such chimeric phosphorylation indicators, as well as kits containing the indicators or the polynucleotides. In addition, a method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.

ASSAY FOR DETECTING THE ENZYMATIC ACTIVITY OF A PHOSPHORYLATION ENZYME USING ENHANCED SIGNAL GENERATION

An assay system is provided for detecting the enzymatic activity of a phosphorylation enzyme, which enzyme may be a phosphatase or protein kinase. The substrate for the enzyme is immobilized on a solid support via covalent or non-covalent binding, through a signal enhancing polymer. The immobilized substrate provides an enhanced signal to background ratio, when compared to a substrate in solution. The methods are easily adapted to high throughput screening systems.

ASSAYS FOR MEASURING PHOSPHATE MODIFICATION ENZYME ACTIVITY

The present invention relates to assays that can measure the activity of enzymes that catalyze phosphate modifications, such as kinases, phosphatases, cyclases and phosphodiesterases. The assays can also be used to identify and screen for substances that modulate the activity of kinases, phosphatases, cyclases and phosphodiesterases.

MEANS FOR USE IN TREATING DISEASES CORRELATED WITH OR CAUSED BY NON-PHYSIOLOGICAL LEVELS OF MICROTUBULE-ASSOCIATED PP2AC

The present invention relates to a method of preventing or treating a disease correlated with or caused by non-physiologically increased intracellular levels of the catalytic subunit of microtubule-associated protein phosphatase 2A (PP2Ac) comprising administering to a subject affected by said disease or in danger of developing said disease a pharmaceutically effective amount of a protein selected from the group of MID1 or MID2 or a nucleic acid encoding said protein. The invention further relates to a method of preventing or treating a disease correlated with or caused by non-physiologically decreased intracellular levels of the catalytic subunit of microtubule-associated protein phosphatase 2A (PP2Ac) comprising administering to a subject affected by said disease or in danger of developing said disease a pharmaceutically effective amount of a peptidic fragment of MID1 or MID2 wherein said peptidic fragment comprises amino acids 108-165 (preferably 110-165) of MID1, amino acids 108-165 ...

METHOD OF SCREENING PTP ZETA ACTIVITY PROMOTER OR INHIBITOR

An object of the present invention is to provide a remedy for dysfunction of central monoamine pathway, a method for screening a PTP.zeta. inhibitor or activator, which is useful as a remedy for gastric ulcer caused by Helicobacter pylori or pleiotrophin which is a heparin-binding secretory protein, and a non-human model animal being hyposensitive to a stimulant drug, VacA which is a toxin of Helicobacter pylori, or pleiotrophin by utilizing the physiological function of PTP.zeta.. After administering a subject material to PTP.zeta. knockout mice and wild-type mice, PTP.zeta. activity in the PTP.zeta. knockout mice and the wild-type mice is compared and evaluated to screen a PTP.zeta. inhibitor or activator. Examples of the comparison and the evaluation of the PTP.zeta. activity include the comparison and the evaluation of the function of central monoamine pathway such as changes in the level of central monoamine metabolism, sensitivity to a stimulant drug, the presence of dysfunction of ...

HYDROLYZABLE FLUORESCENT SUBSTRATES FOR PHOSPHATASES AND ANALYTICAL USE THEREOF

HYDROLYZABLE FLUORESCENT SUBSTRATES FOR PHOSPHATASES AND ANALYTICAL USE THEREOF Hydrolyzable substrates for acid and alkaline phosphatases comprise blocked dye moieties which, when cleaved from the substrate during hydrolysis, provide fluorescent dyes having maximum absorptions above about 530 nm and maximum fluorescent emissions at least about 580 nm. The dyes are blocked prior to hydrolysis with a phosphono or thioxophosphono group or a salt thereof. These substrates can be used in analytical determinations of acid or alkaline phosphatase, or in competitive binding reactions to determine immunologically reactive substances.

MULTISYSTEM TEST MEANS

Docket No. 11694 MULTISYSTEM TEST MEANS Multisystem test means, method of making same and method for determining a constituent in a sample under a plurality of reaction parameters. More particularly, a multisystem test means for the determination of a constituent in a liquid sample comprising a plurality of components associated with at least two reaction systems, said reaction systems being respectively functional under different reaction parameters, at least one of said components being responsive to the presence of said constituent, and at least one other of said components being effective in response to contact with the sample under reaction parameters at which a first of said reaction systems is functional to change said reaction parameters to those under which at least one other of said reaction systems is functional. The test means, or device incorporated therewith, is contacted with a sample to be tested and after a predetermined time at least one component causes a change in reaction ...

PROSTATIC ACID PHOSPHATASE DETERMINATION

PROTEIN PHOSPHATASES

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

MPTENS AS MODIFIERS OF THE PTEN/IGF PATHWAY AND METHODS OF USE

Human MPTEN genes are identified as modulators of the PTEN/IGF pathway, and thus are therapeutic targets for disorders associated with defective PTEN/IGF function. Methods for identifying modulators of PTEN/IGF, comprising screening for agents that modulate the activity of MPTEN are provided.

METHOD AND REAGENT KIT FOR DETERMINING ACTIVITY OF 5'-NUCLEOTIDASE

The invention relates to a method for determining activity of 5'-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a coloured complex by treating it with ammonium molybdate and a reducing agent, colour intensity is measured by a known method, and from the measured value the activity of 5'-nucle- otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5'-nulceotidase, is calculated with a known calculation and/or with the aid of a calibration curve. According to the invention 5'-AMP, 5'-CMP, 5'-UMP, 5'-GMP, 5'-IMP and 5'-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.

ANTIGEN-COUPLED HYBRIDIZATION REAGENTS

The present disclosure provides high-performance hybridization reagents for use in a variety of hybridization assays and other related techniques. The hybridization reagents comprise an oligonucleotide probe and a bridging antigen, wherein the bridging antigen is recognized by a detectable antibody with high affinity. Also provided are compositions comprising panels of hybridization reagents specific for multiple different target nucleic acids and compositions comprising pairs of hybridization reagents and their complementary detectable antibodies. The paired hybridization reagents and detectable antibodies are useful in a variety of hybridization assays, particularly in highly multiplexed assays, where the structure of the bridging antigen is varied in tandem with variation in the detectable antibody, such that a multiplicity of hybridization reagents are provided that are capable of simultaneously detecting a multiplicity of target nucleic acids in a single assay. Also provided are kits ...

A FUNCTIONAL ASSAY OF HIGH-DENSITY LIPOPROTEIN

This invention provides novel assays that are prognostic and/or diagnostic for atherosclerosis or risk of atherosclerosis. It was discovered that high density lipoprotein (HDL) or components thereof can prevent the oxidation of lipids (e.g. lipids present in LDLs) and can also repair (reduce) already oxidized lipids and thereby reduce the inflammatory response associated with and characteristic of atherosclerotic plaque formation. Moreover it was a discovery of the invention that individuals vary in the ability of their HDL to afford such protection. Thus an assay of HDL protective and/or repair activity provides a highly effective assay for risk of atherosclerosis and its associated pathologies and such assays are provided herein.

PHOSPHATASE INHIBITOR SAMPLE COLLECTION SYSTEM

... ²²²A collection container and a method for collecting a biological sample, ²particularly whole blood, includes at least one stabilizing agent in an amount ²effective to stabilize and inhibit protein degradation and/or fragmentation. ²The stabilizing agent is able to stabilize proteases in the biological sample, ²particularly at the point of collection, by inhibiting protein degradation ²and/or fragmentation in the sample when the sample is stored. The stabilizing ²agent comprises or consists of one or more protease inhibitors.² ...

REAGENT COMPOSITIONS FOR ANALYTICAL TESTING

ENZYMATIC METHODS FOR MEASURING PLASMA AND TISSUE SPHINGOMYLELIN AND PHOSPHATIDYLCHOLINE

QUINONE METHIDE ANALOG SIGNAL AMPLIFICATION

Disclosed herein are novel quinone methide analog precursors and embodiments of a method and a kit of using the same for detecting one or more targets in a biological sample. The method of detection comprises contacting the sample with a detection probe, then contacting the sample with a labeling conjugate that comprises an enzyme. The enzyme interacts with a quinone methide analog precursor comprising a detectable label, forming a reactive quinone methide analog, which binds to the biological sample proximally to or directly on the target. The detectable label is then detected. In some embodiments, multiple targets can be detected by multiple quinone methide analog precursors interacting with different enzymes without the need for an enzyme deactivation step.

METHODS FOR DETECTING ORGANISMS AND ENZYMATIC REACTIONS USING RAMAN SPECTROSCOPY AND AROMATIC COMPOUNDS COMPRISING PHOSPHATE

The present disclosure provides systems for the rapid and sensitive detection of organisms and molecules in samples. Reactants that produce Raman-active products are used in combination with Raman light scattering. Such compounds may comprise phosphates permitting the detection of phosphatases. The present disclosure can also be used to measure enzyme-kinetics.

METHODS AND SYSTEMS FOR PREPARING IRREVERSIBLE INHIBITORS OF PROTEIN TYROSINE PHOSPHATASES

Described herein are the preparation and use of novel bromo- phosphonomethylphenylalanine amino acid derivatives (BrPmp) and BrPmp-containing peptides as specific, irreversible protein tyrosine phosphatase inhibitors, which are suitable for application in peptide synthesis. These derivatives are particularly advantageous since their synthesis is both easy and scalable, and they are suitable for peptide synthesis. The BrPmp derivatives described herein can be appropriately protected to allow for solid phase peptide synthesis (SPPS) and incorporation into peptides for preparation of protein tyrosine phosphatase inhibitors and inhibitor libraries. The peptides and peptide libraries can be used to identify new protein tyrosine phosphatase specific sequences and profile protein tyrosine phosphatase activity in cell lysates, diagnostic samples and biopsy samples.

ASSAY

The present invention discloses an assay device (1) for detecting active enzyme in a sample. Said assay device (1) comprises the following components:- (a) a placement region (10) onto which the sample can be placed; (b) a matrix (20) operably connected to said placement region (10) such that the sample when present (such as placed) on said placement region (10) can migrate along said matrix (20); (c) at least one distinct capture location (30) on said matrix (20), wherein each distinct capture location (30) is distanced away from the placement region (10), and wherein the sample can migrate across said distinct capture location (30); (d) capture means (40) being present at or defining each distinct capture location (30), wherein said capture means (40) are capable of binding to said enzyme such that at least a portion of said sample of said enzyme is retained at at least one distinct capture location (30); and (e) selective indication means (50), or at least a component thereof, to provide ...

PTP-D SUBFAMILY OF PROTEIN TYROSINE PHOSPHATASES

... 2141847 9403611 PCTABS00030 A novel subfamily (PTP-D) of protein tyrosine phosphatases is identified. Included in this family are PTP-D proteins or glycoproteins having one, two, or three identified amino acid changes in previously defined consensus sequences in the catalytic phosphatase domains of known protein tyrosine phosphatases. The PTP-D proteins or glycoproteins may be produced by recombinant means. Antibodies to PTP-D proteins or glycoproteins and nucleic acid constructs coding therefor, and methods for screening molecules which can bind to PTP-D proteins or glycoproteins and inhibit or stimulate their enzymatic activity, are provided.

ANTI-MICROBIAL AGENTS AND SCREENING METHOD THEREFOR

A method for determining the anti-microbial activity of a putative anti-microbial agent, the method comprising combining a microbially required nucleotide phosphatase, a nucleoside phosphate and the substance to be tested, and assessing the extent of degradation of the nucleoside phosphate in the presence and absence of the substance. The method thus allows the determination of the extent of inhibition of the nucleotide phosphatase by the substance. Preferably the method determines the degree of inhibition of an RNA helicase such as DbpA, which acts selectively on prokaryotic ribosomal RNA. Suitable DbpA inhibitors are disclosed as well as genetic material encoding for an active form of DbpA.

Methods of diagnosing cervical cancer

The invention provides reagents and methods for detecting pathogen infections in human samples. This detection utilizes specific proteins to detect the presence of pathogen proteins or abnormal expression of human proteins resulting from pathogen infections. Specific methods, compositions and kits are disclosed herein for the detection of oncogenic Human papillomavirus E6 proteins in clinical samples.

Inhibitors of protein tyrosine phosphatases

Disclosed herein are compounds that selectively inhibit members of the PTP family of enzymes. Synthesized compounds demonstrated selective inhibition of TC-PTP. Also provided are methods of using the compounds and formulations containing the compounds. Also described is a fluorescence-tagged combinatorial library synthesis and screening method. And methods of using these compounds to effect enzyme activity both in cells and in vitro as well as method of using these compounds to treat diseases in human and animals.

Analytical applications of enzymatic growth of fluorescent quantum dots

Methods for the detection/quantification of the enzymatic activity of hydrolases in a sample, wherein the hydrolase catalyses a hydrolysis reaction of a substrate which yields either H 2 S or a thiol-containing organic compound that produces a decomposition reaction which generates H 2 S, and methods for the detection/quantification of their substrates in a sample, as well as a method for the preparation of fluorescent CdS quantum dots, the preparation method, comprising first carrying out a enzymatic reaction catalysed by the hydrolase wherein the hydrolase catalyses the hydrolysis reaction of a substrate which yields either H 2 S or a thiol-containing organic compound that produces a decomposition reaction which generates H 2 S, and then reacting the resulting H 2 S with a salt of Cd +2 , thereby fluorescent CdS quantum dots are obtained.

MASS SPECTROMETRY METHOD FOR MEASURING THIAMINE IN BODY FLUID

Provided are methods for determining the amount of total thiamine in a body fluid sample using liquid chromatography and mass spectrometry. Total thiamine is converted to free thiamine by treatment with an acid phosphatase prior to thiamine separation and quantification. 1. A method for determining the amount of total thiamine in a plasma or serum sample , comprising:(i) removing soluble protein from a plasma or serum sample;(ii) incubating the sample from step (i) with an acid phosphatase, for not longer than about 2 hours, to convert phosphorylated thiamine to thiamine;(iii) performing an organic solvent extraction of said sample from step (ii), wherein the result of said extraction is an organic solvent phase and an aqueous phase;(iv) purifying said thiamine from said aqueous phase of step (iii) by liquid chromatography; and(v) determining the amount of thiamine by mass spectrometry, wherein the amount of total thiamine in said sample is determined.2. The method of claim 1 , wherein the incubation of step (ii) is carried out at a pH of about 4.6±0.1.3. The method of claim 1 , wherein said acid phosphatase is carried out at a temperature of about 40° C.4. The method of claim 1 , wherein said incubation is carried out between about 1 and about 2 hours.5. The method of claim 1 , wherein soluble protein is removed in step (i) by treating said sample with an acid.6. The method of claim 1 , wherein said liquid chromatography comprises high performance liquid chromatography (HPLC).7. The method of claim 1 , wherein step (v) comprises ionizing said thiamine to a parent ion having a mass/charge ratio of 265.00±1.0.8. The method of claim 7 , wherein said parent ion is fragmented into one or more daughter ions and wherein the amount of one or more of said daughter ions is determined.9. The method of claim 8 , wherein said one or more daughter ions comprises an ion having a mass/charge ratio of 144.00±1.0 or 121.94±1.0.10. The method of claim 8 , wherein said one or more ...

METHODS FOR DETERMINATION OF PROTEIN PHOSPHATASE ACTIVITY, AND USES IN PREDICTING THERAPEUTIC OUTCOMES

One aspect of the present disclosure encompasses methods for determining a protein kinase or phosphatase activity in a biological sample, comprising: contacting in a reaction mix a first test sample and a fluorescently-labeled peptide substrate capable of being modified by a protein phosphatase or a protein kinase, contacting the reaction mix with a TiOmatrix, thereby partitioning fluorescently-labeled phosphorylated peptide from fluorescently-labeled dephosphorylated peptide; and determining the fluorescence of the fluorescently-labeled dephosphorylated peptide, thereby determining a protein kinase or phosphatase activity. 1. A method of determining calcineurin activity comprising the steps of:i) contacting in a reaction mix a first test sample and a fluorescently-labeled phosphorylated peptide substrate capable of being dephosphorylated by calcineurin, under conditions such that calcineurin dephosphorylates the fluorescently-labeled phosphorylated peptide; and{'sub': '2', 'ii) contacting the reaction mix with a TiOmatrix, thereby partitioning fluorescently-labeled phosphorylated peptide from fluorescently-labeled non-phosphorylated peptide providing a partition with fluorescently-labeled non-phosphorylated peptide; and'}iii) determining calcineurin activity wherein measuring an amount of fluorescence emitted by the partition with fluorescently-labeled non-phosphorylated peptide is an indication of calcineurin activity.2. The method of claim 1 , wherein the fluorescently labeled phosphorylated peptide has an amino acid sequence selected from SEQ ID NO.: 1 and SEQ ID NO.: 2.3. The method of claim 1 , wherein the fluorescently labeled phosphorylated peptide is capable of distinguishing a first isoform of calcineurin from a second isoform.4. The method of claim 2 , wherein the fluorescently labeled phosphorylated peptide has the amino acid sequence according to SEQ ID NO.: 1 claim 2 , is phosphorylated on the Ser-15 position claim 2 , and further comprises an N- ...

Methods for Diagnosis, Prognosis and Methods of Treatment

This invention is directed to methods and compositions for diagnosis, prognosis and for determining methods of treatment. The physiological status of a cell present in a sample (e.g. clinical sample) can be used in diagnosis or prognosis of a condition (e.g. Chronic Lymphocytic Leukemia), in patient selection for therapy, to monitor treatment and to modify or optimize therapeutic regimens. 1. A method for classifying a cell comprisingcontacting said cell with a modulator, wherein the modulator is a tyrosine phosphatase inhibitor;determining the presence or absence of a change in activation level of an activatable element in said cell; andclassifying said cell based on said presence or absence of said change in the activation level of said activatable element.2. The method of claim 1 , wherein said tyrosine phosphatase inhibitor is CD45-associated protein tyrosine phosphatase inhibitor claim 1 , PHPS1 claim 1 , PP1 claim 1 , PP2 claim 1 , Bay U6751 claim 1 , BVT.948 claim 1 , NSC 295642 claim 1 , PRL-3 Inhibitor I claim 1 , Phenylarsine oxide claim 1 , Sodium Stibogluconate claim 1 , Sodium orthovanadate claim 1 , pervanadate claim 1 , bisperoxovanadium claim 1 , phenylarsine oxide claim 1 , alendronate claim 1 , etidronate claim 1 , vanadate claim 1 , gallium nitrate claim 1 , suramin claim 1 , or aplidin.3. The method of claim 1 , wherein said tyrosine phosphatase inhibitor is hydrogen peroxide (HO).4. The method of wherein said change in activation level of an activatable element is an increase in activation level of an activatable element.5. The method of wherein said cell is a cancer cell.6. The method of wherein said presence or absence of a change in activation level of said activatable element is compared to a normal cell contacted with said modulator.7. The method of wherein said cell is a hematopoietically derived cell.8. The method of wherein the presence or absence of a change in the activation levels of a plurality of activatable elements is determined ...

METHODS AND COMPOSITIONS FOR TREATING CANCER

We describe a method of determining whether a cancer cell is likely to be resistant to treatment by an mTOR inhibitor. The method may comprise detecting PPP2R2B (GenBank Accession Number: NM_18167) in or of the cell. It may, alter-natively, or in addition, comprise detecting PDK1 (GenBank Accession Number: NM_002613), in or of the cell. The method may comprise detecting methylation of the PPP2R2B promoter in or of the cell. It may comprise detecting the expression and/or activity of PPP2R2B in or of the cell. It may comprise detecting PDK1 mediated Myc phosphorylation activity. Methods of choosing a treatment for an individual suffering from or suspected to be suffering from a cancer, determining whether an individual suffering from or suspected to be suffering from a cancer will respond to treatment by an mTOR inhibitor, increasing the sensitivity of a cancer cell to treatment by an mTOR inhibitor, for treating or preventing cancer in an individual suffering or suspected to be suffering from cancer are also provided. We further provide for a combination of an inhibitor of PDK1 expression and/or activity and an mTOR inhibitor for use in a method of treatment or prevention of cancer. 1. A method of determining whether a cancer cell is likely to be resistant to treatment by an mTOR inhibitor , the method comprising detecting: (i) PPP2R2B (GenBank Accession Number: NM181678) or (ii) PDK1 (GenBank Accession Number: NM002613) , or both , in or of the cell.2. A method according to claim 1 , which comprises detecting methylation of the PPP2R2B promoter in or of the cell.3. A method according to claim 1 , which comprises detecting expression and/or activity of PPP2R2B or PDK1 claim 1 , or both claim 1 , in or of the cell.4. A method according to claim 1 , wherein a decreased expression and/or activity of PPP2R2B claim 1 , or an increased expression and/or activity of PDK1 claim 1 , or both claim 1 , compared to a cancer cell that is not mTOR inhibitor resistant claim 1 , ...

Methods and Systems for Preparing Irreversible Inhibitors of Protein Tyrosine Phosphatases

Described herein are the preparation and use of novel bromo-phosphonomethylphenylalanine amino acid derivatives (BrPmp) and BrPmp-containing peptides as specific, irreversible protein tyrosine phosphatase inhibitors, which are suitable for application in peptide synthesis. These derivatives are particularly advantageous since their synthesis is both easy and scalable, and they are suitable for peptide synthesis. The BrPmp derivatives described herein can be appropriately protected to allow for solid phase peptide synthesis (SPPS) and incorporation into peptides for preparation of protein tyrosine phosphatase inhibitors and inhibitor libraries. The peptides and peptide libraries can be used to identify new protein tyrosine phosphatase specific sequences and profile protein tyrosine phosphatase activity in cell lysates, diagnostic samples and biopsy samples. 2. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , wherein R is hydrogen and each Ris hydrogen.3. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , wherein R is Fmoc claim 1 , Boc claim 1 , or Cbz and Ris methyl claim 1 , ethyl claim 1 , benzyl claim 1 , dimethylamino (—N(CH)) claim 1 , propylamino (—NHCHCHCH) claim 1 , isopropylamino (—NHCH(CH)) or allyl.423-. (canceled)24. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of claim 1 , which is an L-amino acid derivative.25. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (I) of which is a D-amino acid derivative.2628-. (canceled)29. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (II) of claim 1 , wherein Ris hydrogen claim 1 , Ris the side chain of aspartic acid claim 1 , nis 1 claim 1 , each Ris hydrogen claim 1 , Ris the side chain of leucine claim 1 , Ris hydroxyl claim 1 , and nis 1.30. The bromo-phosphonomethylphenylalanine amino acid derivative of the Formula (II) of claim 1 , wherein ...

In Situ Chemiluminescent Substrates and Assays

Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed. 2. The method of claim 1 , wherein the oxidant is selected from hydrogen peroxide claim 1 , sodium molybdate claim 1 , hydrogen peroxide and sodium molybdate claim 1 , hypochlorite claim 1 , hypochlorite and hydrogen peroxide claim 1 , aryl endoperoxide claim 1 , calcium peroxide peroxyhydrate claim 1 , and combinations thereof.34.-. (canceled)6. The method of claim 1 , wherein Ris alkyl containing 1 to 2 carbon atoms or trifluoalkyl containing 1 to 2 carbon atoms.12. (canceled)13. The method of claim 1 , wherein the enzyme is a hydrolyic enzyme selected from alkaline phosphatase claim 1 , β-galactosidase claim 1 , β-glucosidase claim 1 , β-glucuronidase claim 1 , and neuraminidase.14. (canceled)15. The method of claim 1 , further comprising the step of detecting the light emitted from the reaction mixture after addition of the aqueous solution of the 1 claim 1 ,2-dioxetane enzyme substrate claim 1 , wherein the emission of light is indicative of the presence of the enzyme claim 1 , and the amount of light emitted can be correlated to the amount of the enzyme present in the sample.16. The method of claim 1 , wherein the enzyme moiety is an enzyme-linked antibody comprising a first antibody capable of binding to an antigen and an enzyme; an enzyme-linked antigen comprising an antigen and an enzyme; or an enzyme-linked oligonucleotide comprising an oligonucleotide capable of hydridizing to a nucleic acid claim 1 , wherein the enzyme is capable of cleaving the 1 claim 1 ,2-dioxetane enzyme substrate so that the substrate decomposes and generates light.17. (canceled)18. The method of claim 16 , wherein the first antibody or antigen is covalently linked to a label and the enzyme is ...

Method for Diagnosis of Bladder Cancer and Related Kits

The invention refers to a novel molecular biomarker, namely PTPD1, that is markedly increased in human bladder cancers. PTPD1 expression positively correlated with the grading and invasiveness potential of these tumors. PTPD1 can be detected at high levels in exfoliated bladder cells isolated from urine of bladder cancer patients, while no PTPD1 signal was evident in normal exfoliated bladder cells. Thus, PTPD1 detection in urine samples may represent a novel and reliable marker for non-invasive diagnosis of aggressive bladder cancer. 116-. (canceled)17. A method for diagnosing , or the monitoring control for therapy of , bladder cancer , said method comprising detecting (i) PTPD1 protein or an immunological fragment thereof , (ii) PTPD1 enzymatic activity , or (iii) PTPD1 mRNA in a body sample.18. The method of claim 17 , wherein the detecting of the PTPD1 protein or of an immunological fragment thereof comprises allowing the body sample to react with a PTPD1 protein specific ligand to form a complex; and detecting the complex.19. The method of claim 18 , wherein the PTPD1 protein specific ligand comprises an anti-PTPD1 antibody or a derivative thereof.20. The method of claim 19 , wherein the anti-PTPD1 antibody or a derivative thereof is obtained by using as an immunogen the whole PTPD1 protein of SEQ ID No. 1 or an immunogenic fragment thereof.2120. The method of claim of wherein the immunogenic fragment of the PTPD1 protein of SEQ ID No. 1 comprises between aa 751 and aa 910 of SEQ ID No. 1.22. The method of claim 21 , wherein the immunogenic fragment of the PTPD1 protein of SEQ ID No. 1 is a sequence between aa 751 and aa 910 of SEQ ID No. 1.23. The method of claim 18 , wherein the detecting step comprises detection of a specific fluorescent signal.24. The method of claim 17 , wherein the detecting of the PTPD1 enzymatic activity comprises performance by fluorescent or radiolabeled assays.25. The method of claim 17 , wherein the detecting of the PTPD1 mRNA ...

METHOD FOR LARGE SCALE PREPARATION OF THE ACTIVE DOMAIN OF HUMAN PROTEIN TYROSINE PHOSPHATASE WITHOUT FUSION PROTEIN

The present invention relates to methods for identifying inhibitors or activators of protein tyrosine phosphatase (PTP). In some examples, the methods utilize a PTP active domain with high activity and stability expressed without help of a fusion protein, by using computer based protein structure prediction technique. PTP prepared by the disclosed method may also be used as an antigen protein for the construction of a selective antibody and as a protein for the studies of PTP structure and functions. 1. A method for screening for a protein tyrosine phosphatase (PTP) activity inhibitor or activator in vitro , comprising the following steps: i) investigating homology among subgroups of PTP and selecting a region exhibiting high homology;', 'ii) examining whether the selected region of step i) corresponds to an active domain of a standard protein whose secondary and tertiary structures have already been identified;', 'iii) analyzing the secondary structure of the selected region of step i) if it corresponds to the active domain and then determining a boundary of PTP active domain by the location not containing helix or sheet of the secondary structure;', 'iv) determining 2-3 amino acids of the boundary of N-terminal and C-terminal of the PTP active domain primarily determined in step iii) to be a small amino acid or a charged amino acid by amino acid analysis;', 'v) constructing an expression vector containing a polynucleotide encoding the amino acids included in the inside of the boundary of the PTP active domain determined in step iv);', 'vi) generating a transformant by introducing the expression vector of step v) into a host cell; and,', 'vii) inducing expression of the recombinant PTP active domain by culturing the transformant of step vi) and obtaining the recombinant PTP active domain produced therefrom;, 'a) preparing a recombinant PTP active domain byb) contacting a PTP specific substrate and a candidate inhibitor or activator with the recombinant PTP active ...

METHODS FOR DETECTION OF RARE SUBPOPULATIONS OF CELLS AND HIGHLY PURIFIED COMPOSITIONS OF CELLS

Methods are provided for detection of a target cell type within a cell population, and compositions are provided comprising cells and an indicator that indicates the number of cells of the target cell type in the cell population. Examples are provided in which these methods are used to detect human embryonic stem cells within a differentiated cell population with exquisite sensitivity. Differentiated cells produced from embryonic stem cells can be characterized by these methods before transplantation into a recipient, thereby providing further assurance of safety. 1. A method of detecting the presence of or confirming the absence of target cells in a cell population , comprising:(a) providing a cell population;(b) applying a first stain and a second stain to said cell population, wherein said first stain detects a first marker the expression of which is indicative of the presence of target cells and said second stain detects a second marker the expression of which is also indicative of the presence of the same target cells, and wherein said first stain is detectable under visible light and said second stain is detectable under ultraviolet light;(c) microscopically observing cells of said cell population under visible light to detect any cells that are positive for said first marker;(d) microscopically observing said cells that are positive for said first marker under ultraviolet light and determining whether any of said detected cells are positive for said first marker and said second marker, and(e) identifying any cells that are positive for said first marker and said second marker as target cells.2. A method of detecting the presence of or confirming the absence of target cells in a cell population , comprising:(a) providing a cell population;(b) applying a first stain and a second stain to said cell population, wherein said first stain detects a first marker the expression of which is indicative of target cells and said second stain detects a second marker the ...

SUBSTRATES AND METHODS FOR STAINING LIVE STEM CELLS

The invention relates to novel substrates and methods for staining live stem cells. The stain may be used to identify induced pluripotent stem cell colonies during the process of somatic cell reprogramming. 1. A composition comprising a substrate capable of identifying a live stem cell of interest , wherein the substrate is cell permeable and is capable of being modified by the stem cell into a modified substrate , wherein the modified substrate is both permeable and detectable such that the live stem cell can be identified by detecting the presence of the modified substrate , and wherein the substrate and the modified substrate are non-toxic.2. The composition of wherein the live stem cell of interest is selected from the group consisting of an embryonic stem cell (ESC) claim 1 , an induced pluripotent stem cell (iPSC) claim 1 , a pluripotent cell claim 1 , a progenitor cell claim 1 , a reprogrammed cell claim 1 , and a dedifferentiated cell.3. The composition of wherein the detecting is of a fluorescent signal.4. The composition according to claim 1 , wherein the substrate is chosen from the group consisting of xanthene derivatives claim 1 , coumarin derivatives claim 1 , resorufin derivative claim 1 , triphenylmethanes and dialkylacridinones.5. The composition according to claim 1 , wherein the substrate is a rosamine derivative.6. The composition according to claim 4 , wherein the xanthene derivative is selected from the group consisting of fluorescein derivatives and rhodamine derivatives.7. The composition according to claim 6 , wherein the fluorescein derivative is either a fluorescein monophosphate or a fluorescein diphosphate.10. The composition according to claim 4 , wherein the coumarin derivative is an umbelliferone derivative.12. The composition according to claim 11 , wherein the compound is 6 claim 11 ,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP).14. The composition according to claim 4 , wherein the substrate is reactive with enzymes chosen ...

SMALL MOLECULE ANTAGONISTS OF DUSP5 AND METHODS OF USE

The present invention is directed to compounds that specifically target DUSP5 and act as antagonists of that enzyme. Such compounds are useful in the treatment of various conditions including, but not limited to vascular anomalies, cancer, and macular degeneration. 1. A pharmaceutical composition comprising a compound , or a pharmaceutically acceptable salt , solvate , or hydrate thereof , and a pharmaceutically acceptable carrier , wherein the compound comprises two negatively charged sulfonate groups or bioisosteric sulfonate analog groups tethered by a core scaffold of biphenyl or a planar analog of biphenyl , and wherein the core scaffold separates the negatively charged groups at a distance of about 6 to about 8 Angstroms.2. The composition of claim 1 , wherein the core scaffold comprises one or more of fluorene claim 1 , biphenyl claim 1 , carbazole claim 1 , dibenzofuran claim 1 , calixarene claim 1 , and naphthalene.4. The composition of claim 1 , wherein the negatively charged sulfonate groups or bioisosteric sulfonate analog groups are separated by a distance of about 7 angstroms.5. The composition of claim 1 , wherein the bioisosteric sulfonate analog groups are selected from the group consisting of sulfonamide claim 1 , tetrazole claim 1 , and carboxylic acid.6. The composition according to claim 1 , wherein said compound is a dual specificity phosphatase-5 (DUSP5) inhibitor.7. A medicament for treating claim 1 , preventing claim 1 , or alleviating a vascular anomaly comprising at least one compound as defined in in an effective amount to antagonize DUSP5.8. A method of treating claim 1 , preventing claim 1 , or alleviating a vascular anomaly comprising administering to a subject in need thereof a therapeutically effective amount of a compound or composition defined in wherein the compound antagonizes dual specificity phosphatase-5 (DUSP5).9. The method of claim 7 , wherein the vascular anomaly is associated with a Serine to Proline mutation at amino ...

Measurement of lp-pla2 activity

An object of the present invention is to provide a highly versatile, simple and safe method for measuring Lp-PLA 2 activity. Another object of the present invention is to provide an accurate and highly sensitive method for measuring Lp-PLA 2 activity. Provided is a method for measuring lipoprotein-associated phospholipase A 2 (Lp-PLA 2 ) activity in a sample containing Lp-PLA 2 , the method comprising the following steps (A) to (C): (A) converting PAFs into lyso-PAFs by reacting the PAFs with the Lp-PLA 2 in the sample; (B) hydrolyzing the lyso-PAFs produced in the step (A) with an enzyme (lyso-PAF-PLD) to obtain hydrolysate; and (C) measuring Lp-PLA 2 activity in the sample by utilizing a quantitative change attributable to the hydrolysate obtained in step (B) as an indicator.

Qualitative and quantitative point-of-care assays

Disclosed herein are “equipment-free” flow-through assay devices based on patterned porous media, methods of making same, and methods of using same. The porous, hydrophilic media are patterned with hydrophobic barriers for performing assays on liquids.

METHODS TO ASSAY PHOSPHATASE ACTIVITY

Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes 159-. (canceled)60. A method for a phosphatase assay , comprising:a) incubating free phosphorylated substrate labeled with an element tag with a support having attached thereto metal ion coordination complexes;b) separating free phosphorylated substrate from bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support;c) incubating ADP and at least one phosphatase with the bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support under conditions to enable the phosphatase to dephosphorylate the substrate;d) separating free non-phosphorylated substrate labeled with an element tag from bound phosphorylated substrate attached to the metal ion coordination complexes attached to the support; ande) using elemental analysis to detect the element tag associated with the free non-phosphorylated substrate.61. The method of wherein a multitude of free phosphorylated substrates each labeled with an element tag that is characteristic of the substrate are incubated in step (a) and the tag elements characteristic of the substrate are measured in step (e).62. The method of where in step (c) the conditions to enable the phosphatase to dephosphorylate the substrate include incubation with a phosphatase reaction buffer.63. The method of wherein step (c) comprises incubating antagonists or agonists of phosphatase.64. The method of wherein the metal ion coordination complex is attached to element labeled beads.65. The method of wherein the metal ion coordination complex attached to the support is a ...

ISOLATION OF ADULT MULTIPOTENTIAL CELLS BY TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells. 145-. (canceled)46. An enriched population of TNAP adult multipotential cells , wherein at least 4% of the total cell population are TNAP , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes , osteocytes and chondrocytes , wherein the TNAP marker is a TNAP marker which binds the STRO-3 antibody produced by the hybridoma cell line deposited under ATCC Accession Number PTA-5582.47. An enriched population according to claim 46 , wherein at least 10% of the total cell population are TNAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.48. An enriched population according to claim 46 , wherein at least 20% of the total cell population are TNAP claim 46 , STRO-1 multipotential cells capable of being cultured in vitro to produce adipocytes claim 46 , osteocytes and chondrocytes.49. An enriched population according to claim 46 , wherein the STRO-1 cells are STRO-1.50. An enriched population according to claim 47 , wherein the STRO-1 cells are STRO-1.51. An enriched population according to claim 48 , wherein the STRO-1 cells are STRO-1.52. A method of generating a committed cell population of a specific tissue type claim 46 , the method comprising culturing a population of adult multipotential cells according to in the presence of one or more stimulatory factors; and subjecting said cultured population to conditions biasing differentiation of the adult muitipotential cells to the specific tissue type.53. A method of generating a committed cell population of a specific tissue type claim 47 , the method comprising culturing a population of adult multipotential ...

Compounds for Treating Tauopathies and Restless Leg Syndrome and Methods of Using and Screening for Same

Provided are compounds capable of enhancing the ability of receptor-type tyrosine-protein phosphatase delta (PTPRD) to dephosphorylate a kinase. Also disclosed is a method of treating a tauopathy or restless leg syndrome in a subject comprising administering to the subject an effective amount of a disclosed compound. Also disclosed are kits comprising the compounds together with instructions for treating a condition and/or a compound known for treating the condition. Finally, disclosed herein is a screening method suitable for identifying positive allosteric modulators of the ability of a receptor-type tyrosine-protein phosphatase delta (PTPRD) to dephosphorylate a kinase. 3. The method of claim 1 , wherein the compound is Quercetin.4. The method of claim 1 , wherein the tauopathy is Alzheimer's disease claim 1 , chronic traumatic encephalopathy claim 1 , corticobasal degeneration claim 1 , frontotemporal lobar degeneration claim 1 , behavioral variant frontotemporal dementia claim 1 , language variant frontotemporal dementia claim 1 , right temporal variant frontotemporal dementia claim 1 , Pick disease claim 1 , or progressive supranuclear palsy.5. The method of claim 1 , further comprising administering to the subject an effective amount of a compound known for treating the tauopathy or restless leg syndrome.8. The method of claim 6 , wherein the compound is Quercetin.9. The method of claim 6 , wherein the kinase is glycogen synthase kinase GSK3β claim 6 , glycogen synthase kinase GSK3α claim 6 , cyclin dependent kinase-5 CDK5 claim 6 , or a combination thereof.12. The kit of claim 10 , wherein the compound represented by Formula (I) is Quercetin.13. The kit of claim 10 , wherein the tauopathy is Alzheimer's disease claim 10 , chronic traumatic encephalopathy claim 10 , corticobasal degeneration claim 10 , frontotemporal lobar degeneration claim 10 , behavioral variant frontotemporal dementia claim 10 , language variant frontotemporal dementia claim 10 , right ...

Method of evaluating quality of dephosphorylation reagent and method of detecting target nucleic acid

A method evaluates a quality of a dephosphorylation reagent, the method including the steps of: providing a dephosphorylation reagent containing an alkaline phosphatase and a peptide fragment derived from the alkaline phosphatase; and evaluating the dephosphorylation reagent as having a high quality if a content ratio of the peptide fragment to the alkaline phosphatase is a predetermined reference value or less.

MEDIUM FOR DETECTING STAPHYLOCOCCUS AUREUS, SHEET FOR DETECTING S. AUREUS COMPRISING SAME, AND METHOD FOR DETECTING S. AUREUS USING SAME

The purpose of the present invention is to provide a detection means whereby can be identified at a high accuracy. An aspect of the present invention relates to a medium for detecting which comprises one or more kinds of nutrient components, a color developing agent capable of developing a color in the presence of α-glucosidase, a color developing agent capable of developing a color in the presence of phosphatase and 0.5 mg/cmor more of sodium colistin methanesulfonate. Another aspect of the present invention relates to a sheet for detecting , said sheet comprising the aforesaid medium, and a method for detecting with the use of the medium and sheet as described above. 16-. (canceled)7Staphylococcus aureusStaphylococcus aureus. A microorganism culture substrate used for detection of comprising a substrate and a culture layer provided on an upper surface of the substrate , wherein the culture layer comprises a medium used for detecting comprising one or more nutrient components , a color developer that develops color in the presence of α-glucosidase , a color developer that develops color in the presence of phosphatase , and colistin sodium methanesulfonate at 0.5 mg/cmor more.8. The microorganism culture substrate according to claim 7 , which further comprises the substrate in the form of a sheet and a cover sheet covering the culture layer claim 7 , wherein the culture layer further comprises polyvinylpyrrolidone and one or more gelling agents.9Staphylococcus aureus. A method for detecting comprising:{'claim-ref': {'@idref': 'CLM-00007', 'claim 7'}, 'a step of sample addition comprising adding a microorganism-containing sample to the medium or the culture layer of the microorganism culture substrate according to ;'}a step of colony formation comprising incubating the medium or microorganism culture substrate added with the sample to form a microbial colony; and{'i': 'Staphylococcus aureus', 'a step of strain identification comprising identifying on the basis of the ...

BRET SENSOR MOLECULES FOR DETECTING HYDROLASES

The present invention relates to bioluminescence resonance energy transfer sensor molecules having the structure R-L-R—B or B—R-L-R, wherein Ris a bioluminescent protein, L is a linking element, Ris a non-protein acceptor domain and B is a blocking group, and wherein Rbound to B comprises a hydrolysable bond which produces a change in BRET when hydrolysed. The invention also discloses a method of detecting a hydrolase by contacting a sample with a molecule B—R, then contacting with a compound R-L or L-Runder conditions to cause attaching of Rto L, and detecting a change in the BRET ratio. Specifically exemplified sensors comprise luciferase and fluorescein diacetate, which is hydrolysed by an esterase. The invention also discloses luciferase enzymes derived from RLuc8 by removing cysteine residues. 2. The sensor molecule of claim 1 , wherein the blocking group stabilises the acceptor domain in a low-fluorescent or non-fluorescent state.3. The sensor molecule of or claim 1 , wherein the blocking group comprises a phosphate containing moiety claim 1 , sugar containing moiety claim 1 , amino acid containing moiety claim 1 , nucleotide claim 1 , nucleoside claim 1 , ester or ether.4. The sensor molecule of any one of to claim 1 , wherein the linking element comprises an alkyl chain claim 1 , glycol claim 1 , ether claim 1 , polyether claim 1 , polyamide claim 1 , polyester claim 1 , peptide claim 1 , polypeptide claim 1 , amino acid or polynucleotide.5. The sensor molecule of claim 4 , wherein the linking element comprises a polypeptide.6. The sensor molecule of claim 5 , wherein R-L or L-Rare a single polypeptide.7. The sensor molecule of or claim 5 , wherein the linking element comprises a cysteine residue and/or a lysine residue.8. The sensor molecule of claim 7 , wherein Ris attached to the linking element via the cysteine residue.9. The sensor molecule of any one of to claim 7 , wherein Ris selected from an Alexa Fluor dye claim 7 , Bodipy dye claim 7 , Cy dye ...

MEDICAL DEVICE DESIGN, MANUFACTURE AND TESTING SYSTEMS

Described are methods and systems for testing lumens of cannulated delivery components or assemblies thereof that may be used to deliver cells to a patient. Test cells are contacted with walls of the lumens and/or liquids that contact walls of the lumens, potentially over an incubation period. The test cells are then assessed for an effect of the wall contact, or the liquid contact, on at least one and preferably multiple characteristics of the test cells such as innate immune response, metabolic activity, viability, cytotoxic response, and/or motility. Methods and systems as described can be used in the development and/or manufacture of cannulated delivery devices, for example providing specifications for design or process inputs or outputs, design or process validations, and/or device lot approvals. Also described are devices or products produced in accordance with such methods and systems. 1. A method for testing a region of a medical device , comprising:incubating a liquid medium in contact with a wall of the region;contacting at least one cell with the liquid medium; andassessing the affect of said contacting on the expression of at least one toll-like receptor by the cell.2. The method of claim 1 , wherein the medical device is a medical delivery device claim 1 , and the region is a lumen.3. The method of claim 1 , wherein the at least one cell includes exogenous DNA encoding the at least one toll-like receptor.4. The method of claim 1 , wherein the cell is a mammalian cell.5. The method of claim 1 , wherein the cell is a human cell.69-. (canceled)10. The method of claim 1 , wherein the at least one toll-like receptor includes toll-like receptor 2 claim 1 , toll-like receptor 4 claim 1 , or both.1113-. (canceled)14. A method for testing a region of a medical device claim 1 , comprising:first assessing the region for an effect on innate immune response of a cell; andsecond assessing the region for an effect on at least one other characteristic of a cell, said ...

Detection and quantification of analytes based on signal induced by alkaline phosphate

A general methodology for highly sensitive and selective sensors that can achieve portable, low-cost and quantitative detection of a broad range of targets using only a personal glucose meter (PGM) is disclosed. The method and sensors take advantage of the ability of alkaline phosphatase (ALP) to convert glucose-1-phosphate to glucose, and the ability of PGMs to detect the generated glucose. The disclosed sensors can be part of a lateral flow device. Methods of using such sensors for detecting target agents, for example to diagnose disease, are also provided.

Methods of Identifying Modulators of Dephosphorylation of Histone Deacetylase

The present invention relates to screening methods that make use of a histone deacetylase interacting with a myosin phosphatase for the identification of novel therapeutics useful for inhibiting or inducing apoptosis and for the treatment of pathological conditions, such as smooth muscle cell disorder, cardiac hypertrophy or asthma. Also disclosed are methods for inhibiting or inducing apoptosis and for treatment of a pathological condition by administering to a mammal a therapeutically effective amount of a compound that inhibits or increases the dephosphorylation of a histone deacetylase by a myosin phosphatase or inhibits or increases the binding of a histone deacetylase to a myosin phosphatase. 134-. (canceled)35. An in vitro method for identifying a compound which modulates the dephosphorylation of a histone deacetylase-7 (HDAC7) polypeptide by a myosin phosphatase , the method comprising: i) a purified myosin phosphatase polypeptide comprising an amino acid sequence selected from SEQ NOs: 82, 83, and 84; and', 'ii) a purified HDAC7 polypeptide comprising the amino acid sequence set forth in SEQ ID NO:81, wherein said HDAC7 polypeptide is recognized by one or more of an antibody raised to the peptide FPLRTV[pSer]EPNLKL (SEQ ID NO:39), an antibody raised to the peptide RPLNRTR[pSer]EPLPPS (SEQ ID NO:40), and an antibody raised to the peptide RPLSRTQ[pSer]SPAAPV (SEQ ID NO:41); and, 'a) contacting a candidate compound withb) determining the effect of the candidate compound on dephosphorylation of the HDAC7 polypeptide by the myosin phosphatase, wherein said determining comprises determining the phosphorylation status of the HDAC7 polypeptide using an antibody specific for phosphorylated HDAC7,wherein a candidate compound which modulates the dephosphorylation of the HDAC7 polypeptide, compared to a control in the absence of the candidate compound, is identified as a modulator of dephosphorylation.36. The method of claim 35 , wherein the antibody specific for ...

QUINONE METHIDE ANALOG SIGNAL AMPLIFICATION

Disclosed herein are novel quinone methide analog precursors and embodiments of a method and a kit of using the same for detecting one or more targets in a biological sample. The method of detection comprises contacting the sample with a detection probe, then contacting the sample with a labeling conjugate that comprises an enzyme. The enzyme interacts with a quinone methide analog precursor comprising a detectable label, forming a reactive quinone methide analog, which binds to the biological sample proximally to or directly on the target. The detectable label is then detected. In some embodiments, multiple targets can be detected by multiple quinone methide analog precursors interacting with different enzymes without the need for an enzyme deactivation step. 1. A method of detecting a first target in a biological sample , comprising:contacting the biological sample with a first detection probe specific to the first target;labeling the first target with a first enzyme through the first detection probe;contacting the biological sample with a first quinone methide analog precursor comprising a first enzyme recognition group and a first detectable label, anddetecting the first target by detecting the first detectable label.2. The method of claim 1 , wherein the first enzyme cleaves the first enzyme recognition group claim 1 , thereby converting the first quinone methide analog precursor into a first reactive quinone methide analog which covalently binds to the biological sample proximally to or directly on the first target.3. The method of claim 1 , wherein the first enzyme is a phosphatase claim 1 , phosphodiesterase claim 1 , esterase claim 1 , lipase claim 1 , amidase claim 1 , protease claim 1 , nitroreductase claim 1 , urease claim 1 , sulfatase claim 1 , cytochrome P450 claim 1 , alpha-glucosidase claim 1 , beta-glucosidase claim 1 , beta-lactamase claim 1 , alpha-glucoronidase claim 1 , beta-glucoronidase claim 1 , alpha-galactosidase claim 1 , beta- ...

METHODS OF DIAGNOSING A DISEASE AND METHODS OF MONITORING TREATMENT OF A DISEASE BY QUANTIFYING A NON-REDUCING END GLYCAN RESIDUAL COMPOUND AND COMPARING TO A SECOND BIOMARKER

Provided herein are methods of diagnosing or monitoring the treatment of abnormal glycan accumulation or a disorder associated with abnormal glycan accumulation. 126.-. (canceled)27. A method of determining in an individual the presence , identity , and/or severity of mucopolysaccharidosis (MPS) III , the method comprising: 'wherein prior to enzyme treatment, the first biomarker is not present in abundance in samples from individuals with MPS III relative to individuals without MPS III, and wherein the first biomarker is a non-reducing end (NRE) biomarker;', '(a) generating a first biomarker comprising a glycan residual compound, wherein the first biomarker is generated by treating a population of glycans, in or isolated from a biological sample from the individual, with at least one digesting glycan enzyme,'} wherein prior to enzyme treatment, the second biomarker is not present in abundance in samples from individuals with the MPS III relative to individuals without the MPS III, and wherein:', '1) the second biomarker is a reducing end biomarker,', '2) the second biomarker is an internal glycan biomarker, or', '3) when the MPS III is caused by an abnormal function of a glycan degradation enzyme in the individual, the second biomarker is generated by treating the first biomarker with the glycan degradation enzyme that is functioning abnormally in the individual;, '(b) generating a second biomarker comprising a glycan residual compound, wherein the second biomarker is generated by treating a population of glycans, in or isolated from a biological sample from the individual, with at least one digesting glycan enzyme,'}(c) detecting the presence of and/or measuring the amount of the first and second biomarker produced and displaying or recording the presence of or a measure of a population of the first and second biomarkers by using an analytical instrument; and(d) monitoring and/or comparing the amounts of the first and second biomarkers in a biological sample; ...

Methods for detecting contaminants in solutions containing glucose polymers

The invention relates to a method for detecting contaminants of glucose polymers, said contaminants being capable of acting in synergy with one another so as to trigger an inflammatory reaction, characterized in that it comprises an in vitro inflammatory response test using modified cell lines.

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 18-. (canceled)9. A sample processing system , comprising:one or more operational matrices, wherein at least one of said operational matrices comprises one or more operational states of the sample processing system, and further wherein at least one of said operational states comprises two or more sample processing routines to be sequentially performed by the sample processing system; a sample handling system comprising at least one pipette nozzle for engaging with at least one pipette tip;', 'at least one detector; and', 'a cartridge receiving location;, 'a sample processing device comprising at least one pipette tip;', 'at least one assay unit;', 'at least one reagent unit; and', 'at least one sample collection unit for receiving a sample., 'and the cartridge comprising10. The system of claim 9 , wherein the at least one sample collection unit is configured to receive a sample of no more than about 500 μl.11. A method for restoring an operational state of a sample processing system claim 9 , comprising:{'claim-ref': {'@idref': 'CLM-00009', 'claim 9'}, 'accessing a sample processing catalog of the sample processing system of following a fault condition of the sample processing system; wherein the sample processing catalog comprises a record of operational data of the one or more operational states of the sample processing system;'}identifying a last-in-time operational state of the sample processing system from the sample processing catalog;identifying a last-in-time sample processing ...

USE OF AT LEAST ONE CHROMOGENIC AND/OR FLUOROGENIC PHOSPHATASE SUBSTRATE FOR THE DETECTION AND/OR ENUMERATION OF ENTEROBACTERIA IN A SAMPLE

Use of at least one chromogenic and/or fluorogenic phosphatase substrate for the detection and/or enumeration of enterobacteria in a sample likely to contain them, such as a food sample. 1. Use of at least one chromogenic and/or fluorogenic phosphatase substrate for the detection and/or enumeration of enterobacteria in a sample likely to contain them , such as a food sample , said at least one chromogenic and/or fluorogenic phosphatase substrate being a synthetic substrate comprising two parts , a first part specific to phosphatase activity and a second chromogenic and/or fluorogenic marker part.2. Use according to claim 1 , wherein said phosphatase substrate is selected from:substrates based on indoxyl or a derivative thereof, such as 5-bromo-4-chloro-3-indoxyl-phosphate, 5-bromo-6-chloro-3-indoxyl-phosphate, 6-chloro-3-indoxyl-phosphate, 5-iodo-3-indoxyl-phosphate, 6-bromo-3-indoxyl-phosphate, 5,6-dibromo-3-indoxyl-phosphate, 5-bromo-3-indoxyl-phosphate, Aldol® 458 phosphate, Aldol® 470 phosphate, Aldol® 484 phosphate, Aldol®495 phosphate, Aldol®515 phosphate, Aldol® n phosphate, and3-hydroxyflavone-phosphate.3. Use according to or claim 1 , wherein said phosphatase substrate is a substrate based on indoxyl or a derivative thereof claim 1 , said phosphatase substrate being used in combination with at least one agent promoting oxidative polymerization of the indoxyl derivative claim 1 , such as an ammonium ferric citrate metal complex claim 1 , preferably said phosphatase substrate being chosen from 5-bromo-4-chloro-3-indoxyl-phosphate claim 1 , 5-bromo-6-chloro-3-indoxyl-phosphate claim 1 , 6-chloro-3-indoxyl-phosphate claim 1 , 5-iodo-3-indoxyl-phosphate claim 1 , 6-bromo-3-indoxyl-phosphate claim 1 , 5 claim 1 ,6-dibromo-3-indoxyl-phosphate claim 1 , 5-bromo-3-indoxyl-phosphate.4. Use according to one of to claim 1 , wherein said phosphatase substrate is contained within a composition comprising at least one claim 1 , preferably two claim 1 , and advantageously ...

Method for measuring calcineurin activity

The present invention relates to a method for measuring the activity of calcineurin in a biological sample wherein a kinase inhibitor is present in the assay reaction mix.

Methods for Screening Inhibitors of Tau Phosphorylation By Casein Kinase I

Methods of screening for candidate compounds capable of inhibiting activity of fyn in phosphorylating tau protein at Y394 or binding to fyn to inhibit interaction with tau protein at Y394, including determining whether, and optionally the extent, the candidate compounds have these capabilities under conditions where fyn has these capabilities in the absence of the candidate compound. Methods of screening for substances capable of promoting dephosphorylation of tau protein by a phosphatase at a site of tau protein including contacting a candidate substance, the tau protein and a phosphatase capable of dephosphorylating the tau protein under conditions where the phosphatase is capable of dephosphorylating the site in absence of the candidate substance, where the kinase is fyn; determining whether, and optionally the extent, the candidate substance promotes dephosphorylation of the tau protein at the site; and selecting the candidate substance which promotes dephosphorylation of the tau protein the sites.

Methods for identifying modulators of ras using nonlinear techniques

Provided herein are compositions and methods for identifying and detecting modulators of Ras protein conformational states through the use of second harmonic generation (SHG) technology. Also provided herein are methods for detecting a conformational changes in the three dimensional structure of a protein bound to a supported lipid bilayer.

Flash and Glow 1,2-Dioxetanes

Compounds having chemiluminescent flash and glow properties. Also disclosed are methods using the compounds to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Also disclosed are kits relating to these compounds. 216.-. (canceled)17. The compound of claim 1 , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 25° C. which reaches a maximum in less than about 15 minutes.18. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 10 minutes.19. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 5 minutes.20. The compound of claim 1 , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 37° C. which reaches a maximum in less than about 15 minutes.21. The compound of claim 20 , wherein the generation of light reaches a maximum in less than about 10 minutes.22. The compound of claim 20 , wherein the generation of light reaches a maximum in less than about 5 minutes.24. A method for generating light claim 20 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) providing the compound of ;'}(b) providing an enzyme complex comprising an enzyme moiety which is capable of cleaving the compound;(c) contacting the enzyme complex with the compound to form a reaction mixture; and,(d) allowing the reaction mixture to generate light.2584.-. (canceled)86. An assay method for determining the presence and/or amount of an enzyme in a sample claim 20 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) providing the compound of ;'}(b) providing a sample suspected of comprising the enzyme which is capable of cleaving the compound so that it decomposes and generates light;(c) contacting the sample with the compound to form a reaction mixture; and,(d) detecting the light generated by the reaction mixture after addition of the ...

DATA COLLECTION METHOD TO BE USED FOR CLASSIFYING CANCER LIFE

Blood serum is applied on a support that has been impregnated in a buffer solution, the support is fractionated by electrophoresis at a predetermined liquid temperature to isolate proteins in the blood serum, an ALP isozyme is detected by color-developing with an ALP isozyme staining solution, the mobility, chromosome shape, density, and the like of each isozyme are determined by matching the protein fraction image against the ALP isozyme, development of a minute cancer that is occurring is discovered, and also the risk of tumor and risk of cancer are evaluated by matching against the analysis results of a tumor marker to classify the life of the cancer for early cancer discovery. 1. A data collection method to be used for classifying cancer life to discover development of minute cancer occurring in a human body and to perform data analysis of risk of tumor and risk of cancer in two divided stages of a preclinical cancer stage and a clinical cancer stage , wherein ,for the data used for analysis of a preclinical cancer stage,in order to collect data of an occurrence and a ratio of ALP I and activity value of ALP II and ALP III from patterns of the ALP I to the ALP IV as the ALP isozyme, examine the ratio of the numerical data, and perform data analysis for proliferation activity of cancer cells in view of APA calculated from ALP isozyme angle showing sharpness of the ALP II and the ALP III, andadditionally, for avoiding a cause contributed by an inflammatory disease, to perform an analysis while subtracting numerical data of a related part also occurring in the inflammatory disease from each numerical data obtained by measuring both the C reactive protein value (C inflammatory protein value, CRP value) and sialic acid value so as to perform data analysis for the existence and proliferation status of occurring minute cancer, andfor the data used for analysis of a clinical cancer stage,to collect data from a changed state like a decrease in albumin fraction and an ...

Flash and Glow 1,2-Dioxetanes

Compounds having chemiluminescent flash and glow properties. Also disclosed are methods using the compounds to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Also disclosed are kits relating to these compounds. 4. The compound of claim 1 , wherein Ris a straight chain alkyl comprising 1 to 5 carbon atoms which may be optionally substituted with one or more halogens atoms.5. The compound of claim 1 , wherein Ris a straight chain alkyl comprising 1 to 2 carbon atoms or straight chain trifluoalkyl comprising 1 to 2 carbon atoms.6. The compound of claim 5 , wherein Ris methyl or 1 claim 5 ,1 claim 5 ,1-trifluoroethyl.13. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 20 carbon atoms claim 1 , a branched alkyl comprising 3 to 20 carbon atoms claim 1 , a straight chain alkoxy comprising 1 to 20 carbon atoms claim 1 , or a branched alkoxy comprising 3 to 20 carbon atoms.14. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 20 carbon atoms or straight chain alkoxy comprising 1 to 20 carbon atoms.15. The compound of claim 1 , wherein the electron-donating group is a straight chain alkyl comprising 1 to 5 carbon atoms or straight chain alkoxy comprising 1 to 5 carbon atoms.16. The compound of claim 1 , wherein the electron-donating group is selected from the group consisting of methyl claim 1 , ethyl claim 1 , propyl claim 1 , butyl claim 1 , methoxy claim 1 , ethoxy claim 1 , propyloxy and butyloxy.17. The compound of claim 1 , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 25° C. which reaches a maximum in less than about 15 minutes.18. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 10 minutes.19. The compound of claim 17 , wherein the generation of light reaches a maximum in less than about 5 minutes.20. The compound of claim 1 , ...

SENSOR FOR DETECTING STEM CELL DIFFERENTIATION BASED ON ELECTROCHEMICAL METHODS

This invention relates to a sensor for detecting a stem cell differentiation, including (a) an electrode; and (b) a substrate of an alkaline phosphatase. The phosphorylation or dephosphorylation of the substrate for an alkaline phosphatase as a stem cell undifferentiation marker which dephosphorylates its substrate may be measured using an electrical signal in the present sensor. Therefore, the sensor of the present invention enables to electrically detect a stem cell status in a high-throughput manner and to determine the stem cell differentiation. 1(a) preparing the sensor for detecting the stem cell differentiation, comprising (i) an electrode which comprises gold electrode as the working electrode, a platinum electrode as the counter electrode, and a Ag/AgCl electrode as the reference electrode;and (ii) 1-Naphthyl phosphate (1-NP), a substrate for an alkaline phosphatase;(b) contacting cells of interest to the sensor; and(c) measuring the phosphorylation or dephosphorylation of 1-NP for alkaline phosphatase on the cells through an electrical signal, wherein the stem cell comprises an alkaline phosphatase where the stem cell is an undifferentiated stem cell; and the stem cell comprises no alkaline phosphatase where the stem cell is a differentiated stem cell.. A method for determining a stem cell differentiation, comprising the steps of: This is a continuation application of Application No. 12/579,084 filed on Oct. 14, 2009, which claims under 35 U.S.C. §119(a) the benefit of Korean Application No. 10-2009-0026353 filed Mar. 27, 2009, which applications are incorporated herein by reference.1. Technical FieldThe present invention relates to a sensor for detecting a stem cell differentiation, including: (a) an electrode; and (b) a substrate for an alkaline phosphatase.2. Background ArtCell chip technology is a promising tool for utilization in cell based assays. There are two kinds of cell detection systems for these chips. These systems are based on optical ...

NANOPATTERNED EXTRACELLULAR MATRICES ENABLE CELL-BASED ASSAYS WITH A MASS SPECTROMETRIC READOUT

The present disclosure provides methods in which adherent cells are treated with small molecules, cultured, lysed, and then analyzed by mass spectrometry to measure the activities of endogenous enzymes. The implementation of this method relies on the use of surfaces that are nanopatterned with cell adhesion ligands to mediate cell attachment and a peptide that is a substrate for the desired enzyme activity in the lysate. 1. A method of assaying activity of an intracellular enzyme , comprising: (i) coating a polymer pen lithography (PPL) tip array with a first monolayer reagent and printing the first monolayer reagent at selected positions on the surface to form an array of printed first monolayer reagent,', '(ii) incubating the array of printed first monolayer reagent with a second monolayer reagent such that the second monolayer reagent is adsorbed onto unprinted portions of the surface,, '(a) printing a surface with an array of immobilized cell adhesion ligands and immobilized substrates for the intracellular enzyme by'} (iii) contacting the resulting array of step (ii) with the substrate for the intracellular enzyme under conditions to immobilize the substrate to the surface at the portion of the surface comprising the monolayer reagent for chemical immobilization,', '(iv) contacting the resulting array of step (ii) with the cell adhesion ligand under conditions to immobilize the cell adhesion ligand to the surface at the portion of the surface comprising the monolayer reagent for adsorption of the cell adhesion ligand; wherein steps (iii) and (iv) can be performed in either order;, 'wherein one of the first monolayer reagent and the second monolayer reagent comprises a monolayer reagent for adsorption of the cell adhesion ligand and the other comprises a monolayer reagent for chemical immobilization of the substrate for the intracellular enzyme,'}(b) contacting a cell and the surface of step (a), the contacting resulting in immobilization of the cell via ...

METHODS FOR TREATING DILATED CARDIOMYOPATHY

The presently disclosed subject matter relates to methods of determining the risk of and treating the development of dilated cardiomyopathy (DCM) and to methods of preventing and/or reducing a risk of developing DCM in a dog or canine. In certain embodiments, the method comprises determining biomarkers comprising hematocrit, phosphate, alkaline phosphatase or creatinine. 1. A method of treating dilated cardiomyopathy (DCM) in a dog , the method comprising:a) measuring, in a sample from the dog, an amount of at least one biomarker; andb) administering a treatment or a dietary regimen to treat or prevent DCM.2. The method of claim 1 , wherein the at least one biomarker comprises hematocrit claim 1 , inorganic phosphate claim 1 , alkaline phosphatase claim 1 , creatinine claim 1 , or any combination thereof.3. The method of claim 2 , wherein an amount of the hematocrit is below about 45%.4. The method of claim 2 , wherein an amount of the inorganic phosphate is above about 1.5 mmol/L.5. The method of claim 2 , wherein an amount of the creatinine is below about 100 μmol/L.6. The method of claim 2 , wherein an amount of the alkaline phosphatase is above about 50 U/L.7. The method of claim 2 , wherein the dog is at risk of developing DCM if an amount of hematocrit is below about 45% claim 2 , an amount of the inorganic phosphate is above about 1.5 mmol/L claim 2 , an amount of the creatinine is below about 100 μmol/L claim 2 , and an amount of the alkaline phosphatase is above about 50 U/L.8. The method of claim 2 , wherein the at least one biomarker further comprises a primary bile acid claim 2 , a primary bile salt claim 2 , a secondary bile acid claim 2 , a secondary bile salt claim 2 , alkaline phosphatase claim 2 , amylase claim 2 , total protein claim 2 , BUN or urea level claim 2 , phosphorus claim 2 , calcium claim 2 , urine protein claim 2 , potassium claim 2 , glucose claim 2 , hemoglobin claim 2 , red blood cell (RBC) count claim 2 , red cell distribution width ...

Method and Device for Generating a Tunable Array of Fluid Gradients

Provided herein are devices and methods for generating microfluidic gradients, including an array of unique microfluidic gradients within an array of microchannels. Fluids within conduits are mixed in an intersection region to generate a mixed flow stream in a source reservoir channel that provides a gradient that varies with axial distance from the intersection region. Microchannels having an inlet connected to the source reservoir channel are configured to provide a microfluidic gradient in the microchannel. An outlet end of the microchannel is connected to a sink reservoir channel. By varying the ratio of fluid flow rates from the fluid conduits, the microchannel gradients are tuned. In this manner, a large number of unique gradients or array of microfluidic gradients is provided, wherein the gradient can be any number of physical or chemical parameters, including concentrations and physical fluid properties. 1. A microfluidic gradient generator for tuning dynamic components of fluid comprising:a first fluid conduit;a second fluid conduit,an intersection region that fluidically connects the first fluid conduit and the second fluid conduit, the intersection region comprising an intersection opening between the first fluid conduit and the second fluid conduit and a flow-divider that extends in a downstream direction from the intersection opening;a source reservoir channel fluidically connected to the intersection region and extending downstream from the intersection opening;a sink reservoir channel fluidically connected to the intersection region and extending downstream from the intersection opening;a microchannel array comprising a plurality of microchannels, each microchannel having an inlet end connected to the source reservoir channel and an outlet end connected to the sink reservoir channel, wherein adjacent microchannels are separated from each other by a separation distance, wherein the microchannel array traverses an axial distance along the source ...

METHODS FOR SCREENING VOLTAGE-GATED PROTEINS

In one aspect, the invention relates to a method for identifying a compound which modulates the activity of a voltage-gated protein. In certain embodiments, the voltage gate protein is a voltage-gated ion channel. In certain embodiments, the voltage-gated protein is a voltage sensitive phosphatase. In certain embodiments, the voltage-gated protein used in conjunction with the methods of the invention is modified to altered permeability or voltage sensitivity. 1. A method for identifying a compound which modulates the activity of a voltage-gated ion channel , the method comprising(a) providing a voltage-gated ion channel in a structure that separates a first medium from a second medium, wherein the voltage-gated ion channel exhibits independent ion permeation,(b) contacting the voltage-gated ion channel with a test compound,(c) measuring the amount of said independent ion permeation through the voltage-gated ion channel between the first and second media, and(d) comparing the amount of said independent ion permeation measured for the voltage-gated ion channel contacted with the test compound to the amount of said independent ion permeation measured for the voltage-gated ion channel not contacted with the test compound, wherein an increase or decrease in the amount of said independent ion permeation of the voltage-gated ion channel contacted with the test compound compared to the voltage-gated ion channel not contacted with the test compound indicates that the test compound modulates the activity of the voltage-gated ion channel.2. A method for identifying a compound which modulates the activity of a voltage-sensitive phosphatase , the method comprising(a) providing a voltage-sensitive phosphatase in a structure that separates a first medium from a second medium,(b) contacting the voltage-sensitive phosphatase with a test compound,(c) measuring the activity of the voltage-sensitive phosphatase, and(d) comparing the amount of said activity measured for the voltage- ...

DEGRADABLE CARBON NANOTUBE-CONTAINING BIOSENSORS AND METHODS FOR TARGET CLINICAL MARKER DETECTION

The invention relates to carbon nanotube-containing composites as biosensors to detect the presence of target clinical markers, methods of their preparation and uses in the medical field. The invention is particularly suitable for the detection in patient biological specimens of bone markers and tissue markers. The biosensors of the invention include carbon nanotubes deposited on a substrate, gold nanoparticles deposited on the carbon nanotubes and, binder material and biomolecule deposited on the gold-coated carbon nanotubes. The biomolecule is selected to interact with the target clinical markers. The biosensor can be used as an in-situ or an ex-situ device to detect and measure the presence of the target clinical markers. 1. An ex-situ , biosensor to detect a target clinical marker in a biological fluid sample of a patient , comprising:a substrate;a plurality of carbon nanotubes deposited on the substrate;a plurality of gold nanoparticles electrodeposited on the plurality of carbon nanotubes;a binding material adsorbed on the plurality of gold nanoparticles;a biotinylated biomolecule selected from the group consisting of antibody and aptamer, deposited on the binding material;a target clinical marker selected from the group consisting of a bone marker and a tissue marker,wherein the biotinylated biomolecule binds with the binding material, and interacts with the target clinical marker in the biological fluid sample of the patient; anda mechanism to provide an indication of the presence or absence of the target clinical marker in the biological fluid sample of the patient.2. The biosensor of claim 1 , wherein the target clinical marker is selected from the group consisting of c-terminal telopeptide claim 1 , n-terminal telopeptide claim 1 , alkaline phosphatase claim 1 , Troponin I and myoglobin.3. The biosensor of claim 1 , wherein the biotinylated biomolecule is selected from the group consisting of c-terminal telopeptide antibody claim 1 , n-terminal ...

CHEMICAL TOOLS FOR IMAGING PHOSPHOLIPASE D ACTIVITY

A method for detecting phospholipase D (PLD) activity in a cell, comprising: (i) stimulating endogenous PLD in said cell for said PLD to catalyze a transphosphatidylation reaction between phosphatidylcholine or a derivative thereof and an exogenous functionalized alcohol to form a phosphatidyl alcohol, wherein the functionalized alcohol possesses a first functional group that can react with and form a bond to a functionalized detectable label having a second functional group reactive with the first functional group, and said phosphatidyl alcohol contains said first functional group in available form; (ii) reacting said phosphatidyl alcohol with said functionalized detectable label under conditions where said functionalized detectable label reacts, via its second functional group, with the first functional group to form a linkage between said detectable label and said phosphatidyl alcohol so as to form a labeled phosphatidyl alcohol containing said detectable label; and (iii) detecting said labeled phosphatidyl alcohol. 1. A method for detecting phospholipase D (PLD) activity in a cell , the method comprising:(i) stimulating endogenous PLD in said cell while said cell is in contact with an exogenous functionalized alcohol containing at least one carbon atom in order for said PLD to catalyze a transphosphatidylation reaction between phosphatidylcholine or a derivative thereof and said exogenous functionalized alcohol to form a phosphatidyl alcohol, wherein said exogenous functionalized alcohol possesses a first functional group that can react with and form a bond to a functionalized detectable label having a second functional group reactive with the first functional group, and said phosphatidyl alcohol contains said first functional group in available form, as provided by the exogenous functionalized alcohol;(ii) reacting said phosphatidyl alcohol with said functionalized detectable label under conditions where said functionalized detectable label reacts, via its ...

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 1. A biological sample processing device comprising:a) a sample handling system;b) a detection station;c) a cytometry station comprising an imaging device and a stage for receiving a microscopy cuvette, andd) an assay station configured to support multiple components comprising i) a biological sample and ii) at least a first, a second, and a third fluidically isolated assay unit,wherein the sample handling system is configured to i) transfer at least a portion of the biological sample to the first assay unit, and the second assay unit, and the third assay unit; ii) transfer the first and second assay units containing biological sample to the detection station; and iii) transfer the third assay unit containing biological sample to the cytometry station.28.-. (canceled) This application is a continuation-in-part application of PCT Application No. PCT US2012/057155 and is a continuation-in-part of U.S. patent application Ser. No. 13/244,952 which claims priority to PCT Application No. PCT/US2011/53188, filed Sep. 25, 2011. All of the foregoing applications are incorporated herein by reference in their entirety for all purposes.The majority of clinical decisions are based on laboratory and health test data, yet the methods and infrastructure for collecting such data severely limit the quality and utility of the data itself. Almost all errors in laboratory testing are associated with human or pre-analytic processing errors, and the testing process can take days to weeks to complete. Often ...

Immobilized protein system for rapid and enhanced multiplexed diagnostics

The present invention relates to methods of detecting a neural injury biomarker in a biological sample. The method includes subjecting a biological sample to an assay according to the present invention that produces a measurable signal and detecting the measurable signal. The presence or absence of the measurable signal indicates the presence or absence of the biomarker in the sample. The present invention also relates to methods of determining the state of a subject's neural injury. The present invention also relates to systems and devices useful in carrying out the methods of the present invention.

COMPOUNDS FOR TREATMENT OF INTRACTABLE EPILEPSY AND DOORS SYNDROME

The present invention relates to the field of neurodegenerative diseases, and the prevention and/or treatment of TBC1D24-associated disorders, such as DOORS syndrome, (intractable) epilepsy, and nonsyndromic deafness. In particular, the present invention relates to phosphoinositide phosphatase inhibitors and screening methods for producing such inhibitors, more particularly, Synaptojanin-1 5′ phosphatase inhibitors, to increase phosphatidylinositol 4,5-bisphosphate (PI(4,5)P) levels in neuronal cells. 1. A method of treating a TBC1D24-associated disorder in a subject , the method comprising:{'sub': '2', 'treating the subject with a phosphoinositide phosphatase inhibitor which increases PI(4,5)Plevels in a cell.'}2. The method according to claim 1 , wherein the TBC1D24-associated disorder is selected from the group consisting of severe forms of epilepsy claim 1 , early onset epilepsy claim 1 , familial infantile myoclonic epilepsy (FIME) claim 1 , focal epilepsy claim 1 , dysarthria claim 1 , myoclonic epilepsy with dystonia claim 1 , familial malignant migrating partial seizures of infancy claim 1 , De Flippo malignant migrating partial seizures of infancy claim 1 , variable degrees of intellectual disability claim 1 , hearing loss claim 1 , nonsyndromic deafness claim 1 , autosomal-dominant nonsyndromic hearing loss claim 1 , and dominant nonsyndromic hearing impairment claim 1 , cortical myoclonus and cerebellar ataxia claim 1 , and DOORS syndrome (deafness claim 1 , onychodystrophy claim 1 , osteodystrophy claim 1 , mental retardation and seizures).3. The method according to claim 1 , wherein the phosphoinositide phosphatase is Synaptojanin-1.4. The method according to claim 3 , wherein the phosphoinositide phosphatase is the Synaptojanin-1 5′ phosphatase.5. The method according to claim 1 , wherein the cell is a neuronal cell.6. The method according to claim 1 , wherein the phosphoinositide phosphatase inhibitor is selected from the group consisting of bpV(pic) ...

ASSAY

The present invention discloses an assay device () for detecting active enzyme in a sample. Said assay device () comprises the following components: (a) a placement region () onto which the sample can be placed; (b) a matrix () operably connected to said placement region () such that the sample when present (such as placed) on said placement region () can migrate along said matrix (); (c) at least one distinct capture location () on said matrix (), wherein each distinct capture location () is distanced away from the placement region (), and wherein the sample can migrate across said distinct capture location (); (d) capture means () being present at or defining each distinct capture location (), wherein said capture means () are capable of binding to said enzyme such that at least a portion of said sample of said enzyme is retained at at least one distinct capture location (); and (e) selective indication means (), or at least a component thereof, to provide selective indication of the presence of active enzyme bound to said capture means (). 11. An assay device () for detecting active enzyme in a sample , said device comprising:{'b': '10', '(a) a placement region () onto which the sample can be placed;'}{'b': 20', '10', '10', '20, '(b) a matrix () operably connected to said placement region () such that the sample when present (such as placed) on said placement region () can migrate along said matrix ();'}{'b': 30', '20', '30', '10', '30, '(c) at least one distinct capture location () on said matrix (), wherein each distinct capture location () is distanced away from the placement region (), and wherein the sample can migrate across said distinct capture location ();'}{'b': 40', '30', '40', '30, '(d) capture means () being present at or defining each distinct capture location (), wherein said capture means () are capable of binding to said enzyme such that at least a portion of said sample of said enzyme is retained at least one distinct capture location ();'}{'b': ...

COMPARTMENTALISED SCREENING BY MICROFLUIDIC CONTROL

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development. 1102-. (canceled)103. A method for screening a compound or compounds capable of modulating the activity of a target , comprising the steps of:(a) compartmentalising the compounds into microcapsules, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule;(b) contacting a target having a desired activity with the compound or compounds and monitoring the modulation of an activity of the target by the compound or compounds, wherein one or more of steps a, b and c is performed under microfluidic control; and(c) identifying and sorting the microcapsules which contain the compound(s) having the desired activity using a change in their optical properties.104115-. (canceled)116. The method of claim 103 , wherein the target is compartmentalized together with the compounds in the microcapsules.117. The method of claim 103 , wherein a fluid stream comprising the target is introduced into the microcapsules comprising the compounds.118. The method of claim 103 , further comprising fusing a microcapsule comprising the target with each of the microcapsules comprising the compounds.119. The method of claim 103 , further comprising attaching the repertoire of compounds to microbeads.120. The method of claim 119 , wherein each microbead comprises a detectable tag.121. The method of claim 119 , wherein the ...

ULTRA-HIGH-SENSITIVE ASSAY OF PROTEIN AND NUCLEIC ACID AND KIT, AND NOVEL ENZYME SUBSTRATE

Provided is a ultra-high-sensitivity assay in which the assay can be made on a commonly used assay apparatus such as an absorptiometer and a plate reader or with naked eyes. The high-sensitivity assay in which the assay can be made on a commonly used assay apparatus or with naked eyes can be provided by combining an enzyme cycling method using thio-NAD(P) as a coenzyme, a labeling enzyme and a substrate for the labeling enzyme optimally, and by amplifying thio-NAD(P)H, which is a signaling substance, exponentially and then quantifying the thio-NAD(P)H colorimetrically. 3. The method according to claim 1 , wherein:the enzyme is alkaline phosphatase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a phosphate group, Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}4. The method according to claim 2 , wherein:the enzyme is alkaline phosphatase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a phosphate group, Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}5. The method according to claim 1 , wherein:the enzyme is glucosidase, galactosidase, fructosidase or mannosidase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a sugar moiety, the sugar moiety represents one selected from the group consisting of glucose, galactose, fructose and mannose, and Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative represented by the formula (1).'}6. The method according to claim 2 , wherein:the enzyme is glucosidase, galactosidase, fructosidase or mannosidase, and{'sup': 1', '2, 'sub': 2-6', '1-6, 'X represents a sugar moiety, the sugar moiety represents one selected from the group consisting of glucose, galactose, fructose and mannose, and Yrepresents hydrogen, and Yrepresents a Calkoxy group, or a Calkyl group in the androsterone derivative ...

IMMOBILIZED PROTEIN SYSTEM FOR RAPID AND ENHANCED MULTIPLEXED DIAGNOSTICS

The disclosure relates to methods of detecting a neural injury biomarker in a biological sample. The method includes subjecting a biological sample to an assay disclosed that produces a measurable signal and detecting the measurable signal. The presence or absence of the measurable signal indicates the presence or absence of the neural injury biomarker in the sample, and thereby diagnosing a subject as having a neural injury. The disclosure further relates to methods of determining the state of a subject's neural injury. Further disclosed are systems and devices useful in carrying out the methods disclosed. 1. A method for detecting the presence or absence of a neural injury biomarker in a biological sample , the method comprising:providing a biological sample; [{'sup': +', '+, '(a) an assay to detect S100β, wherein the assay comprises (i) providing aldolase, glyceraldehyde 3-phosphate dehydrogenase (“GAPDH”), and a signal-transducing molecule, each immobilized on one or more supports, (ii) providing fructose 1,6-bisphosphate and NAD, and (iii) contacting the biological sample with the one or more supports, NAD, and fructose 1,6-bisphosphate under conditions effective to cause a cascading biological reaction leading to production of a measurable signal by the signal-transducing molecule if S100β is present in the biological sample;'}, '(b) an assay to detect S100β, wherein the assay comprises (i) providing phosphoglyceromutase (“PGM”), enolase, pyruvate kinase, and a signal-transducing molecule, each immobilized on one or more supports, (ii) providing 3-phosphoglycerate and (iii) contacting the biological sample with the one or more supports and 3-phosphoglycerate under conditions effective to cause a cascading biological reaction leading to production of a measurable signal by the signal-transducing molecule if S100β is present in the biological sample;', {'sub': 2', '2, '(c) an assay to detect glial fibrillary acidic protein (“GFAP”), wherein the assay comprises ( ...

Enzymatically responsive magnetic particles and their use

The invention relates to an enzymatically responsive product that includes an amino acid residue conjugated to a magnetic particle, wherein the amino acid residue is phosphorylated or sulfated or comprises an ester-moiety linked via peptide bond. Compositions containing the enzymatically responsive product, and the use thereof for separating distinct types of mammalian cells (e.g., cancer cells from normal cells), for treating a cancerous condition, and imaging cancer cells are also disclosed.

METHODS FOR SELECTING PHOSPHATASE SELECTIVE AND NON-SELECTIVE PHOSPHATASE INHIBITORS

The present invention discloses a method to discover selective inhibitors of phosphatases. Thus the invention provides a method for screening a test compound to determine whether the compound binds a holophosphatase selectively or non-selectively comprising: i) providing a first holophosphatase wherein said holophosphatase is captured/immobilised; ii) testing a test compound for its ability to bind to the first holophosphatase; iii) providing a second holophosphatase wherein said second holophosphatase is captured/immobilised; iv) testing the same test compound for its ability to bind to the second holophosphatase; v) comparing the binding of the test compound to said first holophosphatase with the binding to said second phosphatase wherein a compound that binds a holophosphatase selectively will bind to said first holophosphatase but not said second holophosphatase; or will bind to said second holophosphatase but not said first; or wherein a compound that binds a holophosphatase non-selectively will bind to both said first holophosphatase and said second holophosphatase. 1. A method for screening a test compound to determine whether the compound binds a holophosphatase selectively or non-selectively comprising:i) providing a first holophosphatase wherein said holophosphatase is captured/immobilised;ii) testing a test compound for its ability to bind to the first holophosphatase;iii) providing a second holophosphatase wherein said second holophosphatase is captured/immobilised;iv) testing the same test compound for its ability to bind to the second holophosphatase;v) comparing the binding of the test compound to said first holophosphatase with the binding to said second phosphatasewherein a compound that binds a holophosphatase selectively will bind to said first holophosphatase but not said second holophosphatase; or will bind to said second holophosphatase but not said first; or wherein a compound that binds a holophosphatase non-selectively will bind to both said ...

METHOD FOR PREVENTING OBESITY-INDUCED FATTY LIVER BY INHIBITING KCTD17

The present invention provides methods for reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that decreases KCTD17 expression in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels. 1. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that decreases KCTD17 expression in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.2. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that prevents PHLPP2 degradation in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.3. The method of claim 2 , wherein the pharmaceutical composition inhibits Glucagon signaling.4. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that inhibits Glucagon signaling in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.5. The method of claim 4 , wherein the pharmaceutical composition reduces PHLPP2 degradation.6. The method of or claim 4 , wherein the pharmaceutical composition increases free Raptor in the liver cells.7. The method of - claim 4 , wherein the pharmaceutical composition decreases PHLPP2 phosphorylation at Serine 1119 and Serine 1210 residues in liver cells.8. The method of claim 6 , wherein the pharmaceutical composition prevents PHLPP2 degradation in liver cells.9. The method of any one of - claim 6 ...

Nanoscale probe structure and application thereof

A nanoscale probe structure, including: a first probe having a tip top end and a second probe having a planar top end, wherein a metallic layer coats the on the tip top end, an insulating layer coats around the tip top end of the first probe; and a metallic layer coats on the planar top end, an insulating layer coats around the planar top end of the second probe. The structure of present invention can applied in atomic force microscopy to measure the electricity physiology signal inside and outside the cell membrane, which can limit the measure region to specific little area for the measure of electricity physiology signal and effectively decrease the miscellaneous noise disturbance from other region.

METHODS FOR ASSESSING TOXICITY

A method for characterizing toxicity of toxic pollutants and a method for characterizing comprehensive toxicity of water bodies. The methods include constructing a reporter gene cell line expressing CHOP gene associated with endoplasmic reticulum stress. 1. A method for constructing a reporter gene cell line expressing CHOP gene associated with endoplasmic reticulum stress , the method comprising:using the CHOP gene associated with endoplasmic reticulum stress as a specific inducing gene to construct a CHOP promoter;ligating the CHOP promoter with a SEAP gene to construct a lentivirus CHOP-SEAP plasmid vector; andtransfecting the CHOP-SEAP plasmid into Hela cells to construct the reporter gene cell line. Pursuant to 35 U.S.C. § 119 and the Paris Convention Treaty, this application claims foreign priority to Chinese Patent Application No. 201710616052.5 filed Jul. 25, 2017, the contents of which are incorporated herein by reference. Inquiries from the public to applicants or assignees concerning this document or the related applications should be directed to: Matthias Scholl P. C., Attn.: Dr. Matthias Scholl Esq., 245 First Street, 18th Floor, Cambridge, Mass. 02142.The present disclosure relates to the field of detection of cytotoxicity caused by contaminants, and more particularly to the use of a reporter gene cell line based on endoplasmic reticulum stress in characterizing toxicity of pollutants.Reporter gene assay is widely used in toxicity evaluation because of its sensitivity, rapidity, and reproducibility. Researches have shown that the reporter gene method can significantly lower the toxicity detection limit, improve the sensitivity, and shorten the response time. The current research based on reporter gene is mainly to monitor the specific target substances, such as DNA damaging substances, endocrine disruptors, dioxins, and other substances through the specific combination of inducible gene and reporter gene. However, different pollutants have different ...

PROTEOGLYCAN IRREGULARITIES IN ABNORMAL FIBROBLASTS AND THERAPIES BASED THEREFROM

Provided herein are methods to identify agents or compounds that specifically modulate the oligomerization and/or functional activities of receptor protein tyrosine phosphatase sigma (RPTPσ) in an abnormal fibroblast cell and therapies based therefrom. 1. A method of treating arthritis in a subject , the method comprising:obtaining a biological sample comprising synovial cells, synovial-like cells and/or synovial fluid from the joint of the subject;contacting the biological sample with a test agent that inhibits clustering and/or promotes PTP activity of receptor protein tyrosine phosphatase sigma (RPTPσ); anddetermining whether (i) there is a change in the clustering and/or biological activity of RPTPσ, or (ii) whether the agent binds to a ligand of the RPTPσ ectodomain;wherein if there is an inhibition of RPTPσ clustering and/or increase in PTP activity, or binding of the test agent to the ligand of the RPTPσ ectodomain the subject is treated with an agent that inhibits RPTPσ clustering or promotes PTP activity and/or RPTPσ biological activity.2. The method of claim 1 , wherein the agent is a soluble extracellular domain of RPTPσ.3. The method of claim 1 , wherein the agent is an RPTPσ Ig1&2 polypeptide.4. The method of claim 1 , wherein the agent is an antibody that specifically interacts with the RPTPσ ectodomain and inhibits clustering.5. The method of claim 1 , further comprising measuring the level of syndecan-4.6. A method of screening a subject having or at risk of having arthritis claim 1 , the method comprising:obtaining fibroblast-like synoviocytes (FLS) from a subject;contacting the FLS cells with an agent that inhibits clustering and/or promotes PTP activity of receptor protein tyrosine phosphatase sigma (RPTPσ), anddetermining whether (i) there is a change in the clustering and/or biological activity of RPTPσ on the FLS cells, or (ii) whether the agent binds to a ligand of the RPTPσ ectodomain on the FLS cells,wherein if there is an inhibition of RPTP ...

MENAINV AND CANCER INVASION AND METASTASIS

Methods and compositions are provided for diagnosing or inhibiting invasion or metastasis of a cancer in a subject based on Mena. 1. A method of identifying an agent as an inhibitor of sarcoma or carcinoma cell invasion or as an inhibitor of metastasis of a carcinoma or sarcoma , comprising contacting a preparation comprising a growth factor receptor and a protein-tyrosine phosphatase 1b (PTP1b) in the presence of an amount of Menaand an amount of Mena and quantifying the association of the growth factor receptor and PTP1b in the presence of the agent and in the absence of the agent , wherein an agent that increases the association of growth factor receptor and PTP1b in the presence of the agent as compared to in the absence of the agent is identified as an inhibitor of sarcoma or carcinoma cell invasion or of metastasis , and an agent that does not increase , or decreases , the association of growth factor receptor and PTP1b in the presence of the agent as compared to in the absence of the agent is not identified as an inhibitor of sarcoma or carcinoma cell invasion or metastasis.2. A method of identifying an agent as a sensitizer of a growth factor receptor-expressing carcinoma cell to a corresponding growth factor or as a dysregulator of a PTP1b-binding growth factor receptor , comprising contacting a preparation comprising a growth factor receptor and a protein-tyrosine phosphatase 1b (PTP1b) in the presence of an amount of Menaand an amount of Mena and quantifying the association of the growth factor receptor and PTP1b in the presence of the agent and in the absence of the agent , wherein an agent that decreases the association of growth factor receptor and PTP1b as compared to the association in the absence of the agent is identified as a sensitizer of a growth factor receptor-expressing carcinoma cell to the corresponding growth factor or as a dysregulator , and an agent that does not change or that increases the association of growth factor receptor and ...

Near-infrared fluorescent probe for detecting alkaline phosphatase and manufacturing method thereof

A near-infrared fluorescent probe for detecting ALP is represented by Chemical 1 . The fluorescent probe capable of detecting ALP can selectively detect ALP only quickly and accurately. In addition, the fluorescent probe allows monitoring of a biological phenomenon occurring in cells and tissues through noninvasive in-vivo imaging and during the early osteogenic differentiation in real time.

BIOMARKERS OF THERAPEUTIC RESPONSIVENESS

The present invention relates to methods of diagnosing breast cancer in a patient, as well as methods of monitoring the progression of breast cancer and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen, The invention also relates to assay methods used in connection with the diagnostic methods described herein. 122-. (canceled)23. A method of administering a treatment regimen to a patient in need thereof for treating breast cancer , comprising:(a) measuring in a first test sample from a patient before said treatment regimen is initiated, baseline levels of a plurality of biomarkers comprising phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta,(b) measuring in an interim test sample from said patient during said treatment regimen for breast cancer interim levels of said plurality of biomarkers in said interim test sample,(c) comparing said interim levels to said baseline levels of said plurality of biomarkers,(d) evaluating from said comparing step (c) whether said patient is responsive to said treatment regimen, wherein if said interim levels of phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta are decreased as compared to said baseline levels, then the patient is responsive to said treatment regimen, and wherein if said interim levels of phosphorylated isoforms of Akt, MEK, mTOR, and GSK3beta are unchanged as compared to said baseline levels, then the patient is not responding to said treatment.24. The method of claim 23 , wherein each measuring step comprises conducting a multiplexed assay measurement of a plurality of said biomarkers in said test sample claim 23 , wherein said multiplexed assay measurement is conducted using one reaction volume comprising said test sample.25. The method of claim 23 , further comprising determining from said interim levels of said plurality of biomarkers the disease progression of breast cancer.26. The method of claim 23 , wherein each measuring step measures said level using ...

Patch-Sized Apparatus And Method For Use With An Apparel To Detect Seminal Fluid

The present invention is a patch-sized apparatus for use with an apparel to detect seminal fluid that includes disposable housing. The disposable housing is removably attached to the apparel. The disposable portion incorporates therein a fluid channel comprising a chemical solution to detect seminal fluid present on the surface of the apparel. The chemical solution includes but not limited to 1-Naphthylphosphate (disodium salt); and prostate-specific antigen (PSA)(A67-B/E3).

Method for quantifying the amount of cholesterol in high-density lipoprotein 3

A method that enables quantification of cholesterol in high-density lipoprotein 3 (HDL3) in a test sample without requiring a laborious operation is disclosed. The method for quantifying cholesterol in HDL3 comprises: Step 1 wherein phospholipase and/or sphingomyelinase is/are allowed to act on a test sample to transfer cholesterol to the outside of the reaction system; and Step 2 wherein cholesterol remaining in the reaction system is quantified. The method enables specific quantification of HDL3 cholesterol in a test sample using an automatic analyzer without requirement of a laborious operation such as ultracentrifugation or pretreatment. Further, quantification of the HDL2 cholesterol level can also be carried out by subtracting the HDL3 cholesterol level from the total HDL cholesterol level obtained by a conventional method for quantifying the total HDL cholesterol in a test sample.

Screening Broths Comprising Esculatin Compounds for the Detection of Specific Microorganisms

The present invention relates to the use of an esculatin compound in a method for detecting the presence or absence of an enzymatic reaction in a sample containing one or more enzymes and one or more first chromogenic indicator of one or more enzymatic reactions, wherein the esculatin compound is a compound of formula 2. The method according to claim 1 , wherein the enzymatically cleavable group is an α or β linked sugar residue.3. The method according to claim 1 , wherein the enzymatically cleavable group is a phosphate group having the formula POW.4. The method according to claim 1 , wherein Rand Rtogether with the carbon atoms to which they are attached form a cyclohexene ring.7. The method according to claim 1 , wherein the one or more first chromogenic indicator of the enzymatic activity is a pH indicator claim 1 , a visible color indicator claim 1 , or a fluorescent indicator.8. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 5-bromo-4-chloroindoxyl phosphate.9. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 4-methylumbelliferyl-β-D glucopyranoside.10. The method according to claim 7 , wherein the one or more first chromogenic indicator of the enzymatic activity is 4-methylumbelliferyl-β-D glucuronide.11. The method according to claim 1 , wherein the dark precipitate masks the fluorescence or color of the one or more first chromogenic indicator.12. The method according to claim 1 , wherein the presence or absence of the dark precipitate is a secondary or confirmatory test after the one or more first chromogenic indicator.13staphylococcus aureus, listeria, salmonella, clostridium, streptococcus, klebsiella, enterobacter, escherichia, citrobacter, proteus, bacillus, pseudomonas, lactobacillus,. The method according to claim 1 , wherein the one or more microorganisms are bacterial microorganisms and are selected from and coliforms ...

DEGRADABLE CARBON NANOTUBE-CONTAINING BIOSENSORS AND METHODS FOR TARGET CLINICAL MARKER DETECTION

The invention relates to carbon nanotube-containing composites as biosensors to detect the presence of target clinical markers, methods of their preparation and uses in the medical field. The invention is particularly suitable for the detection in patient biological specimens of bone markers and tissue markers. The biosensors of the invention include carbon nanotubes deposited on a substrate, gold nanoparticles deposited on the carbon nanotubes and, binder material and biomolecule deposited on the gold-coated carbon nanotubes. The biomolecule is selected to interact with the target clinical markers. The biosensor can be used as an in-situ or an ex-situ device to detect and measure the presence of the target clinical markers.

METHOD FOR DETECTING STREPTOCOCCUS AGALACTIAE USING ESTERASE ACTIVITY

A reaction medium that includes (i) an esterase enzyme substrate that is not capable of using at less than 18 hours after inoculation, (ii) at least one enzymatic substrate selected from β-cellobiosidase substrates, N-acetylglucosaminidase substrates, and a β-glucosidase substrates, and (iii) a phosphatase substrate. 1. A reaction medium , comprising:{'i': 'Streptococcus agalactiae', '(i) an esterase substrate that is not capable of using at less than 18 hours after inoculation;'}(ii) at least one enzymatic substrate selected from β-cellobiosidase substrates, N-acetylglucosaminidase substrates, and β-glucosidase substrates; and(iii) a phosphatase substrate.2. The reaction medium according to claim 1 , wherein the at least one enzymatic substrate is a β-cellobiosidase substrate.3. The reaction medium according to claim 1 , wherein the at least one enzymatic substrate is an N-acetyl-glucosaminidase substrate.4. The reaction medium according to claim 1 , wherein the at least one enzymatic substrate is a β-glucosidase substrate.5. The reaction medium according to claim 4 , wherein the β-glucosidase substrate is an indoxyl substrate.6. The reaction medium according to claim 1 , wherein the esterase substrate is selected from derivatives of indoxyl octanoate claim 1 , indoxyl nonanoate claim 1 , and indoxyl decanoate.7. The reaction medium according to claim 1 , wherein a concentration of each enzymatic substrate is between 10 and 2000 mg/l.8. The reaction medium according to claim 1 , further comprising a phosphate solution.9. The reaction medium according to claim 8 , wherein the phosphate solution is NaHPOor KHPO.10. The reaction medium according to claim 1 , further comprising a mixture of inhibitors for inhibiting or limiting growth of unwanted strains.11. The reaction medium according to claim 10 , wherein the inhibitors are antibiotics.12. The reaction medium according to claim 11 , wherein the antibiotics are aztreonam and amphotericin B. This is a divisional of ...

Systems and methods for sample use maximization

The present invention provides systems, devices, and methods for point-of-care and/or distributed testing services. The methods and devices of the invention are directed toward automatic detection of analytes in a bodily fluid. The components of the device can be modified to allow for more flexible and robust use with the disclosed methods for a variety of medical, laboratory, and other applications. The systems, devices, and methods of the present invention can allow for effective use of samples by improved sample preparation and analysis.

Senescent Cell Biomarkers

The invention relates to senescent cell biomarkers and the uses thereof. The invention also extends to methods and kits for detecting senescence, and drug conjugates and pharmaceutical compositions for killing senescent cells. 1. A method of detecting a senescent cell in a sample , the method comprising detecting the expression , in the sample , of at least one senescent cell biomarker selected from DEP-1 , NTAL , EBP50 , STX4 , VAMP3 , ARMCX-3 , LANCL1 , B2MG , PLD3 and VPS26A , or a variant or fragment thereof , wherein an increased level of expression of the at least one biomarker or a variant or fragment thereof relative to the level of expression detected in a reference sample is an indication of a senescent cell present in the sample.2. A method according to claim 1 , wherein the at least one senescent cell biomarker comprises an amino acid sequence substantially as set out in any one of SEQ ID Nos. 1 to 19 claim 1 , or a variant or fragment thereof.3. A method according to claim 1 , wherein the at least one senescent cell biomarker is DEP-1 claim 1 , NTAL claim 1 , EBP50 claim 1 , STX4 claim 1 , VAMP-3 claim 1 , PLD3 or ARMCX-3 claim 1 , or a variant or fragment thereof.4. (canceled)5. A method according to claim 1 , wherein one or more of DEP-1 claim 1 , NTAL claim 1 , B2MG claim 1 , ARMCX-3 claim 1 , PLD3 and LANCL1 claim 1 , or a variant or fragment thereof claim 1 , is used as an extracellular biomarker.6. (canceled)7. (canceled)8. The method according to claim 1 , wherein the method comprises detecting two claim 1 , three claim 1 , four or more biomarkers claim 1 , or variants or fragments thereof claim 1 , in the sample.9. The method according to claim 1 , wherein the sample is a bodily sample taken from a test subject claim 1 , and wherein the test subject is an experimental animal or a human.10. (canceled)11. (canceled)12. The method according to claim 1 , wherein sample is an ex vivo sample or an in vitro sample.13. A senescent cell detection kit for ...

Senescent Cell Biomarkers

The invention relates to senescent cell biomarkers and the uses thereof. The invention also extends to methods and kits for detecting senescence, and drug conjugates and pharmaceutical compositions for killing senescent cells. 16-. (canceled)7. A method of detecting a senescent cell in a sample , the method comprises detecting the expression , in the sample , of at least one senescent cell biomarker selected from DEP-1 , NTAL , EBP50 , STX4 , VAMP3 , ARMCX-3 , LANCL1 , B2MG , PLD3 and VPS26A , or a variant or fragment thereof , wherein an increased level of expression of the at least one biomarker or a variant or fragment thereof relative to the level of expression detected in a reference sample is an indication of a senescent cell present in the sample.8. The method according to claim 7 , wherein the method comprises detecting two claim 7 , three claim 7 , four or more biomarkers claim 7 , or variants or fragments thereof claim 7 , in the sample.9. The method according to claim 7 , wherein the sample is a bodily sample taken from a test subject.10. The method according to claim 7 , wherein the sample comprises blood claim 7 , plasma claim 7 , serum claim 7 , spinal fluid claim 7 , urine claim 7 , sweat claim 7 , saliva claim 7 , tears claim 7 , breast aspirate claim 7 , prostate fluid claim 7 , seminal fluid claim 7 , vaginal fluid claim 7 , stool claim 7 , cervical scraping claim 7 , cytes claim 7 , amniotic fluid claim 7 , intraocular fluid claim 7 , mucous claim 7 , moisture in breath claim 7 , animal tissue claim 7 , cell lysates claim 7 , tumour tissue claim 7 , hair claim 7 , skin claim 7 , buccal scrapings claim 7 , nails claim 7 , bone marrow claim 7 , cartilage claim 7 , prions claim 7 , bone powder claim 7 , ear wax claim 7 , or combinations thereof.11. The method according to claim 9 , wherein the test subject is an experimental animal or a human.12. The method according to claim 7 , wherein sample is an ex vivo sample or an in vitro sample.13. A ...

DIAGNOSIS AND TREATMENT OF INCIPIENT DIABETES

A method is described for predicting incipient diabetes, metabolic disorders or the metabolic syndrome by developing a personal temporal Phosphatase profile, which is generated by measuring phosphatase concentration in stool at a single time-point or multiple time-points. The phosphatase profile further can be used for diagnosing and determining prognosis of other incipient or overt diseases, such as the metabolic syndrome, coronary heart disease, nonalcoholic fatty liver disease, cancers, other chronic or acute diseases or infectious diseases. Also described is a specific dose of phosphatase for therapeutic use in incipient diabetes and other incipient or overt diseases. 1. A method for developing a temporal profile of stool phosphatase comprising measuring concentration of phosphatase in a stool sample from a subject , wherein said stool sample is collected at a single time point or at multiple time points.2. The method of claim 1 , wherein said multiple time points comprise daily claim 1 , weekly claim 1 , monthly or yearly.3. The method of claim 2 , wherein said subject is a human claim 2 , cattle claim 2 , pig claim 2 , sheep claim 2 , goat claim 2 , cow claim 2 , horse claim 2 , dog claim 2 , cat claim 2 , monkey claim 2 , rabbit claim 2 , rat claim 2 , mouse claim 2 , chicken claim 2 , or turkey.4. The method of claim 3 , wherein said measuring comprises:providing a substrate for phosphatase in a solid or liquid form;contacting said substrate for phosphatase with a stool sample;waiting a specified period of time to allow a color to develop or waiting a specified period of time to allow an enzymatic reaction to occur;optionally, counting pixels of photographs;comparing the developed color with photographs of standards or comparing the number of pixels to standards; andquantifying the concentration of phosphatase in said stool sample or quantifying the concentration of phosphatase in said stool sample using a spectophotometer, a biochemistry analyzer, a high- ...

SYSTEMS AND METHODS FOR SAMPLE USE MAXIMIZATION

The present invention provides systems, devices, and methods for point-of-care and/or distributed testing services. The methods and devices of the invention are directed toward automatic detection of analytes in a bodily fluid. The components of the device can be modified to allow for more flexible and robust use with the disclosed methods for a variety of medical, laboratory, and other applications. The systems, devices, and methods of the present invention can allow for effective use of samples by improved sample preparation and analysis. 111-. (canceled)12. A field testing device for preparing a test sample for an assay process , the field testing device comprising:a sample receptacle configured to receive the test sample, the test sample being a portion of a bodily fluid;a pipette tip configured to extract the test sample from the sample receptacle; and a circular rotor;', 'a motor configured to rotate the circular rotor; and', 'a bucket hanging vertically from the rotor via a pivot point connection, the bucket configured to configured to retain the test sample during the centrifugation process., 'a centrifuge configured to receive the test sample and centrifuge the test sample, the centrifuge including13. The field testing device of claim 12 , wherein the bucket is configured to rotate to a substantially horizontal position during the centrifugation process.14. The field testing device of claim 13 , wherein the bucket rotates via the pivot point connection as a result of centrifugal force caused by a rotation of the rotor.15. The field testing device of claim 12 , wherein the bucket is configured to receive the pipette tip containing the sample.16. The field testing device of claim 12 , wherein the rotor includes a counterbalance configured to offset a weight of the bucket and test sample.17. The field testing device of claim 12 , wherein the rotor further includes a second bucket hanging vertically from the rotor via a second pivot point connection claim 12 , ...

ASSAY FOR SHIP1 EXPRESSION, ACTIVITY AND SEQUENCE ALTERATIONS AS A PREDICTOR OF INFLAMMATORY BOWEL DISEASE RISK

The present disclosure is directed to detecting colon disorders by measuring the expression of SHIP1 in a sample of PBMCs. One method includes the following steps, obtaining a sample including peripheral blood mononuclear cells (PBMCs) from a subject and determining whether SHIP1 is underexpressed in the PBMCs or lacks normal enzymatic activity. The present disclosure is also directed to a method of determining the expression of SHIP1 protein expression and SHIP1 enzyme activity in PBMCs. This method includes the following steps, obtaining a sample comprising PBMCs from a subject and determining the amount of SHIP1 in the PBMCs. 1. A method of detecting a colon disorder in a subject comprising:obtaining a sample comprising peripheral blood mononuclear cells (PBMCs) of the subject; anddetermining levels of expression or enzymatic activity of SH2-containing inositol-5-phosphatase 1 (SHIP1) in the PBMCs, wherein underexpression or lack of a normal enzymatic activity is indicative of the colon disorder.2. The method of claim 1 , wherein underexpression of SHIP1 is indicative of the colon disorder.3. The method of claim 1 , wherein the subject is a human.4. The method of claim 1 , wherein the colon disorder is inflammatory bowel disease.5. The method of claim 4 , wherein the colon disorder is Crohn's Disease.6. The method of claim 1 , wherein the sample is a whole blood sample.7. A method of determining the expression of SH2-containing inositol-5-phosphatase 1 (SHIP1) protein expression and enzyme activity in peripheral blood mononuclear cells (PBMCs) claim 1 , comprising:obtaining a sample comprising PBMCs from a subject;determining the amount of SHIP-1 in the PBMCs; anddetermining the amount of enzyme activity in the PBMCs.8. A method of detecting a colon disorder in a subject comprising:obtaining a sample containing the gene phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase 1(INPP5D) of the subject;determining whether the INPP5D includes a Single Nucleotide ...

Methods for the Treatment of Solid Tumor Cancers Using Illudins and Biomarkers

Methods for determining the likelihood that a subject suffering from a solid tumor cancer will benefit from treatment with an illudin are disclosed herein. Further, there are also methods for treatment based on such determination. In several embodiments, markers prostaglandin reductase 1 (PTGR1), Protein Tyrosine Phosphatase Non-Receptor Type 14 (PTPN14), Aspartate Beta-Hydroxylase (ASPH) together with one or more genes or alone may be used to enhance or guide treatment with an illudin. In certain embodiments, the protein or gene may be expressed or methylated. 2. The method of claim 1 , wherein the solid tumor cancer is prostate cancer claim 1 , ovarian cancer claim 1 , kidney cancer claim 1 , and thyroid cancer.3. The method of claim 1 , wherein the solid tumor cancer is colorectal cancer claim 1 , pancreatic cancer claim 1 , primary liver cancers claim 1 , kidney cancer claim 1 , ovarian cancer claim 1 , uterine cancer claim 1 , or breast cancer.4. The method of claim 1 , wherein the ASPH is methylated.6. The method of claim 5 , further comprising the step of measuring the level of expression of PTGR1 of a combination thereof in the biological sample.7. The method of claim 5 , further comprising the step of measuring the level of expression of PTPN14 in the biological sample.8. The method of claim 5 , further comprising e step of measuring the level of expression of ASPH in the biological sample.9. The method of claim 5 , wherein the cancer is newly diagnosed claim 5 , relapsed claim 5 , or refractory.10. The method of claim 5 , further comprising modifying a targeted drug therapy based on expression of a gene.11. The method of claim 9 , wherein ASPH is methylated.12. The method of claim 1 , wherein the cancer is colorectal cancer claim 1 , pancreatic cancer claim 1 , primary liver cancers claim 1 , kidney cancer claim 1 , ovarian cancer claim 1 , uterine cancer claim 1 , lung cancer claim 1 , breast cancer claim 1 , prostate cancer claim 1 , sarcomas claim 1 , ...

METHODS FOR DETECTING ORGANISMS AND ENZYMATIC REACTIONS USING RAMAN SPECTROSCOPY AND AROMATIC COMPOUNDS COMPRISING PHOSPHATE

The present disclosure provides systems for the rapid and sensitive detection of organisms and molecules in samples. Reactants that produce Raman-active products are used in combination with Raman light scattering. Such compounds may comprise phosphates permitting the detection of phosphatases. The present disclosure can also be used to measure enzyme-kinetics. 1. A method for detecting the activity of at least one enzyme in a sample comprising:a) preparing a mixture comprising the sample and at least one aromatic compound comprising at least one phosphate group; 'i) optionally adding an oxidizing agent; and', 'b) incubating the mixture in the presence of a base to form at least one Raman-active product;'}c) detecting the at least one Raman-active product with Raman spectroscopy,wherein the method does not comprise Surface Enhanced Resonance Raman Scattering.7. The method of claim 1 , wherein the at least one enzyme comprises a phosphatase.8. The method of claim 7 , wherein the phosphatase is alkaline phosphatase.9. The method of claim 8 , wherein the alkaline phosphatase is conjugated to an antibody.10. The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-phenyl-1-phosphate.11. The method of claim 1 , wherein the at least one aromatic compound comprises 4-hydroxy-1-naphthyl-1-phosphate.12. The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-naphthyl-1-phosphate.13. The method of claim 1 , wherein the at least one aromatic compound comprises hydroquinone diphosphate; and wherein the method comprises adding an oxidizing agent.14. The method of claim 1 , wherein the base is sodium hydroxide.15. The method of claim 1 , wherein the oxidizing agent is sodium metaperiodate.16. The method of claim 1 , wherein the Raman spectroscopy is resonant Raman spectroscopy.17. A method for detecting at least one target in a sample comprising:a) preparing a mixture comprising the at least one target;b) incubating the ...

INHIBITORS AND THEIR USES

The present invention relates to inhibitors of PPP1 R15A and PPP1 R15B and their use in therapy, particularly in the treatment of a disease state alleviated by the inhibition of PPP1 R15A and PPP1 R15B, for example a disorder associated with accumulation of misfolded proteins or proteostatsis disorder. Compounds of the invention include compounds having the formula IA or a pharmaceutically acceptable salt thereof, wherein R, R, R, Xand Yare as defined herein. 125-. (canceled)26. A compound of 2-(2 ,3 ,4-trichlorobenzylidene)hydrazine-1-carboximidamide or a salt thereof.27. The compound of claim 26 , wherein the compound is an E-isomer of the compound.28. A pharmaceutical composition comprising (E)-2-(2 claim 26 ,3 claim 26 ,4-trichlorobenzylidene)hydrazine-1-carboximidamide or a pharmaceutically acceptable salt thereof claim 26 , and a pharmaceutically acceptable excipient.29. A method of treating a disorder of a subject claim 27 , the method comprising administering to the subject a therapeutically effective amount of the compound according to claim 27 , wherein the disorder is a disorder associated with accumulation of misfolded proteins or a proteostasis disorder.30. The method of claim 29 , wherein the disorder is selected from Alzheimer's disease claim 29 , Parkinson's disease claim 29 , Huntington's disease claim 29 , ataxias claim 29 , retinal degeneration claim 29 , glaucoma claim 29 , Amyotrophic Lateral Sclerosis claim 29 , tauopathies claim 29 , or a prion disease.31. The method of claim 29 , wherein the disorder is a polyglutamine disorder.32. The method of claim 31 , wherein the polyglutamine disorder is Huntington's disease.33. The method of claim 29 , wherein the disorder is a myelin disorder.34. The method of claim 33 , wherein the myelin disorder is selected from multiple sclerosis claim 33 , Pelizaeus-Merzbacher disease claim 33 , vanishing white matter disease claim 33 , acute disseminated encephalomyelitis claim 33 , periventricular leukomalacia ...

ASSAY FOR SCREENING MODULATORS OF HOLOPHOSPHATASE ACTIVITY

This invention relates to assays for screening test compounds for their ability to modulate holophosphatase activity. 2. A method according to wherein the modulation of holophosphatase activity is inhibition of holophosphatase activity.3. A method according to wherein the modulation of holophosphatase activity is activation of holophosphatase activity.4. A method according to any preceding claim wherein the holophosphatase activity that is modulated by the test compound is selective holophosphatase activity.5. A method according to any preceding claim wherein the holophosphatase is purified.6. A method according to any preceding claim wherein the holophosphatase claim 1 , the catalytic subunit claim 1 , the at least one regulatory subunit and/or the phosphorylated substrate is a recombinant protein.7. A method according to wherein the holophosphatase is synthesised by expressing the catalytic subunit and the at least one regulatory subunit in a cell system so as to generate a functional and selective reconstituted form.8. A method according to any preceding claim wherein the regulatory subunit is a truncated fragment of a naturally occurring regulatory subunit.9. A method according to any preceding claim wherein the catalytic subunit comprises a Ser/Thr phosphoprotein phosphatase (PPP).10. A method according to any preceding claim wherein the catalytic subunit comprises a Ser/Thr protein phosphatase 1 subunit (PP1).11. A method according to any preceding claim wherein the catalytic subunit comprises a PP1c subunit.12. A method according to any preceding claim wherein the regulatory subunit is selected from R15A and R15B claim 6 , or fragments thereof.13. A method according to wherein the fragment of a regulatory subunit is selected from R15Aand R15B.14. A method according to wherein the fragment of a regulatory subunit is selected from R15A(R15A) claim 12 , R15A(R15A) claim 12 , R15B(R15B) or R15B(R15B).15. A method according to any of to wherein the regulatory ...

Fusion peptides or proteins, their use, and systems and kits based thereupon, for the separation and/or detection of plastics, particularly of microplastics

The present invention pertains to a novel fusion protein and/or fusion peptide, preferably for use in the separation from and/or detection in an environment of one or more target polymers or plastics, e.g., one or more target polymer fragments and/M or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics; a method of preparing such novel fusion protein and/or fusion peptide, a system and kit comprising the novel fusion protein and/or fusion peptide and a (polymer or non-polymer) carrier or carrier system, a use of the novel fusion protein and/or fusion peptide or of a system and kit as mentioned in the separation from and/or detection in an environment of one or more target polymers or plastics, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics; a method of separation of one or more target polymers or plastics from an environment, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, and a method of detection of one or more target polymers or plastics in an environment, e.g., one or more target polymer fragments and/or particles or target plastic fragments and/or particles, preferably wherein the one or more target polymer particles or target plastic particles are microplastics.

TREATMENT AND PREVENTION OF MUSCLE LOSS USING L-ORNITHINE IN COMBINATION WITH AT LEAST ONE OF PHENYLACETATE AND PHENYLBUTYRATE

Disclosed herein are methods of treating and preventing muscle loss using ornithine in combination with at least one of phenyl acetate and phenylbutyrate. 1. A method of treating a condition of muscle loss , comprising administering ornithine in combination with at least one of phenylacetate and phenylbutyrate to a subject in need thereof , and thereby relieving the condition.2. The method of claim 1 , further comprising identifying a subject suffering from a condition of muscle loss.3. The method of claim 2 , wherein the subject has received liver transplantation.4. A method of preventing a condition of muscle loss claim 2 , comprising administering ornithine in combination with at least one of phenylacetate and phenylbutyrate to a subject in need thereof claim 2 , and thereby preventing the condition.5. The method of claim 4 , further comprising identifying a subject is at the risk of developing a condition of muscle loss.6. The method of claim 5 , wherein the subject is going to receive liver transplantation.7. The method of claim 1 , further comprising determining muscle weight claim 1 , muscle circumference claim 1 , lean muscle claim 1 , body weight claim 1 , ammonia level claim 1 , function(s) of one or more liver enzymes claim 1 , fat mass claim 1 , lean mass claim 1 , brain water content claim 1 , locomotor activity claim 1 , protein synthesis rate claim 1 , or any combination thereof of the subject.8. The method of claim 7 , wherein the one or more liver enzymes comprise albumin claim 7 , bilirubin claim 7 , aspartate aminotransferase claim 7 , alanine aminotransferase claim 7 , phosphatase alkaline claim 7 , or any combination thereof.9. The method of claim 7 , wherein the brain water content is frontal cortex water content.10. The method of claim 1 , at least one symptom of the condition of muscle loss is skeletal muscle loss or muscle mass loss.11. (canceled)12. The method of claim 1 , wherein the condition of muscle loss is caused by aging claim 1 , ...

METHODS TO ASSAY KINASE ACTIVITY

Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes 1. A method for a kinase assay , comprising:a) Incubating ATP, at least one kinase, and a free non-phosphorylated substrate labeled with an element tag, with a support having attached thereto metal ion coordination complexes under conditions to enable the kinase to phosphorylate the substrate;b) Separating free non-phosphorylated substrate from bound phosphorylated substrate labeled with an element tag to the support;c) Eluting the element tag associated with the resultant phosphorylated substrate into a solution; andd) Performing solution elemental analysis of said solution.2. The method of where in step (a) a multitude of free non-phosphorylated substrates each labeled with a unique element tag are incubated.3. The method of wherein the metal ion coordination complex attached to the support is a titanium oxide bead.4. The method of where in step (a) the conditions to enable the kinase to phosphorylate the substrate include a kinase reaction buffer.5. The method of where in step (a) antagonists or agonists of kinase are incubated.6. The method of wherein the kinase is delivered in the form of a cell lysate.7. A method for a phosphatase assay claim 1 , comprising:a) Incubating free phosphorylated substrate labeled with an element tag with a support having attached thereto metal ion coordination complexes;b) Separating free phosphorylated substrate from bound phosphorylated substrate labeled with an element tag attached to the metal ion coordination complexes attached to the support;c) Incubating ADP and at least one phosphatase with the bound phosphorylated substrate labeled with an element tag attached to the metal ion ...

Methods for measuring enzyme activity useful in determining cell viability in non-purified samples

Methods for detecting microorganisms, in particular detection of bacteria and methods for measuring enzyme activity, such as Deoxyribonucleic acid (DNA) polymerase activity are disclosed. The aforesaid methods include, but are not limited to such methods performed on microbial crude lysates, useful for determining microbial enzyme activities, which can be linked to amplification signal generators such as real-time Polymerase Chain Reaction (PCR) techniques, thereby enabling determination of microbial pathogens in samples such as unpurified blood and other body fluids. Moreover, the disclosed embodiments also relate to reagents for use in such methods, and to test kits comprising such reagents for carrying out the methods.

Constitutively active pyr/pyl receptor proteins for improving plant stress tolerance

The present invention provides methods of regulating plant stress tolerance.

ANTIGEN-COUPLED HYBRIDIZATION REAGENTS

The present disclosure provides high-performance hybridization reagents for use in a variety of hybridization assays and other related techniques. The hybridization reagents comprise an oligonucleotide probe and a bridging antigen, wherein the bridging antigen is recognized by a detectable antibody with high affinity. Also provided are compositions comprising panels of hybridization reagents specific for multiple different target nucleic acids and compositions comprising pairs of hybridization reagents and their complementary detectable antibodies. The paired hybridization reagents and detectable antibodies are useful in a variety of hybridization assays, particularly in highly multiplexed assays, where the structure of the bridging antigen is varied in tandem with variation in the detectable antibody, such that a multiplicity of hybridization reagents are provided that are capable of simultaneously detecting a multiplicity of target nucleic acids in a single assay. Also provided are kits comprising the hybridization reagents, methods of hybridization assay using the hybridization reagents of the disclosure, and methods of preparation of the hybridization reagents. 2. The hybridization reagent composition of claim 1 , wherein the bridging antigen is a peptide.3. The hybridization reagent composition of claim 1 , wherein the bridging antigen comprises a plurality of antigenic determinants.4. The hybridization reagent composition of claim 3 , wherein each antigenic determinant in the plurality of antigenic determinants is the same.5. The hybridization reagent composition of claim 3 , wherein the plurality of antigenic determinants comprises a linear repeating structure.6. The hybridization reagent composition of claim 5 , wherein the linear repeating structure is a linear repeating peptide structure.7. The hybridization reagent composition of claim 3 , wherein the plurality of antigenic determinants comprises at least three antigenic determinants.8. The hybridization ...

METHODS AND COMPOSITIONS FOR SYNTHETIC BIOMARKERS

The present disclosure encompasses embodiments of nucleic acids comprising genetic elements which are useful for the detection of diseased cells. 1. A method for generating a profile of a subject's disease , comprising:(a) contacting at least one cell of said subject with a plurality of genetic constructs, wherein:said plurality of genetic constructs comprises a plurality of disease-activated promoters respectively operably linked to a plurality of barcode molecules or reporter proteins and said plurality of disease-activated promoters drive expression of said corresponding barcode molecules or said reporter proteins in a cell affected by said disease; and(b) quantifying expression levels of said plurality of barcode molecules or said reporter proteins to generate said profile.2. The method of claim 1 , wherein said contacting is ex-vivo.3. The method of claim 1 , wherein said contacting is in vivo.4. The method of claim 1 , further comprising isolating said at least one cell from said subject prior to said contacting.5. The method of claim 1 , comprising quantifying expression levels of said plurality of barcode molecules to generate said profile claim 1 , wherein said plurality of genetic constructs comprise a plurality of disease-activated promoters respectively operably linked to a plurality of barcode molecules claim 1 , said plurality of disease-activated promoters drive expression of said corresponding barcode molecules.6. The method of claim 1 , wherein said plurality of disease-activated promoters comprise a cancer-activated promoter or said plurality of disease-activated promoters comprise a plurality of cancer-specific promoters.7. The method of claim 6 , wherein said cancer-activated promoter or plurality of cancer-activated promoters are activated in Acute Myeloid Leukemia claim 6 , Adrenocortical Carcinoma claim 6 , Bladder Urothelial Carcinoma claim 6 , Breast Ductal Carcinoma claim 6 , Breast Lobular Carcinoma claim 6 , Cervical Carcinoma claim 6 , ...

A BIOMARKER AND TARGET FOR DIAGNOSIS, PROGNOSIS AND TREATMENT OF ANKYLOSING SPONDYLITIS

The present invention relates to a biomarker and target for diagnosis, prognosis and treatment of ankylosing spondylitis (AS). The present invention also relates to a method for producing an animal model for AS, an animal model produced therefrom, and a method for screening for an agent pharmaceutically active in the treatment of A S using such animal model. 1. A method for detecting ankylosing spondylitis (AS) and/or predicting the risk of development of radiographic severity of AS , comprising(i) providing a biological sample from a subject; and(ii) detecting an ALPL gene product as an AS marker in the sample.2. The method of claim 1 , wherein the gene product includes a protein or a RNA transcript.3. The method of claim 1 , wherein the ALPL gene product is a non-specific alkaline phosphatase (TNAP).4. The method of claim 1 , wherein the ALPL gene product is a bone-specific TNAP (BAP).5. The method of claim 1 , wherein the marker is detected with an agent that specifically binds to the ALPL gene product.6. The method of claim 1 , wherein the detection is performed by an immunoassay claim 1 , a mass spectrometric assay claim 1 , a nucleic acid hybridization detection assay claim 1 , and/or a reverse transferase-polymerase chain reaction (RT-PCR).7. The method of claim 1 , wherein the biological sample is a body fluid sample or a tissue sample.8. The method of claim 1 , comprising comparing the results of the detection with a reference level and identifying the subject as having AS and/or at risk of development of radiographic severity of AS claim 1 , if the comparison shows an elevated level of the ALPL gene product.9. The method of claim 8 , further comprising applying a further AS diagnostic assay to the subject to confirm AS occurrence or the risk of development of radiographic severity of AS.10. The method of claim 8 , wherein the radiographic severity includes one or more radiographic features of AS selected from the group consisting of erosion claim 8 , ...

ISOLATION OF ADULT MULTIPOTENTIAL CELLS BY TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

The present invention relates to the use of tissue non-specific alkaline phosphatase (TNAP) as a marker for identifying and/or isolating adult multipotential cells. The present invention also relates to cell populations enriched by methods of the present invention and therapeutic uses of these cells. 1. Use of TNAP as a marker for the identification and/or enrichment of adult multipotential cells.217-. (canceled)18. An enriched population of adult multipotential cells obtained by a method according to .19. An enriched population of TNAP+ adult multipotential cells.20. An expanded cell population obtained by culturing an enriched population of adult multipotential cells according to .2124-. (canceled)25. A composition comprising a population of enriched adult multipotential cells according to .2628-. (canceled)29. A method for generating or repairing tissue in a subject claim 18 , the method comprising administering to the subject an enriched or expanded cell population according to .30. A method for generating or repairing tissue in a subject claim 25 , the method comprising administering to the subject a composition according to .31. A method according to wherein the tissue is bone claim 29 , cardiac or cartilage tissue.3240-. (canceled)41. A STRO-3 hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.42. A STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.43. An isolated antibody which binds to the same epitope on multipotential cells as the STRO-3 antibody produced by the hybridoma cell line deposited with ATCC on 19 Dec. 2005 under the provisions of the Budapest Treaty under deposit accession number PTA-7282.44. A composition comprising an antibody according to or .45. (canceled)46. A method for generating or repairing tissue in a subject claim 19 , the ...

MODULATORS OF PROTEIN TYROSINE PHOSPHATASE AND USES THEREOF

Composition and methods are provided for the specific manipulation of protein tyrosine phosphatase (PTP) activity, including without limitation manipulation of protein tyrosine phosphatase receptor type gamma (PTPRG). The modulation of PTP activity can be performed in vitro or in vivo, and is useful for therapeutic and research purposes. In some embodiments, an effective dose of a PTP modulator is provided to an individual for preventing or treating disease involving dysregulated tyrosine kinase activity and/or signaling mechanisms involving tyrosine phosphorylation and/or tyrosine kinase activity. In other embodiments, a PTP modulator is utilized in the analysis and screening of phosphatase pathways in a cell. 1. A method of increasing protein tyrosine phosphatase receptor type gamma (PTPRG) activity in a targeted cell population , the method comprising:contacting cells in the targeted population with an effective dose of a protein tyrosine phosphatase (PTP) activating agent selected from a PTP wedge domain (WD) polypeptide; a PTP intracellular domain (ICD) polypeptide; a PTP intracellular phosphatase domain (IPD) peptide; and a PTP soluble extracellular domain polypeptide (PTPx); or fusion, variant, or mimetic thereof;wherein PTPRG activity in the cell is increased.2. The method of claim 1 , wherein the effective dose increases PTPRG activity to result in altered tyrosine phosphorylation of one or more target proteins.3. The method of claim 1 , wherein the PTP activating agent is a human polypeptide or a fusion polypeptide thereof claim 1 , optionally fused to a permeant domain.4. (canceled)5. The method of claim 1 , wherein endogenous PTPRG protein in the cell is activated.6. The method of claim 1 , wherein the PTP activating agent is a human PTPRG ICD or IPD polypeptide or fusion polypeptide thereof.7. The method of claim 6 , wherein the ICD or IPD provides phosphatase activity in the absence of endogenous PTPRG.8. The method of claim 1 , wherein the targeted ...

METHODS OF DIAGNOSING A DISEASE AND METHODS OF MONITORING TREATMENT OF A DISEASE BY QUANTIFYING A NON-REDUCING END GLYCAN RESIDUAL COMPOUND AND COMPARING TO A SECOND BIOMARKER

Provided herein are methods of diagnosing or monitoring the treatment of abnormal glycan accumulation or a disorder associated with abnormal glycan accumulation. 126-. (canceled)28. The method of claim 27 , wherein the population of glycans comprises a glycan selected from the group consisting of chondroitin sulfate claim 27 , dermatan sulfate claim 27 , and heparan sulfate.29. The method of claim 27 , wherein the first biomarker is selected from the group consisting of IdoA-GlcNS claim 27 , IdoA-GlcNS6S claim 27 , IdoA-GlcNAc claim 27 , and IdoA-GlcNAc6S.30. The method of claim 29 , wherein the first biomarker is IdoA-GlcNS.31. The method of claim 27 , wherein the second biomarker is selected from the group consisting of ΔUA-GlcN claim 27 , ΔUA-GlcN6S claim 27 , ΔUA2S-GlcN claim 27 , AUAS-GlcN6S claim 27 , ΔUA-GlcNAc claim 27 , ΔUA-GlcNAc6S claim 27 , ΔUA2S-GlcNAc claim 27 , ΔUA2S-GlcNAc6S claim 27 , ΔUA-GlcNS claim 27 , ΔUA-GlcNS6S claim 27 , ΔUA-GlcNS3S claim 27 , ΔUA2S-GlcNS claim 27 , ΔUA2S-GlcNS6S claim 27 , ΔUA2S-GlcNS3S claim 27 , ΔUA-GlcNS6S3S claim 27 , and ΔUA2S-GlcNS6S3S.32. The method of claim 31 , wherein the second biomarker is selected from the group consisting of ΔUA-GlcNAc and ΔUA-GlcNS.33. The method of claim 27 , wherein the first biomarker is selected from the group consisting of IdoA-GlcNS claim 27 , IdoA-GlcNS6S claim 27 , IdoA-GlcNAc claim 27 , and IdoA-GlcNAc6S claim 27 , and the second biomarker is selected from the group consisting of ΔUA-GlcN claim 27 , ΔUA-GlcN6S claim 27 , ΔUA2S-GlcN claim 27 , AUAS-GlcN6S claim 27 , ΔUA-GlcNAc claim 27 , ΔUA-GlcNAc6S claim 27 , ΔUA2S-GlcNAc claim 27 , ΔUA2S-GlcNAc6S claim 27 , ΔUA-GlcNS claim 27 , ΔUA-GlcNS6S claim 27 , ΔUA-GlcNS3S claim 27 , ΔUA2S-GlcNS claim 27 , ΔUA2S-GlcNS6S claim 27 , ΔUA2S-GlcNS3S claim 27 , ΔUA-GlcNS6S3S claim 27 , and ΔUA2S-GlcNS6S3S.34. The method of claim 33 , wherein the first biomarker is IdoA-GlcNS.35. The method of claim 33 , wherein the second biomarker is selected from ...

CELLULAR ASSAYS WITH A MOLECULAR ENDPOINT MEASURED BY SAMDI MASS SPECTROMETRY

The disclosure provides a cell-based, label-free assay compatible with high-throughput screening (HTS) that can report quantitatively on enzyme activities by measuring mass changes of substrates with MALDI-mass spectrometry. 1. A method of assaying activity of an intracellular enzyme , comprising:(a) contacting a cell and a surface, the surface comprising an immobilized cell adhesion ligand and an immobilized substrate for the enzyme, the contacting resulting in immobilization of the cell via interaction between the cell and the immobilized cell adhesion ligand;(b) contacting the cell with a lysing solution to form a cell lysate and release the enzyme, thereby allowing contact between the enzyme and the immobilized substrate to transform the immobilized substrate to a product, the product having a different mass than the substrate; and(c) measuring the amount of the product formed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to assay the activity of the enzyme.2. The method of claim 1 , wherein the surface comprises a multi-well plate.3. The method of or claim 1 , wherein the surface comprises gold claim 1 , silver claim 1 , or copper.4. The method of any one of - claim 1 , wherein more than one cell is applied to the monolayer.5. The method of claim 4 , wherein 2 claim 4 , 5 claim 4 , 10 claim 4 , 20 claim 4 , 50 claim 4 , or 100 cells are applied to the monolayer.6. The method of any one of - claim 4 , wherein at least one of the immobilized substrate and the cell adhesion ligand comprises a peptide.7. The method of claim 6 , wherein the peptide is bound to the surface via a cysteine residue.8. The method of any one of - claim 6 , wherein the cell adhesion ligand comprises a RGD peptide.9. The method of any one of - claim 6 , wherein at least one of the cell adhesion ligand and the immobilized substrate is bound to the surface via a linker.11. The method of claim 9 , wherein the surface comprises a monolayer.12. The method of ...

Formation of immobilized biological layers for sensing

The invention is directed to enzyme immobilization compositions comprising: one or more enzymes, a humectant, an acrylic-based monomer, a water-soluble organic photo-initiator and a water-soluble acrylic-based cross-linker in a substantially homogeneous aqueous mixture. The invention is also directed to methods for forming sensors comprising such compositions and to apparati for forming arrays of immobilized layers on an array of sensors by dispensing such compositions onto a substrate.

Biomarkers of therapeutic responsiveness

The present invention relates to methods of diagnosing breast cancer in a patient, as well as methods of monitoring the progression of breast cancer and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen. The invention also relates to assay methods used in connection with the diagnostic methods described herein.

HYDROLASE ENZYME SUBSTRATES AND USES THEREOF

The present invention provides novel methods for determining the presence or amount of a hydrolytic enzyme in a sample, based on novel substrates for the enzymes, and also provides compositions and methods that provide highly sensitive assay methods for such hydrolytic enzymes. 210-. (canceled)1216-. (canceled)18. The method of claim 17 , wherein the hydrolytic enzyme is an esterase claim 17 , a beta-galactosidase claim 17 , or a glycosidase.19. The method of claim 17 , wherein the step of assessing the presence and/or amount of the aryl alcohol molecule or unsaturated aliphatic alcohol molecule comprises oxidizing the aryl alcohol molecule or unsaturated aliphatic alcohol molecule with an oxidizing reagent.20. The method of claim 19 , wherein the presence and/or amount of the aryl alcohol molecule or unsaturated aliphatic alcohol molecule is assessed by oxidizing the aryl alcohol molecule or unsaturated aliphatic alcohol molecule with an aryl alcohol oxidase or an aliphatic alcohol oxidase in the presence of oxygen to produce HOand assessing the presence and/or amount of the HO.21. The method of claim 17 , wherein the presence and/or amount of the aryl alcohol molecule or unsaturated aliphatic alcohol molecule is assessed by oxidizing the aryl alcohol molecule or unsaturated aliphatic alcohol molecule with an aryl alcohol dehydrogenase or an alcohol dehydrogenase in the presence of NAD or NADP to produce NADH or NADPH claim 17 , and assessing the presence and/or amount of the NAD+ claim 17 , NADP+ claim 17 , NADH or NADPH.22. The method of claim 17 , wherein the presence and/or amount of the aryl alcohol molecule or unsaturated aliphatic alcohol molecule is assessed by:{'sub': 2', '2, 'a) oxidizing the aryl alcohol molecule or unsaturated aliphatic alcohol molecule with an aryl alcohol oxidase or an aliphatic alcohol oxidase in the presence of oxygen to produce aryl aldehyde molecule or unsaturated aliphatic aldehyde molecule and HO;'}{'sub': 2', '2, 'b) reducing ...

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 18-. (canceled)9. A method of performing two or more assays for detecting a plurality of analytes in a biological sample from a subject , comprising:receiving the biological sample and a cartridge in a biological sample processing system, wherein: 'a plurality of vessels; and', 'a) the cartridge comprises a sample handling system;', 'a plurality of assay units for performing said two or more assays to detect a plurality of analytes in the biological sample, and', 'at least two detection units;, 'b) the biological sample processing system comprisesaliquoting at least a diluted portion of the sample into one of the assay unit while the cartridge remains inside the biological sample processing system;decoupling one or more vessels from the cartridge while the cartridge remains inside the biological sample processing system to form one or more decoupled components;using a centrifuge on one of said assay units;performing in the biological sample processing system the two or more assays;detecting the plurality of analytes with at least one of said detection units in the biological sample processing system.10. The method of claim 9 , wherein the biological sample is a blood sample.11. The method of claim 10 , wherein the blood sample obtained from the subject is about 500 μl or less.12. The method of claim 10 , wherein the two or more assays are performed from a single blood sample obtained from the subject.13. The method of claim 10 , wherein the blood sample is a plasma sample.14. The method ...

METHOD FOR DETECTING FLUORESCENCE OR ABSORBANCE, METHOD FOR SUPPRESSING BACKGROUND, METHOD FOR MEASURING ADP, METHOD FOR MEASURING ACTIVITY OF ADP-SYNTHESIZING ENZYME, AND METHOD FOR MEASURING ACTIVITY OF GLUCOSYLTRANSFERASE

In a method for detecting fluorescence or absorbance of the present invention, a diaphorase causes reduction from resazurin to resorufin in the presence of an SH reagent and NADH or NADPH, and the resulting fluorescence intensity or absorbance is measured. A method for measuring ADP of the present invention includes a 2-1 process in which glucose is reacted with ADP and an ADP-dependent hexokinase, a 2-2 process in which the glucose-6-phosphate obtained in the 2-1 process is reacted with NAD or NADP and glucose-6-phosphate dehydrogenase, and a 2-3 process in which resazurin is reacted with the NADH or NADPH obtained in the 2-2 process and a diaphorase in the presence of an SH reagent, and the resulting fluorescence intensity or absorbance is measured. 2. The method for detecting fluorescence or absorbance according to claim 1 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 1 , maleimide or N-(2-sulfoethyl)maleimide.4. The method for suppressing background according to claim 3 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 3 , maleimide claim 3 , or N-(2-sulfoethyl)maleimide.6. (canceled)7. The method for measuring ADP according to claim 5 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 5 , maleimide or N-(2-sulfoethyl)maleimide.9. (canceled)10. The method for measuring activities of ADP-producing enzymes according to claim 8 , wherein the maleimide compound represented by General Formula [1] is N-ethylmaleimide claim 8 , maleimide or N-(2-sulfoethyl)maleimide.11. The method for measuring activities of ADP-producing enzymes according to claim 8 , wherein the ADP-producing enzyme is at least one type selected from the group including kinases claim 8 , ATPases claim 8 , nitrogenases claim 8 , tetrahydrofolate synthases claim 8 , acetyl-CoA carboxylase claim 8 , pyruvate carboxylase claim 8 , and glutathione synthase.13. (canceled)14 ...

METHODS FOR DIAGNOSIS, PROGNOSIS AND METHODS OF TREATMENT

This invention is directed to methods and compositions for diagnosis, prognosis and for determining methods of treatment. The physiological status of a cell present in a sample (e.g. clinical sample) can be used in diagnosis or prognosis of a condition (e.g. Chronic Lymphocytic Leukemia), in patient selection for therapy, to monitor treatment and to modify or optimize therapeutic regimens. 132.-. (canceled)33. A method of correlating and/or classifying an activation state of a CLL cell with a clinical outcome in an individual by:(i) subjecting the CLL cell from the individual to a modulator, where the CLL cell expresses a B-Cell receptor (BCR);(ii) determining the activation levels of a plurality of activatable elements; and(ii) identifying a pattern of the activation levels of the plurality of activatable elements to determine the presence or absence of an alteration in signaling proximal to BCR, where the presence of the alteration is indicative of a clinical outcome.34. The method of claim 33 , further comprising determining the level of one or more cell surface markers on the cell.35. The method of claim 34 , wherein the cell surface marker is selected from the group consisting of CD1 claim 34 , CD2 claim 34 , CD3 claim 34 , CD4 claim 34 , CD5 claim 34 , CD8 claim 34 , CD10 claim 34 , CD14 claim 34 , CD19 claim 34 , CD20 claim 34 , CD22 claim 34 , CD23 claim 34 , CD40 claim 34 , CD52 claim 34 , CD100 claim 34 , CD280 claim 34 , CD281 claim 34 , CD282 claim 34 , CD283 claim 34 , CD284 claim 34 , and CD289.36. The method of claim 35 , wherein the cell surface marker is selected from the group consisting of CD45 claim 35 , CD5 claim 35 , CD14 claim 35 , CD19 claim 35 , CD20 claim 35 , CD22 claim 35 , CD23 claim 35 , CD27 claim 35 , CD37 claim 35 , CD40 claim 35 , CD52 claim 35 , CD79 claim 35 , CD38 claim 35 , CD96 claim 35 , MEW Class I claim 35 , and MEW Class 2.37. The method of claim 34 , wherein the cell surface marker is selected from the group consisting of ...

In Situ Chemiluminescent Substrates and Assays

Methods for generating a chemiluminescent enzyme substrate in situ, in aqueous or other assay conditions. Also disclosed are methods to use the substrates to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Kits relating to these methods are also disclosed. 2. The method of claim 1 , wherein the oxidant is selected from hydrogen peroxide claim 1 , sodium molybdate claim 1 , hydrogen peroxide and sodium molybdate claim 1 , hypochlorite claim 1 , hypochlorite and hydrogen peroxide claim 1 , aryl endoperoxide claim 1 , calcium peroxide peroxyhydrate claim 1 , and combinations thereof.3. The method of claim 2 , wherein the oxidant is hydrogen peroxide claim 2 , and hydrogen peroxide and sodium molybdate.6. The method of claim 1 , wherein Ris alkyl containing 1 to 2 carbon atoms or trifluoalkyl containing 1 to 2 carbon atoms.12. The method of claim 1 , wherein the enzyme moiety comprises a hydrolytic enzyme.13. The method of claim 12 , wherein the hydrolyic enzyme is alkaline phosphatase claim 12 , β-galactosidase claim 12 , β-glucosidase claim 12 , β-glucuronidase claim 12 , or neuraminidase.14. The method of claim 13 , wherein the enzyme moiety is an enzyme.15. The method of claim 14 , further comprising the step of detecting the light emitted from the reaction mixture after addition of the aqueous solution of the 1 claim 14 ,2-dioxetane enzyme substrate claim 14 , wherein the emission of light is indicative of the presence of the enzyme claim 14 , and the amount of light emitted can be correlated to the amount of the enzyme present in the sample.16. The method of claim 13 , wherein the enzyme moiety is an enzyme-linked antibody comprising a first antibody capable of binding to an antigen and an enzyme capable of cleaving the 1 claim 13 ,2-dioxetane enzyme substrate so that the substrate decomposes and generates light.17. The method of claim 16 , wherein the first antibody is covalently or non-covalently linked to the enzyme.18. The ...

INHIBITORS AND THEIR USES

The present invention relates to inhibitors of PPP1 R15A and PPP1 R15B and their use in therapy, particularly in the treatment of a disease state alleviated by the inhibition of PPP1 R15A and PPP1 R15B, for example a disorder associated with accumulation of misfolded proteins or proteostatsis disorder. Compounds of the invention include compounds having the formula IA or a pharmaceutically acceptable salt thereof, wherein R, R, R, Xand Yare as defined herein. 125-. (canceled)26. A compound of 2-(3 ,4 ,5-trichlorobenzylidene)hydrazine-1-carboximidamide or a salt thereof.27. The compound of claim 26 , wherein the compound is an E-isomer of the compound.28. A pharmaceutical composition comprising (E)-2-(3 claim 26 ,4 claim 26 ,5-trichlorobenzylidene)hydrazine-1-carboximidamide or a pharmaceutically acceptable salt thereof claim 26 , and a pharmaceutically acceptable excipient.29. A method of treating a disorder of a subject claim 27 , the method comprising administering to the subject a therapeutically effective amount of the compound according to claim 27 , wherein the disorder is a disorder associated with accumulation of misfolded proteins or a proteostasis disorder.30. The method of claim 29 , wherein the disorder is selected from Alzheimer's disease claim 29 , Parkinson's disease claim 29 , Huntington's disease claim 29 , ataxias claim 29 , retinal degeneration claim 29 , glaucoma claim 29 , Amyotrophic Lateral Sclerosis claim 29 , tauopathies claim 29 , or a prion disease.31. The method of claim 29 , wherein the disorder is a polyglutamine disorder.32. The method of claim 31 , wherein the polyglutamine disorder is Huntington's disease.33. The method of claim 29 , wherein the disorder is a myelin disorder.34. The method of claim 33 , wherein the myelin disorder is selected from multiple sclerosis claim 33 , Pelizaeus-Merzbacher disease claim 33 , vanishing white matter disease claim 33 , acute disseminated encephalomyelitis claim 33 , periventricular leukomalacia ...

SYSTEMS AND METHODS FOR MULTI-ANALYSIS

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample. 18-. (canceled)9. A method for assaying a sample obtained from a subject , comprising: a vessel comprising the biological sample;', 'a reagent unit comprising an inert liquid barrier and a reagent solution for performing an assay for detecting the presence or absence of an analyte in a sample, wherein the inert liquid barrier is immiscible with and has a lower density than the reagent solution, and wherein the inert liquid barrier is capable of having (a) a molten state to prevent fluid from passing through the inert liquid barrier in the absence of an object inserted through the liquid barrier, and (b) a solid state to prevent fluid and the object from passing through the inert liquid barrier; and', 'at least one pipette tip;', 'wherein the vessel and the reagent unit are fluidically isolated and independently moveable;, 'receiving, in a biological sample processing device, a cartridge comprising a biological sample obtained from a subject, wherein the cartridge further comprises a sample handling system comprising a pipette, wherein the pipette is configured to engage with the at least one pipette tip and the reagent unit;', 'a thermal control unit, wherein the thermal control unit is at a temperature such that the inert liquid barrier would be in a molten state; and', 'a detection unit for detecting an analyte in the sample;, 'wherein the biological sample processing device comprisesand wherein prior to insertion of the cartridge, the cartridge was stored at a temperature such that ...

Processes for increasing extraction of enzymes from animal feed and measuring activity of the same

Methods of increasing extraction of enzymes from animal feed and measuring enzyme activity are described herein.

Method of preparation of nanopore and uses thereof

This disclosure provides systems and methods for sequencing nucleic acids using nucleotide analogues and translocation of tags from incorporated nucleotide analogues through a nanopore. In aspects, this disclosure is related to composition, method, and system for sequencing a nucleic acid using tag molecules and detection of translocation through a nanopore of tags released from incorporation of the molecule.

Flash and Glow 1,2-Dioxetanes

Compounds having chemiluminescent flash and glow properties. Also disclosed are methods using the compounds to generate light, detect and/or quantify enzymes, antigens, and/or nucleic acids. Also disclosed are kits relating to these compounds. 193.-. (canceled)951. The compound of claim , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 37° C. which reaches a maximum in less than about 5 minutes.961. The compound of claim , wherein the cleavage of the bond cleavable by an enzyme moiety results in the generation of light at 37° C. which reaches a maximum in 3 minutes or less.100. A method for generating light , comprising the steps of:{'b': '1', '(a) providing a compound of claim ;'}(b) providing an enzyme complex comprising an enzyme moiety which is capable of cleaving the compound;(c) contacting the enzyme complex with the compound to form a reaction mixture; and,(d) allowing the reaction mixture to generate light.101. The method of claim 100 , wherein the cleavage of the bond cleavable by the enzyme moiety results in the generation of light at 37° C. which reaches a maximum in 3 minutes or less.105. The method of claim 100 , wherein the compound has a constant signal-to-noise ratio.106. The method of claim 100 , wherein the compound has improved sensitivity claim 100 , wherein sensitivity is defined as the lowest amount of hIL-6 detect at a signal-to-noise of 2.107. The method of claim 100 , wherein the enzyme moiety is a hydrolytic enzyme chosen from alkaline phosphatase claim 100 , β-galactosidase claim 100 , β-glucosidase claim 100 , β-glucuronidase claim 100 , or neuraminidase.108. A method for determining the presence and/or amount of an enzyme in a sample claim 100 , comprising the steps of:{'b': '1', '(a) providing a compound of claim ;'}(b) providing a sample suspected of comprising the enzyme which is capable of cleaving the compound so that it decomposes and generates light;(c) contacting the sample ...

METHODS FOR MEASURING ENZYME ACTIVITY USEFUL IN DETERMINING CELL VIABILITY IN NON-PURIFIED SAMPLES

An assay kit of reagents including a nucleic acid capable of acting as substrate for polymerase microorganism activity useful in a method of detecting polymerase activity as an indicator of the presence of a micro-organism in a sample are disclosed. The disclosed embodiments also relate to reagents for use in such methods, and to test kits comprising such reagents for carrying out the methods. 1. An assay kit comprised of reagents including a nucleic acid capable of acting as substrate for polymerase microorganism activity useful in a method of detecting polymerase activity as an indicator of the presence of a micro-organism in sample , comprising:(a) a polymerase specific substrate; and(b) differential cell lysis reagents to allow only viable micro-organism derived polymerase activity to modify the polymerase specific substrate.2. A method of detecting polymerase activity as an indicator of the presence of a micro-organism in a sample , comprising:(a) contacting the sample with a nucleic acid molecule which acts as a substrate for polymerase activity in the sample; and(b) determining, specifically, the presence of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule, to thereby indicate the presence of the micro-organism, for screening normally sterile body fluids for the presence or absence of micro-organisms therein and to provide diagnostic patient management information.3. The method of claim 2 , wherein the determining step includes determining the presence and the amount of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule.4. The method of claim 2 , wherein the determining step includes determining the amount of a nucleic acid molecule resulting from the action of the micro-organism polymerase on the substrate nucleic acid molecule. This application is a continuation-in-part of and claims the benefit of pending ...

Phosphate and tensin homolog (PTEN) for the detection of autoimmune diseases or conditions

A method for the detection of impaired responsiveness of CD4+ T-cells to regulatory T-cells (Treg), referred to as Treg resistance. The method includes measuring the expression levels of phosphatase and tension homolog (PTEN) in activated CD4+ T-cells. Furthermore, a screening method for the detection of an autoimmune disease or a condition, may comprise the steps of generating a functional gene expression profile by measuring the expression levels of phosphatase and tension homolog (PTEN) in Treg-resistant CD4+ T-cells from patients suffering of an autoimmune disease or condition, and comparing the obtained gene expression profile with the expression profile from Treg-sensitive CD4+ T-cells from healthy controls. PTEN can be utilized in a screening system for the detection of impaired responsiveness of CD4+ T-cells to Treg. 1. A method for the detection of impaired responsiveness of CD4+ T-cells to regulatory T-cells (Treg) , referred to as Treg resistance , by measuring the expression levels of phosphatase and tensin homolog (PTEN) in activated CD4+ T-cells.2. The method according to claim 1 , wherein the expression levels of PTEN are compared between activated CD4+ T-cells from patients and activated Treg-sensitive CD4+ and CD8+ T-cells from healthy donors claim 1 , wherein a downregulation of PTEN within activated CD4+ T-cells as compared to the activated Treg-sensitive CD4+ T-cells is indicative for Treg resistance.3. The method according to claim 1 , wherein an upregulation of PTEN is correlated with a responsiveness of activated CD4+ T-cells to Treg-mediated suppression.4. The method according to claim 1 , wherein the Treg resistance correlates with an accelerated IL-6 production after T cell receptor (TCR) stimulation claim 1 , enhanced phosphorylation of PKB/c-Akt and/or an increased IL-6 receptor (IL-6R) expression.5. The method according to claim 1 , wherein impaired responsiveness of CD4+ T-cells is restored by normalizing PTEN expression in activated ...