Abstract: 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.

Abstract: A method for analyzing the polypeptide content of animal tissue is described. The method includes the steps of (a) providing an animal tissue specimen; (b) depositing one or more portions of a hydrogel mixture including a protease on spatially discrete portions of the animal tissue specimen; (c) allowing sufficient time to pass for animal tissue under the hydrogel mixture to be form a digested mixture of animal tissue and hydrogel mixture; (d) removing the digested mixture from the animal tissue and extracting the polypeptides from the digested mixture to provide an extract; and (e) analyzing the polypeptide content of the extract by mass spectrometry.

Abstract: Methods of treating subjects with cancer and screening for MDM2 inhibitors that may be effective cancer therapeutics are provided herein. The cancers that may be treated using MDM2 inhibitors using the methods described herein include those that are or may become resistant to treatment with tyrosine kinase inhibitors. Methods of treating subjects with cancers that have, or develop in response to treatment with tyrosine kinase inhibitors, elevated levels of MDM2, Mcl-1 or PP5 or decreased levels of Huwe1 or CAS using MDM2 inhibitors are provided herein. The MDM2 inhibitors may be effective at treating these cancers alone or in combination with a tyrosine kinase inhibitor regardless of p53 status (mutant or wild-type) of the cancer.

Abstract: Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS.

Abstract: The present invention provides an approach for the simultaneous determination of the activation states of a plurality of proteins in single cells. This approach permits the rapid detection of heterogeneity in a complex cell population based on activation states, and the identification of cellular subsets that exhibit correlated changes in activation within the cell population. Moreover, this approach allows the correlation of cellular activities or properties. In addition, the use of potentiators of cellular activation allows for characterization of such pathways and cell populations.

Abstract: A method and kit are provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: 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.

Abstract: The disclosure relates to compositions for use in assays, the compositions comprising at least one latent fluorophore including at least one enzyme-reactive quenching group and a conjugative group; and a support connectable to the latent fluorophore by the conjugative group. The disclosure further relates to methods of measuring the presence and/or concentration of an analyte, as well as methods of measuring the relative activity of at least two enzymes.

Abstract: 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.

Abstract: The invention described herein features methods, compositions, and kits that utilize activators of pyruvate kinase M2 (PKM2) for the treatment or amelioration of disorders related to PKM2 function and characterized by abnormally low levels of serine.

Abstract: 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.

Abstract: The present invention provides methods comprising both genetic and chemical means for the production of hepatocytes from a variety of cell sources, particularly pluripotent stem cells.

Abstract: The subject invention relates in part to the surprising and unexpected discovery that insects that are resistant to Bacillus thuringiensis Cry toxins have measurably altered alkaline phosphatase (ALP) activity as compared to insects that are susceptible to Cry toxins. This and other surprising discoveries reported herein have broad implications in areas such as managing and monitoring the development of insect resistance to B.t. toxins. For example, the subject invention provides a simple and fast assay (enzymatic or otherwise) for detecting ALP activity levels and thereby monitoring the development of resistance by insects to crystal protein insect toxins. There was no prior motivation or suggestion to go about resistance monitoring using this simple and easy approach.

Abstract: A thin film culture device for detecting yeast and mold microorganisms in a sample is provided. The culture device comprises a body comprising a self-supporting substrate having a first major surface and a second major surface; a first adhesive composition disposed on a portion of the first major surface of the substrate; a substantially dry, cold-water-soluble first hydrogel-forming composition adhered to the first adhesive composition; and a plurality of indicator agents. The plurality of indicator agents comprises three indicator agents for detecting distinct glycosidase enzyme activities, an indicator agent for detecting an alkyl esterase enzyme activity, and an indicator agent for detecting a phosphatase enzyme activity, wherein each of the plurality of indicator agents comprises a detectable reporter group. A method of using the culture device is also provided.

Abstract: Methods to assay kinase activity are provided herein. The methods employ elemental analysis, including inductively charged plasma mass spectrometry (ICP-MS). The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by kinase enzymes involved in post-translational modifications.

Abstract: Provided herein are methods for assessing the risk a test subject with heart failure has of experiencing a major adverse cardiac event, requiring revascularization, requiring a heart transplant, requiring unscheduled hospitalization for heart failure, progression of heart failure status, or any combination thereof. Also provided herein are methods for assessing the risk a test subject has of developing heart failure. The present methods comprise determining the levels of paraoxonase 1 activity in the serum, non-chelated plasma, or both in the test subject and comparing the level of PON1 activity in the test subject's sample with a control or baseline value based on levels of PON1 activity in serum, non-chelated plasma, or both samples from a population of control subjects. Also provided herein are kits useful in assessing such risks.

Abstract: The present invention relates to a method for determining the presence or absence of a microorganism, said method including the steps of: 1) providing an enclosure containing a liquid or semi-solid phase consisting of a biological medium capable of containing a living form of said microorganism, nutritional elements, and an enzymatic substrate which is specific to said microorganism and which can be metabolised into at least one VOC metabolite, and a gaseous phase adjacent to said liquid or semi-solid phase; 2) exposing at least said liquid or semi-solid phase to conditions that are propitious for said microorganism to metabolise said enzymatic substrate into a molecule of said VOC metabolite; and 3) determining, by optical transduction, the presence or absence of said VOC metabolite, characterised in that the latter interacts with a nanoporous matrix, said matrix being implemented in a form that is separate from said enzymatic substrate, and in that the detection, by optical transduction, of a change in the

Abstract: In one embodiment of the invention, there is provided a method of selecting a surface chemistry for culturing a given cell line, including the steps of: (i) providing a first substrate having a plasma polymerized surface with first and second regions, said first region having a first concentration of carboxylic acid groups on said plasma polymerized surface and said second region having a second concentration of carboxylic acid groups on said plasma polymerized surface, wherein said first and second concentrations are different; (ii) culturing cells from said cell line on said plasma polymerized surface in each said region; (iii) observing activity of said cultured cells in each said region, said activity selected from the group consisting of alkaline phosphatase activity of the cultured cells, cell growth of the cultured cells, attachment of the cultured cells, and cell spreading of the cultured cells; and iv) selecting, based on a particular observed activity, a secondary substrate for culturing said cells

Abstract: 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 TiO2 matrix, 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.

Abstract: A reaction medium for detecting and/or identifying Staphylococcus aureus bacteria, comprising a combination of two enzymatic substrates for alpha-glucosidase.

Abstract: A Disease Severity Index (DSI) is provided for assessment of chronic liver disease in a patient using non-invasive liver function test results. A DSI was derived from non-invasive liver function test results based on hepatic blood flow. The DSI is used in methods for prediction of clinical outcomes, prediction of response to antiviral treatment, and assessment of progression of chronic liver diseases. Non-invasive methods to diagnose three distinct categories of patients with Primary Sclerosing Cholangitis (PSC) are provided. The methods can be used to diagnose PSC patients as Slow Progressors, Moderate Progressors and Rapid Progressors.

Abstract: The present invention provides a use for alkaline phosphatase for the manufacture of a medicament for the prevention or reduction of toxic LPS influx through a mucosal lining of a mammalian body cavity. A source of alkaline phosphatase is administered for the prophylaxis or treatment of LPS mediated or exacerbated diseases. The invention also provides compositions comprising a source of alkaline phosphatase for the prevention or reduction of (toxic) LPS influx or passage through mucosal layers.

Abstract: The present invention provides fluorogenic substrates and methods of use in detecting and analyzing phospholipase C isozyme (PLC) activity.

Abstract: The present invention provides methods for identifying compounds that selectively and specifically modulate RGS21 gene expression, RGS21 protein expression, and/or the interaction of RGS21 with G proteins in taste signal transduction. In particular, the present invention provides methods for identifying modulators of RGS21 activity for enhancing sweet taste, or other taste perception. Compositions comprising modulators of RGS21 activity for modulating taste signaling transduction are also provided.

Abstract: The present disclosure provides aromatic-cationic peptide compositions and methods of using the same. The methods comprise use of the peptides in electron transport and electrical conductance.

Abstract: The present inventors have surprisingly discovered that phytic acid tenaciously precipitates with soluble metals in food or fuel ethanol-processing fluid, producing insoluble organometallic salt deposit or scale on the processing equipment that must be removed in order to facilitate further ethanol processing. The present invention relates to converting phytic acid salts or phytates to inorganic phosphates to improve metal solubility and reduce deposition within processing equipment.

Abstract: A molecular probe for the luminescent detection of nucleotides (e.g., adenosine nucleotides) is presented. In certain embodiments, the probe can readily distinguish between the three adenosine nucleotides in buffered aqueous conditions at neutral pH, a need for the direct monitoring of enzymatic reactions converting ATP to ADP or AMP. The probe is most efficient under millimolar concentrations of ATP, which are relevant to intracellular conditions. In preferred embodiments, the long luminescence lifetime of the probe readily enables time-gating experiments.

Abstract: A chimeric phosphorylation indicator (CPI) as provided herein can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. Where the phosphorylatable domain is phosphorylatable by protein kinase C (PKC), the CPI is a c-kinase activity reporter (CKAR). Donor and acceptor molecules may be, independently, fluorescent proteins such as non-oligomerizing fluorescent proteins. A CPI can contain a phosphorylatable polypeptide and a fluorescent protein; the phosphorylatable polypeptide may be contained within the sequence of the fluorescent protein, or the fluorescent protein may be contained within the sequence of the phosphorylatable polypeptide. The spatiotemporal properties of the PKC signal pathway may be tested with CKAR, calcium-sensing fluorophores and FRET-based translocation assays. Polynucleotides encoding such CPIs, and kits containing the indicators and/or the polynucleotides, are provided.

Abstract: A single device to collect, transfer, and measure salivary nitric oxide analyte and metabolite, nitrite, a biomarker for nitric oxide, as well as, a method to assess the effects of diet and exercise on changing an individual's nitric oxide status and health.

Abstract: The invention is directed to a composition comprising all or a portion of a beta-tricalcium phosphate (?-TCP) bound to all or a portion of a ?-TCP binding peptide and methods of use thereof.

Abstract: The present invention provides an assay for the identification of agents which can modulate TOR-mediated phosphorylation of substrate proteins. The assays of the invention utilize substrate proteins whose amino acid sequence contains the Ser/Thr motif recognized by TOR. Naturally occurring TOR which may be used in the methods of the invention include TOR isolated from a variety of species, particularly mammalian tissues.

Abstract: The present invention relates to devices and methods for measuring the quantity of multiple analytes in a sample. The device is designed such that each of the analyte sensing elements is configured to measure the quantity of a predetermined analyte and where the machine executable instructions are configured to select the proper analyte sensing element corresponding to the analyte to be measured.

Abstract: Methods and apparatus for evaluating the quality of an environment or process by measuring light emitted from a bioluminescent sample containing ATP, ADP, or alkaline phosphatase. The apparatus comprises a sample collection and analysis system used to collect a sample, mix reagents, react the sample, and collect it in a measurement chamber. The system includes an instrument having a photon detection assembly for use with the sample testing device and one or more probe assemblies that optically cooperate with the instrument. The instrument includes a dark chamber with a reflective interior surface which may be concave or preferably spherical, and a photon detection sensor such as a multi-pixel photon counter sensor. A substantially transparent portion of the probe assembly, and liquid contained therein, focus bioluminescence toward the photon detection sensor.

Abstract: The present invention provide reagents and methods of using the reagents, for example, on automated staining devices, that facilitate detection of two or more antigens in a sample simply and efficiently.

Abstract: 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.

Abstract: 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.

Abstract: The present invention provides a use for alkaline phosphatase for the manufacture of a medicament for the prevention or reduction of toxic LPS influx through a mucosal lining of a mammalian body cavity. A source of alkaline phosphatase is administered for the prophylaxis or treatment of LPS mediated or exacerbated diseases. The invention also provides compositions comprising a source of alkaline phosphatase for the prevention or reduction of (toxic) LPS influx or passage through mucosal layers.

Abstract: The present innovation provides methods and kits that enable rapid and efficient dual end-tagging of RNA to prepare libraries for analysis by applications such as next-generation RNA sequencing, qPCR, microarray analysis, or cloning. The methods do not require time-consuming and inefficient gel-purification steps that are common to methods known in the art. In addition, the present invention provides methods and kits for rapid, high-throughput enzymatic preparation of 5?-activated, 3?-blocked DNA oligonucleotides from standard, single-stranded DNA oligonucleotides.

Abstract: The present invention is directed to a composition comprising a vanadium-containing phosphatase inhibitor and a polyol. In the presence of the polyol the effect of the inhibitor is enhanced, even in the presence of chelating agents or reducing agents. The invention also concerns the use of the inventive composition for inhibiting a phosphatase, as well as kits comprising the composition.

Abstract: Provided herein are methods, test units, systems, assays and kits for qualitative and/or quantitative detection of microorganisms or microbes. Some aspects of the invention utilize a test strip comprising a test area and a self-calibrating control area that permits quantitative measurements to be obtained rapidly at the test site. The test strip is ready-to-use and does not require any preparation. The present systems and methods provide valuable tools for sensitive early detection of microbial contamination.

Abstract: The methods of the disclosure provide fluorescence-based assays for calcineurin activity, especially in isolated T cells. The methods include the stimulation of the T cells with agents that specifically target the TCR with or without influencing co-stimulatory pathways. One TCR agonist is monoclonal antibodies specific for CD3, which more precisely distinguish the inducible activity of calcineurin than does an alternative method targeting the T cell receptor (CD3) combined with CD28 costimulation. This method more accurately distinguishes between the measured level of calcineurin activity of T cells from immunosuppressed transplant recipients and normal individuals, and thus has improved diagnostic accuracy with respect to the response of an individual to immunosuppressant therapy following an organ transplant.

Abstract: The present invention relates to the use of quinolinone derivatives of general formula (I), ligands of the Smo receptor or of related receptors which target a binding site of the receptor which is different from the known ligand binding sites, as research tools for identifying modulators of the Smo receptor or of related receptors, characterizing the Hedgehog signalling pathway and diagnosis; the invention also relates to kits containing said derivatives of general formula (I).

Abstract: A method of detecting cancer cells is disclosed. The method comprises (a) contacting the cell which is suspicious of a cancerous or undifferentiated phenotype with 4-aminophenyl phosphate (p-APP) under conditions wherein alkaline phosphatase of the cell catalyzes a reaction of the cell with the p-APP, so as to generate a product capable of producing an electrical signal; and (b) measuring a level of the electrical signal, wherein a difference in a level of the electrical signal compared to a predetermined threshold is indicative of a cancer cell.

Abstract: An enzyme detection method includes forming a caged substrate; releasing an uncaged substrate by cleaving a caging molecule from the caged substrate; and emitting a light emission from a Bioluminescence Resonance Energy Transfer luminescent nanocrystal conjugate reacting with the uncaged substrate.

Abstract: 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.

Abstract: 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.

Abstract: A single molecule detection platform is disclosed. The single molecule detection platform comprises a light-transmissive substrate, a plurality of spherical particles and a thin film. The surface of the light-transmissive substrate is etched to form a plurality of cone-shaped structures. Each spherical particle is disposed on top of each cone-shaped structure. The sizes of the plurality of spherical particles are suitable to allow only a single protein to be attached to each spherical particle. The thin film is deposited on the surface of the plurality of cone-shaped structures and acts as a reflective layer of one-dimensional waveguide. The plurality of spherical particles is not covered by the thin film.

Abstract: 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.

Abstract: A reaction medium for detecting and/or identifying methicillin-resistant Staphylococcus aureus (MRSA) bacteria includes a chromogenic substrate, a first antibiotic that belongs to the cephalosporin family and a second antibiotic that belongs to the aminoglycoside family.

Abstract: Described herein is the generation of Fah+/? heterozygote pigs by homologous recombination and somatic cell nuclear transfer, and a method for producing Fah?/? homozygote pigs. The Fah-deficient pigs of the disclosure can be used for a variety of research and therapeutic purposes, such as for the expansion of human hepatocytes, and as large animal models of hereditary tyrosinemia type 1, cirrhosis and hepatocellular carcinoma.