Abstract: The invention relates to methods for detecting pyrophosphate by means of bioluminescence detection. In addition, methods for measuring chemical, especially enzyme-catalyzed, reactions in which pyrophosphate is formed or consumed are described. Such reactions are catalyzed for example by guanylyl cyclases, adenylyl cyclases, DNA polymerases or RNA polymerases. The novel methods are distinguished by high sensitivity and low susceptibility to interference, can easily be automated and miniaturized and are additionally suitable for carrying out continuous measurements. The methods can be employed particularly advantageously in the area of medical diagnosis and biomedical research, including pharmaceutical active ingredient research.

Abstract: This application claims a method for detecting biologically active substances, comprising the following steps a) providing a support carrying substances to be tested b) providing a suspension containing luminescent microorganisms, c) coating the support with the suspension of microorganisms, d) detecting the biologically active substances on the support by detecting the change in luminescence of the suspension of microorganisms, and e) stimulating the luminescence of the microorganisms before or during detection, and/or f) extending the period of luminescence of the microorganisms by employing substances for regulating and extending the period of luminescence of the microorganisms.

Abstract: The invention provides methods that employ derivatives of 2-cyano-6-hydroxy- or 2-cyano-6-amino-benzothiazole, for example, in a bioluminogenic reaction. Also provided are novel compounds that can be used in the methods. The invention further provides methods for detecting or determining the presence of molecules and/or enzymes, the modulator activity of such molecules, and/or the activity of such enzymes. The methods are adaptable to high-throughput format.

Abstract: The invention relates to a method for diagnosis of a disease, wherein presence or absence of an anti-endothelin-receptor antibody is determined in a sample from a patient to be diagnosed more in particular an anti-endothelin-receptor-A antibody. The disease according to the invention is in particular selected from diabetes, preferably type I diabetes, graft rejection, pre-eclampsia, hypertension, vasculitis, collagenosis, Raynaud-Syndrom (Morbus Raynaud), and inflammatory rheumatic disease and arteriosclerosis. The invention further relates to the use of an inhibitor of an anti-endothelin-receptor antibody or an inhibitor of an endothelin-receptor for the production of a medicament as well as a method for removing anti-endothelin-receptor antibodies from isolated blood by means of plasmapheresis.

Abstract: The invention provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.

Abstract: Use of a luciferase that has a mutation of at least one amino acid selected from the group consisting of positions 14, 35, 182, 232 and 465, where the numbering is according to the sequence of the luciferase from P. pyralis (SEQ ID NO:1) in a method that is performed at a pH below the optimal pH for the wild-type luciferase during at least part of the time period over which bioluminescence measurements are taken, wherein the specific activity of the mutant luciferase is higher than the specific activity of wild-type at the pH at which the method is carried out.

Abstract: Reagents and compositions for use in reactions catalysed by luciferase enzymes, and in particular for use in luciferase-based gene reporter assays are described. The invention also provides methods and compositions for, inter alia, increasing the sensitivity and/or improving the kinetics of luciferase-catalysed reactions.

Abstract: The full-length nucleic acid sequence of the C1 bacteriophage is disclosed in the present application The specific regions of the C1 genome encoding the PlyC lysin have also been identified and sequenced. The invention relates to the pharmaceutical and diagnostic utility of these sequences and provides for development of pharmaceutical compositions for treating or preventing streptococcal infections in mammals, for compositions for decontamination of inanimate surfaces and for diagnosis of streptococcal infections.

Abstract: The present invention relates generally to assays, methods, and kits that provide reagent mixes and instructions for determining the proliferative status of isolated target cell populations. The methods measure the luminescent output derived from the intracellular ATP content of incubated target cells, and correlate the luminescence with the proliferative status of the cells.

Abstract: The present invention relates to genetic reporters. Specifically, the present invention is directed to a modified gene encoding a luciferase for high level expression in an organism with a bias for cytosine (C) or guanine (G) in the third position of the codon.

Abstract: A sensitive bioluminescent assay to detect proteases including caspases is provided which employs an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a substrate for a caspase or an aminoluciferin or a carboxy-terminal protected derivative thereof covalently linked via a peptide bond to a peptide substrate comprising aspartate that is specifically cleaved by a protease specific for the substrate.

Abstract: Transgenic plants, and a method for making the same, wherein genes encoding the enzyme luciferase and its corresponding substrate luciferin are incorporated into a native plant genome. Once transformed into plant cells, these genes may be regulated such that under certain endogenous or exogenous conditions, their expression in the mature plant results in bioluminescence. Different luciferin/luciferase complexes and/or mechanisms of regulation may be utilized for these transgenic plants, depending on a variety of factors such as plant species and the circumstances under which a bioluminescent reaction is desired. Phototransformation may be utilized to vary the wavelength of light emitted from the mature plant.

Abstract: The present invention relates generally to methods to monitor the transport of proteins through the secretory pathway, and methods to monitor ER stress. In particular, the present invention relates to methods to monitor, in real-time, the processing of protein through the secretory pathway, which can be monitored both at a subcellular level by florescence visualization and quantitatively by detecting the secreted luciferase reporter protein. The present invention also relates to methods to assess biological processes in cells, in particular the secretory pathway and ER stress, as well as methods to identify agents which augment or inhibit the secretory pathway and/or ER stress. The present invention also relates to compositions and nucleic constructs encoding a secreted luciferase-fluorescent protein conjugate for methods to monitor protein trafficking in the cell by simultaneous detection of fluorescence and luciferase secretion.

Abstract: A solid-phase immunoassay for 6-keto-Prostaglandin F1?, the stable hydrolysis product of prostacyclin (Prostaglandin I2) is disclosed. Prostacyclin, a potent vasodilator with anti-platelet and anti-proliferative properties is an effective treatment for primary pulmonary hypertension and pulmonary arterial hypertension associated with scleroderma and scleroderma-like syndrome. Levels of 6-keto-Prostaglandin F1? can be directly correlated with levels of prostacyclin. Therefore, 6-keto-Prostaglandin F1? has become the indicator of choice to measure prostacyclin levels. The single step immunoassay for 6-keto-Prostaglandin F1? uses the bioluminescent protein, aequorin as a label. Analyte-label conjugates were constructed by linking the carboxyl group of 6-keto-Prostaglandin F1? and lysine residues of aequorin by chemical conjugation methods. The binding properties of 6-keto-Prostaglandin F1? towards its antibody and the bioluminescent properties of aequorin are retained in the conjugate.

Abstract: The invention relates to new compounds having heptose synthesis inhibitory properties, of formula (I) or a pharmaceutically acceptable salt, or prodrug thereof, wherein A is an aryl or heterocycle, optionally substituted by one or several identical or different R such as H, C1-C10 alkyl, C1-C10 alkyl-OR1, C1-C10 alkyl-NR1R1, alkoxy, hydroxy, thioalkyl, aryl, heterocycle, halogen, nitro, cyano, CO2R1, NR1R1, NR1C(O)R1, C(O)NR1R1, NR1C(S)R1, C(S)NR1R1, SO2NR1R1, SO2R1, NR1SO2R1, NR1C(O)NR1R1, NR1C(O)OR1, NR1C(S)NR1R1, NR1C(S)OR1, R1C?NOR1, C(O)R1, aryloxy, thioaryl, alkenyl, alkynyl R1 identical or different is H or C1-C10 alkyl B1, B2, B3 identical or not represent C, N, O, S to form a five-membered aromatic ring wherein from one to three carbon atoms are replaced by a heteroatom selected from S, O, N optionally substituted by one or several identical or different R such as defined above B4 is C or N Y is H, C1-C10 alkyl, alkoxy, thio-alkyl, optionally substituted by one or several identical or different R suc

Abstract: An assay method to identify agents that will reduce the inflammation associated with many diseases, providing a method to determine compliance of patients to clinical protocols. The fundamental tool of the inventive method is luminescence. Genetically modified cells are used to express a complex revealing the potential of certain compounds to prevent or reduce adverse effects. More specifically the invention is a method for the determination of inhibition of a chemical compound comprising: culturing genetically modified cells which express an indicator-luminescent complex; placing said complex in the presence of an agent that essentially totally degrades said complex; measuring the luminescence of the resulting reaction; collecting a sample from a mammal consuming a complementary and alternative medicine (CAM); placing said sample in the presence of said complex; and comparing the level of luminescence from step (c) with the luminescence from step (e) to determine the inhibition of said chemical compound.

Abstract: Provided is a method of detecting infection in a wound caused by an infecting organism at a wound site. Also provided is a system for detecting an infection in a wound at a wound site. Additionally, a porous pad comprising luciferase is provided.

Abstract: The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising of the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having a desired activity. The invention enables the in vitro evolution of nucleic acids by repeated mutagenesis and iterative applications of the method of the invention.

Abstract: The objects of the present invention are to provide a new technology platform for quantitation number of copies of nucleic acid molecules of interest by lumonogenic (i.e., enzymatic luminescence) detection. The detection approach of the method of present invention is essentially employing time-resolved approach, e.g., based on detection transient parameters of luminescent signal. The various disclosed embodiments allow DNA and RNA quantification in a broad dynamic range; can be used for detection of DNA, as well as long (mRNA) and short (microRNA) RNA targets. The present invention provides methods, instruments, and kits for fast and highly sensitive identification and measurements of nucleic acids in various life science and biomedical applications.

Abstract: A modified luciferase protein which is a sensor for molecules including cAMP is provided. The modified luciferase protein includes one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with cAMP.

Abstract: A method of testing for sanitization of textiles comprises the steps of cleaning textiles in a water solution and testing the water solution for the presence of contaminants such as adenosine triphosphate (ATP), typically with a luminometer. Typically, the water solution will be drained from a cleaning vessel and tested. Another option is the testing of the water solution extracted after draining such as by a spin cycle. The method provides improved accuracy of test results as to the level of cleanliness. In addition, testing at this early step of the laundering process allows for additional cleaning if needed without having undertaken costly and time-consuming steps such as drying. Moreover, absent re-contamination of the textiles after the cleaning process, drying and finishing procedures may be accomplished without further sanitizing the textiles.

Abstract: Cell potency assays for use with cell-based therapies, and specifically, with stem cell therapies, are provided. Cell potency assays for bone marrow cells, mobilized peripheral blood, and umbilical cord blood are provided.

Abstract: A system and method for integrating microfluidic components in a microfluidic system enables the microfluidic system to perform a selected microfluidic function. A capping module includes a microfluidic element for performing a microfluidic function. The capping module is stacked on a microfluidic substrate having microfluidic plumbing to incorporate the microfluidic function into the system. The microfluidic element may comprise a matrix having an affinity for selected molecules in a sample. The matrix binds, reacts with and/or retains the selected molecules without affecting other molecules in the sample.

Abstract: Provided herein are methods for screening compounds for their ability to modulate the expression of certain isoforms of proteins that are associated with cancer, such as isoforms of proteins that participate in Wnt signaling in cancer cells.

Abstract: A method for detecting the absence or presence of cells of interest in a liquid sample, wherein: (a) the sample: (i) comprises an extracellular medium containing an enzyme with a measurable activity; and (ii) is suspected of containing cells of interest that contain an enzyme with said measurable activity; and (b) the method comprises the steps of: (i) treating the liquid sample with a reagent that inactivates said measurable activity in the extracellular medium, but does not inactivate the measurable activity in said cells of interest; (ii) lysing the cells of interest to release the intracellular enzyme; and (iii) measuring said measurable activity. Thus the intracellular enzyme can be measured without interference from the extracellular enzyme. The invention is particularly useful for treatment of bacterially-infected blood using a detection assay based on adenylate kinase activity.

Abstract: Nucleotide sequences mediating male fertility in plants are described, with DNA molecule and amino acid sequences set forth. Promoter sequences and their essential regions are also identified. The nucleotide sequences are useful in mediating male fertility in plants. In one such method, the homozygous recessive condition of male sterility causing alleles is maintained after crossing with a second plant, where the second plant contains a restoring transgene construct having a nucleotide sequence which reverses the homozygous condition. The restoring sequence is linked with a hemizygous sequence encoding a product inhibiting formation or function of male gametes. The maintainer plant produces only viable male gametes which do not contain the restoring transgene construct. Increase of the maintainer plant is also provided by self-fertilization, and selection for seed or plants which contain the construct.

Abstract: This invention provides novel compounds selected from the group consisting of a 1,5-substituted pyrimidine derivative or analog and substituted furano-pyrimidone analogs. The compounds are useful to treat or prevent diseases such as cancer.

Abstract: The invention concerns methods, agents and kits for qualitative and quantitative detection and identification of pathogens and pathogen spectra based on endotoxins and other pyrogens.

Abstract: The invention provides methods of detecting bacteria in fluids, including blood, platelets and other blood products for transfusion, and urine. The methods are based on lysing the bacteria to release ATP and detecting the ATP. Eukaryotic cell contamination is a problem to be overcome, because eukaryotic cell contain large amounts of ATP. Thus, some of the methods involve separating intact eukaryotic cells (e.g., platelets) from intact bacterial cells before lysing the bacterial cells to release ATP, contacting the ATP with an ATP-consuming enzyme that catalyzes a reaction, and monitoring the enzyme-catalyzed reaction. Typically, the enzyme is luciferin, and the reaction is monitored by detecting light produced by the luciferin. Other methods of the invention involve contacting a fluid sample with a support surface that binds bacterial cells, lysing the bacterial cells to release ATP, contacting the ATP with an ATP-consuming enzyme, and monitoring the enzyme-catalyzed reaction.

Abstract: The present invention provides genes encoding novel luciferases having at least the properties of: being capable of using coelenterazine as their luminescent substrates; and being capable of being recombinantly expressed in a mammal cell as a host and produced to be secreted to the outside of the host cell. Specifically, the gene encoding novel luciferases according to the present invention is a DNA molecule comprising a nucleotide sequence encoding any of the full-length amino acid sequences of two types of luciferase proteins, luciferase 1 and luciferase 2, from M. pacifica, and is, for example, a gene encoding the following full-length amino acid sequence of the luciferase 1.

Abstract: The present invention generally relates to a methods, compositions and assays for real-time monitoring of the progression of a disease, such as a cancer in a subject, by measuring the level of bioluminescence signal in a biological sample obtained from a subject, where the bioluminescence signal is from a secreted luciferase protein expressed by a cell or tissue in the subject. One aspect of the present invention relates to administering to a subject a nucleic acid encoding a secreted luciferase, and in some embodiments, a disease or a diseased tissue such as tumor cells expresses the secreted luciferase protein, which is monitored in a biological sample, such as blood or urine obtained from a subject. One aspect of the invention relates to analysis of a secreted luciferase protein by measuring the level in a biological sample obtained from the subject without the need for invasive monitoring procedures.

Abstract: The invention provides methods and compositions for modulating angiogenesis in a subject. The methods of modulating angiogenesis in a subject include administering to the subject a modulator of N-terminal arginylation activity. The invention also provides a method of identifying such a modulator and a method of in vitro screening for modulators of N-terminal arginylation activity. Additionally, the invention provides a method of treating an angiogenesis-related disorder.

Abstract: The present invention provides tumor cell preparations for use as models of the EMT process for use in the identification of anti-cancer agents, wherein said tumor cell preparations comprise cells of the epithelial tumor cell line H358, which are stimulated by receptor ligands to induce EMT, or which have been engineered to inducibly express a protein that stimulates EMT. The present invention also provides methods of identifying potential anti-cancer agents by using such tumor cell preparations to identify agents that inhibit EMT, stimulate MET, or inhibit the growth of mesenchymal-like cells. Such agents should be particularly useful when used in conjunction with other anti-cancer drugs such as EGFR and IGF-1R kinase inhibitors, which appear to be less effective at inhibiting tumor cells that have undergone an EMT.

Abstract: Sequential reporter enzyme luminescence (SRL) methods are provided. In the subject methods, the activity of a reporter enzyme is evaluated using a secondary reporter system that employs a product of a reporter enzyme mediated reaction as a luminescent substrate, e.g., luciferase substrate. Also provided are kits and other compositions that find use in practicing the subject methods.

Abstract: The invention relates to novel targets in the screening for compounds useful in the treatment and/or prophylaxis of a disease selected from the group comprising cardiovascular diseases, disorders of lipid metabolism or atherosclerosis. The invention relates to novel compounds for use as a medicament for diseases or conditions involving a disease selected from the group comprising cardiovascular diseases, disorders of lipid metabolism or atherosclerosis. The invention especially relates to antagonists and expression-inhibitory compounds that target G-protein coupled receptors (GPCRs), kinases and proteases, and to methods for identifying such compounds. The invention further relates to methods for identifying these antagonists and expression-inhibitory compounds, and methods for diagnosing a disease selected from the group comprising cardiovascular diseases, disorders of lipid metabolism or atherosclerosis or a susceptibility to such a condition.

Abstract: The present invention is directed to novel polypeptides having homology to a polypeptide suppressor of the Drosophila melanogaster fused protein and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

Abstract: Screening assays that allow for the identification of agents that modulate the activity of the arginylation branch of the N-end rule pathway are provided. Also provided are method of using an agent that modulate the activity of the arginylation branch of the N-end rule pathway to increase or decrease protein degradation in a cell, and to modulate physiologic and pathologic associated with N-end rule pathway mediated arginylation.

Abstract: Methods of monitoring enzyme mediated reactions, and particularly nucleic acid synthesis reactions such as pyrosequencing methods that employ enzymatic reporter systems. The methods and systems provide elevated signal levels as compared to conventional pyrosequencing processes, and/or mediate de-phasing of sequencing analyses employing pyrosequencing or other “sequencing by synthesis” methods.

Abstract: The present invention relates to luciferase having resistance to a surfactant and a method for measuring intracellular ATP which is characterized in that the luciferase having resistance to a surfactant is used in this method comprising the steps of: a first step wherein ATP is extracted from cells in a sample; a second step wherein light emission is produced by adding a luminescence reagent containing luciferase to the extracted ATP solution; and a third step wherein the light emission is measured.

Abstract: The invention comprises methods and cell lines for assaying APOBEC3G degradation and methods for identifying inhibitors of APOBEC3G degradation. The invention also provides methods of identifying inhibitors of HIV infection. The methods of the invention are useful for identifying inhibitors of viral infection, in particular, the methods of the invention are useful for treating retroviral infection.

Abstract: The present invention provides an “in vivo and in vitro real-time bioluminescence imaging means,” which can transmit a detection signal promptly in response to an external signal, while taking advantage of a single-molecule-format luminescent probe as a bioluminescent means. The present invention is characterized by using, as a single-molecule-format luminescent probe utilizing the increase and decrease of a second messenger level as an index, a fusion protein including a single-chain protein containing a second messenger recognition protein and optionally a peptide which is capable of binding with the protein, and linked respectively to the N-terminus and the C-terminus thereto, an N-terminal fragment (N-LE) and a C-terminal fragment (C-LE) generated by dissecting a luminescent enzyme (LE).

Abstract: Embodiments of the present disclosure include double-fusion human embryonic stem cells, methods of imaging double-fusion human embryonic stem cells, double-fusion polynucleotides, double-fusion proteins, triple-fusion human embryonic stem cells, methods of imaging triple-fusion human embryonic stem cells, triple-fusion polynucleotides, triple-fusion proteins, methods of monitoring the progression of human embryonic stem cells, methods of making isolated double-fusion human embryonic stem cells, methods of making isolated triple-fusion human embryonic stem cells, and the like.

Abstract: A reagent system comprises a first reagent which includes a high pH phosphate buffer, and a second reagent which includes luciferase, luciferin, a magnesium salt and an enzyme stabilizer. The second reagent has a low pH and a buffer with a pK which is near the optimum pH for activity of luciferase. The reagent system may be used in a process for measuring total adenosine triphosphate (ATP) and/or dissolved extracellular ATP, in a fluid containing microorganisms. The reagent system may also be used in a microbiological remediation or production process.

Abstract: Provided are rapid and sensitive cell-free assay methods for detecting and/or measuring specific bimolecular or higher order interactions via reassembly of a split monomeric reporter protein, and methods of detecting or identifying modulators of such interactions by the effect on the signal provided by the reassembled split reporter protein. This methodology is adaptable to protein-protein, protein-peptide, protein-nucleic acid, protein-methylated or nonmethylated nucleic acid and other small or large molecule ligands and binding proteins.

Abstract: The invention provides assays and methods for determining whether a compound inhibits gamma secretase in a substrate specific manner. The invention provides an isolated cell wherein the cell stably expresses APP and at least one gamma secretase substrate other than APP. The invention provides assays and methods comprising contacting a cell with gamma secretase and detecting production of Abeta, detecting production of intracellular domain (ICD), and detecting a signal from a reporter gene under transcriptional control of the ICD. The invention also provides compounds that inhibit gamma secretase, pharmaceutical compositions comprising such compounds, and methods of treating Alzheimer's disease using such compounds.

Abstract: A promoter comprising the isolated promoter region of the human Thymic Stromal Lymphopoietin (TSLP) gene and functional portions of the promoter region having TSLP promoter activity. The promoters are useful for identifying promoter agonists and antagonists that can be used to prevent or treat allergic conditions and autoimmune diseases.

Abstract: The present invention concerns a method for identifying substances useful for treating inflammation, characterized in that it comprises the following steps: (a) placing said substance in contact with a nucleic acid construct containing the Retinoid-Related Orphan Receptor (ROR) receptor response element of the promoter of the I?B? gene, under conditions suitable for allowing said substance to bind to said response element, (b) measuring the possible binding of said substance to the response element or the transcriptional activity of the response element or of a promoter containing it, (c) optionally comparing the measurement of step (b) with a measurement carried out under conditions similar to those of steps (a) and (b) but with a nucleic acid construct containing the mutated response element.

Abstract: The invention includes methods and kits for rapidly detecting tuberculosis or other mycobacterial infection in a sputum sample inexpensively and within minutes. It includes methods and kits for determining the species or phylogenetic group of mycobacterial infection. It includes methods and kits for determining the drug sensitivity of mycobacteria from a sputum sample inexpensively and within 1-3 days.

Abstract: Methods are provided for sensitive detection of adenosine triphosphate (ATP) in samples using the luciferin-luciferase reaction. Aspects include using a pH composition that maximizes a signal to noise ratio. The maximum signal to noise ratio can be particularly useful with recombinant luciferase including recombinant Coleoptera luciferase.

Abstract: Perforin-2 (P2) molecule is a pore forming protein. The 5? untranslated region of the perforin-2 protein controls translational activity. Compositions include the perforin protein and the 5? untranslated region. Methods of use include high-throughput screening assays for identification of therapeutic compounds in treatment of diseases.