Abstract: Disclosed are luciferase polypeptides with improved light-emitting activity and their encoding nucleic acids. These molecules are useful in a range of assays including luciferase-based gene reporter assays, bioluminescence resonance energy transfer assays, protein complementation assays and other applications in which luciferase enzymes are utilized as detectable and/or quantifiable labels. Also disclosed are methods and compositions for increasing the sensitivity and/or improving the kinetics of luciferase-catalyzed reactions as well as decreasing the impact of undesirable variables.

Abstract: The present invention relates to reagents and methods for influenza virus detection. These reagents and methods disclosed in the present invention enable simple, rapid, specific and sensitive detection of influenza virus types A and B. These reagents are N-acetylneuraminic acid-firefly luciferin conjugates which can be cleaved by influenza virus neuraminidase.

Abstract: The present invention provides novel functional assay for 5-HT2A, histamine H1 or adrenergic alpha 1b receptors, by measuring intracellular cyclic adenosine monophosphate (cAMP) levels utilizing reporter gene driven cell based assay. The novel assay provides both binding affinity as well as mode of action of compounds in a single set. The novel assay of the invention is useful in identification of compounds acting through 5-HT2A, histamine H1 or adrenergic alpha 1b receptors. Furthermore, the assay offers utility in categorizing compounds in to agonist, partial agonist, inverse agonist and antagonist classes. The novel assay can be scaled up to any high throughput format.

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 present invention relates to compound of formula I: and their use as chemiluminescent and/or bioluminescent reagents.

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: According to one embodiment, a first gene encodes a reporter protein. The first gene is disposed at the downstream of the gene promoter. A second gene is disposed at the downstream of the gene promoter and encodes a replication origin-binding protein. An internal ribosome entry site is disposed between the first gene and the second gene. The transcription termination signal sequence encodes a signal for terminating the transcription of the first gene and the second gene. A replication origin sequence is recognized by the replication origin-binding protein.

Abstract: Disclosed is a method and associated device for the rapid identification of viable bacterial contaminants in food products. The method detects viable microbes by using a combined ATP-bioluminescence immunoassay. Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium were selected as target organisms in various matrices including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of well plates in which the sample matrices were incubated. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with a luminometer and signal-to-noise ratios were calculated. The LOD was not affected by the presence of non-target cells. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude.

Abstract: Disclosed herein are methods for determining the amount or activity of one or more luciferases and methods for measuring the luminescent signal generated by one or more luciferases in a sample, the methods comprising incubating the sample with a reactive substrate(s) of the luciferase(s) to be analysed and a reducing agent to inactivate a first luciferase, wherein the first luciferase, in its native form, is a secreted luciferase.

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: 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 minize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: The invention relates to methods, reagents and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel luciferase assay system with reduced background luminescence to allow for increased detection sensitivity. Provided is a method of detecting luciferase activity in a sample using coelenterazine or an analog thereof as a substrate, comprising: (a) initiating luciferase-catalyzed luminescence production by contacting said sample with a luciferase detection reagent to yield a reaction mixture, said reagent comprising coelenterazine and at least one iodide source in an amount sufficient to reduce the autoluminescence of said coelenterazine, (b) incubating said reagent mixture under conditions suitable to produce luminescence, and (c) measuring the luminescence produced. Also provided are detections reagents and kits for use in such a method.

Abstract: The present invention relates to predicting therapeutic response of treating patients suffering from itching and puritis mediated by cutaneous lymphocyte antigen positive T cells in atopic dermatitis. The invention also includes methods of predicting a therapeutically responsive patient population.

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: Provided herein are compositions and methods for monitoring the movement of analytes and/or cellular components across biological membranes (e.g., cell surface internalization). In particular, reporter constructs are provided, the transmembrane movement of which (e.g., by endocytosis) is monitored by methods described herein.

Abstract: The present invention discloses an assay method for the detection of viable microbial cells in a sample, the assay method comprising the steps of: i) adding an ATP degrading enzyme to a sample suspected of containing viable microbial cells to substantially degrade any extracellular ATP in the sample; ii) adding a phosphate containing compound to the sample to substantially halt action of the ATP degrading enzyme; and iii) subjecting the sample to a detection assay to establish the level of undegraded ATP in the sample to provide an indication of the level of viable microbial cells in the sample.

Abstract: A method tor producing multilayered cell sheets, including producing a vascular bed which includes an artery-vein loop and in which a capillary vascular network is constructed; layering cell sheets on the vascular bed; and perfusing a culture medium in vitro to construct a vascular network in the cell sheets. The production method enables vascular networks to he constructed in cell sheets and enables thick multilayered cell sheets to foe easily produced by layering the cell sheets. Such thick multilayered cell sheets are useful as in vivo tissue-like products for regenerative medicine for various tissues and for evaluation of drugs and the like.

Abstract: The present invention provides novel uses of the cAMP/PKA/HDAC5 pathway for the treatment and prevention of myopathies.

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: A codon optimized and stabilized luciferase gene based upon the sequence of the natural luciferase gene isolated from Luciola cruciata (Japanese firefly) and a novel recombinant DNA characterized by incorporating this new gene coding for a novel luciferase into a vector DNA for improved activities in mammalian cells, are disclosed. This new luciferase exhibits long-wavelength light emission, as well as improved thermostability and higher expression levels in mammalian cell systems, compared to native luciferase. Assays using this new enzyme for measuring various biological metabolic functions are described.

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: The present invention provides compounds and methods for assaying redox state of metabolically active cells and methods for assaying enzyme activity and/or metabolite level by coupling to redox defining co-factor NAD(P)/NAD(P)H measurement.

Abstract: The present technology relates to methods and systems for detection of pyrophosphate. As such, disclosed herein are methods and systems that permit improved pyrophosphate detection. Also disclosed herein are methods and systems which utilize improved pyrophosphate detection for nucleotide sequencing.

Abstract: The invention relates to the nucleotide and amino acid sequences, and to the activity and use, of the luciferases LuAL, Lu164, Lu16, Lu39, Lu45, Lu52 and Lu22.

Abstract: The present invention provides compositions and methods to detect and/or determine the amount and/or presence of adenosine monophosphate (AMP) in biological samples. The method comprises converting substantially all AMP in the solution to adenosine diphosphate (ADP) using a first enzyme, suitably polyphosphate:AMP phosphotransferase (PAP), that is capable of converting AMP to ADP; converting the ADP in the solution to adenosine triphosphate (ATP) using a second enzyme, suitably adenylate kinase (AK), that is capable of converting ADP to ATP; determining the amount of the ATP produced using a bioluminescent reaction utilizing a luciferase enzyme and a substrate for the luciferase enzyme; and using the amount of ATP produced to determine the amount of AMP present in the original solution.

Abstract: Bioluminescent endoscopy methods and compounds, wherein an anatomical object is examined by means of filling, perfusing, intubating, injecting, or otherwise administering a solution containing a bioluminescent substance or a mixture of luciferin and luciferase which produces bioluminescence, wherein a color or monochrome image of the object is constituted using the images and information based on bioluminescent emitted by the bioluminescent substance. Procedures are demonstrated which allow bioluminescent solutions to be perfused into certain tissue regions, such as but not limited to the common biliary duct, genitourinary tract, gastro-intestinal tract, cardiovascular system and lymphatic system wherein said structures may be conveniently visualized during surgery to avoid damage to these structures. Such images may also be combined with visual light images. Methods of detection of cancer cells using bioluminescence are provided.

Abstract: Compositions and methods are provided for enhanced expression of light emitting reporters. Such reporters are used in methods for monitoring cultures for production of target compounds.

Abstract: The present invention provides a transgenic Drosophila tumor stem cell model and uses thereof. Also provided is a method to selectively inhibit cancer stem cells in a mammal having refractory intestinal cancer or a relapse of intestinal cancer.

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: Characteristics of a chemical or biological sample are detected using an approach involving light detection. According to an example embodiment of the present invention, an assaying arrangement including a light detector is adapted to detect light from a sample, such as a biological material. A signal corresponding to the detected light is used to characterize the sample, for example, by detecting a light-related property thereof. In one implementation, the assaying arrangement includes integrated circuitry having a light detector and a programmable processor, with the light detector generating a signal corresponding to the light and sending the signal to the processor. The processor provides an output corresponding to the signal and indicative of a characteristic of the sample.

Abstract: A recombinant exosome comprising a fusion protein of a membrane protein and light-emitting protein, and a method of determining an exosome recovery rate by using the recombinant exosome are provided. Use of the method ensures accurate quantification of exosomes in a sample, and thus, improves the efficiency of an exosome-based diagnosis.

Abstract: 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.

Abstract: An object of the present invention is directed to a method for assaying the severity of an illness in real time and is to provide a testing method capable of assessing the severity of an illness in more detail than the conventional APACHE II and SOFA scores. The established method can accurately measure an ATP level in a sample, thereby accurately and quickly deducing the “state of intracellular energy required for living organisms” from the ATP level, and by extension, determining the severity of an illness. The present invention further provides a novel biomarker ATP-lactate energy risk score (A-LES) value that is capable of determining the severity of an illness by the reevaluation, with the ATP concentration as an index (specifically, on the basis of a lactic acid level (mM)/ATP concentration (mM) ratio), of the level of lactic acid that accumulates in the sample due to the breakdown of in vivo metabolic balance accompanied by the increased severity of the illness.

Abstract: Disclosed herein are methods for determining the amount or activity of one or more luciferases and methods for measuring the luminescent signal generated by one or more luciferases in a sample, the methods comprising incubating the sample with a reactive substrate(s) of the luciferase(s) to be analysed and a reducing agent to inactivate a first luciferase, wherein the first luciferase, in its native form, is a secreted luciferase.

Abstract: Provided herein is a large immuno-sorbent surface area assay (ALISSA) for the rapid and sensitive detection of botulinum neurotoxins (BoNTs) and anthrax toxin. This assay is designed to capture a low number of toxin molecules and to measure their intrinsic protease activity via conversion of a fluorogenic or luminescent substrate. Also provided herein are novel peptides that can be specifically cleaved by BoNT and novel peptides that are resistant to cleavage by BoNT. The combination of these cleavable and control peptides can be used for implementation of an exemplary ALISSA used to specifically detect BoNT enzymatic activity. Furthermore, the ALISSA as described herein may also be used in a column based format for use in a high-throughput system for testing large quantities of samples.

Abstract: This disclosure describes methods of screening for compounds that disrupt the interaction between DNMT1 and the gamma-globin promoter or between LSD-1 and the gamma-globin promoter. This disclosure describes methods of screening for compounds that de-repress the gamma-globin gene.

Abstract: The invention relates to methods and kits for determining the toxicity of an agent on a population of eukaryotic cells, particularly human cells. The cells may comprise a nucleic acid construct comprising a DNA damage induced response element operably linked to a sequence encoding a reporter gene. The multiplex methods and kits provide means for distinguishing between genotoxic and cytotoxic agents.

Abstract: This present invention relates to an automatic fluid sample preparation module and method for use thereof, for automatically preparing a fluid sample for detecting micro-organisms and/or biological substances. The invention further relates to an automatic analysis system. The automatic fluid sample preparation module comprises:—an inlet for automatically obtaining a sample directly from a fluid or fluid stream to be analyzed; —preparation means for preparing the fluid sample from the sample, comprising first coupling means for coupling with the inlet; and—an outlet for discharge of prepared fluid to measurement means, wherein the preparation means are provided such that micro-organisms and/or biological substances from one or multiple samples are accumulated in the prepared fluid sample.

Abstract: The present invention provides compounds useful for detection of hydrogen peroxide and methods of using same.

Abstract: According to one embodiment, the present invention relates to luciferase derived from Malaysian Luciola firefly, the luciferase having a maximum luminescent wavelength of 580 nm at pH 8, or the luciferase indicating 23.3 times or more of luminescent intensity in comparison to that of Rhodamine 6G.

Abstract: The present invention provides methods of introducing a polynucleotide into a target cell, wherein the method employs a light generating protein coding sequence acting as a reporter. An important advantage of the methods described herein is that drug resistant target cells or target cells having no useful auxotrophic markers can be effectively transformed. The present invention also includes transformed cells produced by the methods described herein. Also described are light generating protein coding sequence modifications, a variety of vectors, and methods of using the transformed cells of the present invention.

Abstract: Assays using binding studies involving function of BRCA1a protein have use for diagnosis and for evaluation of possible tumorogenicity of agents, particularly estrogenic agents. The assays do not rely on use of a probe for only specific sequences, but on effects of known and unknown or not previously studied sequences (consequence of genetic changes) or posttranslational modification of BRCA1 proteins (as a consequence of epigenetic changes) as seen in hereditary and sporadic cancers.

Abstract: The present invention is one gene construct or a combination of two gene constructs or expression vectors incorporating a Cypridina luciferase gene and a copepod luciferase under the control of distinct promoters. These gene constructs and expression vectors are useful for making a mammalian cell incorporating the Cypridina luciferase gene and the copepod luciferase to be capable of stably expressed and extracellularly secreted under the control of the distinct promoters.

Abstract: The present invention provides methods to concentrate cells onto microparticles, to concentrate the microparticles, and to detect the cells. The present invention also includes unitary sample preparation and detection devices to be used in accordance with the methods.

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 invention describes a method for detecting biomolecular interactions said method comprising: (a) selecting an appropriate reporter molecule selected from the group consisting of a protein, a fluorescent protein, a luminescent protein and a phosphorescent protein; (b) effecting fragmentation of said reporter molecule such that said fragmentation results in reversible loss of reporter function; (c) fusing or attaching fragments of said reporter molecule separately to other molecules; followed by (d) reassociation of said reporter fragments through interactions of the molecules that are fused to said fragments; and (e) detecting said biomolecular interactions by reconstitution of activity of the reporter molecule

Abstract: Described herein are methods for detecting a bacteria in a subject and methods for detecting, imaging or diagnosing a site, disease, disorder or condition in a subject using bacteria. The methods can be used in conjunction with methods for treating a disease, disorder or condition. Such sites, diseases and disorders include sites of cellular proliferation, such as tumors, tumor tissues, metastases, areas of inflammation, immunoprivileged sites or tissues, wounds and infections. Further described are bacteria for use in the methods and compositions, combinations and kits, including diagnostic and pharmaceutical compositions, containing a bacterium. Bacteria described herein include those that bind, sequester or accumulate radiolabeled compounds by expression of an endogenous gene product that binds to the radiolabeled compound. Additional imaging and therapeutic agents and methods also are described.

Abstract: It was discovered that insulin binding to insulin receptors signals the upregulation of expression of the liver enzyme deiodinase 1 (Dio1), which in turn activates the ApoA-1 promoter, thereby thereby increasing ApoA-1 expression (primarily in the liver), that in turn raises the levels of plasma ApoA-1, the major and necessary protein in HDLC. Certain embodiments of the invention are directed to methods for increasing circulating HDLC levels in an animal by administering therapeutically effective amounts of Dio1, or by increasing the level of Dio1 through gene therapy.

Abstract: Bioprinting station (1) comprising:—a Bioprinting device (4) adapted to deposit a pattern of biological material (2) onto an area of interest (3a) of a substrate (3),—an imaging system (15) adapted to acquire an image of the substrate (3) and to reveal on the acquired image the area of interest (3a) with respect to a remaining part (3b) of the substrate (3), the acquired image of the substrate (3) being processed so as to detect the revealed area of interest (3a) on the acquired image and to determine the pattern corresponding to the area of interest (3a) detected on the acquired image.

Abstract: The invention generally features compositions and methods for detecting and regulating cell proliferation, potentiation, and differentiation in a population of cells. In particular, compositions and methods are provided for modulating the activity of Musashi proteins. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided.