Abstract: Methods for production of tumor-specific antibodies are provided. The methods employ microorganisms that are designed to accumulate in immunoprivileged tissues and cells, such as in tumors and other proliferating tissue and in inflamed tissues, compared to other tissues, cells and organs, so that they exhibit relatively low toxicity to host organisms. The microorganisms also are designed or modified to result in leaky cell membranes of cells in which they accumulate, resulting in production of antibodies reactive against proteins and other cellular products and also permitting exploitation of proliferating tissues, particularly tumors, to produce selected proteins and other products.

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. 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: A method is disclosed for determining a target nucleic acid sequence, where the target nucleic acid sequence is comprised in a preparation comprising a non-target nucleic acid sequence, the target nucleic acid sequence and the non-target nucleic acid sequence each having a first region of common sequence upstream of a first region of dissimilar sequence upstream of a second region of dissimilar sequence, and includes the steps of contacting the preparation with an oligonucleotide primer complementary to at least a portion of the first region of common sequence, under conditions to hybridise the primer thereto; contacting the preparation with a first labelled nucleotide bearing a first label, wherein the first labelled nucleotide is complementary to a first template nucleotide comprised in the first region of dissimilar sequence of either the target nucleic acid sequence or the non-target nucleic acid sequence, under conditions to incorporate the first labelled nucleotide either into the primer hybridised to t

Abstract: A method to prepare synthetic nucleic acid molecules having reduced inappropriate or unintended transcriptional characteristics when expressed in a particular host cell.

Abstract: The invention relates to anionic oligosaccharide conjugates that may be used to mimic the structure and/or activity of the anionic bioactive molecules known as glycosaminoglycans (GAGs).

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 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: A method for determining the biological activity of embryonated Trichuris eggs is described, in which at least one of the following determinations is carried out: a) Determination and/or confirmation of the stage of the embryonal development of helminth eggs with the aid of quantitative PCR analysis by using suitable marker sequences for ascertaining the copy number of the genomic DNA, b) Determination of the metabolic activity of embryonated helminth eggs by means of biochemical and/or molecular biological methods, c) Determination of the inducibility of gene expression in embryonated helminth eggs, d) Microscopic determination of the motility of helminth larvae in the egg over long periods of observation after pre-incubation at increased temperatures and/or e) Determination of the hatching rate of Trichuris larvae in a laboratory animal, wherein the intact embryonated eggs recovered from the contents of the intestine are quantified compared to an internal standard.

Abstract: This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins.

Abstract: The technology relates to a nucleic acid expression cassette comprising a TR element encoding an mRNA molecule that is translated in stressed and/or dying cells, and a nucleotide sequence operably linked to the TR element, that is a first open reading frame (ORF) sequence and encodes a polypeptide or a fragment thereof and is co-translated with the TR element. The technology further relates to mammalian cells and a transgenic animal comprising such expression cassette.

Abstract: A system combining a clonogenic differentiation assay with an instrument-based ATP bioluminescence proliferation assay to produce a standardized colony-forming stem and progenitor cell potency assay is provided.

Abstract: An isolated recombinant luciferase having luciferase activity. The recombinant luciferase has an amino acid sequence which differs from the wild-type luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata, Luciola lateralis, Hotaria parvula, Pyrophorus plagiophthalamus, Lampyris noctiluca, Pyrocoelia miyako or Photinus pennsylvanica. In the sequence of the recombinant luciferase, the amino acid residue corresponding to phenylalanine 295 in Photinus pyralis wild-type luciferase or to leucine 297 in Luciola mingrelica, Luciola cruciata or Luciola lateralis wild-type luciferases, is mutated compared to the corresponding amino acid which appears in the corresponding wild-type luciferase sequence. The recombinant luciferase has increased thermostability compared to the corresponding wild-type luciferase.

Abstract: A biological process indicator is provided for validating a treatment process in which the amount or activity of a contaminant in a sample is reduced. The indicator comprises a thermostable kinase covalently linked to a biological component, with the proviso that the biological component is not an antibody. Methods of preparing the indicator, and methods of using the indicator, are also provided.

Abstract: A reagent open mechanism of the luminescence measurement system comprises a triaxial actuator and a reagent dispensing nozzle which is driven by the triaxial actuator. A reagent cartridge where a reagent to be divided by the reagent dispensing nozzle is filled in a concave and the opening of the concave is sealed by an aluminum sheet can be set in. This reagent open mechanism comprises an open needle which is driven by the triaxial actuator and makes a hole in the aluminum sheet and a fixation block between the reagent dispensing nozzle and the open needle which arranges the reagent dispensing nozzle and the open needle in such location that the reagent dispensing nozzle or the open needle does not contact with a structure including the reagent cartridge in a Z-axis operation during opening time or reagent dividing and dispensing time.

Abstract: The present invention relates to an isolated nucleic acid and polypeptide sequence that encodes for a luciferase of Luciola italica, as well as mutants thereof. The luciferase proteins of the present invention have been found to have extended bioluminescence emission that is red- or blue-shifted, and are useful as a bioluminescent marker or as an additive to selected materials.

Abstract: The fusion protein comprising (1) a first region comprising the amino acid sequence of SEQ ID NO: 18 and (2) a second region comprising an amino acid sequence for a polypeptide containing at least one cysteine residue for binding to other useful compound via the thiol group can be modified by chemical modification, and thus has a high catalytic ability for a luminescence activity and is highly available for general purposes.

Abstract: Described are of recombinant adenoviral vectors in vaccination regimens, such as prime/boost set-ups and subsequent vaccinations and applications for gene therapy. Moreover, described are assays to determine the best regimen for applying the most suitable recombinant viral vector in a vaccination or gene therapy setting.

Abstract: An object of the present invention is to develop a probe for measuring in real time the kinetics of CREB or actin closely related to brain functions such as memory formation in live animals.

Abstract: The present invention is directed to methods, kits and compositions for modulating the activity of Stat molecules (e.g., Stat1, Stat3 and Stat5). The compounds of the invention are useful for treating and/or preventing disorders characterized by Stat dysregulation, such as hyperproliferative disorders. Further, the compounds of the present invention are also useful in culturing stem cells and treating ischemic disorders.

Abstract: Novel luciferins, methods of making luciferins, and uses of the same are disclosed.

Abstract: The present invention relates to screening methods for diagnosis, prognosis, or susceptibility to cancer in a subject by means of detecting the presence of serum autoantibodies to specific annexin protein antigens in sera from subjects. The present invention also provides screening methods for diagnosis and prognosis of cancer in a subject by means of detecting increased expression levels of annexin proteins in biological samples of the subject. The method of the invention can also be used to identify subjects at risk for developing cancer. The method of the invention involves the use of subject derived biological samples to determine the occurrence and level of expression of annexin proteins or expression of annexin derived peptides or antigens, and/or the occurrence and level of circulating autoantibodies to specific annexin protein antigens. The present invention further provides for kits for carrying out the above described screening methods.

Abstract: This invention relates to the discovery that midkine modulates pulmonary vasculature development and smooth muscle cell development. Modulation of midkine activity thus alters pulmonary vasculature development and smooth muscle cell development. The invention provides methods of modulating pulmonary disorders, smooth muscle cell related disorders, and pulmonary smooth muscle cell disorders. Disorders of particular interest include, but are not limited to, asthma and pulmonary hyperplasia. Further the invention relates to the discovery that TTF1 and HIF1 -? exhibit midkine modulating activity. The invention pertains to the discovery that midkine modulates myocardin activity.

Abstract: The invention relates to methods and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel assay system with increased light yield for a sensitive and convenient detection of luciferase activity, such as luciferase reporter enzyme activity. Provided is a method of detecting luciferase activity in a sample, comprising incubating the sample in the presence of luciferin and ATP to allow the generation of a light signal, wherein said light signal is enhanced by performing the incubation in a reaction mixture comprising phosphate and ammonium ions, and measuring the light signal. The invention also relates to kits for use in such method.

Abstract: A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided.

Abstract: A method for identifying compounds that inhibit amyloid-beta precursor protein processing in cells, comprising contacting a test compound with a GPCR polypeptide, or fragment thereof, and measuring a compound-GPCR property related to the production of amyloid-beta peptide. Cellular assays of the method measure indicators including second messenger and/or amyloid beta peptide levels. Therapeutic methods, and pharmaceutical compositions including effective amyloid-beta precursor processing-inhibiting amounts of GPCR expression inhibitors, are useful for treating conditions involving cognitive impairment such as Alzheimers Disease.

Abstract: This disclosure provides an intracellular fragment of natriuretic peptide receptor A (NPRA), referred to herein as soluble natriuretic peptide receptor-related fragment (sNRF). It is shown herein that sNRF causes NP resistance. Based on these observations, methods of treating a cardiovascular disorder by inhibiting the activity of sNRF are disclosed. Assays are provided that use sNRF to screen agents for their ability to increase the biological activity of an NPR, for example agents that increase the sensitivity of NPR for NPs (such as atrial natriuretic peptide, ANP), or that decrease growth factor deleterious effects, or combinations thereof. Also provided are agents identified using the disclosed assays, and methods of using the agents, for example to treat or diagnose a cardiovascular disorder, such as heart failure.

Abstract: This invention relates to a new method for improving functional protein expression whereby the folding process is monitored by online measurement and, if required, the protein folding is influenced by adding folding promoting agents and/or co-expression of the periplasmic chaperone (Skp). In this respect, the invention offers a technology to improve the yield of functionally expressed recombinant proteins.

Abstract: The present invention provides a method comprising allowing a reaction of a sample, a reagent containing Factor C, which can be activated by binding with endotoxin, and a synthetic luminescent substrate comprising a luminescent substrate bound to a peptide, for release of the luminescent substrate from the synthetic luminescent substrate, allowing a luminescent enzyme to act on the luminescent substrate released in the luminescent substrate release step, for measurement of the luminescence intensity, and quantifying the level of endotoxin in the sample based on a measured value obtained in the luminescence measuring step, the method enabling endotoxin to be simply and quickly measured at a level that cannot be detected in conventional methods for endotoxin measurement, without use of any dedicated measuring device.

Abstract: A method and apparatus for determining the concentration of an analyte in a sample is provided. This method involves combining enhanced chemiluminescence with microchip capillary electrophoresis or microchip liquid chromatography.

Abstract: Methods and compositions are described for assaying luciferase bioluminescence in vitro and in vivo cells. The compositions provide at least one of enhanced stability of signal or magnitude of signal by varying the composition of the buffer. One or more of the following parameters have been varied: the presence or absence of EDTA, the concentration of NaCI, the concentration of coelenterazine, the evaluation of ionic and non-ionic detergent, the amount of detergent, how the detergent has been added and the time over which the signal has been recorded. Also disclosed are dual reporter systems.

Abstract: The present invention relates to chemiluminescent method and regent to detect analyte. One aspect of the current invention relates to using enzyme substrate that can be cleaved by target enzyme to release chemiluminescent compound giving light signal for the detection of varieties of target enzymes. Another aspect of the current invention relates to use chemiluminescent enzyme coupled with analyte binding molecules to detect specific analyte molecules in a homogenous phase.

Abstract: A method and apparatus for determining the concentration of one or more microbes in a sample is provided. This method involves filtering a sample through a filter inside a sample tube to retain the one or more microbes on the filter. The resulting filtrate, which contains or produces adenosine triphosphate, is passed through the sample tube and enters the reporter region. In the reporter region, the adenosine triphosphate in the filtrate comes in contact with a transparent porous matrix, which includes a luciferin-luciferase complex. The adenosine triphosphate interacts with the luciferin-luciferase complex to provide light response, which is measured by a detector. The light response is compared with a calibration curve to determine the total concentration of one or more microbes in a sample.

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: This invention provides a novel ratiometric bioluminescent sensor and methods of use thereof. The bioluminescent sensor comprises two luminescent proteins that exhibit different characteristics associated a biological condition, and thereby illuminates differently in response to the biological condition. A ratio between the luminescence of the two luminescent proteins indicates a change in the biological condition. The bioluminescent sensor of the invention can be used to image an oxidative stress or its associated conditions, including a programmed cell death.

Abstract: The present invention is drawn to methods for modulating IL-13 expression and/or activity in a mammal comprising administering an effective amount of an agent which modulates the expression and/or activity of IL-9.

Abstract: The present invention provides analytical methods and related kits for detecting the presence of bacteria in a sample, determining the amount of bacteria in a sample, or determining the type of bacteria in a sample. In particular, the invention relates to devices and methods suitable for the rapid detection, quantification, or identification of bacteria in a liquid sample by measuring bacterial ATP activity.

Abstract: This invention relates to: the development of a mutant firefly luciferase in order to use dATP as a DNA polymerase substrate upon pyrosequencing, such luciferase being subjected to substrate specificity modification in a manner such that the dATP-induced activity alone is decreased while the ATP-induced activity is maintained; and a mutant firefly luciferase for which the proportion of activity induced by dATP to activity induced by ATP (dATP/ATP) is lower than that for the wild-type firefly luciferase, in which an amino acid identified based on homology analysis as corresponding with the 421st amino acid (glycine) of the amino acid sequence of the wild-type North American firefly (Photinus pyralis) luciferase has been substituted with a polar amino acid.

Abstract: Nucleotide sequences of a Msca1 gene, critical to 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 impacting male fertility in plants.

Abstract: Novel luciferins, methods of making luciferins, and uses of the same are disclosed.

Abstract: A method for identifying compounds that inhibit amyloid-beta precursor protein processing in cells, comprising contacting a test compound with a GPCR polypeptide, or fragment thereof, and measuring a compound-GPCR property related to the production of amyloid-beta peptide. Cellular assays of the method measure indicators including second messenger and/or amyloid beta peptide levels. Therapeutic methods, and pharmaceutical compositions including effective amyloid-beta precursor processing-inhibiting amounts of GPCR expression inhibitors, are useful for treating conditions involving cognitive impairment such as Alzheimers Disease.

Abstract: As described herein, signaling events occurring in neurons or at neuronal synapses have been identified that involve Cdk5 and various other molecules which bind to, are activated by, and/or activate Cdk5. Of particular relevance are interactions that stimulate calpain cleavage of p35 into p25, which binds Cdk5 in pathologic states. Assays to identify modulators of these interactions are provided.

Abstract: Methods are provided for detection of concentrations of less than 3.4 picomolar of adenosine triphosphate (ATP) using recombinant luciferase in the luciferin-luciferase reaction. Aspects include a low pH composition for use in detecting the presence of ATP. The low pH composition can include low concentrations of detergents and can be used in combination with methods for reading, calculating and interpreting luminescence generated by the ATP-luciferin-luciferase reaction.

Abstract: A protein having luciferase activity and at least 60% similarity to luciferase from Photinus pyralis, Luciola mingrelica, Luciola cruciata or Luciola lateralis.

Abstract: Response element regions, DNA constructs comprising response element regions, host cells comprising response element regions, and methods of using response element regions are provided.

Abstract: A modified luciferase protein which is a sensor for molecules including cAMP, cGMP, calcium, chelators thereof, kinases, or phosphatases is provided. Also provided is a circularly permuted anthozoan luciferase protein and a decapod crustacean luciferase protein, optionally containing one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest. Further provided is a modified anthozoan luciferase protein and a decapod crustacean luciferase protein containing an insertion of one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest.

Abstract: The present invention relates to a compound having a structure analogous to firefly luciferin. In particular, the invention relates to a heterocycle compound which produces a luminescence at a light wavelength different from that of firefly luciferin in nature. The present invention provides a heterocycle compound of following general formula I. In the above general formula, R1, R2 and R3 can be each independently H or C1-4-alky. In the above general formula, X and Y can be each independently C, N, S or O. In the above general formula, the olefin chain part expressed as “n” can be changed to desired length.

Abstract: The present invention relates generally to kits that provide reagent mixes and instructions for the use thereof, in performing high-throughput assay methods that determine 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. The present invention further relates to kits that provide reagent mixes and instructions for high-throughput assays methods for screening compounds that may modulate the proliferative status of a target cell population. The kits of the present invention and methods therein described may be used for determining the proliferative status of any isolated cell line or type. The kits and methods of the present invention address the need for rapid assays that determine the proliferative status of isolated hematopoietic stem and progenitor cells and of subpopulations of differentiated cells thereof.

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: A method to prepare synthetic nucleic acid molecules having reduced inappropriate or unintended transcriptional characteristics when expressed in a particular host cell.

Abstract: The present invention relates to nanostructures for use in luminescent assays as well as methods for the production of nano-sized tube and rods, including arrays of nanotubes and nanorods from a nylon.