Abstract: A cell culture system related to extended in vitro culture of mature neuronal cells and methods for preparing the cell culture system are provided. In a preferred embodiment the invention provides a cell culture system comprising a mixture of mature neuronal retinal cells and cells isolated from a ciliary body. Methods for identifying bioactive agents that alter neurodegeneration of neuronal retinal cells are also provided.

Abstract: Novel human MOAT genes and their encoded proteins are provided herein. The MRP-related ABC transporters encoded by the disclosed nucleic acid sequences play a pivotal role in the efflux of pharmacologically beneficial reagents from tumor cells. MOAT genes and their encoded proteins provide valuable therapeutic targets for the design of anti-cancer agents which inhibit the aberrant growth of malignant cells.

Abstract: A method of producing a binding assay device provides a porous membrane comprising a material enabling capillary movement of a liquid sample from a first area on the membrane on a second area on the membrane. A detection site is disposed on the membrane between the first and second areas in a non-absorbent medium disposed on the membrane between the detection site and the membrane first area is attached by an adhesion with a dry reagent disposed between the medium and the membrane in order to enable mobilization of the reagent by passage of a liquid sample therepast.

Abstract: A simple easy to manufacture analytical device capable of performing membrane based immunoassays on batch of samples within 3 to 10 minutes wherein the method permits focused application of samples, costly labeled immunoassay and signal amplification reagents, said device includes an antibody-immobilized micro porous membrane, breadth corner layer of which is directly attached to a semi-rigid liquid-impervious body with water insoluble adhesive; absorbent body is provided separately and is not attached to analytical device during manufacture, absorbent body is wetted and is placed proximal to the lower surface of the membrane thereby forming networks of capillary channels with the absorbent body; flow of samples or reagents is always kept downwards and focused without application of any force to the absorbent body and the use of disposable adsorbent body permits stepwise addition of signal amplification reagents for ultra sensitive detection of diagnostically important molecules by visual examination of the m

Abstract: Elongated molecules are stretched across a substrate by controlled fluid flow.

Abstract: A method for determining the capability of an arterivirus to replicate in a permissive cell is disclosed. The method includes determining an amino acid at a position that corresponds to an amino acid of a protein of a porcine reproductive and respiratory syndrome virus. The invention also discloses a method for determining the capability of an arterivirus to replicate in a green monkey cell line. The invention further discloses a method for producing arterivirus in a green monkey cell line wherein the virulence of the arterivirus is maintained and the virus yield is increased. Methods for determining the attenuation of an arterivirus and for attenuating the virulence of the arterivirus by changing amino acids are further disclosed.

Abstract: The present invention relates to the use of an alpha complementation assay as a quick, effective, and safe method for the detection of cell fusion mediated by viral proteins. Additionally, the method disclosed herein permits the identification of inhibitors of cell fusion using an alpha complementation assay.

Abstract: The present invention provides a method of screening for a compound that binds to a selected nucleic acid comprising contacting compound fluorescently labeled by a fluorescent protein with a cell having a plurality of copies of the nucleic acid in an array such that the nucleic acid can be directly detected when bound by fluorescently labeled compound; and directly detecting the location of fluorescence within the cell, fluorescence aggregated at the site of the nucleic acid array indicating a compound that binds to the selected nucleic acid. In particular compounds such a transcription factors can be screened. Reagents for such method are provided including a mammalian cell having a plurality of steroid receptor response elements in an array such that the response element can be directly detected when bound by fluorescently labeled steroid receptor and a chimeric protein comprising a fluorescent protein fused to a steroid receptor.

Abstract: The present invention provides compositions and methods for the detection and characterization of mutations associated with cystic fibrosis. More particularly, the present invention provides compositions, methods and kits for using invasive cleavage structure assays (e.g. the INVADER assay) to screen nucleic acid samples, e.g., from patients, for the presence of any one of a collection of mutations in the CFTR gene associated with cystic fibrosis. The present invention also provides compositions, methods and kits for screening sets of CFTR alleles in a single reaction container.

Abstract: The invention provides universal e-tag primers and methods for their use in the sequence specific detection and multiplexed analysis of known, selected target nucleic acid sequences. The universal e-tag primers have sequence specific and universal components for use in e-tag probe-mediated analysis of target nucleic acids.

Abstract: Arrays of oligonucleotide probes which are complementary to a plurality of S. cerevisisae and S. pombe genes are disclosed. The arrays may be used to measure the expression levels of a plurality of genes simultaneously.

Abstract: The present invention provides oligonucleotide primers, compositions, and kits containing the same for rapid identification of viruses that cause viral hemorrhagic fevers by amplification of a segment of viral nucleic acid followed by molecular mass analysis.

Abstract: Tools and methods are provided for determining whether or not a dog is genetically normal, is a carrier of, or is affected with or predisposed to progressive rod-cone degeneration. The method is based on the detection of a transversion from G to A at position corresponding to nucleotide position 1298 of SEQ ID NO: 1.

Abstract: This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as nucleic acids from biological samples, cells from bodily fluids, and nascent proteins from translation reactions. Agents comprise a detectable moiety bound to a photoreactive moiety. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed.

Abstract: The present invention relates to a method for identifying a target nucleotide sequence. This method involves forming a ligation product on a target nucleotide sequence in a ligase detection reaction mixture, amplifying the ligation product to form an amplified ligation product in a polymerase chain reaction (PCR) mixture, detecting the amplified ligation product, and identifying the target nucleotide sequence. Such coupling of the ligase detection reaction and the polymerase chain reaction permits multiplex detection of nucleic acid sequence differences.

Abstract: The present invention relates to the detection of analyte(s) of interest in a test sample using a rupturable microcapsule biosensor. In general, the microcapsule biosensor includes a microcapsule comprising a shell encapsulating a detectable agent and further includes a probe that is joined to the shell of the microcapsule. The present invention contemplates generally using the microcapsule biosensor to determine if a test sample contains an analyte of interest by exposing the microcapsule biosensor to the test sample and allowing the probe to potentially bind to the analyte. External force is applied to the microcapsule to rupture the shell releasing the detectable agent and thereby indicating the presence of the analyte of interest in the test sample.

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

Abstract: An analyte measurement system is provided having sensors with embossed test chamber channels. In one embodiment, the sensors are elongate test strips for in vitro testing, each test strip having a substrate, at least one electrode, an embossed channel in the electrode, and lidding tape covering at least a portion of the embossed channel. Methods of manufacture are also disclosed for filling the sensor channels with reagent, and for trimming the ends of the sensors to eliminate the need for a calibration code during use of the sensors with a meter.

Abstract: A method of characterizing the biological activity of a candidate compound may include exposing cells to the candidate compound, and then exposing the cells to a repetitive application of electric fields so as to set the transmembrane potential to a level corresponding to a pre-selected voltage dependent state of a target ion channel.

Abstract: This invention provides methods of treating defective skeletal muscle function during heart failure and methods for identifying compounds useful in such treatments.

Abstract: The present invention describes synthetic peptide substrates of the metalloproteases, aggrecanase-1 and/or -2 suitable for assays of enzyme activity. The invention also describes methods using these peptides to discover pharmaceutical agents that modulate these proteases.

Abstract: A nanoporous silicon support comprising a plurality of macropores is provided to function as a bioreactor for the maintenance of cells in culture in a differentiated state. Each cell or group of cells is grown in an individual macropore and is provided with nutrients by means such as perfusion of the nanoporous silicon support with fluid. The macropores may be between 0.2 and 200 microns and be coated with a substance that promotes cell adhesion. The support containing cells may be used to used to test compounds for biological activity, metabolism, toxicity, mutagenicity, carcinogenicity or to characterize novel or unknown comounds. The supports are sufficiently robust that they may be assembled into larger reactors to simulate organ function or be used for the production of biomolecules.

Abstract: The present invention relates to the use of an air-liquid interface culture model using asthmatic bronchial epithelial cells, which exhibit impaired epithelial barrier function. It is proposed to use asthmatic epithelial cultures, in the absence of added Th2 or proinflammatory cytokines such as IL-13, as an in vitro model to screen for agents that can act to improve impaired asthmatic epithelial barrier function. Such agents may have therapeutic utility in asthma patients.

Abstract: Novel polyketides and novel methods of efficiently producing both new and known polyketides, using recombinant technology, are disclosed. In particular, a novel host-vector system is described which is used to produce polyketide synthases which in turn catalyze the production of a variety of polyketides.

Abstract: Compositions and methods are provided for the enhanced in vitro synthesis of protein molecules, by optimizing the metabolism of amino acids present in the reaction mix, preferably all amino acids in the reaction mixture. By performing synthesis with extracts from genetically modified microbial strains that are deficient in multiple amino acid metabolizing enzymes reduces the enzymatic activities responsible for catalyzing these reactions and improves the overall yield of synthesis.

Abstract: Methods and compositions for treating ocular disorders such as retinal detachment and macular degeneration and methods and compositions for prognosing or diagnosing retinal detachment or macular degeneration in a subject are disclosed.

Abstract: The present invention relates to a novel human protein called Dendritic Enriched Secreted Lymphocyte Activation Molecule, and isolated polynucleotides encoding this protein. Also provided are vectors, host cells, antibodies, and recombinant methods for producing this human protein. The invention further relates to diagnostic and therapeutic methods useful for diagnosing and treating disorders related to this novel human protein.

Abstract: The subject invention provides novel compositions containing a mixture of (a) an enzyme that possesses substantial 3?–5? exonuclease activity (b) a DNA polymerase with less 3?–5? exonuclease activity than the enzyme with substantial 3?–5? exonuclease activity. Preferably, the DNA polymerase for inclusion in the compositions are DNA polymerases that substantially lack 3?–5? exonuclease activity. A preferred embodiment of the invention is a composition comprising the Taq DNA polymerase (isolated from Thermus aquaticus) and the Pfu DNA polymerase (isolated from Pyrococcus furiosus). Another aspect of the invention is to provide methods for synthesizing polynucleotides, typically DNA, using compositions comprising an enzyme that possesses substantial 3?–5? exonuclease activity and a DNA polymerase with less 3?–5? exonuclease activity than the enzymes possessing substantial 3?–5? exonuclease activity, preferably a DNA polymerase that substantially lacks 3?–5? exonuclease activity.

Abstract: In detecting a nucleic acid by using the nucleic acid amplification method, it is intended to shorten the time required for the measurement so as to systematically and efficiently examine a gene, etc. To achieve this object, use is made of a method of directly amplifying a nucleic acid by treating a collected biological sample wherein a means of inhibiting the degradation activity of the nucleic acid is introduced during or after the step of homogenizing the biological sample and thus the nucleic acid is directly amplified without isolating/purifying the nucleic acid component from the biological sample. In this method, the means of inhibiting the degradation activity of the nucleic acid is introduced at an acidic pH value (more specifically, from pH 2.5 to pH 5) with the use of a salt interacting with a substance inhibiting the nucleic acid amplification reaction and/or the nucleic acid component so as to directly amplify the nucleic acid without isolating/purifying the nucleic acid component.

Abstract: A method for reducing the efficiency of primer extension by polymerase enzymes when the 3? end of a primer or growing nucleic acid chain does not hybridize perfectly with the target, for increasing the selectivity of single nucleotide mutation or gene analyses, for suppressing false positive results and for enhancing the fidelity of the amplification of nucleic acid fragments by avoiding the incorporation of mispairs, the method comprising the steps of: (a) obtaining a nucleic acid sample; (b) hybridizing said nucleic acid sample to a primer; (c) subjecting said nucleic acid sample hybridized to a extension reaction by extending a primer with a polymerizing enzyme; and (d) detecting the presence of extension products; wherein the reaction extension mixture medium contains an intercalating agent such as ethidium bromide, dihydroethidium, ethidium homodimer-1, ethidium homodimer-2, acridine, propidium iodide, YOYO®-1 or TOTO®-1.

Abstract: The present invention relates to a hybrid enzyme comprising at least one carbohydrate-binding module amino acid sequence and at least the catalytic module of a glucoamylase amino acid sequence. The invention also relates to the use of the hybrid enzyme in starch processing and especially ethanol production.

Abstract: An improved method of making an immobilized enzyme comprising (a) treating an immobilization support with an aqueous solution comprising a cross-linking agent and polymeric aldehyde species and active centre species to produce a modified support; (b) isolating the modified support; (c) treating an enzyme solution with the modified support to produce the immobilized enzyme, the improvement comprising treating the aqueous solution of cross-linking agent with an effective amount of a purifying agent to reduce the amount of the polymeric aldehyde species and active centre species.

Abstract: Hydrophobic polymer surfaces whose level of protein binding is less than about 50-80 ng/cm2 are achieved by: (1) applying a coating solution composed of a solvent and a non-ionic surfactant having a HLB number of less than 5 to the surface; and (2) drying the surface to remove the solvent and thereby bring the surfactant into direct contact with the hydrophobic polymer. The combination of a low HLB number and the drying step have been found to produce low binding surfaces which can withstand multiple washes with water and/or protein-containing solutions Alternatively, the low binding surfaces can be produced by applying the non-ionic surfactant to the mold surfaces which contact molten polymer and form the polymer into a desired shape, e.g., into a multi-well plate, a pipette tip, or the like. Further, the low binding surfaces may be produced by incorporating non-soluble, non-ionic surfactants having an HLB number of less than or equal to 10 into a polymer blend prior to molding the article.

Abstract: O-acetylserine, L-cysteine and sulphurous compounds derived therefrom may be produced using a bacterium belonging to the genus Escherichia which harbors a mutant feedback-resistant serine acetyltransferases in which the amino acid sequence corresponding to positions from 89 to 96 in a wild-type serine acetyltransferase is replaced with any one of the amino acid sequences shown in SEQ ID NOS: 4 to 9, and feedback inhibition by L-cysteine in the bacterium is desensitized.

Abstract: The present invention provides a method for identifying a thermostable polymerase having altered fidelity. The method consists of generating a random population of polymerase mutants by mutating at least one amino acid residue of a thermostable polymerase and screening the population for one or more active polymerase mutants by genetic selection. For example, the invention provides a method for identifying a thermostable polymerase having altered fidelity by mutating at least one amino acid residue in an active site O-helix of a thermostable polymerase. The invention also provides thermostable polymerases and nucleic acids encoding thermostable polymerases having altered fidelity, for example, high fidelity polymerases and low fidelity polymerases. The invention additionally provides a method for identifying one or more mutations in a gene by amplifying the gene with a high fidelity polymerase.

Abstract: This invention relates to non-radioactive markers that facilitate the detection and analysis of nascent proteins translated within cellular or cell-free translation systems. Nascent proteins containing these markers can be rapidly and efficiently detected, isolated and analyzed without the handling and disposal problems associated with radioactive reagents. Preferred markers are dipyrrometheneboron difluoride(4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)dyes.

Abstract: The present invention relates to crystals of the ERK2 polypeptide and complexes thereof which are useful, inter alia, for structure assisted drug design.

Abstract: The substrate specificity of a lipolytic enzyme can be modified by making alterations to the amino acid sequence in a defined region of the lipolytic enzyme, so as to increase the level of a desired activity or to decrease the level of an undesired activity. Thus, the inventors have developed lipolytic enzyme variants with a modified amino acid sequence with a substrate specificity which can be tailored for specific uses.

Abstract: The present invention relates to a method of producing a heterologous protein or polypeptide having phytase activity in a yeast system. The invention also provides proteins having phytase activity which have increased thermostability. Yeast strains which produce a heterologous phytase and the vectors used to produce the phytase are also provided.

Abstract: The present invention is based on the development of a dual promoter system (preferably a RNA pol I-pol II system) for the efficient intracellular synthesis of viral RNA. The resultant minimal plasmid-based system may be used to synthesize any RNA virus, preferably viruses with a negative single stranded RNA genome. The viral product of the system is produced when the plasmids of the system are introduced into a suitable host cell. One application of the system is production of attenuated, reassortant influenza viruses for use as antigens in vaccines. The reassortant viruses generated by cotransfection of plasmids may comprise genes encoding the surface glycoproteins hemagglutinin and neuraminidase from an influenza virus currently infecting the population and the internal genes from an attenuated influenza virus. An advantageous property of the present invention is its versatility; the system may be quickly and easily adapted to synthesize an attenuated version of any RNA virus.

Abstract: The present invention relates to Lawsonia intracellularis vaccines and methods for protecting against and diagnosing L. intracellularis infection. The products and processes of the invention are attainable, in part, as the result of an improved method for cultivating large scale supplies of L. intracellularis, including both a novel isolate of L. intracellularis of European origin and a method of preparing a lyophilized product containing the attenuated European isolate as vaccine product.

Abstract: The present invention relates to isolated human and rat nucleic acid molecules encoding a protein or polypeptide that modulates transcriptional activation in a cell with or without collaboration with a nuclear hormone receptor transcriptional co-activator. The present invention also relates to the proteins or polypeptides encoded by those nucleic acid molecules, and antibodies against such proteins or polypeptides. The present invention also relates to a variety of uses for the nucleic acid molecules, proteins or polypeptides, and the related antibodies of the present invention, including methods of: regulating cellular proliferation, differentiation, and development; modulating the activity of a transcriptional co-activator complex and a transcription factor in cells; regulating hormone receptor activity and endocrine function in cells; and treating diabetes and insulin resistance in a subject.

Abstract: The present invention discloses novel isolated strains of lactic acid producing bacteria of the genera Lactobacillus and Pediococcus, and a method for isolation of such bacterial strains, having the ability to colonise and become established in a human vagina, even during menstrual discharge. Furthermore, a composition comprising the bacterial strains and a sanitary article comprising the bacterial strains, such as a tampon, for prophylaxis and/or treatment of infections of the urogenital tract, are described. The present invention also describes a method for prophylaxis and/or treatment of infections of the urogenital tract, wherein at least one of the bacterial strains is administered vaginally.

Abstract: This invention provides improved components (e.g. array “pins”, print head, substrate platen, print head platen, and the like) for microarray printing devices as well as microarray printing devices incorporating such components. In one embodiment, this invention provides a microarray print head comprising a plurality of glass or quartz spotting capillaries disposed in a support that maintains a fixed spacing between the spotting capillaries and that permits the spotting capillaries to move in a direction parallel to the long axis of the capillaries.

Abstract: A sample analyzing method capable of high-precision analysis without being affected by vibration and optical design is provided. In the sample analyzing method, a sample containing a ligand is caused to bind to a receptor that can bind specifically to the ligand, and a change caused by binding of the receptor and the ligand is analyzed by measuring frequency characteristics of a surface plasmon resonance angle while applying external vibration to the receptor. The sample analyzing method is useful in the fields of, for example, biology, medicine, pharmacology, agriculture, and the like.

Abstract: The present invention relates to a ?-catenin oligonucleotide microchip for detecting mutation in the mutational hot spot regions of ?-catenin gene, a manufacturing process thereof and a method for detecting the ?-catenin mutation employing same, wherein specific oligonucleotides are selectively designed to detect various missense mutations and in-frame deletion at the mutational hot spots of ?-catenin gene. The ?-catenin oligo chip of the present invention can be used in studies to detect ?-catenin mutations and unravel the signal transduction mechanism and tumorigenesis related to ?-catenin gene.

Abstract: A device and method for detecting anthrax or other pathogens are disclosed. Individual self-contained monitoring devices of a monitoring system can be portable or stationary (e.g. installed in air ducts or plumbing of buildings) and can be part of a network of devices. Monitoring devices may be used for the detection of a variety of airborne or surface pathogens, including but not limited to anthrax, smallpox, and Salmonella. Bioamplification-coupled proteomics assays provide rapid and reliable detection of pathogens, with self-checking capabilities reducing or eliminating false positives and false negatives. Sample preservation capability allows pathogen samples to be preserved after detection for further testing. The device of the invention can be remotely operated by minimally trained technicians or security personnel.

Abstract: A microorganism culture device comprising a porous matrix layer having a basis weight of 40 to 100 g/m2 and an air permeability of 7 to 24 cm/sec, and at least one layer of a water-soluble polymer layer laminated with the matrix layer.

Abstract: Method for the detection and enumeration of viable microorganisms. A liquid that comprises one or more markers incorporated in a liquid sol-gel precursor, is provided. A transparent slide is coated with a thin uniform layer of the liquid sol-gel precursor composition. The microorganisms are separated from liquid sample to be analyzed by passing the sample through a filter, and then bringing the filter into close contact with the sol-gel coated slide. The filter is co-incubated with the sol-gel coated slide for a period of time and at a temperature suitable to promote uptake of the markers by the microorganisms. The gel-coated slide irradiated with an external energy source, so as to generate detectable signals emitted from the markers uptaken by the microorganisms. Image of the detectable signals emitted from the microorganisms are acquired, and analyzed using a computer system, in order to provide the identification and enumeration of the microorganisms.

Abstract: There is described a method of isolating nucleotide sequences encoding target peptides from DNA libraries using DNA binding proteins to link the peptide to the sequence which encodes it. DNA libraries are prepared from cells encoding the protein of interest, or from synthetic DNA, and inserted into, or adjacent to, a DNA binding protein in an expression vector to create a chimeric fusion protein. Incorporation of the vector DNA into a carrier package, during expression of the chimeric fusion protein, results in the production of a peptide display carrier package (PDCP) displaying the DNA-bound fusion protein on the external surface of the carrier package. Employment of affinity purification techniques results in the PDCP particles containing sequences encoding the desired peptide to be selected and the desired nucleotide sequences obtained therefrom.