Abstract: A method of improving a phenotypic defect in a cell that contains a conformationally defective target protein wherein the conformational defect causes the phenotype defect, comprising contacting a first cell that expresses said conformationally defective target protein with an amount of a protein stabilizing agent that is effective to improve the conformational defect, thereby improving the phenotypic defect of the first cell in comparison with a second cell having the same conformationally defective target protein and phenotypic defect, wherein the second cell is not contacted with the protein stabilizing agent.

Abstract: The present invention provides for novel methods for measuring the levels of cyclic adenosine monophosphate (cAMP) produced by cells. Notably, the methods provided do not require that the cell membranes be disrupted. Specifically, the present invention provides for a methods of detecting and quantifying cAMP extracellularly and for kits useful in employing these methods. Furthermore the present invention also provides for a method of isolating cAMP produced by cell culture.

Abstract: The present invention describes an efficient retroviral or viral based method that allows easy and quick identification of gene transfer in living, transduced mammalian cells. Retroviral and viral vector producer cells were generated containing a gene for an improved humanized red-shifted, Green Fluorescent Protein (hRGFP) which increases the resulting fluorescence yield after excitation. This humanized, red-shifted GFP (hRGFP) gene was cloned into several vectors and transfected into various packaging cell lines to produce vibrant green fluorescence after excitation with blue light at 450-490 nm. These vectors represent a substantial advance over currently available gene transfer marking systems or wild-type GFP marker systems none of which have been stably transfected into cells.

Abstract: The present invention relates to antibodies and other binding molecules specific for hepatitis B viral antigens (HBV), peptides comprising epitopes recognized by such molecules, and cell lines capable of producing antibodies. The invention is further concerned with the use of such molecules in diagnosis of HBV. The invention further relates to a method for the diagnosis of hepatitis B, the method comprising contacting the sample suspected to contain hepatitis B particles or antigens with a specific binding molecule according to the invention. The invention further relates to an assay kit for the detection of a hepatitis B particle or antigen, the kit comprising a specific binding molecule of the invention and means for detecting whether the specific binding molecule is bound to a hepatitis B particle or antigen.

Abstract: A new isolate of Ehrlichia species has been obtained from a patient suffering from ehrlichiosis. The new isolate has been found to be similar, but distinctly different from E. canis. A diagnostic kit and methods for diagnosing ehrlichiosis in humans and for screening drugs toxic to the new isolate have been described.

Abstract: The present invention discloses hybridization assay probes, amplification primers, nucleic acid compositions and methods useful for detecting Neisseria nucleic acids. Hybridization assay probes and amplification primers that selectively detect Neisseria meningitidis and distinguish those Neisseria meningitidis from Neisseria gonorrohoeae are disclosed. Other hybridization probes selectively detect Neisseria gonorrohoeae and not Neisseria meningitidis are also described.

Abstract: The present invention pertains, in general, to the identification, isolation and use of Telomerase Reverse Transcriptase (TERT) genes and the proteins encoded by such genes. In particular, the present invention pertains to the identification, isolation and use of TERT genes and TERT proteins from several genetically diverse and economically important organisms, including two human pathogens, Candida albicans and Plasmodium falciparum and an agronomic crop species, Oryza sativa.

Abstract: The present invention provides an improved system for the rapid and non-destructive identification of chemical compounds attached to solid supports. In general, the invention provides an identification unit comprising a tag attached to the solid support and a binding partner that interacts specifically and detectably with he tag. In preferred embodiments, the identification unit incorporates the advantages of chemically robust tags and a decoding technique capable of amplification for easy detection and analysis. The present invention further provides a kit comprising a collection of tags capable of attachment to a support and binding partners capable of binding selectively and detectably to the tags, to generate an identification unit for the facile determination of the structure of a compound of interest by determining the reaction history and/or structural characteristics of the compounds that are encoded by the identification unit.

Abstract: The present invention provides a method of removing nucleic acid contamination in an amplification reaction which comprises use of a thermolabile DNase and a method of preventing or reducing false positive results due to carry-over in a nucleic acid amplification reaction, said method comprising using a thermolabile DNase to degrade carried-over non-target double-stranded DNA present in the amplification reaction mixture. A thermolabile DNase from the shrimp Pandalus borealis has been identified which is suitable for use in the methods of the invention.

Abstract: A method for assaying a target nucleic acid, comprising: providing an RNA amplification system comprising producing a double-stranded DNA using, as a template, a target RNA containing a specific nucleotide sequence in a sample, said double-stranded DNA having a promoter sequence and being capable of transcribing an RNA comprising the specific nucleotide sequence or a sequence complementary to the specific nucleotide sequence, producing an RNA transcription product comprising the specific nucleotide sequence or a sequence complementary to the specific nucleotide sequence in the presence of an RNA polymerase, and producing the double-stranded DNA using the RNA transcription product as a template, in the presence of a probe labeled with an intercalating fluorochrome having a sequence complementary to the RNA transcription product; measuring the fluorescence intensity in the RNA amplification system with time; calculating a time when the fluorescence intensity satisfies a prescribed criterion based on the measure

Abstract: A method of testing sperm quality including obtaining a sample of sperm to be tested; detecting and measuring the testis-specific HspA2 chaperone protein (or, the chaperone protein homologues to HspA2) in human and animal sperm; and determining a sperm quality parameter based upon the chaperone protein, wherein an increased amount of the chaperone protein species indicates a higher sperm quality. The chaperone protein is detected and measured either by binding one or more antibodies specific to the sperm chaperone protein to the sperm and measuring the antibody content or measuring ATP bound to the sperm chaperone protein. In the case of the latter method, the chaperone protein may be detected and measured by measuring ATP bound to the sperm chaperone protein, and such measuring is by chaperone protein-bound and CK-B generated ATP measurement, or by bioluminescence of the chaperone protein bound-ATP.

Abstract: A method for in vitro construction of a library of recombined homologous polynucleotides from a number of different starting DNA templates and primers by induced template shifts during an polynucleotide synthesis is described, whereby A. extended primers are synthesized by a) denaturing the DNA templates b) annealing primers to the templates, c) extending the said primers by use of a polymerase, d) stop the synthesis, and e) separate the extended primers from the templates, B. a template shift is induced by a) isolating the extended primers from the templates and repeating steps A.b) to A.e) using the extended primers as both primers and templates, or b) repeating steps A.b) to A.e), C. this process is terminated after an appropriate number of cycles of process steps A. and B.a), A. and B.b), or combinations thereof. Optionally the polynucleotides are amplified in a standard PCR reaction with specific primers to selectively amplify homologous polynucleotides of interest.

Abstract: The HIV-1 transactivator protein Tat significantly increases astrocytic expression and release of monocyte chemoattractant protein-1. Monocyte chemoattractant protein-1 (MCP-1) is expressed in the brains of patients with HIV-1-associated dementia, and is present in the cerebrospinal fluid of patients with this condition. This present invention employs compounds, such as MCP-1 antagonists and partial agonists, as well as HIV-1 Tat-inhibitors in methods for treating and/or preventing HIV-1 associated dementia.

Abstract: Disclosed herein are non-endogenous, constitutively activated forms of the human 5-HT2A and human 5-HT2C receptors and uses of such receptors to screen candidate compounds. Further disclosed herein are candidate compounds identified by the screening method which act at the 5HT2A receptors. Yet further disclosed is a new class of compounds which act at the 5HT2A receptors.

Abstract: A process for the preparation of immunogens or diagnostic reagents that mimic an antigen or a pathogenic organism specific to a disease, essentially characterized by the following operations: identification of at least one antibody that reacts with the antigen or pathogenic organism specific to the disease; construction of phage libraries which display on the surface of the capsid oligopeptides, expressed from random sequence oligonucleotidic inserts introduced into a gene coding for a phage capsid protein using genetic manipulation techniques (for example, using a plasmid engineered for the purposes of the invention, the genetic map of which is shown in the figure); selection of the phages that display on the surfaces of the capsid antigenic oligopeptides recognized by said antibody; optional use of the selected phages and/or fragments thereof and/or their derivatives for the formulation of diagnostic kits for the specific pathogenic agent, or in general for the disease, including immunological disorders typ

Abstract: An apparatus and method for synthesizing a combinatorial library comprising a plurality of chemical compounds such that the chemical composition of each compound is easily tracked. The library compounds are synthesized on solid-phase supports, which are spatially arranged in frames during synthesis according to a predetermined protocol, such that each solid-phase support passes through a series of unique spatial 2D or 3D addresses by which the chemical composition of each compound may be determined at any point during synthesis. Solid-phase supports include hollow tubular-shaped lanterns and gears.

Abstract: The invention provides a novel prostate cell-surface antigen, designated Prostate Stem Cell Antigen (PSCA), which is widely over-expressed across all stages of prostate cancer, including high grade prostatic intraepithelial neoplasia (PIN), androgen-dependent and androgen-independent prostate tumors. The invention also provides a method for detecting a Prostate Stem Cell Antigen (PSCA) protein of bone (SEQ ID NO:2), comprising obtaining a sample; contacting the sample with an antibody that recognizes and binds the PSCA protein; and, detecting binding of the antibody with the PSCA protein in the sample.

Abstract: Methods and apparatuses are provided for determining presence and concentration of analytes by exploiting molecular binding reactions and differential diffusion rates. Analyte particles and binding particles are allowed to diffuse toward each other, and slowing of the diffusion front is detected when they meet. From the position of the diffusion front, presence and concentration of analyte particles can be determined. One embodiment provides a competitive immunoassay in a microfluidic format. This diffusion immunoassay (DIA) relies on measuring the concentration of labeled antigen along one dimension of a microchannel after allowing it to diffuse for a short time into a region containing specific antibodies. A simple microfluidic device, the T-Sensor, was used to implement a DIA to measure the concentration of phenytoin, a small drug molecule. Concentrations of analyte over the range of 50 to 1600 nM can be measured in less than a minute.

Abstract: There is disclosed an initial identification of an N-terminally truncated HER-2/neu product. This product is a 95 kDa polypeptide having in vitro kinase activity (as determined by western blotting). Moreover, immunoprecipitations using domain specific antibodies was able to utilize this specific polypeptide from intracellular fragments as a diagnostic and prognostic indicator of adenomacarcinomas without the severe dilution effects encountered by measuring ECD.

Abstract: Disclosed are a method for quantitatively determining mannose, which comprises reacting mannose in a specimen with an enzyme which is capable of oxidizing the mannose by dehydrogenation, in the presence of an electron acceptor, and quantitatively determining a formed reductant of the electron acceptor; and a reagent for the quantitative determination of mannose.

Abstract: A reagent and method for determining the levels of 3-hydroxybutyric acid in a sample are provided. The reagent comprises a ferricyanide salt, a catalytic amount of a first enzyme operative to catalyze the oxidation of 3-hydroxybutyric acid in the sample, a cofactor corresponding to said first enzyme, and a catalytic amount of a second enzyme operative to catalyze the oxidization of the cofactor and the reduction of the ferricyanide. The reagent is incorporated into a test strip that generates an electrical output signal indicative of the level of 3-hydroxybutyric acid when the reagent is contacted with a sample.

Abstract: The present invention provides a novel polypeptide possessing growth activities on hematopoietic stem cells, a gene encoding the polypeptide and an antibody reacting specifically with the polypeptide, as well as a method for isolating the above gene and a vector for use in the method. According to the present invention, it is possible to elucidate the pathogenesis of various hematopoietic diseases due to abnormalities in hematopoietic stem cells and of bone marrow inhibition, and to diagnose and treat such diseases. According to the present invention, it is also possible to amplify hematopoietic stem cells in vitro for bone marrow transplantation necessary for the treatment of such diseases or improve efficiency of a gene transfer into hematopoietic stem cells for utilizing the gene therapy. The vector and the method of gene isolation developed to isolate the SCGF gene of the invention are applicable to search for other novel genes and able to contribute to the technological progress in genetic engineering.

Abstract: The present invention relates to the acid sphingomyelinase gene and to methods of diagnosing Niemann-Pick disease. It is based, at least in part, on the cloning and expression of the full-length cDNA encoding acid sphingomyelinase and on the discovery of mutations in the acid sphingomyelinase gene of Ashkenazi Jewish Niemann-Pick disease patients.

Abstract: Polynucleotide sequences and vectors containing them, for use in gene therapy, the polynucleotide sequences comprising two or more therapeutic genes operably linked to a promoter and encoding a fusion protein product of the therapeutic genes. The fusion protein may be for example a tyrosine hydroxylase (TH)-DOPA decarboxylase (DD) fusion in either TH-DD or DD-TH order, useful for treating Parkinson's disease.

Abstract: The present invention relates to novel parathyroid hormone (PTH) and parathyroid hormone related protein (PTHrP) receptors (PTH1R and PTH3R) isolated from zebrafish. The receptors of the present invention share homology with previously identified parathyroid hormone (PTH)/parathyroid related protein (PTHrP) receptors. Isolated nucleic acid molecules are provided encoding the zebrafish PTH1R and PTH3R receptors. PTH1R and PTH3R receptor polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of PTH1R and PTH3R receptor activity and to diagnostic and therapeutic methods.

Abstract: Expression of an endogenous gene is activated or increased following integration into a cell, by non-homologous or illegitimate recombination, of (1) an enhancer sequence that activates expression of the gene and (2) a sequence that encodes an amplifiable marker. The invention also provides methods for the identification, activation, isolation, and/or expression of genes undiscoverable by current methods since no target sequence is necessary for integration. The invention also provides cells containing the enhancer and amplifiable marker sequence and expressing increased amounts of a desired gene. The invention also provides methods for the isolation of nucleic acid molecules (particularly cDNA molecules) encoding a variety of proteins, including transmembrane proteins, and for the isolation of cells expressing such proteins.

Abstract: The invention provides a Douglas-fir 2S seed-storage promoter (df2SSP; SEQ ID NO: 17) and methods of its use. The promoter is useful for, among other things, directing the tissue-specific expression of transgenes.

Abstract: A method for increasing production yield of viruses, viral proteins, and other related biological materials through enhanced control and stabilization of protein production via stress proteins and the resultant protein products. The present invention is also directed to methods for selection or engineering of cell lines yielding such enhanced stabilized products. More specifically, example embodiments of the present invention are directed to methods for enhancing production of a viral agent, production of cell lines exhibiting permanent genetic modification, production of permissive eucaryotic cell lines, enhancing functional recombinant product yield, and the products of such methods.

Abstract: The invention relates to human TNF delta and TNF epsilon polypeptides, polynucleotides encoding the polypeptides, methods for producing the polypeptides, in particular by expressing the polynucleotides, and agonists and antagonists of the polypeptides. The invention further relates to methods for utilizing such polynucleotides, polypeptides, agonists and antagonists for applications, which relate, in part, to research, diagnostic and clinical arts.

Abstract: The present invention provides a method for generating human monoclonal antibodies, especially those that are specific for surface antigens representative of a particular cell type. The present invention also includes populations of monoclonal antibodies produced by the invention methods, populations of polynucleotides comprising sequences encoding the immunoglobulins or fragments thereof, which are capable of binding to antigens representative of a cell type of interest.

Abstract: A method for specifically cleaving a double-stranded DNA. The method comprises forming a three-stranded portion comprising the double-stranded DNA portion including the position to be cleaved or its vicinity and an oligonucleotide and treating the three-stranded protion with a nuclease.

Abstract: A preparation method of labeled DNA and use thereof are disclosed. More specifically, the invention relates to a preparation method of labeled DNA by preparation of a complex triple-stranded DNA through an deoxyoligonucleotide complementary to the deoxyoligonucleotide sequence of the 3′ terminal sequence of a certain double-stranded DNA followed by replacement of the 3′ terminal region of the plus strand of said double-stranded DNA with at least one labeled dNTP, and also to said immobilization method of a DNA obtained from the preparation method of the labeled (or modified) DNAs onto a solid support. These subject matters are mainly useful for preparation of labeled DNA molecules.

Abstract: A process for purifying and concentrating a gluconic acid derivative, such as 2-keto-L-gulonic acid, comprising introducing a non-viable and/or acidified fermentation medium or an in-vitro reactor medium comprising at least the gluconic acid derivative and/or salt thereof to electrodialysis thereby purifying and concentrating the gluconic acid derivative.

Abstract: Disclosed is an improved process for irradiating cell-containing compositions, especially, red cell-containing compositions, wherein vitamin E or a derivative thereof is added to the cell-containing composition prior to, during or after such irradiation. Addition of vitamin E or a derivative thereof is protective of cells in such compositions, but not of virus. Cells irradiated using the inventive process show a reduced leakage of K+ from cells and also a reduced loss of negative charges from the cell membrane compared to cells subjected to the similar process wherein vitamin E or a derivative thereof are not used. In addition, red blood cells sterilized by this process are better preserved during storage and their life-time in the circulation in vivo is greatly enhanced.

Abstract: A reusable sponge or foam made of a polymer such as polyurethane is prepared containing a plurality of different enzymes or a cross-linked complex of the plurality of enzymes for detoxification of a diverse array of hazardous chemicals such as organophosphorus and/or organosulfur compounds. The plurality of enzymes include enzymes selected from acetylcholinesterase, butyrylcholinesterase, triesterase, pseudocholinesterase, choline oxidase, peroxidase, organophosphate hydrolase, phosphotriesterase, paraoxonase and laccase. A preferred mixture of enzymes contains organophosphate hydrolase and acetylcholinesterase or butyrylcholinesterase. The sponge or foam may additionally contain carbon, an enzyme reactivation compound and/or an indicator for measuring capacity for detoxification. The indicator can be fluorescent, chemiluminescent or visible chromogen or an electrode, and be encapsulated in a liposome or crushable packet.

Abstract: Compositions and methods for introducing a DNA of interest into a genomic target site are provided. In particular, the methods and compositions involve the use of a combination of target sites for two site specific recombinases and expression of a chimeric recombinase with dual target site specificity. Thus, the compositions comprise novel site-specific recombinases with specificities to multiple target sites, and nucleotide sequences and expression cassettes encoding these recombinases or target sites. The methods involve transforming a eukaryotic cell having target sites for the novel recombinase with a DNA of interest that is flanked by corresponding target sites. Expression of the recombinase results in integration of the DNA of interest into the genome of the cell. The compositions and methods of the invention have use in the construction of stably transformed eukaryotic cells, and in particular, plant cells.

Abstract: Purified and isolated IL-1/TNF-&agr;-activated kinase (ITAK), nucleic acids encoding ITAK, processes for production of recombinant forms of ITAK, pharmaceutical compositions containing ITAK, and use of ITAK in therapies and in various assays, including assays to detect antagonists and agonists of ITAK are provided.

Abstract: A &bgr;-Glucanase enzyme capable of hydrolytically cleaving mixed glucans is presented. The &bgr;-Glucanase is sufficiently stable under alkaline conditions for use in industrial cleaning processes, especially in the brewing industry.

Abstract: Novel calcium free subtilisin mutants are taught, in particular subtilisins which have been mutated to eliminate amino acids 75-83 and which retain enzymatic activity and stability. Recombinant methods for producing same and recombinant DNA encoding for such subtilisin mutants are also provided.

Abstract: Novel calcium free subtilisin mutants are taught, in particular subtilisins which have been mutated to eliminate amino acids 75-83 and part or all of amino acids 1-22 (the N-terminal region) and which retain enzymatic activity and stability. Recombinant methods for producing the same and recombinant DNA encoding for such subtilisin mutants are also provided.

Abstract: There are disclosed a protein having an amino acid sequence represented by amino acid numbers 1 to 684 or 49 to 684 shown in SEQ ID NO:2, or a protein having a glutaminase activity in which one or more amino acids is/are deleted from, substituted by, inserted to or added to the amino acid sequence of the above protein; a gene containing DNA encoding the above protein or a gene encoding a protein which hybridizes with the DNA of the above gene under a stringent condition and has a glutaminase activity; a recombinant DNA containing the above gene; a transformant or a transductant containing the above recombinant DNA; and a process for producing glutaminase which comprises culturing the above transformant or the above transductant and collecting glutaminase from a culture medium.

Abstract: A method of preparing 1,5-D-anhydrofructose in large quantities includes treating &agr;-1,4-glucan with a substantially pure &agr;-1,4-glucan lyase, which has been isolated from algae alone, wherein 1,5-D-anhydrofructose is produced directly from the &agr;-1,4-glucan.

Abstract: The present invention relates to the compositions and methods associated with the cloning of the catalytic subunit of cellulose synthase responsible for catalyzing cellulose biosynthesis. The invention relates further to compositions and methods for obtaining host cells containing recombinant cellulose synthase as well as compositions and methods for obtaining cellulose synthase from natural sources. In certain aspects, the present invention provides methods for the cloning of an 83 kd subunit of the cellulose synthase enzyme from Acetobacter xylinum.

Abstract: The present invention relates to a process for the production of 2-keto-L-gulonic acid by fermentative conversion of L-sorbose and/or D-sorbitol. The present invention further relates to novel bacterial strains useful in this process.

Abstract: A microbial process is provided for selective cleavage of only organic C—N bonds while leaving C—C bonds intact which may be used for reducing the nitrogen content of nitrogen-containing organic carbonaceous materials. Microorganisms of Pseudomonas ayucida have been found which have the ability of selective cleavage of organic C—N bonds. A particularly preferred microorganism is Pseudomonas ayucida strain ATCC No PTA-806. Other microorganisms useful in the cleavage of organic C—N bonds are Aneurinibacillus sp, Pseudomonas stutzeri, Yokenella sp. and Pseudomonas nitroreducens.

Abstract: A method of using a paddle in the accelerated remediation of a contaminated material and a paddle, therefore, are provided.

Abstract: Methods for the enzymatic resolution of mixtures of enantiomers, such as &bgr;-lactam compounds, which may be employed as intermediates in the preparation of taxanes bearing a C-13 sidechain containing a heterocyclic or cycloalkyl group, the latter useful in the pharmaceutical field.

Abstract: A perfusion chamber includes a porous oocyte support structure. A continuously sloped top surface and a receiving well in the support structure entrap the underside of the oocyte, thereby localizing the cell in a predetermined fixed position within the reach of dedicated voltage-clamp microelectrodes. A test solution is delivered continuously at the top of the chamber, above the oocyte, and withdrawn from the bottom of the chamber, below the oocyte. The porosity of the support material enables the continuous perfusion of test solution around the membrane of the oocyte, including its bottom portion that is held firmly in contact with the holding well. The geometry of the holding well is selected such as to ensure the automatic and precise placement of the oocyte by gravity and to optimize the pressure distribution over its membrane, thereby minimizing the probability of rupture or other damage to the cell.

Abstract: Disclosed is a nucleic acid construct comprising a murine SOCS-3 promoter sequence having SEQ. ID. NO.: 1, or a non-murine homologue thereof, or an operative fragment or derivative. The construct can also contain, operatively linked to the SOCS-3 promoter, a gene encoding any preselected protein, and optionally contains a reporter gene to facilitate detection and/or selection of successfully transfected cells. Also disclosed are a transgenic vertebrate cell containing the nucleic acid construct and transgenic non-human vertebrates comprising such cells. The nucleic acid construct is useful in methods of treating a growth retardation or growth acceleration disorder in a human subject and in a method of treating an autoimmune disease, immune disease, or inflammatory condition in a human subject. A kit for genetically modifying a vertebrate cell includes a polynucleotide comprising the murine SOCS-3 promoter sequence is also disclosed.

Abstract: The present invention is directed to improved helper vectors and cell lines for the production of pseudoadenoviral (PAV) vectors containing substantially reduced levels of contaminating helper vector. The invention provides for helper vectors for the production of substantially helper vector-free PAV stocks comprising phag C31 recombinase recognition sequences which, depending upon their arrangement within the helper vector, can prevent helper vector packaging. The invention also provides for improved cell lines for the production of substantially helper vector-free PAV stocks comprising a stably introduced novel circular PAV genome into the cell.