Abstract: The present invention provides methods for reducing the complexity of a nucleic acid sample to interrogate a collection of target sequences. Complexity reduction can be accomplished by fragmenting the nucleic acid sample with a restriction enzyme that has at least one variable position in the recognition sequence. In some aspects adaptors that ligate to some but not all possible overhangs generated by digestion are ligated to the fragments. Selective adaptor ligation allows for selective amplification of a subset of the fragments using primers complementary to the adaptor sequence. In another aspect primers that are complementary to a subset of the fragments after adaptor ligation are used for amplification. Reduced complexity samples generated by the disclosed methods may be interrogated for the genotypes of SNPs in the sample.

Abstract: Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers facilitate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R21Z1C(O)R22R28 where R21 is a C1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z1 is either NH, sulfur or oxygen, R22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R28 includes a functional group which attaches the linker to the acceptor dye.

Abstract: Compounds represented by the following structural formulas can be used as photoacid generators: Such compounds are useful, for example, in fabricating arrays of polymers.

Abstract: The invention relates to detectable labels useful for detection of nucleotide sequences. Specifically, the invention relates to labeled-imidazole-PEG compounds, such as nucleosides, nucleotides, and nucleic acids incorporating such compounds, and methods utilizing such compounds. The invention further relates to kits comprising labeled imidazole-PEG compounds.

Abstract: The invention relates, in part, to methods useful in identifying molecules, that bind TRPM4b, which modulate TRPM4b ion channel activity, and/or which alter expression of TRPM4b within cells. The TRPM4b channels as described herein contain TRPM4b polypeptides, which are in turn encoded by TRPM4b nucleic acids. The ion channels described herein are preferably formed in HEK-293 cells from one or more novel TRPM4b polypeptides, which exhibit one or more of the unique TRPM4b properties described herein.

Abstract: The present invention includes antibodies to human GW182 protein and the use of those antibodies in clinical and diagnostic assays.

Abstract: The present invention provides a monoclonal antibody or a fragment thereof binding to the extracellular I-domain of integrin alpha10beta1 and a hybridoma cell line deposited at the Deutsche Sammlung von Microorganismen und Zellkulturen GmbH under the accession number DSM ACC2583. Furthermore, the present invention also provides a monoclonal antibody or a fragment thereof binding to the extracellular I-domain of integrin alpha10beta1 produced by the hybridoma cell line deposited. Method and uses of the antibody or a fragment thereof in identifying and selecting cells of a chondrogenic nature for treatment purposes, in particular for the identification and isolation of chondrocytes, mesenchymal progenitor cells and embryonic stem cells for tissue engineering of cartilage, or for identifying diagnostic and therapeutic tools in studying the biological role and the structural/functional relationships of the integrin alpha10beta1 with its various extracellular matrix ligands are also included.

Abstract: The present invention relates to biomarker polypeptides, polynucleotides, and antibodies that have utility in predicting in vitro and/or in vivo hepatotoxicity of various drugs, compounds, or other therapeutic agents (i.e., test substances). Also related are screens, kits, microarrays, and cell culture systems that employ the polypeptides, polynucleotides, and/or antibodies of the invention. The reagents and methods of the invention are useful for predicting hepatotoxic effects resulting from treatment with one or more test substances, and can be utilized before, after, or concurrently with pre-clinical, clinical, and/or post-clinical testing. In this way, the reagents and methods of the invention can be used to identify test substances or combinations of test substances that cause hepatic injury, including idiosyncratic hepatotoxicity, and thereby prevent medical complications (e.g., liver failure) resulting from such injury.

Abstract: Branched peptide amphiphilic compounds incorporating one or residues providing a pendant amino group for coupling one or more epitope sequences thereto, such compounds and related compositions for enhanced epitope presentation.

Abstract: MCT1 is consistently expressed at high levels in brain microvessel endothelial cells. Disclosed herein are assays for determining whether a test material/molecule is a substrate for, and/or is actively transported by, the MCT1 transporter, and therefore a candidate substrate for crossing the blood brain barrier. The assays are useful in screening for therapeutic, cytotoxic or imaging compounds used in the treatment or diagnosis of neurological diseases.

Abstract: The present invention provides methods of identifying markers indicative of the risk of developing a neurodevelopmental disorder caused in part by antibody- or autoantibody-mediated damage of neural tissue, including autism spectrum disorder (ASD). The invention further provides methods of diagnosing whether an individual has a neurodevelopmental disorder, including an ASD, and methods for determining the risk that a mother's future offspring will develop an a neurodevelopmental disorder, including an ASD.

Abstract: The invention is directed to a method for identifying substances acting as ligands for transfected receptors by using transfected markers to measure receptor/ligand interactions. The present invention also relates to a method of identifying compounds which act as inverse agonists of the 5-HT2A receptor, the method comprising contacting a constitutively active 5-HT2A receptor with at least one test compound and determining any decrease in the amount of basal activity of the receptor so as to identify a test compound which is an inverse agonist of the 5-HT2A receptor. Such inverse agonists may be used in the treatment of schizophrenia and related psychoses.

Abstract: The present invention discloses methods for activating Caspase 9 in such a way that it can be used in assays to discover modulators of Caspase 9.

Abstract: The invention provides isolated nucleic acids molecules, designated collagen XXII nucleic acid molecules, which encode a novel collagen. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing collagen XXII nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a collagen XXII gene has been introduced or disrupted. The invention still further provides isolated collagen XXII proteins, fusion proteins, antigenic-peptides and anti-collagen XXII antibodies. Diagnostic methods utilizing compositions of the invention are also provided.

Abstract: Binding assays for identifying components that modulate human T1R1 polypeptide-associated taste are disclosed. These assays detect the specific binding of compounds to a human T1R1 polypeptide or the modulation of the specific binding of another compound that specifically binds human T1R1 polypeptide. The binding assays may include the use of detectable labels, e.g., radionuclides, enxymes, fluorophores, and the like. Compounds identified in these binding assays have potential application as T1R1 taste modulators and can be used as additives in compositions for human or animal consumption.

Abstract: The present invention describes methods for identifying compounds that inhibit JNK and MLK kinase activity as drugs for treating a mammal susceptible to or having a neurological condition. This invention also discloses methods for preventing neuronal cell death and treating neurological conditions that involve neuronal cell death, particularly neurodegenerative diseases characterized by glutamine or kainate mediated toxicity, such as Huntington's disease and Alzheimer's disease.

Abstract: It is intended to provide a noninvasive method of conveniently detecting mild impaired glucose tolerance and/or insulin hyposecretion at the early stage with the use of an enzyme. Namely, mild impaired glucose tolerance and/or hyposecretion at the early stage are detected by quantifying myoinositol secreted into the urine before loading glucose and after loading glucose for a definite period of time with the use of a reagent and comparing the increase (or the increase ratio) in the myoinositol content thus measured with a characteristic level which has been preliminarily determined in normal subjects.

Abstract: The present invention provides a transformed cell in which a polynucleotide having a nucleotide sequence encoding an amino acid sequence of an osmosensing histidine kinase having no transmembrane region is introduced in a functional form into a cell deficient in at least one hybrid-sensor kinase, a method of assaying the antifungal activity of a test substance using the transformed cell, and a method of searching an antifungal compound using the method, and the like.

Abstract: This document provides methods and materials related to rapid, quantitative determination of TPMT activity in biological samples. Also featured are compositions and kits useful for determination of TPMT activity in biological samples.

Abstract: Methods and reagents are provided for measuring protease activity. The reagent comprises a surface to which is linked an enzyme donor fragment through a protease recognition sequence, where the enzyme donor fragment complexes with an enzyme acceptor fragment to form an active indicator enzyme when the enzyme donor fragment is cleaved from said surface. Conveniently, the surface is a cell membrane surface where the reagent is expressed in the cell. The method comprises bringing together the reagent, the protease, the enzyme acceptor and substrate for the indicator enzyme and measuring the indicator enzyme activity as a measure of the protease activity. The method finds application in screening for compounds modulating the activity of the protease.

Abstract: The present invention provides a method of detecting production of antibiotic-inactivating factor and for determining the antibiotic susceptibility of a microorganism comprising the following steps, a culture of the microorganism suspected of producing inactivating factors and/or whose susceptibility is to be determined is admixed with an antibiotic to which susceptibility is to be assayed, and a permeabilizing agent for the microorganism present in a non-growth-inhibiting microorganism-permeabilizing effective amount to form an assay culture. The assay culture is incubated under appropriate culture conditions and for a time sufficient to determine production of antibiotic-inactivating factors and/or the susceptibility of the microorganism to the antibiotic.

Abstract: Provided is a method for obtaining a macrolide, especially tacrolimus, ascomycin, pimecrolimus, sirolimus, or everolimus, from biomatter.

Abstract: Genetic and biochemical characterization of the leinamycin biosynthesis gene cluster from Streptomyces atroolivaceus S-140 revealed two PKS genes, lnmI and lnmJ, that encode six PKS modules, none of which contains a cognate AT domain. The AT activity is provided in trans by a discrete protein, LnmG, which loads the malonyl coenzyme A extender unit onto the ACP domains of all six PKS modules. This finding provides a basis for methods of engineering modular polyketide synthases and polyketide synthase/nonribosomal peptide synthetases.

Abstract: The invention provides isolated nucleic acid and amino acid sequences of taste cell specific G-protein coupled receptors, antibodies to such receptors, methods of detecting such nucleic acids and receptors, and methods of screening for modulators of taste cell specific G-protein coupled receptors.

Abstract: The invention provides methods of increasing the production of polypeptides, optionally recombinant polypeptides, from mammalian cells using xanthine derivatives or hybrid polar compounds and cultures containing the same. Combinations of inducers including a hybrid polar compound and/or a xanthine derivative and/or an alkanoic acid can also be used, optionally at temperatures less than 37° C.

Abstract: Disclosed is a recombinant plasmid for expressing hepatitis B viral antigens in vivo, comprising an adeno-associated virus (AVV) vector and a replication-competent hepatitis B virus genome fragment. Mice hydrodynamically injected with the recombinant plasmid of the present invention show persistent expression of hepatitis B viral antigens for more than 6 months in the hepatocytes, thus a immuno-competent mouse model for persistent expression of hepatitis B antigens and also for human chronic hepatitis B virus infection is established, which can be applied in evaluation and elucidation of mechanism of chronic hepatitis and anti-viral drug discovery research.

Abstract: Chromatographic processes and systems for purifying a botulinum toxin from an APF fermentation medium.

Abstract: The present invention describes terminal phosphate blocked nucleoside polyphosphates that are stable at high temperature and their use in nucleic acid amplification and analysis. Current invention further describes charge modified terminal phosphate blocked nucleoside polyphosphates for improved incorporation and direct loading of nucleic acid sequencing reactions onto separating media.

Abstract: The present invention is directed to a hairpin nucleic acid structure and its use. In a preferred embodiment, the hairpin nucleic acid structure can be used in a method of amplification of a template nucleic acid sequence that substantially reduces polymerase-induced errors.

Abstract: Methods and compositions for identifying novel pesticidal gene homologues are provided. Specifically, the methods of the invention comprise systematically designing oligonucleotide primers that are specific for a pesticidal gene of interest and performing successive rounds of PCR amplification of nucleic acid material from a microorganism, particularly a Bacillus thuringiensis strain, to identify novel homologues of known pesticidal genes. Oligonucleotide primers that can be used to practice the present methods are further disclosed.

Abstract: Purified ?-amino acids are of considerable interest in the preparation of pharmacologically active compounds and industrial precursors. Although enantiomerically pure ?-amino acids can be produced by standard chemical synthesis, this traditional approach is time consuming, requires expensive starting materials, and results in a racemic mixture which must be purified further. However, DNA molecules encoding lysine 2,3-aminomutase can be used to prepare ?-amino acids by methods that avoid the pitfalls of chemical synthesis. The present invention provides a method of producing enantiomerically pure ?-amino acids from ?-amino acids comprising catalyzing the conversion of an ?-amino acid to a corresponding ?-amino acid by utilizing a lysine 2,3-aminomutase as the catalyst.

Abstract: The present invention is directed to improving productivity of an enzymatic method for making esterified, transesterified or interesterified products. Specifically, a method that can greatly improve the productivity of enzymatic transesterification or esterification by deodorization alone, or by deodorization and purification of the initial substrate to extend the useful life of the enzyme is disclosed.

Abstract: The present invention provides methods for separating isotopes of actinide elements such as uranium using microorganisms, e.g., metal or sulfate reducing bacteria. The microorganisms reduce the actinide element to form a precipitate, which contains a greater proportion of the lighter isotope relative to the heavier isotope than the starting material. The precipitate may be collected, re-oxidized, and subjected to multiple rounds of enrichment. Alternately, separation processes not requiring formation of a precipitate may be used. The invention also features cell-free systems for isotope separation. The invention further provides compositions produced according to the foregoing methods, including compositions comprising enriched uraninite.

Abstract: The object of the present invention is to provide a novel dehydrogenase having a property which is different from that of known dehydrogenases. The present invention provides a dehydrogenase having the following physicochemical properties: (1) effect: to produce N-alkyl-L-alanine from pyruvic acid and alkylamine or dialkylamine using NADPH and/or NADH as coenzyme; (2) substrate specificity: to show activity to alkylamine or dialkylamine but not to ammonium; (3) optimal pH when using phenylpyruvic acid and methylamine as substrates is around 10; and (4) when treated at 30° C. for 30 minutes, the enzyme is stable at around pH 5 to 10.5.

Abstract: A histidine-tagged, deletion mutant of bacteriophage N4-coded, virion RNA polymerase (mini-vRNAP) which is active has been developed. The his-tagged mini-vRNAP has been cloned under the control of the pBAD promoter, is stable and is purified in a single step yielding large amounts (10 mg/liter of E. coli expressing cells). This RNA polymerase uses single-stranded DNA containing 17 bases (the promoter) upstream of the transcribed regions as a template. In the presence of E. coli SSB protein, it transcribes this template efficiently, providing a unique system to synthesize RNAs of the desired sequence using single-stranded DNA templates. The enzyme incorporates derivatized nucleoside triphosphates with high efficiency. A mutant of mini-vRNAP has been generated that incorporates deoxynucleoside triphosphates. In addition, the inventors have developed an in vivo system to express RNAs and proteins under mini vRNA polymerase promoter control.

Abstract: A purified recombinant phytase enzyme derived from Escherichia coli B. The enzyme has a molecular weight of about 47.1 kilodaltons and has phytase activity. The enzyme can be produced from native or recombinant host cells and can be used to aid in the digestion of phytate where desired. In particular, the phytase of the present invention can be used in foodstuffs to improve the feeding value of phytate rich ingredients.

Abstract: Disclosed are a number of homologs and variants of Hypocrea jecorina Cel7A (formerly Trichoderma reesei cellobiohydrolase I or CBH1), nucleic acids encoding the same and methods for producing the same. The homologs and variant cellulases have the amino acid sequence of a glycosyl hydrolase of family 7A wherein one or more amino acid residues are substituted and/or deleted.

Abstract: A novel human cDNA, termed PRSS11-L, was isolated which encodes a polypeptide that belongs to the S2/HtrA serine protease family. The PRSS11-L mRNA is widely expressed in several tissues throughout the body by multi-tissue Northern blotting. The full-length PRSS11-L cDNA, was cloned, expressed and purified. Proteolytic activity was demonstrated using the protein substrate casein. The isolated nucleic acid or polypeptide molecule of the invention can be used in detection assays, gene therapy, and screening assays.

Abstract: Early detection of infectious agents of human prion diseases such as CJD by using an animal model, etc. is needed in order to rapidly determine prion infections in pharmaceuticals such as blood products, foods, or cosmetics. This invention provides a screening method for infectious agents of human or non-human prion diseases in samples, which employs, as an indication, the deposition of the aberrant prion protein in the follicular dendritic cell (FDC) of a non-human animal.

Abstract: The present invention provides an improved method for the biological production of 1,3-propanediol from a fermentable carbon source in a single microorganism. In one aspect of the present invention, an improved process for the conversion of glucose to 1,3-propanediol is achieved by the use of an E. coli transformed with the Klebsiella pneumoniae dha regulon genes dhaR, orfY, dhaT, orfX, orfW, dhaB1, dhaB2, dhaB3, and orfZ, all these genes arranged in the same genetic organization as found in wild type Klebsiella pneumoniae. In another aspect of the present invention, an improved process for the production of 1,3-propanediol from glucose using a recombinant E. coli containing genes encoding a G3PDH, a G3P phosphatase, a dehydratase, and a dehydratase reactivation factor compared to an identical process using a recombinant E. coli containing genes encoding a G3PDH, a G3P phosphatase, a dehydratase, a dehydratase reactivation factor and a 1,3-propanediol oxidoreductase (dhaT).

Abstract: A contact plate for growing cell cultures, bacteria cultures, and the like, comprising a base and a cover. The base has a bottom wall for holding the culture medium and the like, and a circumferential sidewall extending upwardly from the bottom wall and attached thereto. The base further incorporates a flange extending outwardly from the outer periphery of the sidewall. The cover has a top wall, which has a larger diameter than the base sidewall, and a circumferential sidewall extending downwardly from the top wall and attached thereto. The contact plate has a locking means for securely holding the cover to the base of the contact plate in the locked position, wherein the locking means comprises an inter-engaging rib and groove arrangement.

Abstract: A thermal cycling device for thermally cycling samples of biological material contained in a microcard having a top and bottom surface. The thermal cycling device can include a sample block having an upper surface configured for engaging the bottom surface of a microcard, a vacuum device, and a temperature control system operatively connected with the sample block. The upper surface of the sample block may include a plurality of channels, the channels defining spaces between the sample block and the bottom surface of a microcard that may be positioned thereon. The vacuum device may be in fluid communication with the sample block for drawing gas out of the spaces defined by the channels in the sample block. The vacuum device may be configured for substantially maintaining a vacuum between the sample block and microcard so that a retention force is imparted on the microcard to urge the microcard toward the sample block.

Abstract: A process for manufacturing a microfluidic device, including the steps of: forming at least one channel in a semiconductor material body; forming a dielectric diaphragm above the channel, for closing the channel; and forming heating elements for providing thermal energy inside the channel. The heating elements are formed directly on said dielectric diaphragm.

Abstract: A device for storage and transportation of forensic and/or biological material, comprising a sealable container and a material holder for holding the material, which holder can be placed in the container, wherein the container is provided with at least one ventilating hole and with sealing means that shut off the interior of the container from the environment.

Abstract: The expression of the lipidated form of the peptidoglycan-associated protein (PAL) of gram-negative bacteria is achieved through the use of a plasmid containing a tightly regulated promoter. A bacterial host cell is transformed, transduced or transfected with such a plasmid. The host cell is then cultured under conditions such that the lipidated recombinant PAL is expressed. The lipidated recombinant PAL is included in an antigenic composition administered to a mammalian host to immunize against a gram-negative bacterium.

Abstract: Sustained transgene expression will be required for the vast majority of genetic diseases being considered for gene therapy. The initially high levels of expression attained with plasmid DNA (pDNA) vectors containing viral promoters, such as that from cytomegalovirus (CMV), decline precipitously to near background levels within 2 to 3 weeks. We have constructed pDNA vectors containing the human cellular ubiquitin B (Ub) promoter and evaluated their expression in the mouse lung. Cationic lipid-pDNA complexes were instilled intranasally (IN) or injected intravenously (IV) into immunodeficient BALB/c mice. Chloramphenicol acetyltransferase (CAT) reporter gene expression from the Ub promoter was initially very low at day 2 post-administration but by day 35 exceeded the level of expression attained from a CMV promoter vector by 4- to 9-fold.

Abstract: Polynucleotides encoding carboxylesterase enzymes and polypeptides encoded by the polynucleotides which are capable of metabolizing a chemotherapeutic prodrug and inactive metabolites thereof to active drug are provided. Compositions and methods for sensitizing tumor cells to a prodrug chemotherapeutic agent and inhibiting tumor growth with this enzyme are also provided. In addition, screening assay for identification of drugs activated by this enzyme are described.

Abstract: This invention provides a new procedure for generating cardiomyocyte lineage cells from embryonic stem cells for use in regenerative medicine. Differentiating by way of embryoid body formation or in serum is no longer required. Instead, the stem cells are plated onto a solid substrate, and differentiated in the presence of select factors and morphogens. After enrichment for cells with the appropriate phenotype, the cells are allowed to cluster into cardiac bodies™, which are remarkably homogeneous and suitable for the treatment of heart disease.

Abstract: An apparatus and method of using the apparatus to prepare a biocompatible biodegradable matrix capable of supporting cells to form an implantable or engraftable surgical device. A matrix-forming fluid is contained within a chamber defined by top and bottom surfaces of a thermally conductive material and spacers defining the thickness of the matrix. The chamber is then cooled to freeze the solution at a controlled rate, resulting in a matrix with a desired and uniform thickness having symmetric and uniform reticulations. The apparatus and method reproducibly forms such a matrix, which may be populated with cells for transplantation and engraftment into a wound.

Abstract: Methods for the diagnosis of visceral, cutaneous and canine leishmaniasis in a subject suspected of being infected with the parasitic protozoa Leishmania is disclosed. Disclosed are antibody-capture enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to Leishmania parasite soluble antigens and antigen-capture ELISAs for the detection of Leishmania parasite soluble antigens in host samples. Also disclosed are immunodiagnostic kits for the detection of Leishmania parasite circulating antigens or IgM and IgG antibodies in a sample from subject having visceral, cutaneous or canine leishmaniasis. In these methods and kits, detection may be done photometrically or visually. The methods and kits also allow the visualization of Leishmania amastigotes or promastigotes in a sample.