Abstract: A cleavable signal element for use in quantitative and qualitative assay devices and methods is described. Binding of the chosen analyte simultaneously to a first and a second analyte-specific side member of the cleavable signal element tethers the signal-responsive moiety to the signal element's substrate-attaching end, despite subsequent cleavage at the cleavage site that lies intermediate the first and second side members. Assay devices comprising the cleavable signal elements are described, as are analytic methods adapted to their use. The analytic devices of the present invention may be adapted to detection using conventional CD-ROM and DVD readers.

Abstract: Processes are disclosed using the depolymerization of a nucleic acid hybrid to qualitatively and quantitatively analyze for the presence of a predetermined nucleic acid. Applications of those processes include the detection of single nucleotide polymorphisms, identification of single base changes, speciation, determination of viral load, genotyping, medical marker diagnostics, and the like.

Abstract: The present invention relates to a method for detection and identification of at least one microorganism, or for the simultaneous detection of several microorganisms in a sample, involving the steps of: (i) if need be releasing, isolating or concentrating the polynucleic acids present in the sample; (ii) if need be amplifying the 16S-235 rRNA spacer region, or a part of it, with at least one suitable primer pair; (iii) hybridizing the polynucleic acids of step (i) or (ii) with at least one and preferably more than one of the spacer probes as mentioned in table la or equivalents of thereof, under the appropriate hybridization and wash conditions, and/or with a taxon-specific probe derived from any of the spacer sequences as represented in FIGS.

Abstract: The present invention involves a method for characterizing nucleic acid which comprises generating Sanger ladder nucleic acid fragments from a plurality of nucleic acid templates present in the same reaction zone, wherein at least one terminating base is present in the reaction zone. Prior to generating nucleic acid fragments, a labeled primer nucleotide or oligonucleotide is hybridized to each template. The label on each primer is specific to the template to which that primer hybridizes, thereby allowing for identification of the template. The method of the present invention further comprises identifying the length of each nucleic acid fragment produced, the template from which the fragment is derived and the terminating base of the fragment.

Abstract: A denaturation fingerprinting method (dnF) involves subjecting a nucleic acid segment of interest to bidirectional cycle sequencing using oppositely oriented primers and incorporating two different dideoxynucleotides (ddNTPs) in the sequencing reaction. The resulting fragments are separated by denaturing electrophoresis. In one embodiment, designated dnF2R, reactions and electrophoretic separation using the two ddNTPs are conducted separately. In an alternative embodiment, designated dnF1R, one of the ddNTPs has a mobility altering modification such that electorphoretic separation occurs when both ddNTPs are employed in the same reaction. The methods are useful for detecting genetic mutations.

Abstract: A fluorescently labeled nucleic acid having a hairpin structure between the fluorophore label and a point of attachment to a solid phase is useful as a probe to detect nucleic acid from a sample. The solid phase quenches the fluorophore label when the hairpin structure exists but this quenching is relieved by duplex formation between probe and a sample oligonucleotide. Probes for specific nucleic acid sequences can be immobilized as arrays on solid phase surfaces for detection of multiple nucleic acid sequences simultaneously from electrophoresis gels and from aqueous solutions. These probes and methods for their use can be combined with known solid phases, particularly those used for plasmon surface detection and electron transfer detection of nucleic acid. The probes can be washed and reused, and have other advantageous features over known probe methods.

Abstract: Disclosed are the dbp gene and dbp-derived nucleic acid segments from Borrelia burgdorferi, the etiological agent of Lyme disease, and DNA segments encoding dbp from related borrelias. Also disclosed are decorin binding protein compositions and methods of use. The DBP protein and antigenic epitopes derived therefrom are contemplated for use in the treatment of pathological Borrelia infections, and in particular, for use in the prevention of bacterial adhesion to decorin. DNA segments encoding these proteins and anti-(decorin binding protein) antibodies will also be of use in various screening, diagnostic and therapeutic applications including active and passive immunization and methods for the prevention of Borrelia colonization in an animal. These DNA segments and the peptides derived therefrom are contemplated for use in the preparation of vaccines and, also, for use as carrier proteins in vaccine formulations, and in the formulation of compositions for use in the prevention of Lyme disease.

Abstract: The present invention provides a method for the isolation of extrachromosomal amplified nucleic acids that are associated with a cell proliferative disorder. Isolation and further identification of such genes is critical for diagnosis, prognosis, and course of therapy.

Abstract: The present invention relates to methods and compositions for the diagnosis, prevention, and treatment of tumor progression in cells involved in human tumors such as melanomas, breast, gastrointestinal, lung, and bone tumors, various types of skin cancers, and other neoplastic conditions such as leukemias and lymphomas. Genes are identified that are differentially expressed in benign (e.g., non-malignant) tumor cells relative to malignant tumor cells exhibiting a high metastatic potential. Genes are also identified via the ability of their gene products to interact with gene products involved in the progression to, and/or aggressiveness of, neoplastic tumor disease states. The genes and gene products identified can be used diagnostically or for therapeutic intervention.

Abstract: A multistage electromagnetic separator is designed to separate magnetically susceptible materials suspended in fluids. The apparatus includes an upper plate and a lower plate set to a fill position and the fluid samples are filled into upper and lower cuvettes. The upper cuvette rotates into position above the lower cuvette aligning the upper and lower cuvettes. A translating electromagnet energizes to a programmed current level and translates from the bottom of the lower cuvette to the interface of the plates. The translating electromagnet is de-energized, and a holding electromagnet is energized to a programmed current level pulling particles within a specified mobility range into the top of the captured upper collection cuvette. The holding electromagnet is de-energized leaving the permanent holding magnet to keep the collected sample particles in the top cuvette while the upper plate rotates thereby capturing the sample of the collected particles.

Abstract: The present invention relates to a stenographic method for concealing coded messages in DNA. The method of the invention comprises concealing a DNA encoded message within a genomic DNA sample followed by further concealment of the DNA sample to a microdot. The present invention further provides a method for the use of genomic steganography to mark and authenticate objects of interest.

Abstract: A cis-acting posttranscriptional regulatory element (PRE) useful for efficient RNA export of RNA is provided. The element, termed WPRE, is originally derived from woodchuck hepatitis virus. The invention also provides a method for enhancing the expression of transgenes by insertion of the WPRE nucleic acid sequences in operably linkage with the transgene.

Abstract: The present invention provides methods for determining nucleic acid sequences based on known sequences adjacent to the unknown sequences, and reagents for using such methods.

Abstract: The invention provides methods, compositions, and apparatus for performing sensitive detection of analytes, such as biological macromolecules and other analytes, by labeling a probe molecule with an up-converting label. The up-converting label absorbs radiation from an illumination source and emits radiation at one or more higher frequencies, providing enhanced signal-to-noise ratio and the essential elimination of background sample autofluorescence. The methods, compositions, and apparatus are suitable for the sensitive detection of multiple analytes and for various clinical and environmental sampling techniques.

Abstract: The invention relates to a method by which new antigens from vector-borne pathogens may be discovered and analyzed by incubating the viable pathogens in the saliva of their vector host. Three such antigens, proteins with the approximate molecular weights of 19, 22 and 24 kDa, have been discovered and analyzed from a strain of B. burgdorferi T-15. The proteins provide a route for the development of immunodiagnostics for Lyme disease and related disorders. The proteins and related amino acids and DNA sequences may also be used for the immunization, for the detection of B. burgdorfei in human or body fluids, and also for the generation of specific antibodies for use in diagnosis, epidemiology, prevention of and treatment of Lyme disease.

Abstract: The present invention concerns muteins of avidin and streptavidin with a reduced binding affinity for biotin as well as their use as interference elimination reagents in methods for the determination of an analyte e.g. in diagnostic tests such as for example immunoassays and nucleic acid hybridization assays. In addition the invention concerns the use of muteins of avidin and streptavidin as systems capable of regeneration for binding biotin e.g. for the analysis of biotinylated molecules, for examining receptor ligand interactions as well as for the affinity purification of biotinylated molecules.

Abstract: A method of screening a candidate compound for susceptibility to metabolism by a selected enzyme. The method includes the steps of reacting the candidate compound, an indicator compound precursor and the selected enzyme, the enzyme characterized as having a side reaction associated with metabolic activity of the enzyme wherein a chemical species capable of reacting with the indicator compound precursor is produced; and detecting an indicator compound, the indicator compound produced from the indicator compound precursor by reaction with the chemical species produced from the side reaction associated with metabolic activity of the enzyme, the detection of the indicator compound indicating the susceptibility of the candidate compound to metabolism by the enzyme. A preferred example of the selected enzyme is a cytochrome P450 (CYP).

Abstract: There are described a simple examination method of infection with Helicobacter pylori possibly presenting in a gastric mucosa, and a device therefor. The examination is conducted by collecting gas in gastric cavity, and measuring mainly ammonia and additionally organic amines which are generated due to activities of the bacilli. The measurement is carried out by leading the gas in gastric cavity into oral cavity by vomiting-reflex, and sucking the gas by a metering suction pump through a gas detection tube to read-out a length of color-changed area in the gas detection tube.

Abstract: An enzymatic process is presented for reducing the cholesterol level of foods and feeds by converting the cholesterol therein to coprostanol which has a very low intestinal tract absorbability. Cholesterol in meat, egg, milk, seafood and cooked processed foods containing the same, or feeds for animals, poultry, and pisiculture can be reduced by the sequential action of three enzymes isolated from a eubacterium: cholesterol dehydrogenase, 4-cholesten-3-one dehydrogenase and coprostan-3-one dehydrogenase.

Abstract: The invention provides isolated nucleic acid compounds encoding a novel SAM synthetase of Streptomyces fradiae. Also provided are vectors and transformed heterologous host cells for expressing the SAM synthetase and a method for preparing S-adenosylmethionine from recombinant host cells transformed with the SAM synthetase gene.

Abstract: Novel polynucleotides and the proteins encoded thereby are disclosed.

Abstract: The sequences of extended cDNAs encoding secreted proteins are disclosed. The extended cDNAs can be used to express secreted proteins or portions thereof or to obtain antibodies capable of specifically binding to the secreted proteins. The extended cDNAs may also be used in diagnostic, forensic, gene therapy, and chromosome mapping procedures. The extended cDNAs may also be used to design expression vectors and secretion vectors.

Abstract: Compositions and methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form using a yeast host cell as the expression system are provided. Compositions of the invention are nucleotide sequences encoding a signal peptide sequence for a yeast secreted protein, an optional leader peptide sequence for a yeast secreted protein, a native propeptide leader sequence for a mature protein of interest, and a sequence for the mature protein of interest, all operably linked to a yeast promoter. Each of these elements is associated with a processing site recognized in vivo by a yeast proteolytic enzyme. Any or all of these processing sites may be a preferred processing site that has been modified or synthetically derived for more efficient cleavage in vivo. The compositions are useful in methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form.

Abstract: Interferons represent an important class of biopharmaceutical products, which have a proven track record in the treatment of a variety of medical conditions, including the treatment of certain autoimmune diseases, the treatment of particular cancers, and the enhancement of the immune response against infectious agents. The present invention provides a new form of murine interferon-&agr;, which has applications in diagnosis and therapy.

Abstract: Methods and compositions for efficient targeting and modification of target sequences in duplex DNA are provided, utilizing oligonucleotides or oligonucleotide compositions containing two domains. The first domain comprises an entity capable of recognizing a double-stranded DNA sequence. This can be a protein, peptide, antibiotic, minor groove binding agent or a nucleotide sequence capable of triplex formation The second domain, which is covalently joined to the first, is capable of recognizing a single-stranded DNA sequence. This second domain will most often be complementary, in the Watson-Crick sense, to a target sequence in the double-stranded nucleic acid. The second domain can optionally carry one or more modifying groups, capable of causing a mutation, a pre-mutagenic lesion, or some other type of heritable change in the target sequence.

Abstract: The present invention provides an isolated nucleic acid encoding a mammalian capping enzyme. The present invention further provides an isolated nucleic acid encoding a mammalian (Guanine-7-) methyltransferase enzyme. The present invention also provides an isolated mammalian capping enzyme protein or subunit thereof. In addition the present invention provides an isolated mammalian (Guanine-7-) methyltransferase enzyme protein or portion thereof. The present invention further provides methods for catalyzing formation of RNA 5′-terminal GpppN cap complex and for coupled transcription, translation and formation of RNA 5′-terminal GpppN cap complex and methylated RNA 5′-terminal GpppN cap complex. Kits thereto are also provided.

Abstract: Described herein are methods for removing the 3′-untranslated regions from cDNA or mRNA molecules, as well as methods for the use of such products for RNA-protein fusion formation.

Abstract: The present invention is directed to a transcription based amplification method for the amplification of DNA targets. With the method of the present invention an isothermal transcription based amplification method is provided for the amplification of double stranded DNA.

Abstract: The present invention provides compositions and methods for performing an amplification reaction of nucleic acids with internal controls that test the integrity of all aspects of the amplification reaction.

Abstract: The present invention is a method for the detection of a specific target bacteria in a complex sample mixture. The sample mixture may contain a variety of components including non-target or background microorganisms as well as other organic contaminants such as food debris. The method proceeds by first culturing the complex sample mixture in a non-selective growth medium, followed by isolation and detection of target bacteria DNA. Target DNA is detected via a DNA amplification protocol with a primer pair selected to amplify a specific, identifying portion of the target bacteria DNA. A control DNA is amplified concurrently with the target bacteria target DNA. The control DNA is specifically designed to be amplified with a single primer that is identical to one of the primers used in the amplification of the target genomic DNA. Use of this control validates the amplification reaction. Detection of the amplified target DNA and the control is accomplished by gel electrophoresis or by fluorescent means.

Abstract: A method of protecting a biomolecule from substantial degradation while reducing the content of a pathogen or chemical toxin contained by a biologically derived composition is described. The method involves first providing a biologically derived composition containing a pathogen or chemical toxin and at least one biologically active molecule. Then, adding albumin to the composition to create a supplemented composition. Next, subjecting the supplemented composition to polychromatic pulsed light to degrade the pathogen or toxin. The polychromatic light includes at least one high-intensity, short duration pulse of incoherent polychromatic light in a broad spectrum. The light intensity is at least about 0.01 J/cm2, the pulse duration is 10 ns to 100 ms and the light wavelengths are between about 170 nm and about 2600 nm.

Abstract: The invention provides YfiI pseudouridine synthase polypeptides and polynucleotides encoding YfiI pseudouridine synthase polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing YfiI pseudouridine synthase polypeptides to screen for antibacterial compounds.

Abstract: A novel carbonyl reductase useful for producing alcohol, particularly derivatives of (S)-4-halo-3-hydroxybutyrate ester, is provided. A novel carbonyl reductase derived from Kluyveromyces aestuarii and the nucleic acid encoding the enzyme are provided. The carbonyl reductase has excellent reductase activity and stereoselectivity. The carbonyl reductase reduces ketone to produce alcohol. It can be particularly advantageous when used in industrial production of (S)-4-halo-3-hydroxybutyrate ester.

Abstract: Isolated nucleic acid molecules encoding human MEKK proteins, and isolated MEKK proteins, are provided. The invention further provides antisense nucleic acid molecules, recombinant expression vectors containing a nucleic acid molecule of the invention, host cells into which the expression vectors have been introduced and nonhuman transgenic animals carrying a human MEKK transgene. The invention further provides human MEKK fusion proteins and anti-human MEKK antibodies. Methods of using the human MEKK proteins and nucleic acid molecules of the invention are also disclosed, including methods for detecting human MEKK activity in a biological sample, methods of modulating human MEKK activity in a cell, and methods for identifying agents that modulate the activity of human MEKK.

Abstract: The invention relates to the isolation of the prepro form of cathepsin L, of its leader sequence, of cathepsin L and of the affiliated propeptide from ciliates, in particular Paramecium, to the use of these peptides and to a process for preparing cathepsin L from ciliates.

Abstract: Novel protease variants derived from the DNA sequences of naturally-occurring or recombinant non-human proteases are disclosed. The variant proteases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant protease to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor protease. Such variant proteases have properties which are different from those of the precursor protease, such as altered wash performance.

Abstract: The present invention relates to novel metalloproteinase- like proteins. In particular, isolated nucleic acid molecules are provided encoding the human TACE-like and matrilysin-like proteins. TACE-like and matrilysin-like 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 TACE-like and matrilysin-like activity. Also provided are diagnostic methods for detecting cancer and therapeutic methods for cancer and other disorders characterized by an over or under production of these metalloproteinases.

Abstract: DNA encoding a therapeutically suitable glutaminase has been molecularly cloned. This allows one to obtain a polypeptide which is a therapeutically suitable glutaminase free of contaminating endotoxin. It has been found that this polypeptide is a potent anti-viral agent and when coupled to an anti-tumor monoclonal antibody is a potent anti-cancer agent. The glutaminase of the present invention is particularly useful for treating lung, breast and colon cancer cells and in the treatment of HIV-infected cells.

Abstract: Microbial antagonists that will suppress Fusarium head blight (head scab) in cereals, particularly in wheat and barley have been identified. Two superior antagonists include NRRL B-30210 and NRRL B-30211.

Abstract: The invention generally relates to compositions and methods for identifying cytokine, hormone and growth factor signaling pathway agonists and antagonists, and more particularly, methods and compositions for screening compounds and identifying compounds that will modulate the interactions of cytokine, hormone and growth factor receptors with their intracellular ligands, as well as between their intracellular ligands and other members of the signaling pathway.

Abstract: Substances such as chemical substances and biological substances including animal, vegetable and microbial cells are encapsulated using a process and an apparatus wherein a coil through which alternating current flows causes a magnet to vibrate creating vibrations such as in the range of between 300 to 4000 Hz that are transmitted to an encapsulating fluid containing the substance to form small substantially spherical particles containing the substance. The apparatus includes a pulsation chamber containing a movable wall for receiving the encapsulating fluid containing the substance to be encapsulated. A nozzle is spaced downstream from the pulsation chamber for receiving the encapsulating fluid from the pulsation chamber. A permanent magnet is mounted on the movable wall, and a coil is spaced from the permanent magnet by an air gap and is located proximate to the permanent magnet.

Abstract: A sample preparation apparatus for preparing a sample to be used for detecting specimens such as microorganisms isolated on a filter so as to enumerate microorganisms in the sample has a turntable on which multiple sample bases are formed, a filter insertion unit, an extractant spray apparatus, a luminescent reagent spray apparatus and a filter removal unit, disposed in order along a periphery of the turntable in a direction of rotation of the turntable. By eliminating the need to move filters between processing operations as well as any special skill required in the spraying of the extractant and reagent, the sample preparation apparatus can provide accurate microorganism counts quickly and efficiently.

Abstract: The present invention relates, in general, to pneumococcal fimbrial protein A. In particular, the present invention relates to a DNA segment encoding a pneumococcal fimbrial protein A gene; polypeptides encoded by said DNA segment; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing a pneumococcal fimbrial protein A polypeptide; antibodies specific to pneumococcal fimbrial protein A; and a method of measuring the amount of pneumococcal fimbrial protein A in a sample.

Abstract: The present invention provides a human glutamate receptor and related DNA compounds useful not only in assays for potential pharmaceuticals but also in methods for molecular biology techniques.

Abstract: Novel adenovirus-derived viral vectors, the preparation thereof, and their use in gene therapy, are disclosed. In particular, recombinant adenoviruses including an adenovirus genome wherein (i) the E1 region is inactivated, (ii) the genomic organization is modified, and (iii) optional recombination with the producing line genome generates non-viable viral particles, are disclosed.

Abstract: The present invention relates to genes, referred to herein as cell death-protective genes, which protect cells against programmed cell death by antagonizing the activities of genes which cause cell death. As described herein, a cell death-protective gene from the nematode Caenorhabditis elegans, called ced-9, has been identified, sequenced, and characterized. ced-9 is essential for C. elegans development and apparently functions by protecting cells which normally live during development from programmed cell death. Mutations which constitutively activate and inactivate the ced-9 gene are also described. ced-9 was shown to function by antagonizing the activities of the cell death genes, ced-3 and ced-4. As further described, the protein product of the human oncogene bcl-2 was found to have a similar sequence to the Ced-9 protein.

Abstract: The invention relates to a recombinantvector for the cloning and/or expression and/or transfer of an exogenous nucleotide sequence characterized in that it consists of any sequence contained in the ClaI—PvuII fragment comprising nucleotides 7702 to 1527 of the sequence given in FIG. 1 and comprising the LTR sequence included between nucleotides 7842 and 144, the PBS site starting at nucleotides 145, the packaging sequence included in the sequences of 250 nucleotides following the end of the LTR sequence, the said sequence being capable of controlling the cloning and/or expression and/or transfer of the exogenous sequence whatever its transcriptional orientation with respect to the transcriptional orientation of the virus. It relates to the use of this vector for the transfer and/or cloning and/or expression of genes, in particular in the contest of gene therapy.

Abstract: The invention relates to methods and materials involved in the regulation of tyrosine hydroxylase expression as well as the treatment of catecholamine-related diseases. Specifically, the invention provides cells that contain exogenous nucleic acid having a nucleic acid sequence that encodes Nurr1 as well as methods and materials for inducing tyrosine hydroxylase expression, treating catecholamine-related deficiencies, and identifying tyrosine hydroxylase-related deficiencies.

Abstract: There is proposed an apparatus for isolating and recovering an objective cell, which comprises, a filtering device having a filtering member which is capable of setting a porosity thereof to two or more different porosity, a first line for feeding a priming liquid to the filtering device, a second line for feeding a liquid to be treated containing the cell aimed to isolate to the filtering device, and a third line provided with a storage container for storing the priming liquid that has been passed through the filtering device, wherein the priming liquid stored in the storage container is employed for washing the filtering device. There is also proposed a method for isolating and recovering an objective cell using the aforementioned apparatus.

Abstract: There are disclosed nucleotide sequences which can improve expression of recombinant proteins two to eight fold in stable cell pools when present in an expression vector.