Abstract: Stabilized tetrazolium dye reagent compositions and methods for their use in the measurement of an analyte in a sample are provided. The subject reagent compositions include a tetrazolium dye component, e.g., a water soluble tetrazolium salt, and an effective amount of a nitrite stabilizing agent, e.g., a nitrite salt. In many embodiments, the subject reagent compositions include additional members of an analyte oxidizing signal producing system, such as: an analyte oxidizing enzyme, e.g., an analyte dehydrogenase or an analyte oxidase; an electron transfer agent; and an enzyme cofactor. Also provided are test strips that include the subject reagent compositions, as well as systems and kits incorporating the subject test strips. The subject reagent compositions, test strips, systems and kits find use in the detection of a wide variety of analytes in a sample, such as a physiological sample, e.g., blood or a fraction thereof, or ISF (interstitial fluid).

Abstract: The present invention relates to a method for detecting substances specifically inhibiting a type III secretion mechanism and functions of the type III secretory proteins, within short time and large amounts thereof, without depending upon animal infectious experiments. Namely it relates to the method for detection of a type III secretory mechanism inhibitor comprising mixing a bacterium having the type III secretory mechanism and an erythrocyte suspension, adding the type III secretory mechanism inhibitor thereto, and detecting changes in the thus formed hemolytic activity. The method for detecting substances can be treated large amount of samples within short time by exhibiting the substances inhibiting the type III secretion mechanism or the functions of the type III secretory proteins as numerical index of the hemolytic activity of erythrocytes. Consequently, the present invention is useful for development of drugs.

Abstract: A method of chromatographic isolation for non-glyceride components (squalenes, carotenes, vitamin E, sterols and/or the like) from a non-glyceride components-comprising compound by the steps of a. introducing the non-glyceride components-comprising compound onto a selective adsorbent to allow an adsorption of the non-glyceride components, and subsequently b. desorbing the non-glyceride components from the adsorbent, wherein the adsorption and/or desorption of the non-glyceride components is carried out under a supercritical fluid environment.

Abstract: The present invention relates to an isolated DNA sequence encoding an enzyme in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate. Vectors and plasmids including such DNA are also set forth. The invention also includes host cells transformed by such DNAs, or vectors or plasmids containing such DNAs. A process for the production of isoprenoids and carotenoids using such transformed host cells is also provided.

Abstract: The INK4A (MTS1, CDKN2) gene encodes a specific inhibitor (InK4a-p16) of the cyclin D-dependent kinases CDK4 and CDK6. InK4a-p16 can block these kinase from phosphorylating the retinoblastoma protein (pRb), preventing exit from the G1 phase of the cell cycle. Deletions and mutations involving the gene encoding InK4a-p16, INK4A, occur frequently in cancer cells, implying that INK4a-p16, like pRb, suppresses tumor formulation. However, a completely unrelated protein (ARF-p19) arises in major part from an alternative reading frame of the mouse INK4A gene. Expression of an ARF-p19 cDNA (SEQ ID NO:1) in rodent fibroblasts induces both G1 and G2 phase arrest. Economical reutilization of protein coding sequences in this manner is without precedent in mammalian genomes, and the unitary inheritance of INK4a-p16 and ARF-p19 may reflect a dual requirement for both proteins in cell cycle control.

Abstract: Disclosed is the isolation and characterization of EI24, a novel gene whose 2.4 kb mRNA is induced following etoposide treatment. Induction of EI24 mRNA by etoposide required expression of wild-type p53. Overexpression of functional p53 was sufficient to induce expression of the EI24 mRNA. The EI24 mRNA was also induced in a p53-dependent manner by ionizing irradiation of primary murine thymocytes. The invention is thus directed to an isolated EI24 protein, nucleotide sequences coding for and regulating expression of the protein, antibodies directed against the protein, and recombinant vectors and host cells containing the genetic sequences coding for and regulating the expression of the protein sequence. The invention is also directed to genomic DNA, cDNA, and RNA encoding the EI24 protein sequence and to corresponding antisense RNA sequences. Antibodies can be used to detect EI24 in biological specimens, including, for example, human tissue samples.

Abstract: Novel GPCR-like polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated, full-length GPCR-like proteins, the invention further provides isolated GPCR-like fusion proteins, antigenic peptides, and anti-GPCR-like antibodies. The invention also provides GPCR-like 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 in which a GPCR-like gene has been introduced or disrupted. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: The invention provided improved methods of making and producing recombinant proteins in in vitro cultures of host cells using apoptosis inhibitors. The use of one or more apoptosis inhibitors in the methods can reduce apoptosis in the cell cultures and markedly improve yield of the desired recombinant proteins.

Abstract: Methods for producing modified polypeptides containing amino acid analogues are disclosed. The invention further provides purified dihydrofolate reductase polypeptides, produced by the methods of the invention, in which the methionine residues have been replaced with homoallyglycine, homoproparglycine, norvaline, norleucine, cis-crotylglycine, trans-crotylglycine, 2-aminoheptanoic acid, 2-butynylglycine and allylglycine.

Abstract: Suppressor tRNA's are used to regulate expression of transgenes that are toxic, or the expression thereof requires a factor that is toxic, to the host cell.

Abstract: An isolated DNA with a nucleotide sequence encoding a ripening form of a xylanase fungal origin having bread improving activity. Cells are transformed with this DNA and used to produce the ripening form of xylanase which itself is suitable for use in flour and dough.

Abstract: The present invention concerns a novel T1R-like ligand II protein. In particular, isolated nucleic acid molecules are provided encoding the T1R-like ligand II protein. T1R-like ligand II polypeptides are also provided, as are recombinant vectors and host cells for expressing the same.

Abstract: The invention relates to a method for producing polymers, in particular synthetic nucleic acid double strands of optional sequence, comprising the steps: (a) provision of a support having a surface area which contains a plurality of individual reaction areas, (b) location-resolved synthesis of nucleic acid fragments having in each case different base sequences in several of the individual reaction areas, and (c) detachment of the nucleic acid fragments from individual reaction areas.

Abstract: The invention provides a method of obtaining a cellulose material from corn fiber wherein the method comprises the steps of: (a) heating a mixture of corn fiber and a liquid; (b) contacting the mixture of step (a) with a protease enzyme, thereby providing a proteolyzed corn fiber and a liquid; (c) separating the liquid from the proteolyzed corn fiber; (d) contacting the proteolyzed corn fiber at least once with an alkaline extractant, thereby providing an insoluble cellulose material and a first liquid comprising arabinoxylan; (e) separating the insoluble cellulose material from the first liquid comprising arabinoxylan at a temperature of at or above about 60° C.; and (f) rinsing the insoluble cellulose material to remove essentially all alkali, thereby providing a cellulose material having a cellulose content of at least about 50% and consisting essentially of cellulose I. Cellulose esters and ethers are also prepared from the derivatizable cellulose prepared according to the methods herein.

Abstract: A process is disclosed for clarifying an aqueous xanthan gum solution comprising treatment of the xanthan solution with at least one chelating agent, surfactant, organic acid, or a mixture thereof, and with a protease enzyme or a lysozyme and a protease enzyme. Also disclosed is a highly purified solid xanthan gum and compositions containing the same, wherein the gum is obtained from the clarified xanthan gum solution.

Abstract: The present invention is directed to isolated polynucleotides coding for phosphoglucose isomerase (pgi) from coryneform bacteria. In addition, the invention includes methods for increasing the metabolic flux through pentose phosphate cycle of bacteria by reducing or eliminating the activity of pgi. These methods may be used to increase the fermentative production of nucleotides, vitamins and amino acids.

Abstract: The present invention relates to isolated polypeptides having peroxidase activity and isolated nucleic acid sequences encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as methods for producing and using the polypeptides.

Abstract: The present invention is directed to a modified amidotransferase. The amino acid sequence of the modified amidotransferase differs from the amino acid sequence of a native E. coli glutamine PRPP amidotransferase in that one or more amino acid residues of the modified amidotransferase are substituted at positions equivalent to amino acid positions in Bacillus amidotransferase selected from the group consisting of 282, 283, 307, and 347 of SEQ ID NO:1, wherein the modified amidotransferase is less sensitive to end-product inhibition than is the native E. coli glutamine PRPP amidotransferase.

Abstract: Selenophosphate synthetase protein from a mammal, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding this protein. Methods of using these reagents and diagnostic kits are also provided.

Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesised. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesized. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

Abstract: The present invention relates to recombinant DNA encoding the BsaWI restriction endonuclease as well as BsaWI methylase, expression of BsaWI restriction endonuclease and BsaWI methylase in E. coli cells containing the recombinant DNA.

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

Abstract: Methods and materials are disclosed for the production of purified, active recombinant human neutrophil protease, PR-3, via activation of a pro-form herein referred to as proPR-3. Human PR-3 is useful for discovering inhibitors of excessive release of mature, active TNF&agr;. Also disclosed are methods for the identification of inhibitors of the conversion of the pro-form of TNF&agr; to its mature active form.

Abstract: The present invention relates to variants of subtilisin-like proteases having decreased immunogenicity relative to their corresponding wild-type proteases. More particularly, the present invention relates to variants having a modified amino acid sequence of a wild-type amino acid sequence, wherein the modified amino acid sequence comprises a substitution of one or more epitope regions. The invention further relates to mutant genes encoding such variants and cleaning and personal care compositions comprising such variants.

Abstract: The present invention relates to variants of subtilisin-like proteases having decreased immunogenicity relative to their corresponding wild-type proteases. The present invention further relates to such variants additionally having one or more amino acid substitutions in one or more epitope regions or additionally having one or more stabilizing substitutions. The invention further relates to mutant genes encoding such variants and cleaning and personal care compositions comprising such variants.

Abstract: Polynucleotides encoding novel proteases designated “FMH-1” are disclosed. Host cells transformed with such polynucleotides and methods for making FMH-1 proteases are also disclosed. The invention also provides FMH-1 proteases and antibodies that react with them, along with pharmaceutical compositions comprising FMH-1 proteases or polynucleotides encoding FMH-1 proteases. Methods for treating conditions associated with excessive or insufficient apoptosis by administering such pharmaceutical compositions are also disclosed.

Abstract: A method of enriching a solution of an adenovirus comprising applying a mixed solution comprising an adenovirus and at least one undesired type of biomolecule to an anion exchange chromatography resin containing a binding moiety selected from the group consisting of dimethylaminopropyl, dimethylaminobutyl, dimethylaminoisobutyl, and dimethylaminopentyl and eluting the adenovirus from the chromatography resin. Also provided is a method of purifying an adenovirus from adenovirus-infected cells comprising lysing such cells, applying the lysate to a single chromatography resin, eluting the adenovirus from the chromatography resin, and collecting a fraction containing adenovirus that is substantially as pure as triple CsCl density gradient-purified adenovirus.

Abstract: The Porphyromonas gingivalis strain which is deposited at ATCC under accession number 202109. A pharmaceutical composition comprising the Porphyromonas gingivalis strain of the present invention.

Abstract: Nucleic acids encoding mammalian, e.g., primate or rodent receptors, purified receptor proteins and fragments thereof. Antibodies, both polyclonal and monoclonal, are also provided. Methods of using the compositions for both diagnostic and therapeutic utilities are provided.

Abstract: Bacterial strains transformed with the pcu genes are useful for the production of para-hydroxybenzoate (PHBA). Applicant has provided the p-cresol utilizing (pcu) and tmoX gene sequences from Pseudomonas mendocina KR-1, the proteins encoded by these sequences, recombinant plasmids containing such sequences, and bacterial host cells containing such plasmids or integrated sequences. Method for the use of these materials to produce PHBA are also disclosed.

Abstract: Novel human polynucleotide and polypeptide sequences are disclosed that can be used in therapeutic, diagnostic, and pharmacogenomic applications.

Abstract: A novel antibiotic-producing Bacillus sp. is provided that exhibits antifungal activity only on certain specific plant pathogens and no antibacterial activity is provided by this invention as well as a biologically pure culture of a strain having all the identifying characteristics of this strain. Also provided is a method of treating or protecting plants, fruit and roots from fungal infections by applying an effective amount of these strains, supernatants produced by these strains or a metabolite isolated from these strains.

Abstract: A mass biosensor uses an intermediate avidin layer to facilitate binding of a biotinylated antibody to a measurement surface of the biosensor. The avidin layer can be added by the manufacturer of the biosensor, while the biotinylated layer can be added by the user. This two-phase method of chemically modifying the measurement surface significantly reduces the user time required to customize the measurement surface to render it capable of binding selected compounds. An organosilane coupling agent attached to the surface provides sites to which avidin is bound. Avidin acts as a universal receptor of biotinylated compounds with specific binding affinities. Biotinylated antibodies or other biotinylated compounds are added and bind to the immobilized avidin. Surface adsorption is reduced by washing the modified surface with biotin to block potential sites of weak bond formation, electrostatic and hydrophobic interactions.

Abstract: A polymerase chain reaction system provides an upper temperature zone and a lower temperature zone in a fluid sample. Channels set up convection cells in the fluid sample and move the fluid sample repeatedly through the upper and lower temperature zone creating thermal cycling.

Abstract: The present invention relates to the detection of specific nucleic acid sequences after an amplification process, or directly without amplification. In particular, the invention provides for the automation of the amplification and detection process, the amplification and detection of one or more specific nucleic acid sequences, the use of internal controls, reduced potential for contamination caused by the manual manipulation of reagents, and improved reagent compositions to better control assay performance and provide for further protection against contamination.

Abstract: A cell culture device for inducing shear stress and on substrate strain on cells. The device includes a cell culture membrane and a flow pathway for moving fluid across cells growing on the membrane to apply shear stress on the cells.

Abstract: A modified filamentous phage that contains a gene for a wild type phage coat protein and a gene for a synthetic phage coat protein is provided. Uses of the modified phage and kits containing the phage are also provided.

Abstract: Compositions comprising two or more linear DNA molecules capable of self-replicating when circularized, the linear DNA molecules comprising a single overhanging nucleotide at each 3′ end, wherein a pair of overhanging nucleotides of a linear DNA molecule of the composition are different than a pair of overhanging nucleotides of another linear DNA molecule of the composition.

Abstract: An enzymatic nucleic acid molecule comprising a 5′- and/or a 3′-cap structure, wherein said structure is not a 5′-5′-linked inverted nucleotide or a 3′-3′-linked inverted nucleotide.

Abstract: A method is provided for obtaining a cell population enriched in microglial cells by contacting a composition containing microglial cells with immunoglobulin immobilized on a matrix such as a polystyrene matrix before or after contact with the cells, allowing the cells to bind to the with immunoglobulin, and removing non-adherent cells to obtain a cell population containing preferably at least 95% microglial cells. The immunoglobulin may be Fc domain-containing immunoglobulin G, and Fc receptors of the microglial cells bind to the Fc domain of immunoglobulin G. Purified Fc fragments from immunoglobulin G may be used in place of immunoglobulin G. The microglial cells may be from brain tissue, and from tissue of a normal animal or tissue of an animal having a neurological disorder.

Abstract: A method for visualizing the location and movement of a specific RNA of interest in a living cell, in real time, is disclosed. The method includes the following steps: (a) providing a DNA encoding the RNA, which RNA includes a protein-binding site; (b) providing a nucleic acid encoding a fusion protein, which fusion protein comprises a fluorescent domain and an RNA-binding domain; (c) introducing the DNA encoding the RNA, and the nucleic acid encoding the fusion protein, into a eukaryotic cell so that the DNA encoding the RNA and the nucleic acid encoding the fusion protein are expressed in the cell; and (d) detecting fluorescence outside the nucleus or inside the nucleus of the cell, with the fluorescence being from the fusion protein bound to the RNA. In some embodiments, the fusion protein also includes an intracellular localization domain.

Abstract: The present invention relates to methods for identifying a compound capable of modulating mitochondrial function, comprising contacting a eukaryotic cell with one or more candidate compounds, and detecting a change in the mitochondrial redox state of the cell. The methods further relates to such methods wherein endogenous fluorescence of the cell mitochondria is indicative of a change of redox state.

Abstract: Compositions and methods are provided for potentiating the activity of the mitogen-activated protein kinase p38. In particular the mitogen-activated protein kinase kinase MEK6, and variants thereof that stimulate phosphorylation of p38 are provided. Such compounds may be used, for example, for therapy of diseases associated with the p38 cascade and to identify antibodies and other agents that inhibit or activate signal transduction via p38.

Abstract: A method is described whereby dendritic cells derived from the CD34+ and CD 34− hematopoietic cell lineages are directed to become programmable antigen presenting cells. The programmed cells may be pulsed with tumor cell RNA or tumor cell RNA expression products. The protocol provides for directing the maturation of dendritic cells to become antigen presenting cells. The protocol further provides for isolating tumor cell RNA from biopsy material that has been prepared in paraffin block storage. The directed dendritic cell is provided with a plurality of tumor markers by using tumor RNA in toto, the poly A+RNA fraction or the expression product of such RNA. Once activated the dendritic cells are incubated with T4 and T8 lymphocytes to stimulate and sensitize the T lymphocytes which upon introduction either into a donor host or a nondonor recipient will provide immune response protection.

Abstract: Inhibitors of laminin5beta3 are provided that reduce the expression or biological activities of laminin5beta3 or the expression of laminin5beta3 mRNA in a mammalian cell. Laminin5beta3 inhibitors include antisense molecules, ribozymes, antibodies and antibody fragments, proteins and polypeptides as well as small molecules. Laminin5beta3 inhibitors find use in compositions and methods for decreasing laminin5beta3 gene expression as well as methods for inhibiting the proliferation of mammalian cells, including tumor cells of epithelial origin, and methods for treating neoplastic diseases.

Abstract: Antisense compounds, compositions and methods are provided for modulating the expression of CD40 ligand. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding CD40 ligand. Methods of using these compounds for modulation of CD40 ligand expression and for treatment of diseases associated with expression of CD40 ligand are provided.

Abstract: Three-dimensional porous biodegradable and biocompatible, polymeric scaffolds for tissue engineering are prepared by a method involving effervescing of an effervescent salt in a gel to result in a porous structure. A polymer is dissolved in an organic solvent to prepare a polymer solution of high viscosity. Optionally, the polymer solution is mixed with an organic solvent that does not dissolve the polymer to concentrate the solution. An effervescent salt is homogeneously mixed with the polymer solution to give a polymer/salt/organic solvent mixed gel. The organic solvent is removed from the mixed gel to produce an organic solvent-free polymer/salt gel slurry. The gel slurry is submerged in a hot aqueous solution or acidic solution to cause the salt to effervesce at room temperature to form a porous three-dimensional polymeric structure. The polymeric structure is washed with distilled water and freeze-dried to yield a scaffold that is suitable for cell or tissue culture.

Abstract: A medium for cultivating an animal embryo that is capable of readily culturing a fertilized ovum can be provided. A cultivation medium is separated into two parts, i.e., one is a neutral aqueous solution and the other is an acidic aqueous solution. The two parts are mixed before use. The neutral aqueous solution contains a nutrient such as lactic acid or a lactic salt and another inorganic component and an indicator that indicates a state of a development environment with its color, and the acidic aqueous solution contains a nutrient such as pyruvic acid and an antibiotic. The pH change in the medium according to cultivation of animal embryo can be determined by the change of the color of the indicator.

Abstract: The invention provides a process for the induction of normal rooting on nodal and internodal stem segments without using hormones and/or other chemical treatments in Mentha species, which comprises oblique excision of shoots of Mentha species from the plants, immediately dipping the cut end of twigs or stems into water and keeping in complete dark for up to one week with by varying the temperature cycle for each set of 24 hours duration of the treatment.