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

Abstract: The invention provides DNA sequences encoding novel members of the TGF-&bgr; family of proteins. The TGF-&bgr; family comprises proteins which function as growth and/or differentiation factors and which are useful in medical applications. Accordingly, the invention also describes the isolation of the above-mentioned DNA sequences, the expression of the encoded proteins, the production of said proteins and pharmaceutical compositions containing said proteins.

Abstract: Novel SPOIL-1 polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated, full-length SPOIL-1 proteins, the invention further provides isolated SPOIL-1 fusion proteins, antigenic peptides and anti-SPOIL-1 antibodies. The invention also provides SPOIL-1 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 non-human transgenic animals in which a SPOIL-1 gene has been introduced or disrupted. Diagnostic, screening and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: A process for preparing 2,6-diaminopurine-2′-deoxyriboside and 2′-deoxyguanosine. These compounds may be used as materials for pharmaceuticals, such as antiviral agents and the like, and particularly as starting materials for antisense oligonucleotides.

Abstract: A process is disclosed for the large scale isolation and purification of plasmid DNA from large scale microbial fermentations. The process exploits a rapid heating method to induce cell lysis and precipitate genomic DNA, proteins and other debris while keeping the plasmid in solution. Suspending the microbial cells in buffer and then heating the suspension to about 70-100° C. in a flow-through heat exchanger results in excellent lysis. Continuous flow or batch-wise centrifugation of the lysate effects a pellet that contains the cell debris, protein and most of the genomic DNA while the plasmid remains in the supernatant. This invention offers a number of advantages including higher product recovery than by chemical lyses, inactivation of Dnases, operational simplicity and scaleability.

Abstract: The present invention provides a fast, simple and specific method for generating a complete full-length cDNA library from single cells. The first reverse transcription of intracellular mRNAs with an oligo(dT)n-promoter primer introduces a recognition site for following transcription of newly reverse-transcribed cDNAs. The poly-nucleotide tailing of above cDNAs in addition to aforementioned promoter region further forms binding templates for specific PCR amplification. After repeating the reverse transcription, transcription, reverse transcription and PCR procedure, we can multiply a single copy of mRNA to two billion folds by calculation based upon the comparison between the amount of a synthesized cDNA library and that of theoretically presumed mRNAs within a cell (0.1 pg). In conjunction with a cell fixation and permeabilization step, the complete full-length cDNA library can be directly generated from few single cells without mRNA degradation.

Abstract: A nucleoside/tide compound having the structure NUC-L-S-LB/LG is described wherein NUC is a nucleoside/tide having a nucleobase portion B, L is a rigid linkage, S is a spacer; and LB/LG is a member of a linkage pair or a label. NUC is attached to L through B such that when B is a purine, L is attached to the 8-position of the purine, when B is 7-deazapurine, L is attached to the 7-position of the 7-deazapurine, and when B is pyrimidine, L is attached to the 5-position of the pyrimidine. In an important aspect of the invention, L has the structure wherein each of n, o and p are integers ranging from 0 to 3, and the sum of n, o and p is at least 2, and each of W, X, Y and Z is selected from the group consisting of carbon and nitrogen. The invention further includes polynucleotide compounds comprising the nucleoside/tide, and primer extension methods utilizing the nucleoside/tide, particularly when used in combination with certain mutant polymerase enzymes.

Abstract: Methods and compositions expand the options for making primers for use in amplifying nucleic acid segments. The invention eliminates the step of custom synthesis of primers for Polymerase Chain Reactions (PCR). Instead of being custom-synthesized, a primer is replaced by a combination of several oligonucleotide modules selected from a pre-synthesized library. A modular combination of just a few oligonucleotides essentially mimics the performance of a conventional, custom-made primer by matching the sequence of the priming site in the template. Each oligonucleotide module has a segment that matches one of the stretches within the priming site.

Abstract: Disclosed are a number of methods that can be used in a variety of embodiments, including, creation of a nucleic acid terminated at one or more selected bases, sequence analysis of nucleic acids, mapping of sequence motifs within a nucleic acid, positional mapping of nucleic acid clones, and analysis of telomeric regions. The methods utilize double-stranded templates, and in most aspects involve a strand replacement reaction initiated at one or more random or specific locations created in a nucleic acid molecule, and in certain aspects utilizing an oligonucleotide primer.

Abstract: A process for making natural and unnatural amino acids which comprises reacting a first amino acid, a keto acid and a transaminase enzyme under conditions appropriate to produce a second amino acid and pyruvate; reacting the pyruvate with acetolactate synthase under conditions appropriate to produce a compound that does not react with the transaminase enzyme and separating the second amino acid.

Abstract: A method for producing L-glutamic acid which comprises culturing a microorganism belonging to the genus Klebsiella, Erwinia or Pantoea and having an ability to produce L-glutamic acid in a culture medium, and collecting produced L-glutamic acid from the culture medium. The microbial strain used is preferably a strain which decreases in or is deficient in an activity of an enzyme catalyzing a reaction branching from a pathway for L-glutamic acid biosynthesis and producing a compound other than L-glutamic acid, or a strain which increase in an activity of an enzyme catalyzing a reaction for L-glutamic acid biosynthesis.

Abstract: A method for producing lovastatin by a microorganism in a fermentation process having a seed culture stage and a main fermentation stage, including a) cultivating a microorganism biomass in the seed culture stage to produce an inoculum; b) transferring the inoculum into a fermentation medium in the main fermentation stage; and, c) maintaining steady stage conditions in the main fermentation stage, thereby producing a fermentation broth containing lovastatin. Preferably, the steady state conditions are maintained in the main fermentation stage by one or more of feeding of organic carbon sources; controlling glucose and/or total reducing sugar content; feeding of organic and/or inorganic nitrogen sources; controlling pH; controlling foam level; controlling the mass of the fermentation broth by withdrawals and feedings; and, controlling the dissolved oxygen level.

Abstract: This invention relates to newly identified polynucleotides and polypeptides, variants and derivatives of same; methods for making the polynucleotides, polypeptides, variants, derivatives and antagonists. In particular the invention relates to polynucleotides and polypeptides of the phytate metabolic pathway.

Abstract: A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.

Abstract: The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.

Abstract: A DNA construct encoding an enzyme(s) exhibiting xylanase activity, which enzyme is immunologically reactive with antibody raised against a purified xylanase derived from Aspergillus aculealus, CBS 101.43, recombinant vectors and cells comprising said construct and a method for producing said enzyme using said cell comprising said construct.

Abstract: The invention relates to a variant of a parent Termamyl-like &agr;-amylase, comprising mutations in two, three, four, five or six regions/positions. The variants have increased stability at high temperatures (relative to the parent). The invention also relates to a DNA construct comprising a DNA sequence encoding an &agr;-amylase variant of the invention, a recombinant expression vector which carries a DNA construct of the invention, a cell which is transformed with a DNA construct of the invention, the use of an &agr;-amylase variant of the invention for washing and/or dishwashing, textile desizing, starch liquefaction, a detergent additive comprising an &agr;-amylase variant of the invention, a manual or automatic dishwashing detergent composition comprising an &agr;-amylase variant of the invention, a method for generating a variant of a parent Termamyl-like &agr;-amylase, which variant exhibits increased.

Abstract: An isolated polypeptide having &agr;-galactosidase activity and characterized as having a pH optimum in the range of 5.0-7.0, and a temperature optimum within the range of 50-70° C. The &agr;-galactosidase is derived from Aspergillus niger.

Abstract: Enzymes produced by mutating the genes for a number of subtilisin proteases and expressing the mutated genes in suitable hosts are presented. The enzyme exhibit improved wash performance in comparison to their wild type parent enzymes. The enzymes are well-suited for use in detergent compositions.

Abstract: Methods and compositions for the isolation, diagnosis and treatment of microorganisms such as flaviviruses and other hemorrhagic fever viruses are based on the sulfated polyanion-dependent interaction of flaviviruses and hemorrhagic fever viruses, in particular dengue virus, with target cells. The cellular receptors targeted by these viruses have been identified as sulfated polyanionic glycoproteins, that include highly sulfated heparan sulfate glycosaminoglycans for some target cell types, and as a sulfated mucin on vascular endothelium. Compounds such as heparin, highly sulfated heparan sulfate, and synthetic polyanions such as Suramin, inhibit the interaction between the microorganisms and target cells, thereby disrupting the infective process.

Abstract: The invention provides a novel phospholipase A2 enzyme, polynucleotides encoding such enzyme and methods for screening unknown compounds for anti-inflammatory activity mediated by the arachidonic add cascade.

Abstract: The present invention relates to Nocardia sp. CYKS2 (KCTC 0432 Br) capable of selective removal of organically bound sulfurs from carbonaceous fossil fuel such as petroleum and coal by cleaving bonds between carbon and sulfur atoms in the said sulfur-containing organic compounds, and a method for biological desulfurization using this strain at the room temperature and atmospheric pressure. Since Nocardia sp. CYKS2 (KCTC 0432 Bp) utilizes various organic sulfur compounds in fossil fuel besides dibenzothiophene as a sole sulfur source, the method for biological desulfurization employing the Nocardia strain has advantages over the conventional chemical methods as followings: The desulfurization can be carried out at a mild condition; the cost for installation and operation of equipments can be reduced; and, the desulfurization of highly complex organic sulfur compounds can be realized.

Abstract: The present invention relates to preparation of protein polysaccharide 0041 obtained by extracting a dried product of cultured mycelia of Agaricus blazei Murrill 0041 with hot water, and protein polysaccharide 0041 shows immunopotentiating antitumor activity when administered, so that it is useful as an antitumor agent against various tumors, and as an immunopotentiating substance against other diseases.

Abstract: This invention involves a thermal cycler capable of performing amplification of nucleic acids. This thermal cycler provides for the fully automatic positioning of its movable lid, which can be placed in two positions. The lid is also suitable for heating a plurality of reaction vessels and for shielding them from light from the environment. In addition, the thermal cycler also includes a thermal block having a plurality of chambers each of which receives the lower part of a reaction vessel. Finally, the thermal cycler has a lid carrier for moving the lid from the first position to the second position and vice versa.

Abstract: The present invention provides an improved solid state fermentation device that combines all of the operations of microorganism cultivation (sterilization, inoculation, cultivation, extraction, and post extraction treatment). This solid state fermentation device is modular in nature and operates in a contained manner so that the live microorganisms from the reactor cannot come into contact with the environment and pollute the environment and also so that the environment inside the bioreactor is aseptic. Another aspect of this invention allows fermentation of microorganisms without inhibiting the growth of the microorganism. Specifically, the bioreactor is designed to remove heat that accumulates inside the bioreactor during fermentation by conduction. Additionally, there is a mechanism to add fluid to the interior of the bioreactor that permits equal distribution and precise control of a variety of environmental parameters.

Abstract: An object of the present invention is to offer a microorganism-detecting apparatus where the microorganism causing a food poisoning can be detected/identified by a simple operation and also a disposal treatment can be conducted safely and surely. An apparatus for achieving the object is as follows: The apparatus for detecting microorganism as mentioned below is portable and enables one to selectively incubate and detect the microorganism (particularly those which are a cause of food poisoning) without skillfulness and the detecting apparatus containing pathogenic microorganism (such as that which is a cause of food poisoning) can be safely and surely disposed.

Abstract: Systems including (1) a micromatrix and perfusion assembly suitable for seeding and attachment of cells within the matrix and for morphogenesis of seeded cells into complex, hierarchical tissue or organ structures, wherein the matrix includes channels or vessels through which culture medium, oxygen, or other nutrient or body fluids can be perfused while controlling gradients of nutrients and exogenous metabolites throughout the perfusion path independently of perfusion rate, and (2) sensor means for detecting changes in either cells within the matrix or in materials exposed to the cells, have been developed. Methods for making the micromatrices include micromachining, micromolding, embossing, laser drilling, and electro deposition machining. Cells can be of one or more types, either differentiated or undifferentiated.

Abstract: An instrument to detect the presence of live microorganisms is described. The instrument is capable of providing simultaneous optical readings of multiple test vials containing different samples. Spectral variations due to metabolic activity of microorganisms are continuously recorded. An automated calibration scheme compensates for the parametric differences among the test vial locations.

Abstract: A device and method allow for detecting the presence of microorganisms in clinical and non-clinical specimens. The device, a sensor, provides an environment to culture microbial organism colonies from a fluid sample, and a means to facilitate microbial detection and quantification, either manually or with an instrument. The sensor has a microorganism immobilization matrix layer and a sensor layer. Detected microbial colonies are immediately available for further testing. The sensor provides an area for accepting a fluid sample, a mechanism to immobilize the fluid sample on an interior surface of the plate, nutrients to facilitate growth of microorganisms in the sample, and a sensor for allowing the detection and/or enumeration of microorganism colonies within the sample.

Abstract: The susceptibility to human immunodeficiency virus (HIV) infection depends on the cell surface expression of the human CD4 molecule and a human fusion accessory factor associated with HIV infection (CXCR4). CXCR4 is a member of the 7-transmembrane segment superfamily of G-protein-coupled cell surface molecules. CXCR4 plays an essential role in the membrane fusion step of HIV infection. The establishment of stable cell lines that coexpress human CD4 and CXCR4 provides valuable tools for the continuing research of HIV infection and the development of more effective anti-HIV therapeutics.

Abstract: Disclosed is a cell washing device and method of washing cells that utilizes the device. The device is particularly designed for sterile transfer of cells from a primary centrifuge tube and for maintaining the cells in a sterile environment during subsequent washing steps. The device is configured to provide for decantation of a supernatant by inversion without appreciable loss of a selected population of lower density cells from the pellet during the washing procedure.

Abstract: Regulatory elements responsible for tissue-specific transcriptional regulation of the human &bgr;3-adrenergic receptor (&bgr;3-AR) were identified. A region localized between −6.50 and −6.30 kb of the proximal promoter contained three sequences that act synergistically to achieve full transcriptional activity. One segment, termed segment A, contains an Sp 1 binding site. Another of the sequences, termed segment B, is a binding site for a trans-acting factor present in cells that constitutively express &bgr;3-AR. In a specific embodiment, the trans-acting factor is expressed in neuroblastoma (SK-N-MC) and brown adipose tissue cells, but little or not at all in CV-1, HeLa, or white adipose tissue cells. The third segment, C, is an S1 nuclease-sensitive site having CCTT repeats.

Abstract: There is provided a novel adenylate cyclase enzyme, the nucleotide sequence of which is set out in SEQ ID NO: 98. The activity of the adenylate cyclase is uniquely regulated by calcineurin, a protein phosphatase. The calcineurin binding site has been identified at amino acids 503 to 610 of the novel adenylate cyclase. Regulation of the novel adenylate cycalse may be useful in treating certain disorders. Suitable regulators include agents which bind to the 503-610 amino acid site and include calcineurin, its activators, inhibitors and competitors and antibodies specific to that site. Specific disorders include neurological disorders such as Parkinsons' disease, cardiovascular disorders such as angina pectoris and tumors, especially ovarian tumors.

Abstract: The present invention provides a trioma cell which does not produce any antibody obtained by fusing a hetermomyeloma cell which does not produce any antibody with a human lymphoid cell, wherein the heteromyeloma cell is designated B6B11. The invention also provides a tetroma cell capable of producing a monoclonal antibody having specific binding affinity for an antigen obtained by fusing a trioma cell which does not produce any antibody with a human lymphoid cell capable of producing antibody having specific binding affinity for the antigen. The invention also provides methods for generating trioma cells and tetroma cells, and the cells generated by the methods.

Abstract: The invention relates to a biologically-active peptide fragment of the Nef protein of human immunodeficiency virus, to pharmaceutical compositions comprising these peptides or biologically-active analogues thereof, to antagonists of the peptides and to pharmaceutical compositions comprising these antagonists, and to therapeutic and screening methods utilizing compounds and compositions of the invention.

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

Abstract: The invention relates to a method for producing human cell lines and cell and cell-lines produced by such a method. The method comprising the use of precursor or undifferentiated cells treated with an immortalising agent which is susceptible to environmental conditions so as to provide for selective activation/deactivation of said immortalising agent and so selective activation of differentiation.

Abstract: Fibroblast cells are treated with a chemical inhibitor of protein kinase C such as staurosporine, in conjunction with functionally hypoxic micromass culture so as to be induced into chondrogenic differentiation. Such fibroblast-derived, chondrocyte-like cells may be seeded onto three-dimensional polymer scaffolds for use in the repair of articular cartilage lesions, and thus can obviate the need for invasive techniques to harvest autologous chondrocytes from a limited supply of existing articular cartilage, or to avoid the need for obtaining allogeneic chondrocytes from non-biocompatible donor tissues.

Abstract: The present invention relates to improved methods of (i) inducing somatic embryogenesis from cacao tissue explants and (ii) regenerating cacao plants from somatic embryos. The invention further relates to cacao somatic embryos and plants obtained according to the methods of the invention. Novel tissue culture media adapted for use in the above-identified methods are also within the scope of the invention. The novel media of the invention include primary callus growth medium, secondary callus growth medium, embryo development medium, primary embryo conversion medium, secondary embryo conversion medium and plant regeneration medium.

Abstract: A polypeptide amino acid sequence capable of targeting the plastid inner envelope membrane of a plant, and a nucleotide sequence therefor, are described, as well as a chimaeric gene comprising a gene promoter, a nucleotide sequence encoding a polypeptide capable of targeting the plastid inner envelope membrane of a plant, or a variant, derivative or homologue thereof, a coding sequence and a terminator sequence. A method of transforming plants, such as potato and tobacco, using the targeting sequence so as to increase starch production is also described. The targeting sequence can be used in many other applications.

Abstract: This invention relates to proteases with improved application properties in cleaners and detergents. The improvement's achieved by substituting positively charged or uncharged amino acids in the substrate binding region of the wild type subtilisin protease.

Abstract: The present invention relates to a process for integration of a chosen gene or of a specific DNA sequence in a DNA sequence such as the chromosome or episome of a bacterium, wherein: a) the said chosen gene or the chosen DNA sequence is cloned inside a defective transposon outside the essential parts of the transposon, b) the said transposon is integrated in the DNA sequence such as the chromosome or the episome of the said bacterium, and also the bacterium strains obtained by implementation of this process.

Abstract: The present invention is directed to compositions and methods for producing avermectins, and is primarily in the field of animal health. The present invention relates to the identification and characterization of two novel genes, herein referred to as the aveR1 and aveR2 genes, that are involved in regulating avermectin polyketide synthase (PKS) expression and avermectin biosynthesis in Streptomyces avermitilis. The present invention is based on the discovery that inactivation of these genes results in an increase in the amount of avermectin produced by S. avermitilis.

Abstract: Control materials containing specified concentrations of the osteoporosis markers deoxypyridinoline, pyridinoline, C-telopeptide and N-telopeptide in a matrix that is substantially the same as that of human urine are prepared by selecting amounts of human urine that contain sufficient quantities of the markers to achieve the target levels upon concentration, lyophilizing the selected urine, optionally after clarification by filtering, freezing and thawing, and pH adjustments, then reconstituting the lyophilized material, and combining and/or diluting the lyophilized material, either before or after reconstitution, to adjust the marker contents to the target levels. Surprisingly, the markers survive this process sufficiently intact to serve as reliable control materials of known composition and concentration.

Abstract: A method for enumerating and distinguishing between blood cell populations in a biological sample includes the steps of contacting a biological sample with a cell membrane-permeant, red-excited, nucleic acid binding dye, without significantly disrupting the integrity of the cells; exciting this sample with light in one red wavelength; and measuring fluorescence emitted from different cell populations in the sample. This method is particularly desirable for enumerating different WBC subpopulations using flow cytometry. This method is also useful for enumerating reticulocytes or NRBC or mature RBC. This method is enhanced by pretreating the sample with a nucleic acid-specific blocking agent. Dyes useful in this method include, without limitation, SYTO®17 dye, SYTO®159 dye, SYTO®60 dye, SYTO®61 dye, SYTO®62 dye, SYTO®63 dye, and SYTO®64 dye.

Abstract: The present invention concerns a method for the detection of an analyte in a sample liquid by luminescence measurement according to the principle of a ligand-receptor assay, e.g. an immunoassay or a hybridization assay or a combination thereof, wherein a sample liquid is incubated with at least one receptor which carries a luminescent label and the presence or/and the amount of the analyte to be detected is determined in the sample liquid by measuring the luminescence in a measuring medium containing dispersed components.

Abstract: The present invention provides a miniaturized integrated nucleic acid diagnostic device and system. The device of the invention is generally capable of performing one or more sample acquisition and preparation operations, in combination with one or more sample analysis operations. For example, the device can integrate several or all of the operations involved in sample acquisition and storage, sample preparation and sample analysis, within a single integrated unit. The device is useful in a variety of applications, and most notably, nucleic acid based diagnostic applications and de novo sequencing applications.

Abstract: The present invention relates to a method to determine antibody-mediated autoimmune diseases in a patient, which comprises the steps of: a) determining the amount of clusterin present in a serum, saliva or tissue sample of the patient with an anti-clusterin antibody; b) comparing the amount of clusterin in step a) with normal serum, saliva or tissue sample, wherein a lower than normal amount is indicative of active antibody-mediated autoimmune disease.

Abstract: The invention concerns a solid-phase determination method and equipment and an adapter for use in these. In the method a sample is allowed to react with a separating reagent bound to the outer surface of a separate solid phase body, whereafter the body is removed from the vessel and is taken to a measurement vessel, if required through one or several intermediate step vessels. At least one vessel contains a medium needed in a determination step to be performed therein when the phase body is brought into this vessel. The invention is especially suitable for use in automatic immunodetermination systems.

Abstract: A device for the separation of the liquid portion of blood from the cellular components of blood comprises: (1) a pad of porous material permeable to the liquid portion of blood but capable of trapping the cellular components of blood; (2) a substrate supporting the pad; and (3) means, attached to the pad, for facilitating the flow of the liquid portion of the blood: (i) through interstices around the trapped cellular components of the blood and (ii) from the pad of porous material. The separation of the liquid portion of blood from the cellular components of the blood occurs by flow through the pad of porous material without significant hemolysis. The device can be incorporated into a device for the performance of specific binding assays such as immunoassays. The pad of porous material can contain an agglutinating agent such as a lectin or an anti-blood cell antibody, or a carbohydrate such as mannitol.