Abstract: A process for the preparation of nucleoside diphosphate and triphosphate sugars wherein hydroxyl protective groups are removed enzymatically, with acetylesterase and a process for the preparation of these sugars, which comprises coupling a nucleotide with a sugar-1-phosphate activated with a carbonyl bisazole and then removing the hydroxyl protective groups enzymatically with acetylesterase.

Abstract: The present invention relates to processes for the enzymatic synthesis of nucleotide-6-deoxy-D-xylo-4-hexuloses starting from a nucleoside monophosphate (NMP). These processes comprise simultaneous incubation of the following substances in a buffer solution:(a) substrates comprising a nucleoside monophosphate, phosphoenolpyruvate, adenosine triphosphate, and sucrose; and(b) enzymes comprising pyruvate kinase, nucleoside-monophosphate kinase, sucrose synthase and deoxythymidine-D-glucose 4,6dehydratase.

Abstract: A preparation of a labelled nucleotide comprising at least one compound having a Mg.sup.2+ association constant between 1.times.10.sup.-11 to 1.times.10.sup.-2, inclusive. The compound is preferably selected from the group consisting of citrate, isocitrate, phosphate, EGTA, EDTA, and CDTA. The concentration of the compound is preferably at least 5 mM.

Abstract: Nucleotide linked 2-deoxy-2-fluoroglycosides are employed as potent competitive inhibitors of glycosyltransferases. More particularly, uridine-5'-diphospho-2-deoxy-2-fluoro-galactose (UDP-2F-Gal), guanidine-5'-diphospho-2-deoxy-2-fluoro-L-fucose (GDP-2F-Fuc), uridine-51-diphospho-2-deoxy-2-fluoro-D-glucose (UDP-2F-Glu), guanosine-5'-diphospho-2-deoxy-2-fluoro-D-mannose (GDP-2F-Man), cytosine-5'-monophospho-2-deoxy-2-fluoro-D-sialic acid, and cytosine-5'-monophospho-2-deoxy-2-KDO may be employed as inhibitors of .beta.-1,4-galactosyltransferase, .alpha.-1,3-fucosyltransferase, glucosyltransferases, N-acetylglucosaminyltransferases, (.alpha.-mannosyltransferases, .alpha.-sialyltransferases, and KDO-transferases, respectively. Synthesis of nucleotide-linked-2-deoxy-2-fluoroglycosides is achieved using either chemoenzymatic or chemical methodologies.

Abstract: Substituted propargylethoxyamido nucleosides are disclosed having the structure ##STR1## wherein X is selected from the group consisting of amino alkanoic acid, alkylamino benzoic acid, .alpha.-amino acid, and 4-amino-2-butynoic acid. R.sub.1 and R.sub.2 taken separately are selected from the group consisting of --H, lower alkyl, protecting group, and label; R.sub.3 is selected from the group consisting of --H and lower alkyl. B is a 7-deazapurine, purine, or pyrimidine nucleoside base. When B is purine or 7-deazapurine, the sugar moiety is attached at the N.sup.9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N.sup.1 -position of the pyrimidine.

Abstract: The present invention relates to an inosine-guanosine kinase which catalyzes the reaction of forming 5'-inosinic acid (5'-IMP) from inosine and adenosine triphosphate (ATP) or deoxyadenosine triphosphate (dATP) and the reaction of forming 5'-guanylic acid (5'-GMP) from guanosine and ATP or dATP.

Abstract: Nucleotide sugars, especially UDP, ADP, CDP or TDP saccharoses can be enzymatically obtained by the reaction of nucleoside diphosphates with di or trisaccharides with a saccharose synthase in which the virtual absence of nucleoside phosphatases (0.1% or less) can be ensured by special purification methods and sensitive detection. The purification of the raw extract, obtained preferably from rice grains, comprises especially the application of the ultra-filtered extract containing 50 mM KCl with a pH 8 on a sepharose Q column and a gradient elution out of the column at a pH 8 with 50 to 500 mM KCl.

Abstract: What is described is a recombinant poxvirus, such as vaccinia virus, fowlpox virus and canarypox virus, containing foreign DNA from flavivirus, such as Japanese encephalitis virus, yellow fever virus and Dengue virus. In a preferred embodiment, the recombinant poxvirus generates an extracellular particle containing flavivirus E and M proteins capable of inducing neutralizing antibodies, hemagglutination-inhibiting antibodies and protective immunity against flavivirus infection. What is also described is a vaccine containing the recombinant poxvirus for inducing an immunological response in a host animal inoculated with the vaccine.

Abstract: Process for preparing hepatitis A (HAV) antigens and vaccines.The HAV virus is multiplied on competent cells, the infected cells are lysed, the supernatant is recovered and the purification is carried out by a chromatographic procedure on an anion-exchange support and a gel filtration procedure, the purification procedures being carried out in the presence of a detergent, and the chromatographic procedure being carried out under conditions which retain the virions or viral capsids, which are then eluted.

Abstract: A novel cyclic nucleotide phosphodiesterase, which is different from known isozyme families, is isolated and purified from rat cerebrum. This enzyme has the following physicochemical properties (1) to (3). (1) The enzyme acts on cAMP to form 5'-AMP and on cGMP to form 5'-GMP; (2) The Km values for cAMP and cGMP are 0.11 .mu.M and 1.78 .mu.M, respectively, as indicating substrate specificity; and (3) the molecular weight is approximately 298000.

Abstract: Novel 2'-methylidenenucleotide compounds of the formula (I) ##STR1## wherein R is a hydrogen or a halogen, R.sup.1 and R.sup.2 are the same or different and each is a fatty acid residue or a hydrocarbon residue, and R.sup.3 and R.sup.4 are the same or different and each is a hydrogen, a halogen or an alkyl; salts thereof; methods for production thereof; and pharmaceutical use thereof. The compounds and salts thereof show an excellent antitumor effect in mammals. More specifically, they show a remarkable activity of inhibiting growth of mouse tumors, cultured human tumor cells, and human tumors transplanted to nude mice, and are useful for the treatment and prevention of recurrence of lung cancer, gastrointestinal cancer, breast cancer, cervical cancer, gynecological cancer, urinological cancer, leukemia, melanoma, lymphogenous metastatic tumor and the like in mammals. They are also useful as antitumor agents since they have an increased bioavailability and low toxicity.

Abstract: A method for producing uridine diphosphate N-acetylglucosamine comprising culturing osmo-tolerant yeasts in aerobic conditions in a medium having inorganic salt concentration of about 2-8%.

Abstract: Fluorescently labelled dideoxynucleoside triphosphates are widely used as chain-terminators in Sanger dideoxy sequencing operations. But these unincorporated dye-terminators migrate in an electrophoresis gel and obscure the desired sequence ladder. This invention provides a method and a kit for modifying the unincorporated dye-terminators, e.g. by removal of a 5'-triphosphate group by chemical or enzymatic means e.g. by use of a phosphatase enzyme.

Abstract: Disclosed are purified and isolated DNA sequences encoding eukaryotic proteins possessing biological properties of inosine 5'-monophosphate dehydrogenase ("IMPDH"). Illustratively, mammalian (e.g., human) IMPDH-encoding DNA sequences are useful in transformation or transfection of host cells for the large scale recombinant production of the enzymatically active expression products and/or products (e.g., GMP) resulting from IMPDH catalyzed synthesis in cells. Vectors including IMPDH-encoding DNA sequences are useful in gene amplification procedures. Recombinant proteins and synthetic peptides provided by the invention are useful as immunological reagents and in the preparation of antibodies (including polyclonal and monoclonal antibodies) for quantitative detection of IMPDH.

Abstract: A reagent such as a heat resistant enzyme is entrapped in a material such as wax or a liposome that releases the reagent when heated so the reagent is available for reaction. In a preferred embodiment, wax beads containing the reagent are prepared by injecting the reagent into beads of molten wax and cooling to solidify the wax. In another embodiment, droplets of a solution of the reagent are dropped through a layer of molten wax to coat the droplets with the wax and the coated droplets are cooled to solidify the wax. The entrapped reagents have application in nucleic acid hybridizations, polymerase chain reactions (PCR), reverse transcriptase reactions (RTR), nucleic acid sequencing, and product generating reactions such as colorimetic, fluorometric and chemiluminescent enzyme labeled immunoassays.

Abstract: Therapeutic oligonucleotide analogues which have improved nuclease resistance and improved cellular uptake are provided. Replacement of the normal phosphorodiester inter-sugar linkages found in natural oligomers with four atom linking groups forms unique di- and poly- nucleosides and nucleotides useful in regulating RNA expression and in therapeutics. Methods of synthesis and use are also disclosed.

Abstract: A personal communications apparatus using a garment-based audio interface. A garment member is worn on the upper torso of a person, wherein the garment member includes a neck opening which allows extension therethrough of the neck of the person. An audio output device capable of producing hi-fidelity spatialized 3-D sound aiming in selected directions is located adjacent the neck opening of the garment member. A receiver capable of receiving at least one transmitted signal and producing an audio signal based thereupon is coupled to the audio output device. An audio input device capable of capturing spatialized 3-D sound from selected directions is located adjacent the neck opening of the garment member. The audio signal from the audio input device is provided to a transmitter capable of transmitting a signal in dependence upon the audio signal. Embodiments of the garment member include a shirt and a necklace.

Abstract: Crystals of an amino acid, a nucleic acid or a derivative thereof can be efficiently isolated and purified at low cost from a solution containing the crystals, bacterial cells and medium components by using a liquid cyclone.

Abstract: A one-pot glycosylation reaction is disclosed in which a N-acetylgalactosamine (GalNAc) or N-acetylglucosamine (GlcNAc) group is enzymatically transferred to an acceptor molecule. The starting glycoside is a N-acetylamino monosaccharide 1-phosphate that is enzymatically converted to its UDP derivative via UTP and a pyrophorylase. The formed UDP derivative is epimerized, and the epimerized UDP derivative is used in the enzyme-catalyzed glycosyl transfer. That enzyme-catalyzed glycosyl transfer to an acceptor releases UDP that is enzymatically converted to UTP for further conversion of the N-acetylamino monosaccharide 1-phosphate into its UDP derivative.

Abstract: Flavine nucleotides, which are useful as ingredients of nutrient compositions, raw materials for various pharmaceutical products, biochemical research reagents and so on, are produced from flavine nucleotide precursors and ATP by utilizing cells or a culture of a microorganism which belongs to the genus Escherichia, Enterobactor or Pseudomonas and harbors a recombinant DNA comprising a vector DNA and a DNA fragment carrying the genetic information relevant to the synthesis of FMN and/or FAD, or treated cells or a treated culture of the microorganism.

Abstract: The present invention concerns a process for producing nucleosides by carrying out the reaction of a base donor, a saccharide residue donor and a phosphoric acid donor by the use of an enzyme preparation containing nucleoside phosphorylase, thereby forming an N-glycosidic bond between the base moiety of the base donor and the saccharide moiety of the saccharide residue donor, which comprises using, as the enzyme preparation containing nucleoside phosphorylase, a preparation derived from the cells of one or more kinds of microorganisms belonging to thermophiles of the genus Bacillus and having high nucleoside phosphorylase activity per unit cell weight.

Abstract: This invention relates to a process for the production of fructose 2,6-bisphosphate which comprises effecting reactions among (i) fructose 6-phosphate, (ii) glucose, (iii) fructose or (iv) glucose 6-phosphate, ATP and a phosphate donor, in the presence of (i) fructose 6-phosphate 2-kinase (PFK 2) and an enzyme which converts ADP into ATP (ADP/ATP converting enzyme), (ii) PFK 2, an ADP/ATP converting enzyme, hexokinase or glucokinase and glucose 6-phosphate isomerase, (iii) PFK 2, an ADP/ATP converting enzyme and hexokinase or glucokinase, or (iv) PFK 2, an ADP/ATP converting enzyme and glucose 6-phosphate isomerase; to a process for the production of fructose 2,6-bisphosphate which comprises allowing diesterase to coexist in a solution containing fructose 1,2-cyclic, 6-bisphosphate; and to a process for the purification of fructose 2,6-bisphosphate which comprises adding zinc salt to a solution containing fructose 2,6-bisphosphate, removing formed precipitate of impurities and adding a zinc salt to the result

Abstract: Compounds of formula (I): ##STR1## (wherein R represents a hydrogen atom or an acetyl group), which we have named "the adenophostins", and, have the ability to increase intracellular calcium ion concentrations by acting on the inositol 1,4,5-trisphosphate (InsP.sub.3) receptors which exist in the endoplasmic reticulum. The adenophostins are useful as hypertensive agents. They can be prepared by cultivation of a microorganism of the genus Penicillium, e.g. Penicillium brevicompactum SANK 11991 (FERM BP-3499) or Penicillium brevicompactum SANK 12177 (FERM BP-3500).

Abstract: A method for producing 3'-phosphoadenosine 5'-phosphosulfate, which comprises (1) reacting adenosine 5'-triphosphate with a sulfate donor in the presence of heat-stable adenosine 5'-triphosphate sulfurylase to produce adenosine 5'-phosphosulfate, and (2) reacting said adenosine 5'-phosphosulfate with adenosine 5'-triphosphate in the presence of heat-stable adenosine 5'-phosphosulfate kinase; and another method for producing 3'-phosphoadenosine 5'-phosphosulfate which comprises (1) reacting adenosine 5'-triphosphate with a sulfate donor in the presence of adenosine 5'-triphosphate sulfurylase to produce adenosine 5'-phosphosulfate, (2) reacting said adenosine 5'-phosphosulfate with adenosine 5'-triphosphate in the presence of adenosine 5'-phosphosulfate kinase to produce 3'-phosphoadenosine 5'-phosphosulfate and adenosine 5'-diphosphate, and (3) converting said adenosine 5'-diphosphate into adenosine 5'-triphosphate in the presence of a phosphate donor and an enzyme capable of converting adenosine 5'-diphospha

Abstract: The invention relates to a process for the production of cytidine 5'-monophosphosialic acids which comprises reacting a sialic acid with cytidine 5'-triphosphate in the presence of a cell extract of a naturally occurring microorganism having cytidine 5'-monophospho-N-acetylneuraminic acid synthetase activity, the extract optionally having been subjected to one purification step.

Abstract: The present invention provides a process of quantifying the number of viable cells in an aqueous suspension of cells using an energy-emitting non-hazardous probe and a probe-trigger. The process provides quantification data in short periods of time without the use of hazardous materials. A process of the present invention can also be used to quantify negatively charged particle number, assay for cytotoxicity, assay for cell proliferation and assay for cell differentiation. Still further, the present invention provides an assay kit for quantification of cells or negatively charged particles.

Abstract: Compositions and methods for producing the activating moiety of a site-directed catalytic antibody are provided. The activating moiety serves to enhance the rate of chemical reactions involving the conversion of the prodrug to one or more active substrates or drugs. The activating moiety typically comprises a catalytic antibody. Compositions and methods for producing the catalytic antibodies, as well as the haptens which are used to generate the catalytic antibodies, are provided. Compositions and methods for producing the prodrugs are also provided.

Abstract: A method of assaying L-carnitine in a specimen comprises reacting a specimen containing L-carnitine with:a) L-carnitine dehydrogenase having coenzymes of the thio-NAD group and of the NAD group, and which catalyzes a reversible reaction forming dehydrocarnitine from a substrate of carnitine,b) A.sub.1 andc) B.sub.1to effect a cycling reaction of the formula ##STR1## wherein A.sub.1 is thio-NAD group or NAD group, A.sub.2 is a reduced form of A.sub.1, when A.sub.1 is thio-NAD group, B.sub.1 is reduced NAD group and when A.sub.1 is NAD group, B.sub.1 is reduced thio-NAD, and wherein B.sub.2 is an oxidized form of B.sub.1 ; and measuring an amount of A.sub.2 or B.sub.1 generated or consumed by the cycling reaction. A composition for performing the assay comprises the above L-carnitine dehydrogenase, as well as the above components A.sub.1 and B.sub.1.

Abstract: Unsaturated esters can be converted into unsaturated polymerizable monomers using a biocatalyst derived from Corynebacterium oxydans. The method involves the step of reacting an unsaturated ester with an organic compound having a primary or secondary hydroxy group in a substantially organic environment in the presence of the noted biocatalyst. A transacylase has been isolated from the microorganism and at least partially purified.

Abstract: DNA coding for at least one enzyme that causes the accumulation of a pyrimidine deoxyribonucleoside is used, in conjunction with metabolic mutations or heterologous DNA coding for metabolic enzymes that also increase pyrimidine deoxyribonucleoside production, to engineer cultured cells to express a pyrimidine deoxyribonucleoside (PdN) in recoverable quantities, providing a commercially useful fermentation source for PdNs.

Abstract: An enzymatic process for extracting a nucleoside, such as thymidine, from a biomass without substantial thymine production.

Abstract: Polynucleotides are labeled with chemiluminescent acridine esters or luminescent lanthanides. Labelling is preferably carried out by incorporating a functional group into a nucleotide or polynucleotide at the C4 position of the pyrimidine portion or the C6 position of the purine portion and bonding a chemiluminescent acridine ester or a luminescent lanthanide to the functional group. The labeled polynucleotides are useful for direct detection of homologous polynucleotide sequences.

Abstract: A galactose transfer product is prepared by a process of allowing a microorganism capable of producing a galactose transfer product of the formula: (Gal).sub.n --R, wherein Gal represents a galactose residue, n represents an integer of 1 to 4 and R represents a galactose receptor to act on a combination of lactose or a galactose donor and a galactose receptor; and collecting the galactose transfer product produced.

Abstract: An enzyme preparation is obtained containing a nuclease that is produced by a fungus such as Trichoderma, Aspergillus and Fusarium and which remains active even after heating at 100.degree. C. for 30 minutes. This enzyme preparation may be effectively used when it is necessary to decompose nucleic acids at elevated temperature over a prolonged period.

Abstract: A process for producing fructose-1,6-diphosphate comprising the steps of: (a) enzymatically converting adenosine 5'-diphosphate to adenosine 5'-triphosphate using an acetate kinase-containing microorganism or an extract of the microorganism and phosphate donor; and (b) enzymatically converting a substrate capable of being converted to glucose or fructose to fructose-1,6-diphosphate using the adenosine 5'-triphosphate resulting from step (a) and the acetate kinase-containing microorganism or the extract of the microorganism.

Abstract: A process for the fermentative production of the deoxyribonucleoside thymidine and/or its corresponding base thymine by aerobically cultivating a strain of the genus Brevibacterium, in particular one of the strains NCIMB 40117 and 40116. The produced thymidine may be used as an intermediate in the production of azidothymidine and active ingredient in a composition for use in the treatment of auto imune deficiency syndrome (AIDS). Biologically pure cultures of strain NCIMB 40014 and variants and mutants derived therefrom are claimed per se.

Abstract: A process for the production of .sup.32 P-labeled nucleotides in accordance with an enzymatic pathway utilizing phosphotransacetylase and acetate kinase.

Abstract: The liposome compositions entrapping a drug are prepared by constituting the liposomal membrane with a saturated phospholipid and a glycolipid having sulfo group. Thus obtained compositions circulate stably in blood for a long time after intravenous administration.

Abstract: A process for synthesizing nucleosides is disclosed. The process includes the reaction of an alkylated nucleoside (such as 7-methylguanosine or 7-methylinosine) with a heterocyclic base (such as adenine, 3-deazaadenine or 1,2,4-triazole-3-carboxamide) in the presence of a nucleoside-forming enzyme to form a nucleoside that includes a glycosyl component, donate by the alkylated nucleoside, bonded to the heterocyclic base.

Abstract: A method of preparing a mixture of ribonucleotides consisting in hydrolysis f yeast nucleic acid with pancreatic ribonuclease at a pH 4.5-5.5. Thereafter, separating the ribonucleotide fraction from the obtained hydrolyzate is effective on membranes with pores sized 50-150 .ANG. with subsequent purification and isolation of the end product.

Abstract: This invention relates to a 5'-phosphodiesterase enzyme preparation which exhibits excellent storage stability. It is obtained by a special extraction process from rapidly proliferating parts of the germinating seeds. The enzyme is useful for the hydrolysis of RNA to form 5'-nucleotides.

Abstract: A process for producing a physiologically active substance by a combined enzymatic method is disclosed.

Abstract: A new method for releasing sample nucleic acids from cells, bacteria and viruses comprises non-invasively sonicating the sample contained within a sample container brought into physical contact with the vibrating element of a sonicator tuned to resonate at a frequency of 40 KHz or greater.

Abstract: A method of producing dideoxyinosine involving contacting as a substrate 2',3'-dideoxyadenosine with a microorganism which is capable of converting the substrate into 2',3'-dideoxyinosine.

Abstract: A process for converting AMP into ATP which comprises (a) using an enzyme which converts AMP into ADP and has been produced from microorganisms having an optimum growth temperature of 50.degree. C. to 85.degree. C. and an enzyme which converts ADP into ATP and has been produced from microorganisms having an optimum growth temperature of 50.degree. C. to 85.degree. C. is disclosed. In addition, there is disclosed a process for producing a physiologically active substance by a multienzyme process which comprises forming ATP from AMP by the step (a), (b) synthesizing a physiologically active substance with the resulting ATP, converting AMP resulting from the reaction in step (b) into ATP by the reaction in step (a), and repeatedly utilizing the converted ATP for synthesis of the physiologically active substance in step (b). By using the process it is possible to stably and efficiently carry out conversion of AMP into ATP over a long period of time.

Abstract: A process for producing diadenosine tetraphosphate or derivatives thereof in very high yields with little formation of undesired by-products is disclosed. The process comprises reacting adenosine-5'-triphosphate or a derivative thereof with an amino acid under the catalytic action of an aminoacyl-tRNA synthetase in the presence of an enzyme that converts adenosine-5'-diphosphate to adenosine-5'-triphosphate. The formation of by-products can be substantially completely inhibited if the last-mentioned enzyme is used in combination with an enzyme that converts adenosine-5'-monophosphate to adenosine-5'-diphosphate.

Abstract: A process for converting AMP into ATP which comprises (a) using an enzyme which converts AMP into ADP and has been produced from microorganisms having an optimum growth temperature of 50.degree. C. to 85.degree. C. and an enzyme which converts ADP into ATP and has been produced from microorganisms having an optimum growth temperature of 50.degree. to 85.degree. C. is disclosed. In addition, there is disclosed a process for producing a physiologically active substance by a multienzyme process which comprises forming ATP from AMP by the step (a), (b) synthesizing a physiologically active substance with the resulting ATP, coverting AMP resulting from the reaction in step (b) into ATP by the reaction in step (a), and repeatedly utilizing the converted ATP for synthesis of the physiologically active substance in step (b). By using the process it is possible to stably and efficiently carry out conversion of AMP into ATP over a long period of time.

Abstract: A biological process for producing a 2',3'-dideoxyncleoside from 2',3'-dideoxyuridine is disclosed. The 2',3'-dideoxynucleoside can be purified readily using a porous nonpolar resin adsorbent.

Abstract: Disclosed are an apparatus and a method for the poration and fusion of cells using radiofrequency electrical pulses. The electrodes of the apparatus can be hand held or part of integrated equipment with special containers for cells. The electrodes, which are positioned equal distance from each other, are attached to a power function generator. The power function generator can apply a continuous AC electrical field and/or a pulsed radiofrequency electrical field across the electrodes. The alternating electrical field induces cell dielectrophoresis. The pulsed radiofrequency electrical field porates or fuses the cells. The method can be used to fuse or porate a variety of cells including erythrocyte ghosts, liposomes, vesicles, isolated cells and cultured cells. During the poration or fusions a variety of chemical agents including antibodies, proteins, drugs, molecular probes, hormones, growth factors, DNA, RNA, enzymes, organic chemicals and inorganic chemicals can be introduced into these cells.

Abstract: A process for producing a primary or secondary alcohol derivative of a phosopholipid which comprises reacting the phospholipid with a primary or secondary alcohol in the presence of phospholipase DM.