Abstract: A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridizations, antibody/antigen reactions, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micro-machining techniques. The device can electronically control the transport and attachment of specific binding entities to specific micro-locations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific micro-locations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.

Abstract: Compounds having structure (1) ##STR1## wherein R.sup.1 is --H a protecting group, a linker or a binding partner; and R.sup.2 and R.sup.34 are as defined in the specification. The invention also provides intermediates and methods make the structure (1) compounds, as well as methods to use the compounds as labels in diagnostic assays and to enhance binding to complementary bases.

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 protein, Ancylostoma Secreted Protein (ASP), that is released by hookworm larvae following infection, and which is highly immunogenic in experimental animals, is disclosed. Nucleic acids encoding ASP, antibodies recognizing ASP, and methods of detecting ASP, nucleic acids encoding ASP, and antibodies recognizing ASP, in a sample are also disclosed. ASP is useful in a vaccine for hookworm as well as other soil-transmitted human and veterinary nematodiases. ASP is also useful as a target for specific treatment of hookworm, and can be used in a diagnostic assay for hookworm, using standard protein detection techniques, especially those based on antibodies. DNA encoding ASP is useful both for producing ASP recombinantly and in a diagnostic assay for hookworm. Antibodies recognizing ASP are useful in diagnostic assays to detect protein produced during hookworm infection.

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: 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 self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridization, antibody/antigen reaction, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micromachining techniques. The device can electronically control the transport and attachment of specific binding entities to specific micro-locations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.

Abstract: Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction chamber, having at least one cross-sectional dimension of about 0.1 to 1000 .mu.m. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.

Abstract: The present invention relates to a process for the electrochemical regeneration of pyridine cofactors.The process of the invention is characterized by the use, in a reaction medium subjected to electrolysis, of a cytoplasmic hydrogenase enzyme.

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: A method for producing a dinucleoside polyphosphate, a nucleoside polyphosphate or a derivative thereof which comprises using adenosine-5'-triphosphate, polyphosphate or a derivative thereof and a sulfate as reaction substrates and forming a dinucleoside polyphosphate, nucleoside polyphosphate or derivatives thereof via a two-stage reaction through the use of two enzymes, namely, adenosine-5'-triphosphate sulfurylase and diadenosine tetraphosphate phosphorylase as catalysts.

Abstract: A conjugated enzyme electrode comprising diaphorase and an amino-acid dehydrogenase immobilized on a conductive support by a polyfunctional aldehyde is disclosed, by which amino acids can be rapidly determined with high sensitivity.

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: The present invention relates to a method for determining a slight amount of NADH or XTP which is present in a solution, with high sensitivity by the use of an NADH kinase specific for NADH by utilizing a cycling reaction, and this method permits highly sensitive determination of NADH without any influence of NAD.sup.+ and hence is useful for diagnoses of diseases and the like.

Abstract: Polypeptides that bind to CD2, the receptor on the surface of T-lymphocytes. Most preferably, the polypeptides are soluble. DNA sequences that code on expression and/or secretion in appropriate unicellular hosts for those polypeptides.

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: 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: A catalytic RNA (ribozyme) derived from an intervening sequence (IVS) RNA of Tetrahymena thermophila will catalyze an RNA polymerization reaction in which pentacytidylic acid (C.sub.5) is extended by the successive addition of mononucleotides derived from a guanylyl-(3',5')-nucleotide (GpN). Cytidines or uridines are added to C.sub.5 to generate chain lengths of 10 to 11 nucleotides; longer products are also generated but at reduced efficiency. The reaction is analogous to that catalyzed by a replicase with C.sub.5 acting as the primer, GpNs as the nucleoside triphosphates, and a sequence in the ribozyme providing a template.

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: The invention relates to a process for the enzymatic preparation or regeneration of coenzymes selected from the group consisting of ATP, acetyl-CoA, acetyl-phosphate, NAD, NADP, NADH, and NADPH from their known oxidized and reduced forms, in which alkanals, such as acetaldehyde, are used as an oxidizing agent or alkanals such as acetaldehyde and alcohols, such as ethanol, are used as a reducing agent and a cell lysate of Clostridium kluyveri is used as a biocatalyst.

Abstract: 3'-deoxyadenosine 5'-triphosphate is oligomerized to form (2'-5')-oligo (3'-deoxyadenylate) by incubation with adenosine triphosphate: (2'-5')-oligo adenosine adenyl transferase, in the presence of an inert support carrying a double straded polynucleotide. The (2'-5')-oligo (3'-deoxyadenylate) is digested with a suitable phosphatase to remove the terminal phosphate groups. The thus produced corresponding 3'-deoxyadenosine compound is an anti-viral material effective against Herpes Simplex infection and effective in inhibiting the transformation of cells infected with Epstein Barr virus.

Abstract: A method for producing a reduced-form coenzyme by reacting malic acid with either nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate in the presence of malate dehydrogenase to obtain the corresponding reduced product. The reaction is preferably carried out under reduced pressure, while removing carbon dioxide, formed during the reaction, from the reaction system.

Abstract: DNA cloning and expression vectors capable of replication in a microbial host comprising from upstream to downstream (a) at least one promoter; (b) a translation start codon; (c) a cloning segment which provides a means for inserting nucleic acid sequences and (d) a sequence coding for a detectable gene product, out of translational phase with the translation start codon but capable of being readjusted to the translational phase of said start codon by insertion into the cloning segment of nucleic acid sequences containing the proper number of nucleotides for readjustment, said gene product providing a means for detecting expression of inserted nucleic acid sequences.

Abstract: Nicotinamide cofactors are prepared in a process of reacting ribose -5- phosphate with a basic material selected from the group consisting of ammonia, primary and secondary amines in a polar non-aqueous solvent, reacting the resultant 1-ribosylamine -5- phosphate with a pyridinium salt and reacting the resultant nicotinamide mononucleotide with adenosine triphosphate in the presence of nicotinamide adenine dinucleotide pyrophosphorase to produce nicotinamide adenine dinucleotide which can be used directly in crude form without further purification in co-factor - requiring enzymatic reactions. The nicotinamide adenine dinucleotide pyrophosphorase may be immobilized on a solid support.

Abstract: ATP and other nucleoside triphosphates labeled in the .gamma.-phosphate with .sup.32 P are prepared from L-.alpha.-glycerophosphate and their corresponding nucleoside diphosphates by a series of enzymatic reactions in the presence of NAD.sup.+, preferably regenerated by lactate dehydrogenase and pyruvate. The resulting [.gamma.-.sup.32 P]nucleotides are useful as reagents for analytical determinations.